Search results for: chemiluminescence

Authors: Igor Izosimov

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This work is devoted to applications of the Time-resolved laser-induced luminescence (TRLIF) spectroscopy and time-resolved laser-induced chemiluminescence spectroscopy for detection of lanthanides and actinides. Results of the experiments on Eu, Sm, U, and Pu detection in solutions are presented. The limit of uranyl detection (LOD) in urine in our TRLIF experiments was up to 5 pg/ml. In blood plasma LOD was 0.1 ng/ml and after mineralization was up to 8pg/ml – 10pg/ml. In pure solution, the limit of detection of europium was 0.005ng/ml and samarium, 0.07ng/ml. After addition urine, the limit of detection of europium was 0.015 ng/ml and samarium, 0.2 ng/ml. Pu, Np, and some U compounds do not produce direct luminescence in solutions, but when excited by laser radiation, they can induce chemiluminescence of some chemiluminogen (luminol in our experiments). It is shown that multi-photon scheme of chemiluminescence excitation makes chemiluminescence not only a highly sensitive but also a highly selective tool for the detection of lanthanides/actinides in solutions.

Keywords: actinides/lanthanides detection, laser spectroscopy with time resolution, luminescence/chemiluminescence, solutions

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Authors: Sonu Gandhi, Neena Capalash, Prince Sharma, C. Raman Suri

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A sensitive chemiluminescence immunoassay (CIA) for heroin and its major metabolites is reported. The method is based on the competitive reaction of horseradish peroxidase (HRP)-labeled anti-MAM antibody and free drug in spiked urine samples. A hapten-protein conjugate was synthesized by using acidic derivative of monoacetyl morphine (MAM) coupled to carrier protein BSA and was used as an immunogen for the generation of anti-MAM (monoacetyl morphine) antibody. A high titer of antibody (1:64,0000) was obtained and the relative affinity constant (Kaff) of antibody was 3.1×107 l/mol. Under the optimal conditions, linear range and reactivity for heroin, mono acetyl morphine (MAM), morphine and codeine were 0.08, 0.09, 0.095 and 0.092 ng/mL respectively. The developed chemiluminescence inhibition assay could detect heroin and its metabolites in standard and urine samples up to 0.01 ng/ml.

Keywords: heroin, metabolites, chemiluminescence immunoassay, horse radish peroxidase

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Authors: Sewon Kim, Chang Yeop Lee, Minjun Kwon

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This study is aiming at establishing the relationship between the optical signal of flame and an equivalent ratio of flame. In this experiment, flame optical signal in a furnace is measured using photodiode. The combustion system which is composed of metal fiber burner and vertical furnace and flame chemiluminescence is measured at various experimental conditions. In this study, the flame chemiluminescence of laminar premixed flame is measured by using commercially available photodiode. It is experimentally investigated the relationship between equivalent ratio and photodiode signal. In addition, The strategy of combustion control method is proposed by using the optical signal and fuel pressure. The results showed that certain relationship between optical data of photodiode and equivalence ratio exists and this leads to the successful application of this system for instantaneous measurement of equivalence ration of the combustion system.

Keywords: flame chemiluminescence, photo diode, equivalence ratio, combustion control

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Authors: Nasiruddin Khan

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The action of (-)-epicatechin, a cocoa (Theobroma cacao) flavanol that modulates redox/oxidative stress are contributed mainly to their antioxidant properties. The present study investigates the concentration and time dependent effect of (-)-epicatechin metabolites 3MeEc, 4MeEc, and 4SulEc on the production of ROS on BAEC using L-012, Lucigenin as chemiluminescence dye and XO/HX system. Our result demonstrates that 3MeEc shows significant (P <0.05) lowering effect of ROS production in BAEC with increasing concentration of metabolite while L-012 was used as chemiluminescence dye but not in the case of Lucigenin. In XO/HX system, using L-012 as chemiluminescence dye, 3MeEc and 4MeEc showed significant lowering effect on ROS production with increasing concentration from 100-500nM as compared to the positive control (SOD). When Lucigenin was used as chemiluminescence dye, 3MeEc exerted significant lowering effect with increasing concentration when compared to the positive control (SOD) whereas 4MeEc showed significant lowering effect in ROS production from 250 nM on as compared to positive control. For 4SulEc, a significant lowering effect of ROS production was only observed at 100 and 250 nM. Overall, although each metabolite shows considerable effect, 3MeEc exhibited more pronounced effect on decreasing the production of ROS as compared to other two metabolites.

Keywords: epicatechin metabolites, HO-1, Nrf2, ROS

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Authors: Chinh Nguyen Nhu Bao, Hien To Thi, Norimichi Takenaka

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Formaldehyde (HCHO) concentrations in rainwater including in tropical storms in Sakai City, Osaka, Japan have been measured continuously during rain event by developed chemiluminescence method. The level of formaldehyde was ranged from 15 µg/L to 500 µg/L. The high concentration of HCHO in rainwater is related to the wind direction from the south and west sides of Sakai City where manufactures related to chemicals, oil-refinery, and steel. The in-situ irradiated experiment on rainwater sample was conducted to prove the aqueous phase photo-production of HCHO and the degradation of HCHO. In the daytime, the aqueous phase photolysis is the source of HCHO in rainwater (4.52 ± 5.74 µg/L/h for UV light source in-situ condition, 2.84-8.96 µg/L/h under sunlight). However, in the night time, the degradation is the function of microorganism.

Keywords: chemiluminescence, formaldehyde, rainwater, typhoon

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Authors: Minh-Phuong Ngoc Bui, Abdennour Abbas

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Microbial spoilage of food/drug has been a constant nuisance and an unavoidable problem throughout history that affects food/drug quality and safety in a variety of ways. A simple and rapid test of fungi and bacteria in food/drugs and environmental clinical samples is essential for proper management of contamination. A number of different techniques have been developed for detection and enumeration of foodborne microorganism including plate counting, enzyme-linked immunosorbent assay (ELISA), polymer chain reaction (PCR), nucleic acid sensor, electrical and microscopy methods. However, the significant drawbacks of these techniques are highly demand of operation skills and the time and cost involved. In this report, we introduce a rapid method for detection of bacteria and fungi in food/drug products using a specific interaction between a reducing agent (tris(2-carboxylethyl)phosphine (TCEP)) and the microbial surface proteins. The chemical reaction was transferred to a transduction system using gold nanoplates-enhanced chemiluminescence. We have optimized our nanoplates synthetic conditions, characterized the chemiluminescence parameters and optimized conditions for the microbial assay. The new detection method was applied for rapid detection of bacteria (E.coli sp. and Lactobacillus sp.) and fungi (Mucor sp.), with limit of detection as low as single digit cells per mL within 10 min using a portable luminometer. We expect our simple and rapid detection method to be a powerful alternative to the conventional plate counting and immunoassay methods for rapid screening of microorganisms in food/drug products.

