Search results for: interferon gamma release assay

Authors: Zhdanova A. O., Volkov R. S., Kuznetsov G. V., Strizhak P. A.

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Fires in different types of facilities are a serious problem worldwide.It is still an unaccomplished science and technology objective to establish the minimum number and type of sensors in automatic systems of compartment fire suppression which would turn the fire-extinguishing agent spraying on and off in real time depending on the state of the fire, minimize the amount of agent applied, delay time in fire suppression and system response, as well as the time of combustion suppression. Based on the results of experimental studies, the conclusion was made that it is reasonable to use a gas analysis system and heat sensors (in the event of their prior activation) to determine the effectiveness of fire suppression (fire-extinguishing composition interacts with the fire). Thus, the concentration of CO in the interaction of the firefighting liquid with the fire increases to 0.7–1.2%, which indicates a slowdown in the flame combustion, and heat sensors stop responding at a gas medium temperature below 80 ºC, which shows a gradual decrease in the heat release from the fire. The evidence from this study suggests that the information received from the video recording equipment (video camera) should be used in real time as an additional parameter confirming fire suppression. Research was supported by Russian Science Foundation (project No 21-19-00009, https://rscf.ru/en/project/21-19-00009/).

Keywords: compartment fires, fire suppression, continuous control of fire behavior, feedback systems

Procedia PDF Downloads 129

Authors: Katawut Kaewbanjong

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We analyzed a volume of data and found significant user satisfaction in software project factors. A statistical significance analysis (logistic regression) and collinearity analysis determined the significance factors from a group of 71 pre-defined factors from 191 software projects in ISBSG Release 12. The eight prediction models used for testing the prediction potential of these factors were Neural network, k-NN, Naïve Bayes, Random forest, Decision tree, Gradient boosted tree, linear regression and logistic regression prediction model. Fifteen pre-defined factors were truly significant in predicting user satisfaction, and they provided 82.71% prediction accuracy when used with a neural network prediction model. These factors were client-server, personnel changes, total defects delivered, project inactive time, industry sector, application type, development type, how methodology was acquired, development techniques, decision making process, intended market, size estimate approach, size estimate method, cost recording method, and effort estimate method. These findings may benefit software development managers considerably.

Keywords: prediction model, statistical analysis, software project, user satisfaction factor

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Authors: Michael Shandalov, Tzvi Templeman, Michael Schmidt, Itzhak Kelson, Eyal Yahel

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We present a new method to produce a model system for the study of radiation damage in non-radioactive materials. The method is based on homogeneously incorporating 228Th ions in PbS thin films using a small volume chemical bath deposition (CBD) technique. The common way to alloy metals with radioactive elements is by melting pure elements, which requires considerable amounts of radioactive material with its safety consequences such as high sample activity. Controlled doping of the thin films with (very) small amounts (100-200ppm) of radioactive elements such as thorium is expected to provide a unique path for studying radiation damage in materials due to decay processes without the need of sealed enclosure. As a first stage, we developed CBD process for controlled doping of PbS thin films (~100 nm thick) with the stable isotope (t1/2~106 years), 232Th. Next, we developed CBD process for controlled doping of PbS thin films with active 228Th isotope. This was achieved by altering deposition parameters such as temperature, pH, reagent concentrations and time. The 228Th-doped films were characterized using X-ray diffraction, which indicated a single phase material. Film morphology and thickness were determined using scanning electron microscopy (SEM). Energy dispersive spectroscopy (EDS) mapping in the analytical transmission electron microscope (A-TEM), X-ray photoelectron spectroscopy (XPS) depth profiles and autoradiography indicated that the Th ions were homogeneously distributed throughout the films, suggesting Pb substitution by Th ions in the crystal lattice. The properties of the PbS (228Th) film activity were investigated by using alpha-spectroscopy and gamma spectroscopy. The resulting films are applicable for isochronal annealing of resistivity measurements and currently under investigation. This work shows promise as a model system for the analysis of dilute defect systems in semiconductor thin films.

Keywords: thin films, doping, radiation damage, chemical bath deposition

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Authors: Livy Febria Tedjamulia, Aas Nurasyiah, Ivana Josephin Purnama, Monika Diah Maharani Kusumastuti, Achmad Ridwan Ariyantoro

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Papua's candy is one of Indonesia’s indigenous consumption consisting of areca nut (Areca catechu), forest betel fruit (Piper aduncum), and CaCO3. This research aims to determine the concentration of tannins in areca nut, alkaloids in areca nut, flavonoids in forest betel fruit; detect their interaction and CaCO3; also toform a standardize consumption recommendation. The research methodwas includingDPPH assay for papua’s candy mixture, which resulted in IC50 value. Data analysis used is mathematical linear regression for each experiment. The test result of alkaloid is a Rf value of 0.773, while concentration of tannin and flavonoidare 0.603 mgGAE/g and 125.402 gQE/g, respectively. The IC50 value shows number of 3.0403, showing high antioxidant capacity.Other antioxidant assays were being studied using literature review, namely trolox and oxygen radical absorbance capacity, to figure out interaction among the bioactive compounds. It turned out that the interaction detected is antagonistic, which means the compound that is joined already has a stable molecular structure so that could reduce free radicals by donating hydrogen atoms. The recommendation consumptions given are 4 areca nuts, 5 forest betels, and 1 gram of lime betel. Therefore, papua's candy has its potential to be developed into functional food.

