Search results for: serum urea nitrogen
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2172

Search results for: serum urea nitrogen

552 Isolation and Characterization of Bio-surfactant Producing Alcaligenes sp YLA1 and Its Diesel Degradation Potentials

Authors: Abdulrahman Abdulhamid Arabo, Raji Arabi Bamanga, Mujiburrahman Fadilu, Musa Abubakar, Fatima Abdullahi Shehu, Hafeez Muhammad Yakasai, Nasiru Abdullahi

Abstract:

The aim of this study was to isolate and identify biosurfactant-producing and diesel alkanes degrading bacteria. For this reason, bacteria isolated from the diesel-contaminated site were screened for their potential to produce biosurfactants and degrade diesel alkanes. Primary selection of diesel degraders was carried out by using the conventional enrichment culture technique, where 12 bacterial strains were isolated based on their ability to grow on minimal media supplemented with diesel as the sole carbon source, which was followed by qualitative screening methods for potential biosurfactant production. Isolate B11 was the only candidate that showed positive signs for drop collapse, foaming, hemolytic test, oil displacement of more than 22 ± 0.05 mm, and emulsification (E24) of 14 ± 0.30%. The effect of various culture parameters (incubation time, diesel concentration, nitrogen source, pH and temperature) on the biodegradation of diesel was evaluated. The optimum incubation time was confirmed to be 120 days for isolate B11, and the optimum PH was confirmed as 8.0 for the isolate; similarly, the optimum temperature was confirmed as 35oC. In addition, diesel oil was used as the sole carbon source for the isolates. The favorable diesel concentration was 12.5 % (v/v) for the isolate. The isolate has shown degradative ability towards Tridecane (C13), dodecane, 2, 6, 10-trimethyl- (C15), Tetradecane (C14), 2,6,10-Trimethyltridecane (C16), Pentadecane (C15). It degraded between 0.27% - 9.65% of individual diesel oil alkanes. The strain has exhibited the potential of degrading diesel oil n-alkanes and was identified as Alcaligenes species strain B11 (MZ027604) using the 16S rRNA. Sequencing.

Keywords: diesel oil, biosurfactant, Alcaligenes sp, biodegradation

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551 Chromosomal Damage in Human Lymphocytes by Ultraviolet Radiation

Authors: Felipe Osorio Ospina, Maria Adelaida Mejia Arango, Esteban Onésimo Vallejo Agudelo, Victoria Lucía Dávila Osorio, Natalia Vargas Grisales, Lina María Martínez Sanchez, Camilo Andrés Agudelo Vélez, Ángela Maria Londoño García, Isabel Cristina Ortiz Trujillo

Abstract:

Excessive exposure to ultraviolet radiation, has shown to be a risk factor for photodamage, alteration of the immune mechanisms to recognize malignant cells and cutaneous pro-inflamatorios States and skin cancers. Objective: Identify the time of exposure to ultraviolet radiation for the production of chromosomal damage in human lymphocytes. Methodology: We conducted an in vitro study serial, in which samples were taken from heparinized blood of healthy people, who do not submit exposure to agents that could induce chromosomal alterations. The samples were cultured in RPMI-1640 medium containing 10% fetal bovine serum, penicillin and streptomycin antibiotic. Subsequently, they were grouped and exposed to ultraviolet light for 1 to 20 seconds. At the end of the treatments, cytology samples were prepared, and it was colored with Giemsa (5%). Reading was carried out in an optical microscope and 100 metaphases analysed by treatment for posting chromosomal alterations. Each treatment was conducted at three separate times and each became two replicas. Results: We only presented chromosomal alterations in lymphocytes exposed to UV for a groups 1 to 3 seconds (p<0.05). Conclusions: Exposure to ultraviolet radiation generates visible damage in chromosomes from human lymphocytes observed in light microscopy, the highest rates of injury was observed between two and three seconds, and above this value, the reduction in the number of mitotic cells was evident.

Keywords: ultraviolet rays, lymphocytes, chromosome breakpoints, photodamage

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550 Ultraviolet Radiation and Chromosomal Damage in Human Lymphocytes

Authors: Felipe Osorio Ospina, Maria Adelaida Mejia Arango, Esteban Onésimo Vallejo Agudelo, Victoria Lucía Dávila Osorio, Natalia Vargas Grisales, Lina María Martínez Sanchez, Camilo Andrés Agudelo Vélez, Ángela Maria Londoño García, Isabel Cristina Ortiz Trujillo

Abstract:

Excessive exposure to ultraviolet radiation, has shown to be a risk factor for photodamage, alteration of the immune mechanisms to recognize malignant cells and cutaneous pro-inflamatorios states and skin cancers. Objective: To identify the time of exposure to ultraviolet radiation for the production of chromosomal damage in human lymphocytes. Methodology: We conducted an in vitro study serial, in which samples were taken from the heparinized blood of healthy people, who do not submit exposure to agents that could induce chromosomal alterations. The samples were cultured in RPMI-1640 medium containing 10% fetal bovine serum, penicillin, and streptomycin antibiotic. Subsequently, they were grouped and exposed to ultraviolet light for 1 to 20 seconds. At the end of the treatments, cytology samples were prepared, and it was colored with Giemsa (5%). Reading was carried out in an optical microscope and 100 metaphases analysed by treatment for posting chromosomal alterations. Each treatment was conducted at three separate times and each became two replicas. Results: We only presented chromosomal alterations in lymphocytes exposed to UV for groups 1 to 3 seconds (p < 0.05). Conclusions: Exposure to ultraviolet radiation generates visible damage in chromosomes from human lymphocytes observed in light microscopy, the highest rates of injury was observed between two and three seconds, and above this value, the reduction in the number of mitotic cells was evident.

Keywords: chromosome breakpoints, lymphocytes, photodamage, ultraviolet rays

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549 The Hepatoprotective Effects of Aquatic Extract of Levesticum Officinale against Paraquat Toxicity of Hepatocytes

Authors: Hasan Afarnegan, Ali Shahraki, Jafar Shahraki

Abstract:

Paraquat is widely used as a strong nitrogen-based herbicide for controlling of weeds in agriculture. This poison is extremely toxic for humans which induces several – organ failure by accumulation in cells and many instances of death occurred due to its poisoning. Paraquat metabolized primarily in the liver. The purpose of this study was to assess the effects of aquatic extract of levisticum officinale on oxidative status and biochemical factors in hepatocytes exposed to paraquat. Our results determined that hepatocytes destruction induced by paraquat is mediated by reactive oxygen species (ROS) production, lipid peroxidation and decrease of mitochondrial membrane potential were significantly (P<0.05) prevented by aquatic extract of Levisicum officinale (100, 200 and 300 µg/ml). These effects of paraquat also prevented via antioxidants and ROS scavengers (α-tocopherol, DMSO, manitol), mitochondrial permeability transition (MPT) pore sealing compound (carnitine).MPT pore sealing compound inhibited the hepatotoxicity, indicating that paraquat induced cell death via mithochondrial pathway. Pretreatment of hepatocytes with aquatic extracts of Levisticum officinale, antioxidants and ROS scavengers also blocked hepatic cell death caused by paraquat, suggesting that oxidative stress may be directly induced decline of mithochondrial membrane potential. In conclusion, paraquat hepatotoxicity can be attributed to oxidative stress and continued by mithochondrial membrane potential disruption. Levisticum officinale aquatic extract, presumably due to its strong antoxidant properties, could protect the destructive effects of paraquat on rat hepatocytes.