Keywords: microorganism testing, gold nanoplates, chemiluminescence, reducing agents, luminol

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Authors: Mohammad Jamil Abd AlGhani

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The natural native antioxidants N,N-P-methyl phenyl acetone and N,N-phenyl acetone were isolated from the Iraqi crude oil region of Kirkuk by ion exchange and their structure was characterized by spectral and chemical analysis methods. Tetraline was used as a liquid hydrocarbon to detect the efficiency of isolated molecules at elevated temperature (393 K) that it has physicochemical specifications and structure closed to hydrocarbons fractionated from crude oil. The synthesized universal antioxidant 2,6-ditertiaryisobutyl-p-methyl phenol (Unol) with known stochiometric coefficient of inhibition equal to (2) was used as a model for comparative evaluation at the same conditions. Modified chemiluminescence method was used to find the amount of absorbed oxygen and the induction periods in and without the existence of isolated antioxidants molecules. The results of induction periods and quantity of absorbed oxygen during the oxidation process were measured by manometric installation. It was seen that at specific equal concentrations of N,N-phenyl acetone and N, N-P-methyl phenyl acetone in comparison with Unol at 393 K were with (2) and (2.5) times efficient than do Unol. It means that they had the ability to inhibit the formation of new free radicals and prevent the chain reaction to pass from the propagation to the termination step rather than decomposition of formed hydroperoxides.

Keywords: antioxidants, chemiluminescence, inhibition, Unol

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Authors: Afsal Mohammed Kadavilpparampu, Haider A. J. Al Lawati, Fakhr Eldin O. Suliman, Salma M. Z. Al Kindy

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In this work, we evaluated the appropriateness of various oxidants that can be used potentially with Ru(bipy)32+ CL system while performing CL detection in a microfluidic device using eight common active pharmaceutical ingredients- ciprofloxacin, hydrochlorothiazide, norfloxacin, buspirone, fexofenadine, cetirizine, codeine, and dextromethorphan. This is because, microfludics have very small channel volume and the residence time is also very short. Hence, a highly efficient oxidant is required for on-chip CL detection to obtain analytically acceptable CL emission. Three common oxidants were evaluated, lead dioxide, cerium ammonium sulphate and ammonium peroxydisulphate. Results obtained showed that ammonium peroxydisulphate is the most appropriate oxidant which can be used in microfluidic setup and all the tested analyte give strong CL emission while using this oxidant. We also found that Ru(bipy)33+ generated off-line by oxidizing [Ru(bipy)3]Cl2.6H2O in acetonitrile under acidic condition with lead dioxide was stable for more than 72 hrs. A highly sensitive microbore HPLC- CL method using ammonium peroxydisulphate as an oxidant in a microfluidic on-chip CL detection has been developed for the analyses of fixed-dose combinations of pseudoephedrine (PSE), fexofenadine (FEX) and cetirizine (CIT) in biological fluids and pharmaceutical formulations with minimum sample pre-treatment.

Keywords: oxidants, microbore High Performance Liquid Chromatography, chemiluminescence, microfluidics

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Authors: Vishnu Raj, Chockalingam Prathap

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The objective of this study was to explore the impact of variation in the exit annular area on the local flow field features and the flame stability of an annular premixed swirl burner (unconfined) operated with premixed n-butane air mixture at equivalence ratio (ϕ) = 1, 1 bar, and 300K. A swirl burner with an axial swirl generator having a swirl number of 1.5 was used. Three different burner heads were chosen to have the exit area increased from 100%, 160%, and 220% resulting in inner and outer diameters and cross-sectional areas as (1) 10mm&15mm, 98mm2 (2) 17.5mm&22.5mm, 157mm2 and (3) 25mm & 30mm, 216mm2. The bulk velocity and Reynolds number based on the hydraulic diameter and unburned gas properties were kept constant at 12 m/s and 4000. (i) Planar PIV with TiO2 seeding particles and (ii) OH* chemiluminescence were used to measure the velocity fields and reaction zones of the swirl flames at 5Hz, respectively. Velocity fields and the jet spreading rates measured at the isothermal and reactive conditions revealed that the presence of a flame significantly altered the flow field in the radial direction due to the gas expansion. Important observations from the flame measurements were: the height and maximum width of the recirculation bubbles normalized by the hydraulic diameter, and the jet spreading angles for the flames for the three exit area cases were: (a) 4.52, 1.95, 28ᵒ, (b) 6.78, 2.37, 34ᵒ, and (c) 8.73, 2.32, 37ᵒ. The lean blowout was also measured, and the respective equivalence ratios were: 0.80, 0.92, and 0.82. LBO was relatively narrow for the 157mm2 case. For this case, particle image velocimetry (PIV) measurements showed that Turbulent Kinetic Energy and turbulent intensity were relatively high compared to the other two cases, resulting in higher stretch rates and narrower lean blowout (LBO).

Keywords: chemiluminescence, jet spreading rate, lean blowout, swirl flow

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Authors: Benghezel Hichem

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Thyroid dysfunction in hemodialysis subjects is represented mainly by hypothyroidism. The objective of our work is to determine the thyroid profile of our hemodialysis patients and to highlight the prevalence of different thyroid disorders. Methods: This is a retrospective study performed on a mono centric 2 months (February and March 2013) on 42 hemodialysis patients (11 male and 31 female). We made the dosage of thyroid hormones Thyrotropin (TSH) ((free thyroxin ) FT4 and free Triodothyronin ) FT3) by chemiluminescence immunoassay method on cobas 6000 Roche Diagnostics. The results: The prevalence of biological hypothyroidism was 18% (7% with a high TSH isolated and a mean +/- SD 9.44 +/- 6.29, 5% with high TSH, and with low FT4 a mean +/- SD is 8.18 +/- 0.53 for TSH and 9.69 +/- 0.22 for FT4, One patient with a high TSH, and low FT4, FT3. 4% of patients with a low T3 syndrome with a mean +/- SD of 3.93 +/- 0,3 for FT3), we notice that 5% of patients with hyperthyroidism TSH collapsed and mean +/- SD of TSH is 0.017 +/- 0,001. Conclusion: The biological Hypothyroidism is a common endocrine disorder in chronic hemodialysis.