Keywords: antioxidant, bioactive compounds interaction, free radical, papua’s candy

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Authors: Shu-Chuan Liao, Chia-Ti Chang, Ko-Shao Chen

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Titanium and its alloy are widely used in many fields such as dentistry or orthopaedics. Due to their high strength low elastic modulus that chemical inertness and bio inert. The micro-arc oxidation used to formation a micro porous ceramic oxide layer film on Titanium surface and also to improve the resistance corrosion. For improving the biocompatibility, micro-arc oxidation surfaces bio-inert need to introduce reactive group. We introduced boundary layer by used plasma enhanced chemical vapor deposition of hexamethyldisilazane (HMDS) and organic active layer by UV light graft reactive monomer acrylic acid (AAc) therefore we can immobilize Chondroitin sulphate on surface easily by crosslinking EDC/NHS. The surface properties and composition of the modified layer were measured by scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS) and X-ray diffraction (XRD) and water contact angle. Water contact angle of the plasma-treated Ti surface decreases from 60° to 38°, which is an indication of hydrophilicity. The results of electrochemical polarization analysis showed that the sample plasma treated at micro-arc oxidation after plasma treatment has the best corrosion resistance. The result showed that we can immobilize chondroitin sulfate successful by a series of modification and MTT assay indicated the biocompatibility has been improved in this study.

Keywords: MAO, plasma, graft polymerization, biomedical application

Procedia PDF Downloads 258

Authors: Monika Szymańska-Czerwińsk, Kinga Zaręba-Marchewka, Krzysztof Niemczuk

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Chlamydiaceae are Gram-negative bacteria distributed worldwide in animals and humans. One of them is Chlamydia gallinacea recently discovered. Available data show that C. gallinacea is dominant chlamydial agent found in poultry in European and Asian countries. The aim of the studies was screening of poultry flocks in order to evaluate frequency of C. gallinacea shedding and genetic diversity. Sampling was conducted in different regions of Poland in 2019-2020. Overall, 1466 cloacal/oral swabs were collected in duplicate from 146 apparently healthy poultry flocks including chickens, turkeys, ducks, geese and quails. Dry swabs were used for DNA extraction. DNA extracts were screened using a Chlamydiaceae 23S rRNA real-time PCR assay. To identify Chlamydia species, specific real-time PCR assays were performed. Furthermore, selected samples were used for sequencing based on ompA gene fragments and variable domains (VD1-2, VD3-4). In total, 10.3% of the tested flocks were Chlamydiaceae-positive (15/146 farms). The presence of Chlamydiaceae was confirmed mainly in chickens (13/92 farms) but also in turkey (1/19 farms) and goose (1/26 farms) flocks. Eleven flocks were identified as C. gallinacea-positive while four flocks remained unclassified. Phylogenetic analysis revealed at least 16 genetic variants of C. gallinacea. Research showed that Chlamydiaceae occur in a poultry flock in Poland. The strains of C. gallinacea as dominant species show genetic variability.

Keywords: C. gallinacea, emerging agent, poultry, real-time PCR

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Authors: Raana Babadi Fathipour

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Food packaging plays a crucial role in protecting food from unwanted external factors. Antibacterial edible films are a promising option for food packaging due to their biodegradability, environmental friendliness, and safety. This paper reviews recent research progress on antimicrobial edible films, focusing on those made from polysaccharides, proteins, and lipids. Polysaccharides and proteins are the primary components of antimicrobial edible films, while lipids primarily serve as plasticizers and carriers for active substances in composite films. For instance, second-generation liposomes have shown great potential as carriers for antimicrobial substances and other bioactive compounds due to their exceptional stability. Furthermore, this paper analyzes recent advancements and future trends in antimicrobial edible films. One promising direction is the integration of antimicrobial edible film materials with delivery systems, such as nanoemulsion and microencapsulation technologies, to ensure stable loading of bioactive substances. Another emerging area of interest is the development of smart and active packaging that allows consumers to assess the freshness of food products without opening the package. pH-sensitive films and smart fluorescent "on-off" sensors for humidity are currently being explored as materials for smart and active packaging to monitor food product freshness, with further exploration anticipated in the future.

Keywords: antimicrobial edible film, biopolymer, antimicrobial agent, encapsulation, antimicrobial assay

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Authors: T. E. Tshikalange, B. C. Mophuting

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Traditional medicine often consists of complex ingredients prepared from a mixture of plant species. These herbal mixtures are used in the treatment of various ailments such as sexually transmitted diseases including HIV. The present study was carried out to determine the biological activities of the herbal mixture used traditionally in the treatment of sexually transmitted diseases. This herbal mixture consists of four plant species from families Asteraceae, Bignoniaceae, Fabaceae, and Myrtaceae. Five crude extracts (hexane, dichloromethane, methanol, water and boiled) of the herbal mixture were investigated for anti-gonococcal, anti-inflammatory, and reverse transcriptase activities. The anti-inflammatory activity of the plant extracts was determined by measuring the extract inhibitory effect on the pro-inflammatory enzyme lipoxygenase. The extracts were also tested for anti-HIV activity against recombinant HIV-1 enzyme using non-radioactive HIV-RT colorimetric assay. The boiled extract exhibited good anti-inflammatory activity with an IC₅₀ of 87 µg/ml compared to that of the positive control quercetin (IC₅₀= 92 µg/ml). All the other extracts showed little or no activity. Hexane extract was the only extract that showed reverse transcriptase extract inhibitory effect with an IC₅₀ of 74 µg/ml. Anti-gonococcal and cytotoxicity investigations are underway. The preliminary results support the use of herbal mixture by traditional healers.