Keywords: hepatocyte protection, levisticum officinale, oxidative stress, paraquat

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548 Clay Hydrogel Nanocomposite for Controlled Small Molecule Release

Authors: Xiaolin Li, Terence Turney, John Forsythe, Bryce Feltis, Paul Wright, Vinh Truong, Will Gates

Abstract:

Clay-hydrogel nanocomposites have attracted great attention recently, mainly because of their enhanced mechanical properties and ease of fabrication. Moreover, the unique platelet structure of clay nanoparticles enables the incorporation of bioactive molecules, such as proteins or drugs, through ion exchange, adsorption or intercalation. This study seeks to improve the mechanical and rheological properties of a novel hydrogel system, copolymerized from a tetrapodal polyethylene glycol (PEG) thiol and a linear, triblock PEG-PPG-PEG (PPG: polypropylene glycol) α,ω-bispropynoate polymer, with the simultaneous incorporation of various amounts of Na-saturated, montmorillonite clay (MMT) platelets (av. lateral dimension = 200 nm), to form a bioactive three-dimensional network. Although the parent hydrogel has controlled swelling ability and its PEG groups have good affinity for the clay platelets, it suffers from poor mechanical stability and is currently unsuitable for potential applications. Nanocomposite hydrogels containing 4wt% MMT showed a twelve-fold enhancement in compressive strength, reaching 0.75MPa, and also a three-fold acceleration in gelation time, when compared with the parent hydrogel. Interestingly, clay nanoplatelet incorporation into the hydrogel slowed down the rate of its dehydration in air. Preliminary results showed that protein binding by the MMT varied with the nature of the protein, as horseradish peroxidase (HRP) was more strongly bound than bovine serum albumin. The HRP was no longer active when bound, presumably as a result of extensive structural refolding. Further work is being undertaken to assess protein binding behaviour within the nanocomposite hydrogel for potential diabetic wound healing applications.

Keywords: hydrogel, nanocomposite, small molecule, wound healing

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547 Supplementation of Annatto (Bixa orellana)-Derived δ-Tocotrienol Produced High Number of Morula through Increased Expression of 3-Phosphoinositide-Dependent Protein Kinase-1 (PDK1) in Mice

Authors: S. M. M. Syairah, M. H. Rajikin, A. R. Sharaniza

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Several embryonic cellular mechanism including cell cycle, growth and apoptosis are regulated by phosphatidylinositol-3-kinase (PI3K)/Akt signaling pathway. The goal of present study is to determine the effects of annatto (Bixa orellana)-derived δ-tocotrienol (δ-TCT) on the regulations of PI3K/Akt genes in murine morula. Twenty four 6-8 week old (23-25g) female balb/c mice were randomly divided into four groups (G1-G4; n=6). Those groups were subjected to the following treatments for 7 consecutive days: G1 (control) received tocopherol stripped corn oil, G2 was given 60 mg/kg/day of δ-TCT mixture (contains 90% delta & 10% gamma isomers), G3 was given 60 mg/kg/day of pure δ-TCT (>98% purity) and G4 received 60 mg/kg/day α-TOC. On Day 8, females were superovulated with 5 IU Pregnant Mare’s Serum Gonadotropin (PMSG) for 48 hours followed with 5 IU human Chorionic Gonadotropin (hCG) before mated with males at the ratio of 1:1. Females were sacrificed by cervical dislocation for embryo collection 48 hours post-coitum. About fifty morula from each group were used in the gene expression analyses using Affymetrix QuantiGene Plex 2.0 Assay. Present data showed a significant increase (p<0.05) in the average number (mean + SEM) of morula produced in G2 (26.0 + 0.45), G3 (23.0 + 0.63) and G4 (25.0 + 0.73) compared to control group (G1 – 16.0 + 0.63). This is parallel with the high expression of PDK1 gene with increase of 2.75-fold (G2), 3.07-fold (G3) and 3.59-fold (G4) compared to G1 (1.78-fold). From the present data, it can be concluded that supplementation with δ-TCT(s) and α-TOC induced high expression of PDK1 in G2-G4 which enhanced the PI3K/Akt signaling activity, resulting in the increased number of morula.

Keywords: delta-tocotrienol, embryonic development, nicotine, vitamin E

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546 Transport Medium That Prevents the Conversion of Helicobacter Pylori to the Coccoid Form

Authors: Eldar Mammadov, Konul Mammadova, Aytaj Ilyaszada

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Background: According to many studies, it is known that H. pylori transform into the coccoid form, which cannot be cultured and has poor metabolic activity.In this study, we succeeded in preserving the spiral shape of H.pylori for a long time by preparing a biphase transport medium with a hard bottom (Muller Hinton with 7% HRBC (horse red blood cells) agar 5ml) and liquid top part (BH (brain heart) broth + HS (horse serum)+7% HRBC+antibiotics (Vancomycin 5 mg, Trimethoprim lactate 25 mg, Polymyxin B 1250 I.U.)) in cell culture flasks with filter caps. For comparison, we also used a BH broth medium with 7% HRBC used for the transport of H.pylori. Methods: Rapid urease test positive 7 biopsy specimens were also inoculated into biphasic and BH broth medium with 7% HRBC, then put in CO2 Gaspak packages and sent to the laboratory. Then both mediums were kept in the thermostat at 37 °C for 1 day. After microscopic, PCR and urease test diagnosis, they were transferred to Columbia Agar with 7% HRBC. Incubated at 37°C for 5-7 days, cultures were examined for colony characteristics and bacterial morphology. E-test antimicrobial susceptibility test was performed. Results: There were 3 growths from biphasic transport medium passed to Columbia agar with 7% HRBC and only 1 growth from BH broth medium with 7% HRBC. It was also observed that after the first 3 days in BH broth medium with 7%, H.pylori passed into coccoid form and its biochemical activity weakened, while its spiral shape did not change for 2-3 weeks in the biphase transport medium. Conclusions: By using the biphase transport medium we have prepared; we can culture the bacterium by preventing H.pylori from spiraling into the coccoid form. In our opinion, this may result in the wide use of culture method for diagnosis of H.pylori, study of antibiotic susceptibility and molecular genetic analysis.