Keywords: hypothyroidism, hemodialysis, thyréostimulin, free thyroxin, triodothyronin

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Authors: R. I. You, C. L. Ho, M. S. Dai, H. M. Hung, S. F. Yen, C. S. Chen, T. Y. Chao

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Neutrophils are the most abundant innate immune cells to against invading microorganisms. Numerous data shown neutrophils have plasticity in response to physiological and pathological conditions. Tumor-associated neutrophils (TAN) exist in distinct types of tumor and play an important role in cancer biology. Different transcriptomic profiles of neutrophils in tumor and non-tumor samples have been identified. Several miRNAs have been recognized as regulators of gene expression in neutrophil, which may have key roles in neutrophil activation. However, the miRNAs expression patterns in TAN are not well known. To address this question, magnetic bead isolated neutrophils from tumor-bearing mice were used in this study. We analyzed production of reactive oxygen species (ROS) by luminol-dependent chemiluminescence assay. The expression of miRNAs targeting NADPH oxidase, ROS generation and autophagy was explored using quantitative real-time polymerase chain reaction. Our data suggest that tumor environment influence neutrophil develop to differential states of activation via miRNAs regulation.

Keywords: tumor-associated neutrophil, miRNAs, neutrophil, ROS

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Authors: Sam Bartlett, Thomas Bacquart, Arul Murugan, Abigail Morris

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Fuel cell electric vehicles provide the potential to decarbonise road transport, create new economic opportunities, diversify national energy supply, and significantly reduce the environmental impacts of road transport. A potential issue, however, is that the catalyst used at the fuel cell cathode is susceptible to degradation by impurities, especially sulphur-containing compounds. A recent European Directive (2014/94/EU) stipulates that, from November 2017, all hydrogen provided to fuel cell vehicles in Europe must comply with the hydrogen purity specifications listed in ISO 14687-2; this includes reactive and toxic chemicals such as ammonia and total sulphur-containing compounds. This requirement poses great analytical challenges due to the instability of some of these compounds in calibration gas standards at relatively low amount fractions and the difficulty associated with undertaking measurements of groups of compounds rather than individual compounds. Without the available reference materials and analytical infrastructure, hydrogen refuelling stations will not be able to demonstrate compliance to the ISO 14687 specifications. The hydrogen purity laboratory at NPL provides world leading, accredited purity measurements to allow hydrogen refuelling stations to evidence compliance to ISO 14687. Utilising state-of-the-art methods that have been developed by NPL’s hydrogen purity laboratory, including a novel method for measuring total sulphur compounds at 4 nmol/mol and a hydrogen impurity enrichment device, we provide the capabilities necessary to achieve these goals. An overview of these capabilities will be given in this paper. As part of the EMPIR Hydrogen co-normative project ‘Metrology for sustainable hydrogen energy applications’, NPL are developing a validated analytical methodology for the measurement of speciated sulphur-containing compounds in hydrogen at low amount fractions pmol/mol to nmol/mol) to allow identification and measurement of individual sulphur-containing impurities in real samples of hydrogen (opposed to a ‘total sulphur’ measurement). This is achieved by producing a suite of stable gravimetrically-prepared primary reference gas standards containing low amount fractions of sulphur-containing compounds (hydrogen sulphide, carbonyl sulphide, carbon disulphide, 2-methyl-2-propanethiol and tetrahydrothiophene have been selected for use in this study) to be used in conjunction with novel dynamic dilution facilities to enable generation of pmol/mol to nmol/mol level gas mixtures (a dynamic method is required as compounds at these levels would be unstable in gas cylinder mixtures). Method development and optimisation are performed using gas chromatographic techniques assisted by cryo-trapping technologies and coupled with sulphur chemiluminescence detection to allow improved qualitative and quantitative analyses of sulphur-containing impurities in hydrogen. The paper will review the state-of-the art gas standard preparation techniques, including the use and testing of dynamic dilution technologies for reactive chemical components in hydrogen. Method development will also be presented highlighting the advances in the measurement of speciated sulphur compounds in hydrogen at low amount fractions.

Keywords: gas chromatography, hydrogen purity, ISO 14687, sulphur chemiluminescence detector

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Authors: Andreas Latz, Daniela Heinz, Fatima Hashemi, Melek Baygül

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Introduction: Feline leukemia virus (FeLV), feline immunodeficiency virus (FIV), and feline coronavirus (FCoV) are serious infectious diseases affecting cats worldwide. Transmission of these viruses occurs primarily through close contact with infected cats (via saliva, nasal secretions, faeces, etc.). FeLV, FIV, and FCoV infections can occur in combination and are expressed in similar clinical symptoms. Diagnosis can therefore be challenging: Symptoms are variable and often non-specific. Sick cats show very similar clinical symptoms: apathy, anorexia, fever, immunodeficiency syndrome, anemia, etc. Sample volume for small companion animals for diagnostic purposes can be challenging to collect. In addition, multiplex diagnosis of diseases can contribute to an easier, cheaper, and faster workflow in the lab as well as to the better differential diagnosis of diseases. For this reason, we wanted to develop a new diagnostic tool that utilizes less sample volume, reagents, and consumables than multiplesingleplex ELISA assays Methods: The Multiplier from Dynextechonogies (USA) has been used as platform to develop a Multiplex diagnostic tool for the detection of antibodies against FIV and FCoV/FIP and antigens for FeLV. Multiplex diagnostics. The Dynex®Multiplier®is a fully automated chemiluminescence immunoassay analyzer that significantly simplifies laboratory workflow. The Multiplier®ease-of-use reduces pre-analytical steps by combining the power of efficiently multiplexing multiple assays with the simplicity of automated microplate processing. Plastic beads have been coated with antigens for FIV and FCoV/FIP, as well as antibodies for FeLV. Feline blood samples are incubated with the beads. Read out of results is performed via chemiluminescence Results: Bead coating was optimized for each individual antigen or capture antibody and then combined in the multiplex diagnostic tool. HRP: Antibody conjugates for FIV and FCoV antibodies, as well as detection antibodies for FeLV antigen, have been adjusted and mixed. 3 individual prototyple batches of the assay have been produced. We analyzed for each disease 50 well defined positive and negative samples. Results show an excellent diagnostic performance of the simultaneous detection of antibodies or antigens against these feline diseases in a fully automated system. A 100% concordance with singleplex methods like ELISA or IFA can be observed. Intra- and Inter-Assays showed a high precision of the test with CV values below 10% for each individual bead. Accelerated stability testing indicate a shelf life of at least 1 year. Conclusion: The new tool can be used for multiplex diagnostics of the most important feline infectious diseases. Only a very small sample volume is required. Fully automation results in a very convenient and fast method for diagnosing animal diseases.With its large specimen capacity to process over 576 samples per 8-hours shift and provide up to 3,456 results, very high laboratory productivity and reagent savings can be achieved.