Keywords: sexually transmitted diseases, lipoxygenase, anti-inflammatory, herbal mixture

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Authors: Mostosi C., Stéphenne J., Kempeneers E.

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Introduction: Refuellers from Brussels South Charleroi Airport (BSCA) expressed concerns about the risks involved in handling JET-A1 fuel. The HSE Manager of BSCA, in collaboration with the occupational physician and the industrial hygiene unit of the External Service of Occupational Medicine, decided to assess the toxicological exposure of these workers. Materials and methods: Two measurement methods were used. The first was to assay three types of metabolites in urine to highlight the exposure to xylenes, toluene, and benzene in aircraft fuels. Out of 32 refuellers in the department, 26 participated in the sampling, and 23 samples were exploited. The second method targeted the assessment of environmental exposure to certain potentially hazardous substances that refuellers are likely to breathe in work areas at the airport. It was decided to carry out two ambient air measurement campaigns, using static systems on the one hand and, on the other hand, using individual sensors worn by the refuellers at the level of the respiratory tract. Volatile organic compounds and diesel particles were analyzed. Results: Despite the fears that motivated these analyzes, the overall results showed low levels of exposure, far below the existing limit values, both in air quality and in urinary measurements. Conclusion: These results are comparable to a study carried out in several French airports. The staff could be reassured, and then the medical surveillance was modified by the occupational physician. With the aviation development at BSCA, equipment and methods are evolving. Their exposure will have to be reassessed.

Keywords: refuelling, airport, exposure, fuel, occupational health, air quality

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Authors: Nawel Ouennoughi, Kamel Daoud

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During recent years, new pharmaceutical dosage forms were developed in the research laboratories and which consists of encapsulating one or more active molecules in a polymeric envelope. Several techniques of encapsulation allow obtaining the microparticles or the nanoparticles containing one or several polymers. In the industry, microencapsulation is implemented to fill the following objectives: to ensure protection, the compatibility and the stabilization of an active matter in a formulation, to carry out an adapted working, to improve the presentation of a product, to mask a taste or an odor, to modify and control the profile of release of an active matter to obtain, for example, prolonged or started effect. To this end, we focus ourselves on the encapsulation of the antidiabetic. It is an oral hypoglycemic agent belonging to the second generation of sulfonylurea’s commonly employed in the treatment of type II non-insulin-dependent diabetes in order to improve profile them dissolution. Our choice was made on the technique of encapsulation by complex coacervation with two types of polymers (gelatin and the gum Arabic) which is a physicochemical process. Several parameters were studied at the time of the formulation of the microparticles and the nanoparticles: temperature, pH, ratio of polymers etc. The microparticles and the nanoparticles obtained were characterized by microscopy, laser granulometry, FTIR and UV-visible spectrophotometry. The profile of dissolution obtained for the microparticles showed an improvement of the kinetics of dissolution compared to that obtained for the active ingredient.

Keywords: coacervation, gum Arabic, microencapsulation, gelatin

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Authors: Ali Heidari

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This paper presents the principles for improving flood mitigation operation in multipurpose dams and maximizing reservoir performance during flood occurrence with a focus on the real-time operation of gated spillways. The criteria of operation include the safety of dams during flood management, minimizing the downstream flood risk by decreasing the flood hazard and fulfilling water supply and other purposes of the dam operation in mid and long terms horizons. The parameters deemed to be important include flood inflow, outlet capacity restrictions, downstream flood inundation damages, economic revenue of dam operation, and environmental and sedimentation restrictions. A simulation model was used to determine the real-time release of the Dez dam located in the Dez rivers in southwest Iran, considering the gate regulation curves for the gated spillway. The results of the simulation model show that there is a possibility to improve the current procedures used in the real-time operation of the dams, particularly using gate regulation curves and early flood forecasting system results. The Dez dam operation data shows that in one of the best flood control records, % 17 of the total active volume and flood control pool of the reservoir have not been used in decreasing the downstream flood hazard despite the availability of a flood forecasting system.

Keywords: dam operation, flood control criteria, Dez dam, Iran

Procedia PDF Downloads 225

Authors: Mansour F. Hussein, Mohammed A. Alshaikh, Riyadh S. Al-Jumaah, A. GarelNabi, I. Al-Khalifa, Osama B. Mohammed

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Serum samples from 489 male and female camels were tested for antibodies against C. burnetii using indirect enzyme-linked immunosorbent assay (ELISA). Antibodies to C. burnetii were recorded in sera of 252 (51.64%) camels. Significant differences in prevalence were found between male and female camels, juvenile and adult camels, different ecotypes and different sampling locations. 307 camels were simultaneously tested for C. burnetii antibodies by ELISA and indirect immunofluorescence (IFA). Close agreement was found between the results of the two tests. A high prevalence of C. burnetii antibodies was also recorded in milk samples tested by ELISA. Clinical samples from serologically positive camels were subjected to PCR analysis using primers which amplify the repetitive transposon-like and transposase gene regions of C. burnetii. Positive DNA amplification was obtained from both regions, with highest shedding of C. burnetii in faecal samples (27.59%) followed, in descending order, by urine (23.81%), blood (15.85%) and milk (6.5%). The present results indicate that camels are a major reservoir of C. burnetii in Saudi Arabia. The high prevalence of infection in camels, the poor sanitary standards under which the animals are kept and the consumption of raw camel milk indicate that camels could also be a major source of transmission of Q fever to humans in Saudi Arabia.