Keywords: clinical trial, H.pylori, coccoid form, transport medium

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545 Construction of Submerged Aquatic Vegetation Index through Global Sensitivity Analysis of Radiative Transfer Model

Authors: Guanhua Zhou, Zhongqi Ma

Abstract:

Submerged aquatic vegetation (SAV) in wetlands can absorb nitrogen and phosphorus effectively to prevent the eutrophication of water. It is feasible to monitor the distribution of SAV through remote sensing, but for the reason of weak vegetation signals affected by water body, traditional terrestrial vegetation indices are not applicable. This paper aims at constructing SAV index to enhance the vegetation signals and distinguish SAV from water body. The methodology is as follows: (1) select the bands sensitive to the vegetation parameters based on global sensitivity analysis of SAV canopy radiative transfer model; (2) take the soil line concept as reference, analyze the distribution of SAV and water reflectance simulated by SAV canopy model and semi-analytical water model in the two-dimensional space built by different sensitive bands; (3)select the band combinations which have better separation performance between SAV and water, and use them to build the SAVI indices in the form of normalized difference vegetation index(NDVI); (4)analyze the sensitivity of indices to the water and vegetation parameters, choose the one more sensitive to vegetation parameters. It is proved that index formed of the bands with central wavelengths in 705nm and 842nm has high sensitivity to chlorophyll content in leaves while it is less affected by water constituents. The model simulation shows a general negative, little correlation of SAV index with increasing water depth. Moreover, the index enhances capabilities in separating SAV from water compared to NDVI. The SAV index is expected to have potential in parameter inversion of wetland remote sensing.

Keywords: global sensitivity analysis, radiative transfer model, submerged aquatic vegetation, vegetation indices

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544 A Retrospective Analysis of the Use of Vancomycin by Continous Infusion in the Critical Care Setting, Edinburgh

Authors: Sonia Nemakallu, Pota Kalima

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Introduction: Vancomycin is a glycopeptide antibiotic, commonly used to treat gram-positive bacteraemia. It has been increasingly used in the critical care setting due to an increased awareness of resistant gram positive organisms. In Edinburgh both tertiary hospitals, The Western General Hospital and The Royal Infirmary Of Edinburgh, commonly use Vancomycin for a variety of infections. Administration of Vancomyicn in these hospitals is by continuous infusion as it is thought to maintain serum concentrations easier and is a simpler monitoring system. Purpose: The aim of the study was to evaluate the efficacy and reliability in which Vancomycin is used. Material and Methods: A retrospective study, over a 6-month period from January 2014 to June 2014. 91 admissions were included, all received Vancomycin by continuous infusion during their critical care stay. Results: The number one use for Vancomycin in critical care settings was in the treatment of ventilator or hospital-acquired pneumonia. Only 3% of population had MRSA. 49% of admissions were not therapeutic on day 1 post loading dose. Of those that were therapeutic on day 1 post loading dose, 39% of admissions showed no organisms in any cultures taken, 42% had organisms sensitive to Vancomycin and 19% had only organisms resistant to Vancomycin. Those that were not therapeutic on day 1 showed similar organism sensitivities. 15% of admissions had Vancomycin levels above 25 (levels should be maintained between 15-25). An increase in creatinine was proportionally seen with an increase in Vancomycin levels. Conclusion: Within Edinburgh Vancomycin is being overused in the critical care setting with only 3% of the population having highly resistant organisms. Continuous infusion have not ruled out the complexity of maintaining therapeutic levels, with a large proportion of patients not being therapeutic on day 1. Further research is also required into the nephrotoxic effects of using higher doses of Vancomycin.

Keywords: Vancomycin, continuous infusion, multi resistant organisms, sepsis, renal toxicity

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543 The Taxonomic and Functional Diversity in Edaphic Microbial Communities from Antarctic Dry Valleys

Authors: Sean T. S. Wei, Joy D. Van Nostrand, Annapoorna Maitrayee Ganeshram, Stephen B. Pointing

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McMurdo Dry Valleys are a largely ice-free polar desert protected by international treaty as an Antarctic special managed area. The terrestrial landscape is dominated by oligotrophic mineral soil with extensive rocky outcrops. Several environmental stresses: low temperature, lack of liquid water, UV exposure and oligotrophic substrates, restrict the major biotic component to microorganisms. The bacterial diversity and the putative physiological capacity of microbial communities of quartz rocks (hypoliths) and soil of a maritime-influenced Dry Valleys were interrogated by two metagenomic approaches: 454 pyro-sequencing and Geochp DNA microarray. The most abundant phylum in hypoliths was Cyanobacteria (46%), whereas in solils Actinobacteria (31%) were most abundant. The Proteobacteria and Bacteriodetes were the only other phyla to comprise >10% of both communities. Carbon fixation was indicated by photoautotrophic and chemoautotrophic pathways for both hypolith and soil communities. The fungi accounted for polymer carbon transformations, particularly for aromatic compounds. The complete nitrogen cycling was observed in both communities. The fungi in particular displayed pathways related to ammonification. Environmental stress response pathways were common among bacteria, whereas the nutrient stress response pathways were more widely present in bacteria, archaea and fungi. The diversity of bacterialphage was also surveyed by Geochip. Data suggested that different substrates supported different viral families: Leviviridae, Myoviridae, Podoviridae and Siphoviridiae were ubiquitous. However, Corticoviridae and Microviridae only occurred in wetter soils.

Keywords: Antarctica, hypolith, soil, dry valleys, geochip, functional diversity, stress response

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542 Mode of Action of Surface Bound Antimicrobial Peptides Melimine and Mel4 against Pseudomonas aeruginosa

Authors: Muhammad Yasir, Debarun Dutta, Mark Willcox

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Biomaterial-associated infections are a multi-billion dollar burden globally. Antimicrobial peptide-based coatings may be able to prevent such infections. The aim of this study was to investigate the mechanism of action surface bound peptides (AMPs) against Pseudomonas aeruginosa 6294. Melimine and Mel4 were covalently attached to glass coverslips using azido-benzoic acid. Attachment was confirmed using X-ray photoelectron spectroscopy. P. aeruginosa was allowed to attach to AMP-coated glass for up to 6 hours. The effect of the surface-bound AMPs on bacterial cell membranes was evaluated using the dyes DiSC3-(5), Sytox green, SYTO 9 and propidium iodide with fluorescence microscopy. Release of cytoplasmic materials ATP and DNA/RNA were determined in the surrounding fluid. The amount of cell death was estimated by agar plate counts. The AMPs were successfully covalently bound to the glass as demonstrated by increases in %nitrogen of 3.6% (melimine) and 2.3% (Mel4) compared to controls. Immobilized peptides disrupted the cytoplasmic membrane potential of P. aeruginosa within 10 min. This was followed by the release of ATP after 2 h. Membrane permeabilization started at 3 h of contact with glass coated AMPs. There was a significant number of bacteria (59% for melimine; 36% for Mel-4) with damaged membranes after 4 h of contact. At the 6 h time point, release of DNA occurred with melimine releasing 2 times the amount of DNA/RNA than Mel4 surfaces (p < 0.05). Surface bound AMPs were able to disrupt cell membranes with subsequent release of cytoplasmic materials, and ultimately resulting in bacterial death.