Keywords: Multiplex, FIV, FeLV, FCoV, FIP

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Authors: Wooseok Song, Sunjung Park, Jongkwon Lee, Jaye Koo

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The objective of this paper is to observe the effects of injection conditions on flame structures in gas-centered swirl coaxial injector. Gaseous oxygen and liquid kerosene were used as propellants. For different injection conditions, two types of injector, which only differ in the diameter of the tangential inlet, were used in this study. In addition, oxidizer injection pressure was varied to control the combustion chamber pressure in different types of injector. In order to analyze the combustion instability intensity, the dynamic pressure was measured in both the combustion chamber and propellants lines. With the increase in differential pressure between the propellant injection pressure and the combustion chamber pressure, the combustion instability intensity increased. In addition, the flame structure was recorded using a high-speed camera to detect CH* chemiluminescence intensity. With the change in the injection conditions in the gas-centered swirl coaxial injector, the flame structure changed.

Keywords: liquid rocket engine, flame structure, combustion instability, dynamic pressure

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Authors: Toshiyuki Sato, Kenta Iitani, Koji Toma, Takahiro Arakawa, Kohji Mitsubayashi

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A 2-dimensional imaging system (Sniff-camera) for gaseous ethanol emissions from a human palm skin was constructed and demonstrated. This imaging system measures gaseous ethanol concentrations as intensities of chemiluminescence (CL) by luminol reaction induced by alcohol oxidase and luminol-hydrogen peroxide system. A conversion of ethanol distributions and concentrations to 2-dimensional CL was conducted on an enzyme-immobilized mesh substrate in a dark box, which contained a luminol solution. In order to visualize ethanol emissions from human palm skin, we developed highly sensitive and selective imaging system for transpired gaseous ethanol at sub ppm-levels. High sensitivity imaging allows us to successfully visualize the emissions dynamics of transdermal gaseous ethanol. The intensity of each pixel on the palm shows the reflection of ethanol concentrations distributions based on the metabolism of oral alcohol administration. This imaging system is significant and useful for the assessment of ethanol measurement of the palmar skin.

Keywords: sniff-camera, gas-imaging, ethanol vapor, human body gas

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Authors: S. A. Khan, B. Mishra, O. Sherchan

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Background and Aims: Thyroid disorder is the most common endocrine disorder after diabetes mellitus. Females are more vulnerable to this disease, and old age is an important risk factor. This study was undertaken to investigate the burden of thyroid disorder in Nepalese postmenopausal women. Methods: In the present cross-sectional study, we included 271 post-menopausal women. Three ml of blood was collected following standard protocol after taking the written consent. Serum was separated and analyzed for free T3, free T4, and Thyroid Stimulating Hormone (TSH) by Chemiluminescence Immunoassay (CLIA) method in Snibe Maglumi 1000 analyzer. Data obtained was analyzed in SPSS Version 21. P < 0.05 was set for statistical significant at 95% Confidence Interval (CI). Results: Majority of the participants belong to Janjati (46.5%) ethnicity, followed by Brahmin/Chhetri (41.7%), residing either in urban or suburban locality. Most of them were non-vegetarian, non-smoker, and non-alcoholic. Subjects were divided into hyperthyroid (TSH < 0.3 uIU/ml), hypothyroid (TSH > 4.5 uIU/ml), and euthyroid (TSH=0.3-4.5 uIU/ml) based on TSH value. We reported 10.3% hyperthyroid and 29.2% hypothyroid cases. TSH was significantly correlated with T3 (r=-0.244; p < 0.001) T4 (r=-0.398; p < 0.001); age (r=-0.138; p=0.023) and BMI (r=0.123; p=0.043). Multiple linear regression model for TSH reveals only T3 and T4 were significantly associated with TSH (p < 0.001; p=0.001). Conclusion: To conclude, nearly 39.5% of the postmenopausal women had thyroid disorder. Postmenopausal women are vulnerable to thyroid disorder; therefore, requires regular thyroid monitoring.

Keywords: thyroid stimulating hormone, TSH, T3, T4, thyroid disorder

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Authors: Eva Tvrdá, Hana Greifová, Peter Ivanič, Norbert Lukáč

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The aim of this study was to evaluate the dose- and time-dependent in vitro effects of berberine (BER), a natural alkaloid with numerous biological properties on bovine spermatozoa during three time periods (0 h, 2 h, 24 h). Bovine semen samples were diluted and cultivated in physiological saline solution containing 0.5% DMSO together with 200, 100, 50, 10, 5, and 1 μmol/L BER. Spermatozoa motility was assessed using the computer assisted semen analyzer. The viability of spermatozoa was assessed by the metabolic (MTT) assay, production of superoxide radicals was quantified using the nitroblue tetrazolium (NBT) test, and chemiluminescence was used to evaluate the generation of reactive oxygen species (ROS). Cell lysates were prepared and the extent of lipid peroxidation (LPO) was evaluated using the TBARS assay. The results of the movement activity showed a significant increase in the motility during long term cultivation in case of concentrations ranging between 1 and 10 μmol/L BER (P < 0.01; P < 0.001; 24 h). At the same time, supplementation of 1, 5 and 10 μmol/L BER led to a significant preservation of the cell viability (P < 0.001; 24 h). BER addition at a range of 1-50 μmol/L also provided a significantly higher protection against superoxide (P < 0.05) and ROS (P < 0.001; P < 0.01) overgeneration as well as LPO (P < 0.01; P<0.05) after a 24 h cultivation. We may suggest that supplementation of BER to bovine spermatozoa, particularly at concentrations ranging between 1 and 50 μmol/L, may offer protection to the motility, viability and oxidative status of the spermatozoa, particularly notable at 24 h.