Keywords: Arabian camel, Camelus dromedarius, Coxiella brunetii, ELISA, immunofluoresence, PCR

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Authors: Afsoun Nikravan, Parisa Babaei, Gulen Gullu

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We aimed to study the association between house dust endotoxin, β-(1→3)-D-glucan, and asthma in a sample representative of the Turkish population. We analyzed data from 240 participants. The house dust was collected from the homes of 110 asthmatics and 130 control (without asthma) school-aged children (6-11 years old). House dust from the living room and from bedroom floors were analyzed for endotoxin and beta-glucan contents. House dust was analyzed for endotoxin content by the kinetic limulus amoebocyte lysate assay and for β-(1→3)-D-glucan by the inhibition enzyme immunoassay. The parents answered questions regarding potential determinants. We found geometric means 187.5 mg/m² for dust. According to statistical values, the endotoxin geometric mean was 13.86×103 EU/g for the control group and 6.16×103 EU/g for the asthma group. As a result, the amount of bacterial endotoxin was measured at a higher level in the homes of children without asthma. The geometric mean for beta-glucan was 46.52 µg/g and 44.39 µg/g for asthma and control groups, respectively. No associations between asthma and microbial agents were observed in Turkish children. High correlations (r > 0.75) were found between floor dust and endotoxin loads, while endotoxin and β-(1→3)-D-glucan concentrations were not correlated. The type of flooring (hard-surface or textile) was the strongest determinant for loads of floor dust and concentrations of endotoxin. Water damage and dampness at home were determinants of β-(1→3)-D-glucan concentrations. Endotoxin and β-(1→3)-D-glucan concentrations in Turkish house dust might lower than concentrations seen in other European countries.

Keywords: indoor air quality, asthma, microbial pollutants, case-control

Procedia PDF Downloads 124

Authors: Kye Gilder, Kevin Shan, Amy Halseth, Steve Smith

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This study assessed the effect of prolonged-release naltrexone 32 mg/bupropion 360 mg (NB) on cardiovascular (CV) events in overweight/obese participants at elevated CV risk. Participants must lose ≥2% body weight at 16 wks, without a sustained increase in blood pressure, to continue drug. Only serious adverse events (SAE) and adverse events leading to discontinuation of study drug (AELDSD) were collected. The study was terminated early after second interim analysis with 50% of all CV events. Data on CV endpoints has been published. Current analyses focused on AEs in participants on antidepressants at baseline, as these individuals were excluded from Phase 3 trials. Intent-to-treat (ITT) population (placebo [PBO] N=4450, NB N=4455) was 54.5% female, 83.5% white, mean age of 61 yrs, mean BMI 37.3 kg/m2, 22.8% with a history of depression, 23.1% on antidepressants, including 15.4% on an SSRI. SAEs in participants receiving antidepressants was similar between NB (10.7%) and PBO (9.9%) and also similar to overall population (9.5% NB, 8.1% PBO). SAEs in those on SSRIs were similar, 10.1% NB and PBO 9.4%. For those on SSRIs or other antidepressants, AELDSDs were similar to overall population and were primarily GI disorders. Obesity increases the risk of developing depression. For participants taking NB and antidepressants, including SSRIs, there is a similar AE profile as the overall population and data revealed no evidence of an additional health risk with combined use.

Keywords: antidepressant, Contrave, Mysimba, obesity, pharmacotherapy

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Authors: Behic Mert

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This study reports a facile approach to structure soft solids from microfluidizer lycopene-rich plant based structure and oil. First carotenoid-rich plant material (pumpkin was used in this study) processed with high-pressure microfluidizer to release lycopene molecules, then an emulsion was formed by mixing processed plant material and oil. While, in emulsion state lipid soluble carotenoid molecules were allowed to dissolve in the oil phase, the fiber material of plant material provided the network which was required for emulsion stabilization. Additional hydrocolloids (gelatin, xhantan, and pectin) up to 0.5% were also used to reinforce the emulsion stability and their impact on final product properties were evaluated via rheological, textural and oxidation studies. Finally, water was removed from emulsion phase by drying in a tray dryer at 40°C for 36 hours, and subsequent shearing resulted in soft solid (ole gel) structures. The microstructure of these systems was revealed by cryo-scanning electron microscopy. Effect of hydrocolloids on total lycopene and surface lycopene contents were also evaluated. The surface lycopene was lowest in gelatin containing oleo gels and highest in pectin-containing oleo gels. This study outlines the novel emulsion-based structuring method that can be used to encapsulate lycopene without the need of separate extraction of them.