Keywords: biomaterials, immobilized antimicrobial peptides, P. aeruginosa, mode of action

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541 Low Impact Development Strategies Applied in the Water System Planning in the Coastal Eco-Green Campus

Authors: Ying Li, Zaisheng Hong, Weihong Wang

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With the rapid enlargement of the size of Chinese universities, newly built campuses are springing up everywhere in recent years. It is urged to build eco-green campus because the role of higher education institutions in the transition to a more sustainable society has been highlighted for almost three decades. On condition that a new campus is usually built on an undeveloped site, where the basic infrastructure is not completed, finding proper strategies in planning and design of the campus becomes a primary concern. Low Impact Development (LID) options have been proposed as an alternative approach to make better use of rainwater in planning and design of an undeveloped site. On the basis of analyzing the natural circumstance, geographic condition, and other relative information, four main LID approaches are coordinated in this study of Hebei Union University, which are ‘Storage’, ‘Retaining’, ‘Infiltration’ and ‘Purification’. ‘Storage’ refers to a big central lake in the campus for rainwater harvesting. ‘Retaining’ means rainwater gardens scattered in the campus, also being known as bioretention areas which mimic the naturally created pools of water, to decrease surface flow runoff. ‘Infiltration’ is designed of grassed swales, which also play a part of floodway channel. ‘Purification’ is known as either natural or artificial wetland to reduce pollutants such as nitrogen and phosphorous in the waterbody. With above mentioned measures dealing with the synthetic use of rainwater in the acid & alkali area in the coastal district, an eco-green campus construction and an ecological sustainability will be realized, which will give us more enlightenment and reference.

Keywords: newly built campus, low impact development, planning design, rainwater reuse

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540 Contribution of Different Farming Systems to Soil and Ecological Health in Trans Nzoia County, Kenya

Authors: Janeth Chepkemoi, Richard Onwonga, Noel Templer, Elkana Kipkoech, Angela Gitau

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Conventional agriculture is one of the leading causes of land degradation, threatening the sustainability of food production. Organic farming promotes practices that have the potential of feeding the world while also promoting ecological health. A study was therefore carried out with the aim of conceptualizing how such farming systems are contributing to ecological health in Trans Nzoia County. 71 farmers were interviewed and data was collected on parameters such as land preparation, agroforestry, soil fertility management, soil and water conservation, and pests and diseases. A soil sample was also collected from each farm for laboratory analysis. Data collected were analyzed using Microsoft Excel and SPSS version 21. Results showed that 66% of the respondents practiced organic farming whereas 34% practiced conventional farming. Intercropping and crop rotations were the most common cropping systems and the most preferred land preparation tools among both organic and conventional farmers were tractors and hand hoes. Organic farms fared better in agroforestry, organic soil amendments, land and water conservation, and soil chemical properties. Pests and disease, however, affected organic farms more than conventional. The average nitrogen (%), K (Cmol/ kg and P (ppm) of organic soils were 0.26, 0.7 and 26.18 respectively, conventional soils were 0.21, 0.66 and 22.85. Soil organic carbon content of organic farms averaged a higher percentage of 2.07% as compared to 1.91 for the conventional. In conclusion, most farmers in Trans Nzoia County had transitioned into ecologically friendly farming practices that improved the quality and health of the soil and therefore promoted its sustainability.

Keywords: organic farming, conventional farming, ecological health, soil health

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539 Effects of Microbial Biofertilization on Nodulation, Nitrogen Fixation, and Yield of Lablab purpureus

Authors: Benselama Amel, Ounane S. Mohamed, Bekki Abdelkader

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A collection of 20 isolates from fresh Nodules of the legume plant Lablab purpureus was isolated. These isolates have been authenticated by seedling inoculation grown in jars containing sand. The results obtained after two months of culture have revealed that the 20 isolates (100% of the isolates) are able to nodulate their host plants. The results obtained were analyzed statistically by ANOVA using the software statistica and had shown that the effect of the inoculation has significantly improved all the growth parameters (the height of the plant and the dry weight of the aerial parts and roots, and the number of nodules). We have evaluated the tolerance of all strains of the collection to the major stress factors as the salinity, pH and extreme temperature. The osmotolerance reached a concentration up to 1710mm of NaCl. The strains were also able to grow on a wide range of pH, ranging from 4.5 to 9.5, and temperature, between 4°C and 40°C. Also, we tested the effect of the acidity, aluminum and ferric deficit on the Lablab-rhizobia symbiosis. Lablab purpureus has not been affected by the presence of high concentrations of aluminum. On the other hand, iron deficiency has caused a net decrease in the dry biomass of the aerial part. The results of all the phenotypic characters have been treated by the statistical Minitab software, the numerical analysis had shown that these bacterial strains are divided into two distinct groups at a level of similarity of 86 %. The SDS-PAGE was carried out to determine the profile of the total protein of the strains. The coefficients of similarity of polypeptide bands between the isolates and strains reference (Bradyrhizobium, Mesorizobium sp.) confirm that our strain belongs to the groups of rhizobia.

Keywords: SDS-PAGE, rhizobia, symbiosis, phenotypic characterization, Lablab purpureus

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538 Acute Phase Proteins, Proinflammatory Cytokines and Oxidative Stress Biomarkers in Sheep with Pneumonic Pasteurellosis

Authors: Wael M. El-Deeb

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The aim of this study was to assess the pathophysiological importance of lipid profile, acute phase proteins, proinflammatory cytokines and oxidative stress markers in sheep with pneumonic pasteurellosis. Blood samples were collected from 36 Pasteurellamultocida-infected sheep, together with 20 healthy controls. Samples for bacteriological examination (nasal swabs, bronchoalveolar lavage) were collected from all animals and subjected to bacteriological examinations. Moreover, heart blood and lung samples were collected from the dead pneumonic sheep and subjected also to bacteriological examinations. A lipid profile was determined, along with a blood picture and other biochemical parameters. The acute phase proteins (fibrinogen, haptoglobin, serum amyloid A), the proinflammatory cytokine tumour necrosis factor-alpha, interleukins (IL-1α, IL-1β, IL-6), interferon-gamma and the oxidative stress markers malondialdehyde, super oxide dismutase, glutathione and catalase were also measured. The examined biochemical parameters were increased in the pneumonic sheep, except for cholesterol and high-density lipoprotein cholesterol (HDL-c), which were significantly lower than control group. Acute phase proteins and cytokines were significantly higher in the pneumonic sheep when compared to the healthy sheep. There was a significant increase in the levels of malondialdehyde; however, a significant decrease in the levels of super oxide dismutase, glutathione and catalase was observed. The present study shed the light on the possible pathphysiological role of lipid profile, acute phase proteins (APPs), proinflammatory cytokines and oxidative stress markers in pneumonic pasteurelosis in sheep.