Keywords: berberine, bulls, motility, oxidative profile, spermatozoa, viability

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Authors: Niki K. Burns, James R. Pearson, Paul G. Stevenson, Xavier A. Conlan

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In Australian state Victoria, synthetic cannabinoids have recently been made illegal under an amendment to the drugs, poisons and controlled substances act 1981. Identification of synthetic cannabinoids in popular brands of ‘incense’ and ‘potpourri’ has been a difficult and challenging task due to the sample complexity and changes observed in the chemical composition of the cannabinoids of interest. This study has developed analytical methodology for the targeted extraction and determination of synthetic cannabinoids available pre-ban. A simple solvent extraction and solid phase extraction methodology was developed that selectively extracted the cannabinoid of interest. High performance liquid chromatography coupled with UV‐visible and chemiluminescence detection (acidic potassium permanganate and tris (2,2‐bipyridine) ruthenium(III)) were used to interrogate the synthetic cannabinoid products. Mass spectrometry and nuclear magnetic resonance spectroscopy were used for structural elucidation of the synthetic cannabinoids. The tris(2,2‐bipyridine)ruthenium(III) detection was found to offer better sensitivity than the permanganate based reagents. In twelve different brands of herbal incense, cannabinoids were extracted and identified including UR‐144, XLR 11, AM2201, 5‐F‐AKB48 and A796‐260.

Keywords: electrospray mass spectrometry, high performance liquid chromatography, solid phase extraction, synthetic cannabinoids

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Authors: Stela Papa, Klementina Puto, Migena Pllaha

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

Keywords: CLIA, ELISA, Hepatitis C virus, immunoassay

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Authors: Harumi Katayama, Mina Suzuki, Taeko Muramatsu, Yui Shimogawa, Yoshimi Mizushima, Mitsuo Hiramatsu, Kimitsugu Nakamura, Takeshi Suzue

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Purpose: There are very often footbaths in the hot-spring area as culture from old days in Japan. This culture moderately supported mental and physical health among people. In Japanese hospitals, nurses provide footbath for severe patients to mental comfortable. However, there are only a few evidences effect of footbath for mental comfortable. In this presentation, we show the effect of stress relief of the footbath using biomarker among 35 college students in volunteer. Methods: The experiment was designed in two groups of the footbath group and the simple relaxation group randomly. As mental load, Kraepelin test was given to the students beforehand. Ultra-weak chemiluminescence (UCL) in saliva and self-administered liner scale measurable emotional state were measured on four times concurrently; there is before and after the mental load, after the stress relief, and 30 minutes after the stress relief. The scale that measured emotional state was consisted of 7 factors; there is excitement, relaxation, vigorous, fatigue, tension, calm, and sleepiness with 22 items. ANOVA was calculated effect of the footbath for stress relief. Results: The level of UCL (photons/100sec) was significantly increased in response on both groups after mental load. After the two types of stress relief, UCL (photons/100sec) of footbath group was significantly decreased compared to simple relaxation group. Score of sleepiness and relaxation were significantly increased after the stress relief in the footbath group than the simple relaxation group. However, score of excitement, vigorous, tension, and calm were exhibit the same degree of decrease after the stress relief on both group. Conclusion: It was suggested that salivary UCL may be a sensitive biomarker for mild stress relief as nursing care. In the future, we will measure using UCL to evaluate as stress relief for inpatients, outpatients, or general public as the subjects.

Keywords: bio-marker, footbath, Japan, stress relief

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Authors: Amar Ranjan, Julieana Durai, Pranay Tanwar

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Background &Introduction: Ovarian cancer is one of the most common malignant tumor in the female. 70% of the cases of ovarian cancer are diagnosed at an advanced stage. The five-year survival rate associated with ovarian cancer is less than 30%. The early diagnosis of ovarian cancer becomes a key factor in improving the survival rate of patients. Presently, CAl25 (carbohydrate antigen125) is used for the diagnosis and therapeutic monitoring of ovarian cancer, but its sensitivity and specificity is not ideal. The introduction of HE4, human epididymis protein 4 has attracted much attention. HE4 has a sensitivity and specificity of 72.9% and 95% for differentiating between benign and malignant adnexal masses, which is better than CA125 detection.  Methods: Serum HE4 and CA -125 were estimated using the chemiluminescence method. Our cases were 40 epithelial ovarian cancer, 9 benign ovarian tumor, 29 benign gynaecological diseases and 13 healthy individuals. This group include healthy woman those who have undergoing family planning and menopause-related medical consultations and they are negative for ovarian mass. Optimal cut off values for HE4 and CA125 were 55.89pmol/L and 40.25U/L respectively (determined by statistical analysis). Results: The level of HE4 was raised in all ovarian cancer patients (n=40) whereas CA125 levels were normal in 6/40 ovarian cancer patients, which were the cases of OC confirmed by histopathology. There is a significant decrease in the level of HE4 with comparison to CA125 in benign ovarian tumor cases. Both the levels of HE4 and CA125 were raised in the nonovarian cancer group, which includes cancer of endometrium and cervix. In the healthy group, HE4 was normal in all patients except in one case of the rudimentary horn, and the reason for this raised HE4 level is due to the incomplete development of uterus whereas CA125 was raised in 3 cases. Conclusions: Findings showed that the serum level of HE4 is an important indicator in the diagnosis of ovarian cancer, and it also distinguishes between benign and malignant pelvic masses. However, a combination of HE4 and CA125 panel will be extremely valuable in improving the diagnostic efficiency of ovarian cancer. These findings of our study need to be validated in the larger cohort of patients.