Keywords: lycopene, encapsulation, fiber, oleo gel

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Authors: Ilham Ihoume, Rachid Tadili, Nora Arbaoui

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This study examines the effects of a solar-based heating system, in a north-‎south oriented agricultural greenhouse on the development of strawberry ‎plants during winter. This system relies on the circulation of water as a heat ‎transfer fluid in a closed circuit installed on the greenhouse roof to store heat ‎during the day and release it inside at night. A comparative experimental ‎study was conducted in two greenhouses, one experimental with the solar ‎heating system and the other for control without any heating system. Both ‎greenhouses are located on the terrace of the Solar Energy and Environment ‎Laboratory of the Mohammed V University in Rabat, Morocco. The devel-‎oped heating system consists of a copper coil inserted in double glazing and ‎placed on the roof of the greenhouse, a water pump circulator, a battery, and ‎a photovoltaic solar panel to power the electrical components. This inexpen-‎sive and environmentally friendly system allows the greenhouse to be heated ‎during the winter and improves its microclimate system. This improvement ‎resulted in an increase in the air temperature inside the experimental green-‎house by 6 °C and 8 °C, and a reduction in its relative humidity by 23% and ‎‎35% compared to the control greenhouse and the ambient air, respectively, ‎throughout the winter. For the agronomic performance, it was observed that ‎the production was 17 days earlier than in the control greenhouse.‎

Keywords: sustainability, solar energy, thermal energy storage.‎, greenhouse heating

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Authors: Azzam H., Abousamra N. K., Wafa A. A., Hafez M. M., El-Gilany A. H.

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Platelet activation occurs in peripheral blood of patients with rheumatic mitral stenosis (MS) and atrial fibrillation (AF) and could be related to abnormal thrombogenesis. The CD40/CD40 ligand (CD40L) which reflects platelet activation, mediate a central role in thrombotic diseases. However, the role of CD40/CD40L system in rheumatic MS with or without AF remains unclear. Expressions of CD40 on monocytes and CD40L on platelets were determined by whole blood flow cytometry and serum levels of soluble CD40L were measured by enzyme-linked immunosorbent assay in group 1 (19 patients with MS) and group 2 (20 patients with MS and AF) compared to group 3 (10 controls). Patients with groups 1 and 2 had a significant increase in expression of CD40 on monocytes (P1 and P2 = 0.000) and serum levels of sCD40L (P1 = 0.014 and P2 = 0.033, respectively), but nonsignificant increase in expression of CD40L on platelets (P1 = 0.109 and P2 = 0.060, respectively) as compared to controls. There were no significant difference in all the parameters in group 1 compared to group 2. Correlation analysis demonstrated that there was a significant direct relationship between the severity of MS and serum levels of sCD40L (r = -0.469, p = 0.043). In conclusion, rheumatic MS patients with or without AF had upregulation of the CD40/CD40L system as well as elevated sCD40L levels. The levels of sCD40L had a significantly direct relationship with the severity of MS and it was the stenotic mitral valve, not AF, that had a significant impact on platelet activation.

Keywords: CD40, CD40L, mitral stenosis, atrial fibrillation

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Authors: Nastaran Hemmati, Farhad Nikkhahi, Amir Javadi, Sahar Eskandarion, Seyed Mahmuod Amin Marashi

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Neisseria meningitidis, Escherichia coli K, Streptococcus agalactiae, and Streptococcus pneumoniae cause 90% of bacterial meningitis. Almost all infected people die or have irreversible neurological complications. Therefore, it is essential to have a diagnostic kit with the ability to quickly detect these fatal infections. The project involved 212 patients from whom cerebrospinal fluid samples were obtained. After total genome extraction and performing multiplex quantitative polymerase chain reaction (qPCR), the presence or absence of each infectious factor was determined by comparing with standard strains. The specificity, sensitivity, positive predictive value, and negative predictive value calculated were 100%, 92.9%, 50%, and 100%, respectively. So, due to the high specificity and sensitivity of the designed primers, they can be used instead of bacterial culture that takes at least 24 to 48 hours. The remarkable benefit of this method is associated with the speed (up to 3 hours) at which the procedure could be completed. It is also worth noting that this method can reduce the personnel unintentional errors which may occur in the laboratory. On the other hand, as this method simultaneously identifies four common factors that cause bacterial meningitis, it could be used as an auxiliary method diagnostic technique in laboratories particularly in cases of emergency medicine.

Keywords: cerebrospinal fluid, meningitis, quantitative polymerase chain reaction, simultaneous detection, diagnosis testing

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Authors: Ergun Sakalar, Kubra Bilgic

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In the last decades, real-time PCR has become a reliable tool preferred to use in many laboratories for pathogen detection. This technique allows for monitoring target amplification via fluorescent molecules besides admit of quantitative analysis by enabling of convert outcomes of thermal cycling to digital data. Sensitivity and traceability of real-time PCR are based on measuring of fluorescence that appears only when fluorescent reporter dye bound to specific target DNA.The fluorescent reporter systems developed for this purpose are divided into two groups. The first group consists of intercalator fluorescence dyes such as SYBR Green, EvaGreen which binds to double-stranded DNA. On the other hand, the second group includes fluorophore-labeled oligonucleotide probes that are separated into three subgroups due to differences in mechanism of action; initial primer-probes such as Cyclicons, Angler®, Amplifluor®, LUX™, Scorpions, and the second one hydrolysis probes like TaqMan, Snake assay, finally hybridization probes, for instance, Molecular Beacons, Hybprobe/FRET, HyBeacon™, MGB-Eclipse, ResonSense®, Yin-Yang, MGB-Pleiades. In addition nucleic acid analogues, an increase of probe affinity to target site is also employed with fluorescence-labeled probes. Consequently, abundant real-time PCR detection chemistries are chosen by researcher according to the field of application, mechanism of action, advantages, and proper structures of primer/probes.