Keywords: acute phase proteins, sheep, pasteurella, interleukins, stress

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537 Clouds Influence on Atmospheric Ozone from GOME-2 Satellite Measurements

Authors: S. M. Samkeyat Shohan

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This study is mainly focused on the determination and analysis of the photolysis rate of atmospheric, specifically tropospheric, ozone as function of cloud properties through-out the year 2007. The observational basis for ozone concentrations and cloud properties are the measurement data set of the Global Ozone Monitoring Experiment-2 (GOME-2) sensor on board the polar orbiting Metop-A satellite. Two different spectral ranges are used; ozone total column are calculated from the wavelength window 325 – 335 nm, while cloud properties, such as cloud top height (CTH) and cloud optical thick-ness (COT) are derived from the absorption band of molecular oxygen centered at 761 nm. Cloud fraction (CF) is derived from measurements in the ultraviolet, visible and near-infrared range of GOME-2. First, ozone concentrations above clouds are derived from ozone total columns, subtracting the contribution of stratospheric ozone and filtering those satellite measurements which have thin and low clouds. Then, the values of ozone photolysis derived from observations are compared with theoretical modeled results, in the latitudinal belt 5˚N-5˚S and 20˚N - 20˚S, as function of CF and COT. In general, good agreement is found between the data and the model, proving both the quality of the space-borne ozone and cloud properties as well as the modeling theory of ozone photolysis rate. The found discrepancies can, however, amount to approximately 15%. Latitudinal seasonal changes of photolysis rate of ozone are found to be negatively correlated to changes in upper-tropospheric ozone concentrations only in the autumn and summer months within the northern and southern tropical belts, respectively. This fact points to the entangled roles of temperature and nitrogen oxides in the ozone production, which are superimposed on its sole photolysis induced by thick and high clouds in the tropics.

Keywords: cloud properties, photolysis rate, stratospheric ozone, tropospheric ozone

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536 Nitrification Efficiency and Community Structure of Municipal Activated Sewage Sludge

Authors: Oluyemi O. Awolusi, Abimbola M. Enitan, Sheena Kumari, Faizal Bux

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Nitrification is essential to biological processes designed to remove ammonia and/or total nitrogen. It removes the excess nitrogenous compound in wastewater which could be very toxic to the aquatic fauna or cause a serious imbalance of such aquatic ecosystem. Efficient nitrification is linked to an in-depth knowledge of the structure and dynamics of the nitrifying community structure within the wastewater treatment systems. In this study, molecular technique was employed for characterizing the microbial structure of activated sludge [ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB)] in a municipal wastewater treatment with intention of linking it to the plant efficiency. PCR-based phylogenetic analysis was also carried out for. The average operating and environmental parameters, as well as specific nitrification rate of a plant, was investigated during the study. During the investigation, the average temperature was 23±1.5oC. Other operational parameters such as mixed liquor suspended solids and chemical oxygen demand inversely correlated with ammonia removal. The dissolved oxygen level in the plant was constantly lower than the optimum (between 0.24 and 1.267 mg/l) during this study. The plant was treating wastewater with the influent ammonia concentration of 31.69 and 24.47 mg/l. The influent flow rates (ML/day) was 96.81 during the period. The dominant nitrifiers include: Nitrosomonas spp. Nitrobacter spp. and Nitrospira spp. The AOB had a correlation with nitrification efficiency and temperature. This study shows that the specific ammonia oxidizing rate and the specific nitrate formation rates can serve as a good indicator of the plant overall nitrification performance.

Keywords: Ammonia monooxygenase α-subunit gene, amoA, ammonia-oxidizing bacteria, AOB, nitrite-oxidizing bacteria, NOB, specific nitrification rate

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535 Development of a Web-Based Application for Intelligent Fertilizer Management in Rice Cultivation

Authors: Hao-Wei Fu, Chung-Feng Kao

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In the era of rapid technological advancement, information technology (IT) has become integral to modern life, exerting significant influence across diverse sectors and serving as a catalyst for development in various industries. Within agriculture, the integration of IT offers substantial benefits, notably enhancing operational efficiency. Real-time monitoring systems, for instance, have been widely embraced in agriculture, effectively improving crop management practices. This study specifically addresses the management of rice panicle fertilizer, presenting the development of a web application tailored to handle data associated with rice panicle fertilizer management. Leveraging the normalized difference red edge index, this application optimizes the quantity of rice panicle fertilizer used, providing recommendations to agricultural stakeholders and service providers in the agricultural information sector. The overarching objective is to minimize costs while maximizing yields. Furthermore, a robust database system has been established to store and manage relevant data for future reference in rice cultivation management. Additionally, the study utilizes the Representational State Transfer software architectural style to construct an application programming interface (API), facilitating data creation, retrieval, updating, and deletion for users via the HyperText Transfer Protocol methods. Future plans involve integrating this API with third-party services to incorporate it into larger frameworks, thus catering to the diverse requirements of various third-party services.

Keywords: application programming interface, HyperText Transfer Protocol, nitrogen fertilizer intelligent management, web-based application

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534 The Connection between Body Composition and Blood Samples Results in Aesthetic Sports

Authors: Réka Kovács, György Téglásy, Szilvia Boros

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Introduction: Aim of the Study: Low body fat percentage frequently occurs in aesthetic sports. Because of the unrealistic expectations, their quantity and quality of nutrition intake are inadequate. This can be linked to several health issues which appear in blood samples (iron, ferritin, creatine kinase, etc.). Our retrospective study aimed to investigate the connection between body composition (InBody 770 monitor) and blood samples test results among elite adolescent (14-18 years) and adult (19-28 years) aesthetic athletes. Methods: Data collection happened between 01.08.2022. and 15.08.2022 in National Institute for Sports Medicine, Budapest. The final group consisted of 111 athletes (n=111; adolescents: n=68, adults: n=43). We used descriptive statistics, a two-sample t-test, and correlation analysis with Microsoft Office Excel 2007 software. Our results were considered significant if p<0,05. Results: In 33,3% (37/111) we found low body fat percentage (girls and women: <12%, boys and men: <8%) and in 64% (71/111) high creatine kinase levels. Differences were found mainly in the adolescent group. We found a correlation between body weight and total cholesterol, visceral fat and triglyceride, hematocrit and iron-linking capacity, moreover body fat percentage and ferritin levels. Discussion: It is important to start education about sports nutrition at an early age. The connection between low body fat percentage, serum iron, triglyceride, and ferritin levels refers to the fact that the nutrition intake of the athletes is inadequate. High blood concentrations of creatine kinase may show a lack of proper recovery, which is essential to improve health and performance.