Keywords: human epididymis protein 4, ovarian cancer, diagnosis, benign lesions

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Authors: Ambrish Tiwari, Sudhasini Panda, Archana Singh, Kalpana Luthra, S. K. Sharma

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Introduction: On the basis of available literature we know that various host factors play a role in outcome of Tuberculosis (TB) infection by modulating innate immunity. One such factor is Inducible Nitric Oxide Synthase enzyme (iNOS) which help in the production of Nitric Oxide (NO), an antimicrobial agent. Expression of iNOS is in control of various host factors in which Vitamin D along with its nuclear receptor Vitamin D receptor (VDR) is one of them. Vitamin D along with its receptor also produces cathelicidin (antimicrobicidal agent). With this background, we attempted to investigate the levels of Vitamin D and NO along with their associated molecules in tuberculosis patients and household contacts as compared to healthy controls and assess the implication of these findings in susceptibility to tuberculosis (TB). Study subjects and methods: 100 active TB patients, 75 household contacts, and 70 healthy controls were taken. VDR and iNOS mRNA levels were studied using real-time PCR. Serum VDR, cathelicidin, iNOS levels were measured using ELISA. Serum Vitamin D levels were measured in serum samples using chemiluminescence based immunoassay. NO was measured using colorimetry based kit. Results: VDR and iNOS mRNA levels were found to be lower in active TB group compared to household contacts and healthy controls (P=0.0001 and 0.005 respectively). The serum levels of Vitamin D were also found to be lower in active TB group as compared to healthy controls (P =0.001). Levels of cathelicidin and NO was higher in patient group as compared to other groups (p=0.01 and 0.5 respectively). However, the expression of VDR and iNOS and levels of vitamin D was significantly (P < 0.05) higher in household contacts compared to both active TB and healthy control groups. Inference: Higher levels of Vitamin D along with VDR and iNOS expression in household contacts as compared to patients suggest that vitamin D might have a protective role against TB which prevents activation of the disease. From our data, we can conclude that decreased vitamin D levels could be implicated in disease progression and we can use cathelicidin and NO as a biomarker for early diagnosis of pulmonary tuberculosis.

Keywords: vitamin D, VDR, iNOS, tuberculosis

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Authors: Junaid Mahmood Alam

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Revalidating and harmonizing clinical chemistry analytical principles and optimizing methods through quality control programs and assessments is the preeminent means to attain optimal outcome within the clinical laboratory services. Present study reports revalidation of our existing IFCC regularized analytical methods, particularly hepatic and thyroid function tests, by optimization of precision analyses and processing through external and internal quality assessments and regression determination. Parametric components of hepatic (Bilirubin ALT, γGT, ALP), cardiac (LDH, AST, Trop I) and thyroid/pituitary (T3, T4, TSH, FT3, FT4) function tests were used to validate analytical techniques on automated chemistry and immunological analyzers namely Hitachi 912, Cobas 6000 e601, Cobas c501, Cobas e411 with UV kinetic, colorimetric dry chemistry principles and Electro-Chemiluminescence immunoassay (ECLi) techniques. Process of validation and revalidation was completed with evaluating and assessing the precision analyzed Preci-control data of various instruments plotting against each other with regression analyses R2. Results showed that: Revalidation and optimization of respective parameters that were accredited through CAP, CLSI and NEQAPP assessments depicted 99.0% to 99.8% optimization, in addition to the methodology and instruments used for analyses. Regression R2 analysis of BilT was 0.996, whereas that of ALT, ALP, γGT, LDH, AST, Trop I, T3, T4, TSH, FT3, and FT4 exhibited R2 0.998, 0.997, 0.993, 0.967, 0.970, 0.980, 0.976, 0.996, 0.997, 0.997, and R2 0.990, respectively. This confirmed marked harmonization of analytical methods and instrumentations thus revalidating optimized precision standardization as per IFCC recommended guidelines. It is concluded that practices of revalidating and harmonizing the existing or any new services should be followed by all clinical laboratories, especially those associated with tertiary care hospital. This is will ensure deliverance of standardized, proficiency tested, optimized services for prompt and better patient care that will guarantee maximum patients’ confidence.

Keywords: revalidation, standardized, IFCC, CAP, harmonized

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Authors: Bela Siska Afrida, Wisnu Barlianto, Desy Wulandari, Ery Olivianto

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Background: Allergic asthma, caused by IgE-mediated allergic reactions, remains a global health issue with high morbidity and mortality rates. Immunotherapy is the only etiology-based approach to treating asthma, but no standard biomarkers have been established to evaluate the therapy’s effectiveness. This study aims to determine the correlation between the ratios of serum levels of HDM-specific IgE/total IgE and Asthma Control Test (ACT) score as a biomarker of the response to immunotherapy in pediatric allergic asthma patients. Patient and Methods: This retrospective cohort study involved 26 pediatric allergic asthma patients who underwent HDM-specific subcutaneous immunotherapy for 14 weeks at the Pediatric Allergy Immunology Outpatient Clinic at Saiful Anwar General Hospital, Malang. Serum levels of HDM-Specific IgE and Total IgE were measured before and after immunotherapy using Chemiluminescence Immunoassay and Enzyme-linked Immunosorbent Assay (ELISA) method. Changes in asthma control were assessed using the ACT score. The Wilcoxon Signed Ranked Test and Spearman correlation test were used for data analysis. Results: There were 14 boys and 12 girls with a mean age of 6.48 ± 2.54 years. The study showed a significant decrease in serum HMD-specific levels before immunotherapy [9.88 ± 5.74 kuA/L] compared to those of 14 weeks after immunotherapy [4.51 ± 3.98 kuA/L], p = 0.000. Serum Total IgE levels significant decrease before immunotherapy [207.6 ± 120.8IU/ml] compared to those of 14 weeks after immunotherapy [109.83 ± 189.39 IU/mL], p = 0.000. The ratios of serum HDM-specific IgE/total IgE levels significant decrease before immunotherapy [0.063 ± 0.05] compared to those of 14 weeks after immunotherapy [0.041 ± 0.039], p = 0.012. There was also a significant increase in ACT scores before and after immunotherapy (each 15.5 ± 1.79 and 20.96 ± 2.049, p = 0.000). The correlation test showed a weak negative correlation between the ratios of HDM-specific IgE/total IgE levels and ACT score (p = 0.034 and r = -0.29). Conclusion: In conclusion, this study showed that a decrease in HDM-specific IgE levels, total IgE levels, and HDM-specific IgE/total IgE ratios, and an increase in ACT score, was observed after 14 weeks of HDM-specific subcutaneous immunotherapy. The weak negative correlation between the HDM-specific IgE/total IgE ratio and the ACT score suggests that this ratio can serve as a potential biomarker of the effectiveness of immunotherapy in treating pediatric allergic asthma patients.