Keywords: fluorescent dye, food safety, molecular probes, nucleic acid analogues

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Authors: Sandra Tapin-Lingua, Katia Ruel, Jean-Paul Joseleau, Daouia Messaoudi, Olivier Fahy, Michel Petit-Conil

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The preservative efficacy of organic biocides is strongly related to their capacity of penetration and retention within wood tissues. The specific detection of the pyrethroid insecticide is currently obtained after extraction followed by chemical analysis by chromatography techniques. However visualizing the insecticide molecule within the wood structure requires specific probes together with microscopy techniques. Therefore, the aim of the present work was to apply a new methodology based on antibody-antigen recognition and electronic microscopy to visualize directly pyrethroids in the wood material. A polyclonal antibody directed against cypermethrin was developed and implement it on Pinus sylvestris wood samples coated with technical cypermethrin. The antibody was tested on impregnated wood and the specific recognition of the insecticide was visualized in transmission electron microscopy (TEM). The immunogold-TEM assay evidenced the capacity of the synthetic biocide to penetrate in the wood. The depth of penetration was measured on sections taken at increasing distances from the coated surface of the wood. Such results correlated with chemical analyzes carried out by GC-ECD after extraction. In addition, the immuno-TEM investigation allowed visualizing, for the first time at the ultrastructure scale of resolution, that cypermethrin was able to diffuse within the secondary wood cell walls.

Keywords: cypermethrin, insecticide, wood penetration, wood retention, immuno-transmission electron microscopy, polyclonal antibody

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Authors: Adetunji Olusegun Aderigibigbe

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Shales, especially black shales, are of great geological significance, in the study of heavy/trace metal contamination. This is due to their abundance in occurrence and high concentration of heavy metals embedded which are released during their weathering. Heavy metals constitute one of the most dangerous pollution known to human because they are toxic (i.e., carcinogenic), non-biodegradable and can enter the global eco-biological circle. In the past, heavy metal contamination in aquatic environment and agricultural top soil has been attributed to industrial wastes, mining extractions and pollution from traffic vehicles; only a few studies have focused on weathering of shale as possible source of heavy metal contamination. Based on the above background, this study attempts to establish weathering of shale as possible source of trace/heavy metal contaminations. This was done by carefully selecting fresh and their corresponding weathered shale samples from selected localities in Bida and Anambra Basins. The samples were analysed in Activation Laboratories Ltd; Ontario, Canada for trace/heavy metal. It was observed that some major and trace metals were released during weathering, i.e., some were depleted and some enriched. By this contamination of water zones and agricultural top soils are not only traceable to biogenic processes but geogenic inputs (weathering of shale) as well.

Keywords: contamination, fresh samples, heavy metals, pollution, shales, trace metals, weathered samples

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Authors: Shubham Dwivedi, Raghavender Medishetti, Rita Rani, Aarti Sevilimedu, Pushkar Kulkarni, Yogeeswari Perumal

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Autism Spectrum Disorder (ASD) is a complex neurodevelopmental disorder of early onset, characterized by impaired sociability, cognitive function and stereotypies. There is a significant urge to develop and establish new animal models with ASD-like characteristics for better understanding of underlying mechanisms. The aim of the present study was to develop a cost and time effective zebrafish model with quantifiable parameters to facilitate mechanistic studies as well as high-throughput screening of new molecules for autism. Zebrafish embryos were treated with valproic acid and a battery of behavioral tests (anxiety, inattentive behavior, irritability and social impairment) was performed on larvae at 7th day post fertilization, followed by study of molecular markers of autism. This model shows a significant behavioural impairment in valproic acid treated larvae in comparison to control which was again supported by alteration in few marker genes and proteins of autism. The model also shows a rescue of behavioural despair with positive control drugs. The model shows robust parameters to study behavior, molecular mechanism and drug screening approach in a single frame. Thus we postulate that our 7 days zebrafish larval model for autism can help in high throughput screening of new molecules on autism.

Keywords: autism, zebrafish, valproic acid, neurodevelopment, behavioral assay

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Authors: Asif Jamal, Samia Sakindar, Ramla Rehman

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Biosurfactants are amphiphilic surface active compounds obtained from natural resources such as plants and microorganisms. Because of their diverse physicochemical characteristics biosurfactant are replacing synthetic compounds in various commercial applications. In present study, a strain of P. aeruginosa was isolated from crude oil contaminated soil as efficient biosurfactant producers. The biosurfactant production was analyzed as a function of surface tension reduction, oil spreading capacity, emulsification index and hemolysis assay. This bacterial strain showed excellent emulsion activity of EI24 85%, surface tension reduction up to 28.6 mNm-1 and 7.0 mm oil displacement zone. Physicochemical and biological properties of extracted rhamnolipid were also investigated in current study. The chemical composition of product from strain PSS was analyzed by FTIR spectroscopy. The results revealed that extracted biosurfactant was rhamnolipid type in nature having RL-1 and RL-2 homologues. The surface behavior of rhamnolipid in aqueous phase was investigated varying extreme pH, temperature, salt conditions and with various hydrocarbons. The results indicated that biosurfactant produced by strain PSS Which showed stability during high temperature up to 121 C, salt concentrations up to 20% and pH range between (4—14). The emulsification activity with different hydrocarbons was also remarkable. It was concluded that rhamnolipid biosurfactant produced by strain PSS has excellent potential as emulsifying/remediation agent for broad range of hydrophobic pollutants.