Keywords: body fat percentage, creatine kinase, recovery, sports nutrition

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533 Enhancement in Bactericidal Activity of Hydantoin Based Microsphere from Smooth to Rough

Authors: Rajani Kant Rai, Jayakrishnan Athipet

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There have been several attempts to prepare polymers with antimicrobial properties by doping with various N-halamines. Hydantoins (Cyclic N-halamine) is of importance due to their stability rechargeable chloroamide function, broad-spectrum anti-microbial action and ability to prevent resistance to the organisms. Polymerizable hydantoins are synthesized by tethering vinyl moieties to 5,5,-dialkyl hydantoin sacrificing the imide hydrogen in the molecule thereby restricting the halogen capture only to the amide nitrogen that results in compromised antibacterial activity. In order to increase the activity of the antimicrobial polymer, we have developed a scheme to maximize the attachment of chlorine to the amide and the imide moieties of hydantoin. Vinyl hydantoin monomer, (Z)-5-(4-((3-methylbuta-1,3-dien-2-yl)oxy)benzylidene)imidazolidine-2,4-dione (MBBID) was synthesized and copolymerized with a commercially available monomer, methyl methacrylate, by free radical polymerization. The antimicrobial activity of hydantoin is strongly dependent on their surface area and hence their microbial activity increases when incorporated in microspheres or nanoparticles as compared to their bulk counterpart. In this regard, smooth and rough surface microsphere of the vinyl monomer (MBBID) with commercial monomer was synthesized. The oxidative chlorine content of the copolymer ranged from 1.5 to 2.45 %. Further, to demonstrate the water purification potential, the thin column was packed with smooth or rough microspheres and challenged with simulated contaminated water that exhibited 6 log kill (total kill) of the bacteria in 20 minutes of exposure with smooth (25 mg/ml) and rough microsphere (15.0 mg/ml).

Keywords: cyclic N-halamine, vinyl hydantoin monomer, rough surface microsphere, simulated contaminated water

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532 A Three-Dimensional Investigation of Stabilized Turbulent Diffusion Flames Using Different Type of Fuel

Authors: Moataz Medhat, Essam E. Khalil, Hatem Haridy

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In the present study, a numerical simulation study is used to 3-D model the steady-state combustion of a staged natural gas flame in a 300 kW swirl-stabilized burner, using ANSYS solver to find the highest combustion efficiency by changing the inlet air swirl number and burner quarl angle in a furnace and showing the effect of flue gas recirculation, type of fuel and staging. The combustion chamber of the gas turbine is a cylinder of diameter 1006.8 mm, and a height of 1651mm ending with a hood until the exhaust cylinder has been reached, where the exit of combustion products which have a diameter of 300 mm, with a height of 751mm. The model was studied by 15 degree of the circumference due to axisymmetric of the geometry and divided into a mesh of about 1.1 million cells. The numerical simulations were performed by solving the governing equations in a three-dimensional model using realizable K-epsilon equations to express the turbulence and non-premixed flamelet combustion model taking into consideration radiation effect. The validation of the results was done by comparing it with other experimental data to ensure the agreement of the results. The study showed two zones of recirculation. The primary one is at the center of the furnace, and the location of the secondary one varies by changing the quarl angle of the burner. It is found that the increase in temperature in the external recirculation zone is a result of increasing the swirl number of the inlet air stream. Also it was found that recirculating part of the combustion products back to the combustion zone decreases pollutants formation especially nitrogen monoxide.

Keywords: burner selection, natural gas, analysis, recirculation

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531 Ghrelin, Obestatin and Ghrelin/Obestatin Ratio: A Postprandial Study in Healthy Subjects of Normal Weight

Authors: Panagiotis T. Kanellos, Vaios T. Karathanos, Andriana C. Kaliora

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Introduction: The role of ghrelin and obestatin in appetite regulation has been investigated. However, data on ghrelin and obestatin changes after food ingestion are negligible. Objective: We aimed at assessing the appetite-regulating hormones, ghrelin, and obestatin, and furthermore calculate ghrelin/obestatin ratio in healthy normal-weight subjects after consumption of raisins. This survey is a comparative study of a glucose control with raisins containing fructose and glucose in similar concentrations as well as fibers. Methodology: Ten apparently healthy subjects who reported no history of glucose intolerance, diabetes, gastrointestinal disorders, or recent use of any antibiotics were enrolled in the study. The raisins used (Vitis vinifera) originate in Greece and are distributed worldwide as Corinthian raisins. In a randomized crossover design, all subjects after an overnight fast consumed, either 50g of glucose diluted in 240 mL of water (control) or 74 g of raisins (sugar content 50 g) with a 5-day interval between individual trials. Vein blood samples were collected at baseline and at 60, 120 and 180 min postprandially. In blood samples ghrelin and obestatin were measured applying specific enzyme linked immuno absorbent assays. Results: The subjects were of mean age 26.3 years, with BMI of 21.6 kg/m2, waist circumference of 77.7 cm, normal serum lipidemic parameters and normal HbA1c levels. Ghrelin levels were significantly lower after raisin consumption compared to glucose at 120 and at 180 min post-ingestion (p= 0.011 and p= 0.035, respectively). However, obestatin did not reach statistical significance between the two interventions. The ghrelin/obestatin ratio was found significantly lower (p=0.020) at 120 min after raisin ingestion compared to control. Conclusion: Two isocaloric foods containing equal amounts of sugars, however with a different composition, have different effects on appetite hormones ghrelin and obestatin in normal-weight healthy subjects.

Keywords: appetite, ghrelin, obestatin, raisins

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530 The Effect of Vitamin D Supplementation on Prostate Cancer: A Systematic Review and Meta-Analysis of Clinical Trials

Authors: Simin Shahvazi, Sepideh Soltani, Seyed Mehdi Ahmadi, Russell J. De Souza, Amin Salehi-Abargouei

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Background and Objectives: Vitamin D has received attention for its potential to disrupt cancer processes such as attenuating cell proliferation and exacerbating differentiation and apoptosis. However, whether there exists a role for vitamin D in the treatment of prostate cancer specifically remains controversial. We systematically review the literature to assess whether supplementation with vitamin D influences PSA response and overall survival in patients with prostate cancer. Methods: We searched PubMed, Scopus, ISI Web of Science and Google scholar from inception through up to 10 September 2017 for both before-and-after and randomized trials that evaluated the effect of vitamin D supplementation on the prostate specific antigen (PSA) response rate in participants with prostate cancer. The DerSimonian and Laird, inverse-weighted random-effects model was used to pool effect estimates from the studies. Heterogeneity and potential publication bias were evaluated. Subgroup analyses were also performed. Results: Twenty-two studies (16 before-after and 6 randomized controlled trials) were found and included in meta-analysis. The analysis on controlled clinical trials revealed that PSA change from baseline [weighted mean difference (WMD) = -1.66 ng/ml, 95%CI: -0.69, 0.36, P= 0.543)], PSA response (RR=1.18, 95%CI: 0.97, 1.45, P=0.104) and mortality rate (risk ratio (RR) = 1.05, 95% CI: 0.81-1.36; P=0.713) was not significantly different between vitamin D supplementation and placebo groups. Single arm trials revealed that vitamin D supplementation had had a modest effect on PSA response rate: 19% of those enrolled had at least a 50% reduction in PSA by the end of treatment (95% CI: 7% to 31%; p=0.002). Conclusion: We found that vitamin D modestly increases the PSA response rate in single arm studies. No effect on serum PSA levels, PSA response and mortality was seen in randomized controlled clinical trials. It does not seem patients with prostate cancer benefit from vitamin D supplementation.