Keywords: HDM-specific IgE/total IgE ratio, ACT score, immunotherapy, allergic asthma

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Authors: Binaya Tamang, Buddhhi Raj Pokhrel, Narayan Gautam

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Background: There may be a connection between thyroid function and vitamin D status since both bind to similar nuclear hormone receptors and have similar response regions on gene promoters. The purpose of the current study was to investigate the relationship between thyroid hormones and vitamin D levels in females who were attending a tertiary care center in western Nepal and were either hypothyroid or euthyroid. Methods: This hospital-based cross-sectional study was carried out between March 2020 and March 2021 by the Biochemistry department of the Universal College of Medical Sciences (UCMS), Bhairahawa, Province No. 5, Nepal, in cooperation with Internal medicine. Prior to the study, institutional review committee approval (UCMS/IRC/008/20) was acquired from UCMS. Women who visited the Internal Medicine OPD of UCMS and were advised to get a thyroid function test (TFT) were included in the study population. Only those who were willing to participate in the study were enrolled after the goals and advantages of the study had been explained to them. Participants who had recently used vitamin D supplements and medications that affected thyroid hormones were excluded. The participants gave their consent verbally and in writing. After getting the consent, a convenient sample technique was applied. Serum was isolated after drawing 3 ml of blood in a plain vial. Chemiluminescence assay was used to analyze vitamin D and thyroid hormones (MAGLUMI 2000). SPSS version 16.0 for Windows was used to conduct the statistical analysis. Statistical significance was defined as a P-value < 0.05. Results: Majority of the study population (n=214, 71%) had insufficient serum vitamin D levels. Among the thyroid groups, the median Vitamin D levels were significantly lower in hypothyroid (16.88 ng/ml) as compared to the euthyroid groups (25.01 ng/ml) (P<0.001). Similarly, serum Vitamin D levels were considerably lower in the obese population (16.86 ng/ml) as compared to the normal BMI group (24.90 ng/ml) (P<0.001) as well as in the vegetarian (15.43 ng.ml) than mixed diet consumer (24.89 ng/ml) (P<0.01). Even after the adjustment for these variables, the Vitamin D levels were significantly lower in the hypothyroid population than in the euthyroid group (P<0.001). Conclusion: Comparing the hypothyroid population to the euthyroid, the median serum vitamin D levels were considerably lower. We were alarmed to see that the majority of euthyroid participants also had low levels of vitamin D. Therefore if left untreated, low vitamin D levels in hypothyroid patients could worsen their health further.

Keywords: vitamin D, thyroid hormones, euthyroid, hypothyroid, Nepal

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Authors: David X. Dong, Qingming Zhang, Meng Lu

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Nitrites enter waterways as runoff from croplands and are discharged from many industrial sites. Excessive nitrite inputs to water bodies lead to eutrophication. On-site rapid detection of nitrite is of increasing interest for managing fertilizer application and monitoring water source quality. Existing methods for detecting nitrites use spectrophotometry, ion chromatography, electrochemical sensors, ion-selective electrodes, chemiluminescence, and colorimetric methods. However, these methods either suffer from high cost or provide low measurement accuracy due to their poor selectivity to nitrites. Therefore, it is desired to develop an accurate and economical method to monitor nitrites in environments. We report a low-cost optical sensor, in conjunction with a machine learning (ML) approach to enable high-accuracy detection of nitrites in water sources. The sensor works under the principle of measuring molecular absorptions of nitrites at three narrowband wavelengths (295 nm, 310 nm, and 357 nm) in the ultraviolet (UV) region. These wavelengths are chosen because they have relatively high sensitivity to nitrites; low-cost light-emitting devices (LEDs) and photodetectors are also available at these wavelengths. A regression model is built, trained, and utilized to minimize cross-sensitivities of these wavelengths to the same analyte, thus achieving precise and reliable measurements with various interference ions. The measured absorbance data is input to the trained model that can provide nitrite concentration prediction for the sample. The sensor is built with i) a miniature quartz cuvette as the test cell that contains a liquid sample under test, ii) three low-cost UV LEDs placed on one side of the cell as light sources, with each LED providing a narrowband light, and iii) a photodetector with a built-in amplifier and an analog-to-digital converter placed on the other side of the test cell to measure the power of transmitted light. This simple optical design allows measuring the absorbance data of the sample at the three wavelengths. To train the regression model, absorbances of nitrite ions and their combination with various interference ions are first obtained at the three UV wavelengths using a conventional spectrophotometer. Then, the spectrophotometric data are inputs to different regression algorithm models for training and evaluating high-accuracy nitrite concentration prediction. Our experimental results show that the proposed approach enables instantaneous nitrite detection within several seconds. The sensor hardware costs about one hundred dollars, which is much cheaper than a commercial spectrophotometer. The ML algorithm helps to reduce the average relative errors to below 3.5% over a concentration range from 0.1 ppm to 100 ppm of nitrites. The sensor has been validated to measure nitrites at three sites in Ames, Iowa, USA. This work demonstrates an economical and effective approach to the rapid, reagent-free determination of nitrites with high accuracy. The integration of the low-cost optical sensor and ML data processing can find a wide range of applications in environmental monitoring and management.