Keywords: P. aeruginosa, bioremediation, rhamnolipid, surfactants

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Authors: Shirin Jalili, Sadegh Hasannia, Hadi Shirzad, Afshin Khara

Abstract:

Morphine is one of the most medicinally important analgesics and narcotics. Structurally, it is classified as an alkaloid because of the presence of nitrogen. Its structure is similar to that of codeine, thebaine, and heroin. An immunoassay to accurately discriminate between these analogous alkaloids would be highly beneficial. A key factor for such an assay is specificity with high sensitivity, which is totally dependent on the antibody employed. However, most antibodies against haptens are polyclonal serum antibodies that exhibit significant cross-reactivities with closely related compounds. The camel-derived single-chain antibody fragments (VHH) are the smallest molecules with antigen-binding capacity, possessing unique properties compared to other conventional antibodies. In this study, a library containing the VHH genes of a camel immunized with with morphine conjugated BSA following phage display technology was generated. By screening the camel-derived variable region of the heavy chain cDNA phage display library with the ability to bind the desired hapten, we obtained some nanobodies that recognize this hapten. Phage display expression of the Nbs from this library and pannings against this hapten resulted in a clear enrichment of four distinct Nb-displaying phages with specificity for morphine that could be a potential target site for the development of new strategies for the development of a biosensor for detecting illicit drug.

Keywords: phage display, nanobody, Morphine-3, glucuronide, ELISA, biosensor

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Authors: Khater N., Rahmine S., Bougoffa C., Bouguenna T., Ouanes H.

Abstract:

Moringa oleifera L. species is considered one of the most useful trees in the world, possessing many interesting properties that make it of great scientific interest. It has been described as the miracle tree, the tree of a thousand virtues, the tree of life and God's gift to man. The present study aims to introduce, produce, and develop Moringa Oleifera as a species with high ecological potential (resistance to biotic and abiotic stresses and productivity), high added value, and multiple virtues. The aim of this work is to study the germination potential of this species under different temperature conditions. In this study, the germination assay was tested in two different temperature ranges: internal (laboratory ambient temperature between 22°c and 25°c) and external (seasonal temperature between 4°c and 8°c). Morphological and physiological analyses were carried out by Shoot length (SL), root length (RL), diameter at the crown (DC), fresh weight of shoots (FWS), fresh weight of roots (FWR), dry weight of shoots (DWS) and dry weight of roots (DWS). For all these variables, the results of the study reveal a significant difference between the two temperature intervals, with a high germination rate of 81. 81% and plant growth was rapid (7cm during 24h) in the laboratory temperature; in contrast to the external temperatures, a germination rate value of around 27% was recorded, and germination took place after 20 days of sowing, with slower plant growth. The results obtained show that a temperature greater than or equal to 25° is the ideal temperature for the germination and growth of moringa seeds and has a positive influence on the speed and percentage of germination.

Keywords: moringa oleifera, temperature, germination rate, growth, biomass

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Authors: Arfan Ali, Idrees Ahmad Nasir

Abstract:

Potato (Solanum tuberosum) is an important cash crop and popular vegetable in Pakistan and throughout the world. Cold storage of potatoes accelerates the conversion of starch into reduced sugars (glucose and fructose). This process causes dry mass and bitter taste in the potatoes that are not acceptable to end consumers. In the current study, the phosphofructokinase B gene was cloned into the pET-30 vector for protein expression and the pCambia-1301 vector for plant expression. Amplification of a 930bp product from an E. coli strain determined the successful isolation of the phosphofructokinase B gene. Restriction digestion using NcoI and BglII along with the amplification of the 930bp product using gene specific primers confirmed the successful cloning of the PfkB gene in both vectors. The protein was expressed as a His-PfkB fusion protein. Western blot analysis confirmed the presence of the 35 Kda PfkB protein when hybridized with anti-His antibodies. The construct Fani-01 was evaluated transiently using a histochemical gus assay. The appearance of blue color in the agroinfiltrated area of potato leaves confirmed the successful expression of construct Fani-01. Further, the area displaying gus expression was evaluated for PfkB expression using ELISA. Moreover, PfkB gene expression evaluated through transient expression determined successful gene expression and highlighted its potential utilization for stable expression in potato to reduce sweetening due to long-term storage.

Keywords: potato, Solanum tuberosum, transformation, PfkB, anti-sweetening

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Authors: Worakrit Worananthakij, Kamonchanok Doungtadum, Nattagan Mingkwan, Supatsorn Chupong

Abstract:

Natural products from herb have been used in different aspects of life as a result of their various biological activities. Generally, plant growth and production of secondary compounds largely depend on environmental conditions. To better understand this correlation, study on biological activity and soil parameter is necessary. This research aims to study the soil parameters which affect the efficiency of the antioxidant activity of essential oils extracted from the Zingiber zerumbet in three areas of Thailand, including Min Buri district, Bangkok province; Muang district, Chiang Mai province and Kaeng Sanam Nang district, Nakhon Ratchasima province. The soil samples in each area were collected and analyzed in the laboratory. The essential oil of Z. zerumbet in each province was extracted and tested for antioxidant activity by hydrodistillation method and DPPH (2,2-diphenyl-1-picrylhydrazyl radical) assay, respectively. The results showed that, the soil parameters such as pH, nitrogen, potassium and phosphorus elements and exchange of cations of soil specimen from Nakhon Ratchasima province were the highest (P<0.05) (6.10 ±0.03, 0.15 ± 0.04 percent of total nitrogen, 16.67 ± 0.46 mg/L, 3.35 ± 0.65 mg/kg and 12.87 ± 0.11 cmol/kg, respectively). In addition, IC50 (Inhibition Concentrtion of antioxidant at 50%) of Z. zerumbet essential oil collected from Nakhon Ratchasima showed the highest value (P<0.05) (1,400 µg/mL). In conclusion, the soil parameters are once important factor for the efficiency of essential oils extract from Z. zerumbet.