Keywords: mortality, prostatic neoplasms, PSA response, vitamin D

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529 Review of Sulfur Unit Capacity Expansion Options

Authors: Avinashkumar Karre

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Sulfur recovery unit, most commonly called as Claus process, is very significant gas desulfurization process unit in refinery and gas industries. Explorations of new natural gas fields, refining of high-sulfur crude oils, and recent crude expansion projects are needing capacity expansion of Claus unit for many companies around the world. In refineries, the sulphur recovery units take acid gas from amine regeneration units and sour water strippers, converting hydrogen sulfide to elemental sulfur using the Claus process. The Claus process is hydraulically limited by mass flow rate. Reducing the pressure drop across control valves, flow meters, lines, knock-out drums, and packing improves the capacity. Oxygen enrichment helps improve the capacity by removing nitrogen, this is more commonly done on all capacity expansion projects. Typical upgrades required due to oxygen enrichment are new burners, new refractory in thermal reactor, resizing of 1st condenser, instrumentation changes, and steam/condensate heat integration. Some other capacity expansion options typically considered are tail gas compressor, replacing air blower with higher head, hydrocarbon minimization in the feed, water removal, and ammonia removal. Increased capacity related upgrades in sulfur recovery unit also need changes in the tail gas treatment unit, typical changes include improvement to quench tower duty, packing area upgrades in quench and absorber towers and increased amine circulation flow rates.

Keywords: Claus process, oxygen enrichment, sulfur recovery unit, tail gas treatment unit

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528 Effect of Different Levels of Fibrolytic Enzyme on Feed Digestibility and Production Performance in Lactating Dairy Cows

Authors: Hazrat Salman Sidique, Muhammad Tahir Khan, Haq Aman Ullah, Muhammad Mobashar, Muhammad Ishtiaq Sohail Mehmood

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The poor quality conventional feed for the livestock production in Pakistan are wheat straw, tops of sugar cane and tree leaves. To enhance the nutritive value of feed, this study focused on investigating the effects of fibrolytic enzyme (Fibrozyme®, Alltech Inc. Company, USA) at different levels i.e. 0, 5, 10, and 15g/kg of total mix ration on feed intake, digestibility, milk yield and composition, and economics of the ration in Holstein Friesians cows. Twelve Holstein Friesians cows of almost the same age, and lactation stage were randomly allocated into 4 equal groups i.e. A, B, C, and D. Four experimental rations supplemented with Fibrozyme® 0g, 5g, 10g, and 15g/Kg of total mix ration were assigned to these sets correspondingly. The dry matter intake was linearly and significantly (P<0.05) improved. A significant effect of Fibrozyme® was observed for organic matter digestibility, ether extract digestibility, crude fiber digestibility, nitrogen free extract digestibility, and acid detergent fiber digestibility while the results were statistically non-significant for crude protein digestibility, neutral detergent fiber digestibility, and ash digestibility. Milk yield and composition except fat were significantly (P<0.05) increased in all Fibrozyme® treated groups. This study concludes that supplementation of Fibrozyme® at the rate of 15g/Kg total mix ration improved the dry matter intake, nutrients digestibility, and milk production and constituents like protein, lactose, and solid not fat. Therefore, treatment of total mix ration with Fibrozyme® was desirably reasonable and profitable.

Keywords: digestibility, fibrozyme, TMR, digestibility, lactating cow

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527 Urine Neutrophil Gelatinase-Associated Lipocalin as an Early Marker of Acute Kidney Injury in Hematopoietic Stem Cell Transplantation Patients

Authors: Sara Ataei, Maryam Taghizadeh-Ghehi, Amir Sarayani, Asieh Ashouri, Amirhossein Moslehi, Molouk Hadjibabaie, Kheirollah Gholami

Abstract:

Background: Acute kidney injury (AKI) is common in hematopoietic stem cell transplantation (HSCT) patients with an incidence of 21–73%. Prevention and early diagnosis reduces the frequency and severity of this complication. Predictive biomarkers are of major importance to timely diagnosis. Neutrophil gelatinase associated lipocalin (NGAL) is a widely investigated novel biomarker for early diagnosis of AKI. However, no study assessed NGAL for AKI diagnosis in HSCT patients. Methods: We performed further analyses on gathered data from our recent trial to evaluate the performance of urine NGAL (uNGAL) as an indicator of AKI in 72 allogeneic HSCT patients. AKI diagnosis and severity were assessed using Risk–Injury–Failure–Loss–End-stage renal disease and AKI Network criteria. We assessed uNGAL on days -6, -3, +3, +9 and +15. Results: Time-dependent Cox regression analysis revealed a statistically significant relationship between uNGAL and AKI occurrence. (HR=1.04 (1.008-1.07), P=0.01). There was a relation between uNGAL day +9 to baseline ratio and incidence of AKI (unadjusted HR=.1.047(1.012-1.083), P<0.01). The area under the receiver-operating characteristic curve for day +9 to baseline ratio was 0.86 (0.74-0.99, P<0.01) and a cut-off value of 2.62 was 85% sensitive and 83% specific in predicting AKI. Conclusions: Our results indicated that increase in uNGAL augmented the risk of AKI and the changes of day +9 uNGAL concentrations from baseline could be of value for predicting AKI in HSCT patients. Additionally uNGAL changes preceded serum creatinine rises by nearly 2 days.

Keywords: acute kidney injury, hemtopoietic stem cell transplantation, neutrophil gelatinase-associated lipocalin, Receiver-operating characteristic curve

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526 A Handheld Light Meter Device for Methamphetamine Detection in Oral Fluid

Authors: Anindita Sen

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Oral fluid is a promising diagnostic matrix for drugs of abuse compared to urine and serum. Detection of methamphetamine in oral fluid would pave way for the easy evaluation of impairment in drivers during roadside drug testing as well as ensure safe working environments by facilitating evaluation of impairment in employees at workplaces. A membrane-based point-of-care (POC) friendly pre-treatment technique has been developed which aided elimination of interferences caused by salivary proteins and facilitated the demonstration of methamphetamine detection in saliva using a gold nanoparticle based colorimetric aptasensor platform. It was found that the colorimetric response in saliva was always suppressed owing to the matrix effects. By navigating the challenging interfering issues in saliva, we were successfully able to detect methamphetamine at nanomolar levels in saliva offering immense promise for the translation of these platforms for on-site diagnostic systems. This subsequently motivated the development of a handheld portable light meter device that can reliably transduce the aptasensors colorimetric response into absorbance, facilitating quantitative detection of analyte concentrations on-site. This is crucial due to the prevalent unreliability and sensitivity problems of the conventional drug testing kits. The fabricated light meter device response was validated against a standard UV-Vis spectrometer to confirm reliability. The portable and cost-effective handheld detector device features sensitivity comparable to the well-established UV-Vis benchtop instrument and the easy-to-use device could potentially serve as a prototype for a commercial device in the future.