Keywords: optical sensor, regression model, nitrites, water quality

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Authors: Pauline Nyssen, Ange Mouithys-Mickalad, Maryse Hoebeke

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Inflammation is a complex physiological phenomenon involving chemical and enzymatic mechanisms. Polymorphonuclear neutrophil leukocytes (PMNs) play an important role by producing reactive oxygen species (ROS) and releasing myeloperoxidase (MPO), a pro-oxidant enzyme. Released both in the phagolysosome and the extracellular medium, MPO produces during its peroxidase and halogenation cycles oxidant species, including hypochlorous acid, involved in the destruction of pathogen agents, like bacteria or viruses. Inflammatory pathologies, like rheumatoid arthritis, atherosclerosis induce an excessive stimulation of the PMNs and, therefore, an uncontrolled release of ROS and MPO in the extracellular medium, causing severe damages to the surrounding tissues and biomolecules such as proteins, lipids, and DNA. The treatment of chronic inflammatory pathologies remains a challenge. For many years, MPO has been used as a target for the development of effective treatments. Numerous studies have been focused on the design of new drugs presenting more efficient MPO inhibitory properties. However, some designed inhibitors can be toxic. An alternative consists of assessing the potential inhibitory action of clinically-known molecules, having antioxidant activity. Propofol, 2,6-diisopropyl phenol, which is used as an intravenous anesthetic agent, meets these requirements. Besides its anesthetic action employed to induce a sedative state during surgery or in intensive care units, propofol and its injectable form Diprivan indeed present antioxidant properties and act as ROS and free radical scavengers. A study has also evidenced the ability of propofol to inhibit the formation of the neutrophil extracellular traps fibers, which are important to trap pathogen microorganisms during the inflammation process. The aim of this study was to investigate the potential inhibitory action mechanism of propofol and Diprivan on MPO activity. To go into the anti-inflammatory action of propofol in-depth, two of its oxidative derivatives, 2,6-diisopropyl-1,4-p-benzoquinone (PPFQ) and 3,5,3’,5’-tetra isopropyl-(4,4’)-diphenoquinone (PPFDQ), were studied regarding their inhibitory action. Specific immunological extraction followed by enzyme detection (SIEFED) and molecular modeling have evidenced the low anti-catalytic action of propofol. Stopped-flow absorption spectroscopy and direct MPO activity analysis have proved that propofol acts as a reversible MPO inhibitor by interacting as a reductive substrate in the peroxidase cycle and promoting the accumulation of redox compound II. Overall, Diprivan exhibited a weaker inhibitory action than the active molecule propofol. In contrast, PPFQ seemed to bind and obstruct the enzyme active site, preventing the trigger of the MPO oxidant cycles. PPFQ induced a better chlorination cycle inhibition at basic and neutral pH in comparison to propofol. PPFDQ did not show any MPO inhibition activity. The three interest molecules have also demonstrated their inhibition ability on an important step of the inflammation pathway, the PMNs superoxide anions production, thanks to EPR spectroscopy and chemiluminescence. In conclusion, propofol presents an interesting immunomodulatory activity by acting as a reductive substrate in the peroxidase cycle of MPO, slowing down its activity, whereas PPFQ acts more as an anti-catalytic substrate. Although PPFDQ has no impact on MPO, it can act on the inflammation process by inhibiting the superoxide anions production by PMNs.

Keywords: Diprivan, inhibitor, myeloperoxidase, propofol, spectroscopy

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Authors: Tom S. Owens, Tom A. Calverley, Benjamin S. Stacey, Christopher J. Marley, George Rose, Lewis Fall, Gareth L. Jones, Priscilla Williams, John P. R. Williams, Martin Steggall, Damian M. Bailey

Abstract:

Introduction and aims: Sports-related concussion (SRC) represents a significant and growing public health concern in rugby union, yet remains one of the least understood injuries facing the health community today. Alongside increasing SRC incidence rates, there is concern that prior recurrent concussion may contribute to long-term neurologic sequelae in later-life. This may be due to an accelerated decline in cerebral perfusion, a major risk factor for neurocognitive decline and neurodegeneration, though the underlying mechanisms remain to be established. The present study hypothesised that recurrent concussion in current professional rugby union players would result in elevated systemic oxidative-nitrosative stress, reflected by a free radical-mediated reduction in nitric oxide (NO) bioavailability and impaired cerebrovascular and cognitive function. Methodology: A longitudinal study design was adopted across the 2017-2018 rugby union season. Ethical approval was obtained from the University of South Wales Ethics Committee. Data collection is ongoing, and therefore the current report documents result from the pre-season and first half of the in-season data collection. Participants were initially divided into two subgroups; 23 professional rugby union players (aged 26 ± 5 years) and 22 non-concussed controls (27 ± 8 years). Pre-season measurements were performed for cerebrovascular function (Doppler ultrasound of middle cerebral artery velocity (MCAv) in response to hypocapnia/normocapnia/hypercapnia), cephalic venous concentrations of the ascorbate radical (A•-, electron paramagnetic resonance spectroscopy), NO (ozone-based chemiluminescence) and cognition (neuropsychometric tests). Notational analysis was performed to assess contact in the rugby group throughout each competitive game. Results: 1001 tackles and 62 injuries, including three concussions were observed across the first half of the season. However, no associations were apparent between number of tackles and any injury type (P > 0.05). The rugby group expressed greater oxidative stress as indicated by increased A•- (P < 0.05 vs. control) and a subsequent decrease in NO bioavailability (P < 0.05 vs. control). The rugby group performed worse in the Ray Auditory Verbal Learning Test B (RAVLT-B, learning, and memory) and the Grooved Pegboard test using both the dominant and non-dominant hands (visuomotor coordination, P < 0.05 vs. control). There were no between-group differences in cerebral perfusion at baseline (MCAv: 54 ± 13 vs. 59 ± 12, P > 0.05). Likewise, no between-group differences in CVRCO2Hypo (2.58 ± 1.01 vs. 2.58 ± 0.75, P > 0.05) or CVRCO2Hyper (2.69 ± 1.07 vs. 3.35 ± 1.28, P > 0.05) were observed. Conclusion: The present study identified that the rugby union players are characterized by impaired cognitive function subsequent to elevated systemic-oxidative-nitrosative stress. However, this appears to be independent of any functional impairment in cerebrovascular function. Given the potential long-term trajectory towards accelerated cognitive decline in populations exposed to SRC, prophylaxis to increase NO bioavailability warrants consideration.

Keywords: cognition, concussion, mild traumatic brain injury, rugby

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