Keywords: antioxidant, essential oil, herb, soil parameter, Zingiber zerumbet

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Authors: Aleksandra Twarda, Dobrosława Krzemień, Grzegorz Dubin, Tad A. Holak

Abstract:

Stabilin-2 belongs to the group of scavenger receptors and plays a crucial role in clearance of more than 10 ligands from the bloodstream, including hyaluronic acid, products of degradation of extracellular matrix and metabolic products. The Link domain, a defining feature of stabilin-2, has a sequence similar to Link domains in other hyaluronic acid receptors, such as CD44 or TSG-6, and is responsible for most of ligands binding. Present knowledge of signal transduction by stabilin-2, as well as ligands’ recognition and binding mechanism, is limited. Until now, no experimental structures have been solved for any segments of stabilin-2. It has recently been demonstrated that the stabilin-2 knock-out or blocking of the receptor by an antibody effectively opposes cancer metastasis by elevating the level of circulating hyaluronic acid. Moreover, loss of expression of stabilin-2 in a peri-tumourous liver correlates with increased survival. Solving of the crystal structure of stabilin-2 and elucidation of the binding mechanism of hyaluronic acid could enable the precise characterization of the interactions in the binding site. These results may allow for designing specific small-molecule inhibitors of stabilin-2 that could be used in cancer therapy. To carry out screening for crystallization of stabilin-2, we cloned constructs of the Link domain of various lengths with or without surrounding domains. The folding properties of the constructs were checked by nuclear magnetic resonance (NMR). It is planned to show the binding of hyaluronic acid to the Link domain using several biochemical methods, i.a. NMR, isothermal titration calorimetry and fluorescence polarization assay.

Keywords: stabilin-2, Link domain, X-ray crystallography, NMR, hyaluronic acid, cancer

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Authors: Alireza Barari, Saeed Shirali

Abstract:

The etiology of obesity is heterogeneous with several factors, and the pathophysiology of obesity has recently related to leptin, oxidative damage, and inflammation. Silybum marianum have a health-promoting perspective and has shown that bioactive molecules of silymarin have the antioxidant and antitumor properties and can affect secretion of hormones and enzyme activity in animal. This study aimed to evaluate the antioxidant effects and changes in hormonal levels and body composition after silymarin consumption. Forty-five healthy untrained colleges male take part in the 4-week investigation. The subjects were assigned to 5 groups: endurance training, Silymarin with endurance training, strength training with placebo, Silymarin with strength training or placebo. Body fat percentage and Blood sample analysis were measured before and after the intervention to assay leptin, adiponectin and paraoxonase in the sample of subject's serum. There was a considerable decrease in body fat percent and a significant increase in VO2 max in 'Strength training' and 'Strength training with Silymarin' groups. But, no significant changes in levels of leptin, adiponectinin, and paraoxanase (PON) that were observed between exercise and exercise with Silymarin in these groups. We observed reduction in body fat% and increase in adiponectin induced by exercise for 4 weeks in untrained healthy men. Silybin, could not effectively improve all parameters and don’t prevent the progression of cell damage by antioxidant activity of PON.

Keywords: anti-inflammatory activity, antioxidant activity, silymarin, body composition, paraoxonase (PON)

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Authors: Sina Mahdavi

Abstract:

Background and Objective: Multiple sclerosis (MS) is an inflammatory autoimmune disease of the CNS that affects the myelination process in the central nervous system (CNS). Complex interactions of various "environmental or infectious" factors may act as triggers in autoimmunity and disease progression. The association between viral infections, especially Human Herpesvirus 6 (HHV-6), and MS is one potential cause that is not well understood. In this study, we aim to summarize the available data on HHV-6 infection in MS disease progression. Materials and Methods: For this study, the keywords "Multiple sclerosis", " Human Herpesvirus 6 ", and "central nervous system" in the databases PubMed and Google Scholar between 2017 and 2022 were searched, and 12 articles were chosen, studied, and analyzed. Results: HHV 6 tends towards TCD 4+ lymphocytes and enters the CNS due to the weakening of the blood-brain barrier due to inflammatory damage. Following the observation that the HHV-6 U24 protein has a seven amino acid sequence with myelin basic protein, which is one of the main components of the myelin sheath, it could cause a molecular mimicry mechanism followed by cross-reactivity. Reactivation of HHV-6 in the CNS can cause the release of proinflammatory cytokines, including TNF-α, leading to immune-mediated demyelination in patients with MS. Conclusion: There is a high expression of endogenous retroviruses during the course of MS, which indicates the relationship between HHV-6 and MS, and that this virus can play a role in the development of MS by creating an inflammatory state. Therefore, measures to modulate the expression of HHV-6 may be effective in reducing inflammatory processes in demyelinated areas of MS patients.

Keywords: multiple sclerosis, human herpesvirus 6, central nervous system, autoimmunity

Procedia PDF Downloads 111