Keywords: aptasensors, colorimetric gold nanoparticle assay, point-of-care, oral fluid

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525 Effect of Segregation on the Reaction Rate of Sewage Sludge Pyrolysis in a Bubbling Fluidized Bed

Authors: A. Soria-Verdugo, A. Morato-Godino, L. M. García-Gutiérrez, N. García-Hernando

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The evolution of the pyrolysis of sewage sludge in a fixed and a fluidized bed was analyzed using a novel measuring technique. This original measuring technique consists of installing the whole reactor over a precision scale, capable of measuring the mass of the complete reactor with enough precision to detect the mass released by the sewage sludge sample during its pyrolysis. The inert conditions required for the pyrolysis process were obtained supplying the bed with a nitrogen flowrate, and the bed temperature was adjusted to either 500 ºC or 600 ºC using a group of three electric resistors. The sewage sludge sample was supplied through the top of the bed in a batch of 10 g. The measurement of the mass released by the sewage sludge sample was employed to determine the evolution of the reaction rate during the pyrolysis, the total amount of volatile matter released, and the pyrolysis time. The pyrolysis tests of sewage sludge in the fluidized bed were conducted using two different bed materials of the same size but different densities: silica sand and sepiolite particles. The higher density of silica sand particles induces a flotsam behavior for the sewage sludge particles which move close to the bed surface. In contrast, the lower density of sepiolite produces a neutrally-buoyant behavior for the sewage sludge particles, which shows a proper circulation throughout the whole bed in this case. The analysis of the evolution of the pyrolysis process in both fluidized beds show that the pyrolysis is faster when buoyancy effects are negligible, i.e. in the bed conformed by sepiolite particles. Moreover, sepiolite was found to show an absorbent capability for the volatile matter released during the pyrolysis of sewage sludge.

Keywords: bubbling fluidized bed, pyrolysis, reaction rate, segregation effects, sewage sludge

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524 A Structure-Switching Electrochemical Aptasensor for Rapid, Reagentless and Single-Step, Nanomolar Detection of C-Reactive Protein

Authors: William L. Whitehouse, Louisa H. Y. Lo, Andrew B. Kinghorn, Simon C. C. Shiu, Julian. A. Tanner

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C-reactive protein (CRP) is an acute-phase reactant and sensitive indicator for sepsis and other life-threatening pathologies, including systemic inflammatory response syndrome (SIRS). Currently, clinical turn-around times for established CRP detection methods take between 30 minutes to hours or even days from centralized laboratories. Here, we report the development of an electrochemical biosensor using redox probe-tagged DNA aptamers functionalized onto cheap, commercially available screen-printed electrodes. Binding-induced conformational switching of the CRP-targeting aptamer induces a specific and selective signal-ON event, which enables single-step and reagentless detection of CRP in as little as 1 minute. The aptasensor dynamic range spans 5-1000nM (R=0.97) or 5-500nM (R=0.99) in 50% diluted human serum, with a LOD of 3nM, corresponding to 2-orders of magnitude sensitivity under the clinically relevant cut-off for CRP. The sensor is stable for up to one week and can be reused numerous times, as judged from repeated real-time dosing and dose-response assays. By decoupling binding events from the signal induction mechanism, structure-switching electrochemical aptamer-based sensors (SS-EABs) provide considerable advantages over their adsorption-based counterparts. Our work expands on the retinue of such sensors reported in the literature and is the first instance of an SS-EAB for reagentless CRP detection. We hope this study can inspire further investigations into the suitability of SS-EABs for diagnostics, which will aid translational R&D toward fully realized devices aimed at point-of-care applications or for use more broadly by the public.

Keywords: structure-switching, C-reactive protein, electrochemical, biosensor, aptasensor.

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523 Properties Optimization of Keratin Films Produced by Film Casting and Compression Moulding

Authors: Mahamad Yousif, Eoin Cunningham, Beatrice Smyth

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Every year ~6 million tonnes of feathers are produced globally. Due to feathers’ low density and possible contamination with pathogens, their disposal causes health and environmental problems. The extraction of keratin, which represents >90% of feathers’ dry weight, could offer a solution due to its wide range of applications in the food, medical, cosmetics, and biopolymer industries. One of these applications is the production of biofilms which can be used for packaging, edible films, drug delivery, wound healing etc. Several studies in the last two decades investigated keratin film production and its properties. However, the effects of many parameters on the properties of the films remain to be investigated including the extraction method, crosslinker type and concentration, and the film production method. These parameters were investigated in this study. Keratin was extracted from chicken feathers using two methods, alkaline extraction with 0.5 M NaOH at 80 °C or sulphitolysis extraction with 0.5 M sodium sulphite, 8 M urea, and 0.25-1 g sodium dodecyl sulphate (SDS) at 100 °C. The extracted keratin was mixed with different types and concentrations of plasticizers (glycerol and polyethylene glycol) and crosslinkers (formaldehyde (FA), glutaraldehyde, cinnamaldehyde, glyoxal, and 1,4-Butanediol diglycidyl ether (BDE)). The mixtures were either cast in a mould or compression moulded to produce films. For casting, keratin powder was initially dissolved in water to form a 5% keratin solution and the mixture was dried in an oven at 60 °C. For compression moulding, 10% water was added and the compression moulding temperature and pressure were in the range of 60-120 °C and 10-30 bar. Finally, the tensile properties, solubility, and transparency of the films were analysed. The films prepared using the sulphitolysis keratin had superior tensile properties to the alkaline keratin and formed successfully with lower plasticizer concentrations. Lowering the SDS concentration from 1 to 0.25 g/g feathers improved all the tensile properties. All the films prepared without crosslinkers were 100% water soluble but adding crosslinkers reduced solubility to as low as 21%. FA and BDE were found to be the best crosslinkers increasing the tensile strength and elongation at break of the films. Higher compression moulding temperature and pressure lowered the tensile properties of the films; therefore, 80 °C and 10 bar were considered to be the optimal compression moulding temperature and pressure. Nevertheless, the films prepared by casting had higher tensile properties than compression moulding but were less transparent. Two optimal films, prepared by film casting, were identified and their compositions were: (a) Sulphitolysis keratin, 20% glycerol, 10% FA, and 10% BDE. (b) Sulphitolysis keratin, 20% glycerol, and 10% BDE. Their tensile strength, elongation at break, Young’s modulus, solubility, and transparency were: (a) 4.275±0.467 MPa, 86.12±4.24%, 22.227±2.711 MPa, 21.34±1.11%, and 8.57±0.94* respectively. (b) 3.024±0.231 MPa, 113.65±14.61%, 10±1.948 MPa, 25.03±5.3%, and 4.8±0.15 respectively. A higher value indicates that the film is less transparent. The extraction method, film composition, and production method had significant influence on the properties of keratin films and should therefore be tailored to meet the desired properties and applications.

Keywords: compression moulding, crosslinker, film casting, keratin, plasticizer, solubility, tensile properties, transparency

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