Search results for: lactate oxidase

Authors: Antonella Cartoni, Mattea Carmen Castrovilli

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A biosensor for lactate detection has been developed using an environmentally friendly approach. The biosensor is based on lactate oxidase (LOX) and has remarkable capabilities for reuse and storage at room temperature. The manufacturing technique employed is ambient electrospray deposition (ESD), which enables efficient and sustainable immobilization of the LOX enzyme on a cost-effective com-mercial screen-printed Prussian blue/carbon electrode (PB/C-SPE). The study demonstrates that the ESD technology allows the biosensor to be stored at ambient pressure and temperature for extended periods without affecting the enzymatic activity. The biosensor can be stored for up to 90 days without requiring specific storage conditions, and it can be reused for up to 24 measurements on both freshly prepared electrodes and electrodes that are three months old. The LOX-based biosensor exhibits a lin-ear range of lactate detection between 0.1 and 1 mM, with a limit of detection of 0.07±0.02 mM. Ad-ditionally, it does not exhibit any memory effects. The immobilization process does not involve the use of entrapment matrices or hazardous chemicals, making it environmentally sustainable and non-toxic compared to current methods. Furthermore, the application of a electrospray deposition cycle on previously used biosensors rejuvenates their performance, making them comparable to freshly made biosensors. This highlights the excellent recycling potential of the technique, eliminating the waste as-sociated with disposable devices.

Keywords: green friendly, reuse, storage performance, immobilization, matrix-free, electrospray deposition, biosensor, lactate oxidase, enzyme

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Authors: Zoltan Horvath, Dora Nagy

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In this research, it was examined whether individual COVID variants cause differences in the lactate curve of cyclists. After all, the virus variants attacked different organs in our body during the infections. During our tests, we used a traditional lactate step test, the results of which were compared with the values before the infection. In the tests, it has been proven that different virus variants show unique lactate curves. In this way, based on the lactate curve, it is possible to identify which variant caused the disease. Thanks to this, it has been shorten the return time, because we can apply the best return protocol after infection to the competitors.

Keywords: COVID-Sars19, lactate, virus mutation, lactate profile

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Authors: Khaled S. Al Salhen

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Aldehyde oxidase is molybdo-flavoenzyme involved in the oxidation of hundreds of endogenous and exogenous and N-heterocyclic compounds and environmental pollutants. Uncharged N-heterocyclic aromatic compounds such phenanthridine are commonly distributed pollutants in soil, air, sediments, surface water and groundwater, and in animal and plant tissues. Phenanthridine as uncharged N-heterocyclic aromatic compound was incubated with partially purified aldehyde oxidase from rainbow trout fish liver. Reversed-phase HLPC method was used to separate the oxidation products from phenanthridine and the metabolite was identified. The 6(5H)-phenanthridinone was identified the major metabolite by partially purified aldehyde oxidase from fish liver. Kinetic constant for the oxidation reactions were determined spectrophotometrically and showed that this substrate has a good affinity (Km = 78 ± 7.6 µM) for hepatic aldehyde oxidase, coupled with a relatively high oxidation rate (0.77± 0.03 nmol/min/mg protein). In addition, the kinetic parameters of hepatic fish aldehyde oxidase towards the phenanthridine substrate indicate that in vitro biotransformation by hepatic fish aldehyde oxidase will be a significant pathway. This study confirms that partially purified aldehyde oxidase from fish liver is indeed the enzyme responsible for the in vitro production 6(5H)-phenanthridinone metabolite as it is a major metabolite by mammalian aldehyde oxidase.

Keywords: aldehyde oxidase, fish, phenanthridine, specificity

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Authors: Sherif Sabri, Suzy Fawzi, Sanaa Abdelshafy, Ayman Nagah

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Introduction: Static derangements in lactate homeostasis during ICU stay have become established as a clinically useful marker of increased risk of hospital and ICU mortality. Lactate indices or kinetic alteration of the anaerobic metabolism make it a potential parameter to evaluate disease severity and intervention adequacy. This is an inexpensive and simple clinical parameter that can be obtained by a minimally invasive means. Aim of work: Comparing the predictive value of dynamic indices of hyperlactatemia in the first twenty four hours of intensive care unit (ICU) admission with other static values are more commonly used. Patients and Methods: This study included 40 critically ill patients above 18 years old of both sexes with Hyperlactamia (≥ 2 m mol/L). Patients were divided into septic group (n=20) and low oxygen transport group (n=20), which include all causes of low-O2. Six lactate indices specifically relating to the first 24 hours of ICU admission were considered, three static indices and three dynamic indices. Results: There were no statistically significant differences among the two groups regarding age, most of the laboratory results including ABG and the need for mechanical ventilation. Admission lactate was significantly higher in low-oxygen transport group than the septic group [37.5±11.4 versus 30.6±7.8 P-value 0.034]. Maximum lactate was significantly higher in low-oxygen transport group than the septic group P-value (0.044). On the other hand absolute lactate (mg) was higher in septic group P-value (< 0.001). Percentage change of lactate was higher in the septic group (47.8±11.3) than the low-oxygen transport group (26.1±12.6) with highly significant P-value (< 0.001). Lastly, time weighted lactate was higher in the low-oxygen transport group (1.72±0.81) than the septic group (1.05±0.8) with significant P-value (0.012). There were statistically significant differences regarding lactate indices in survivors and non survivors, whether in septic or low-oxygen transport group. Conclusion: In critically ill patients, time weighted lactate and percent in lactate change in the first 24 hours can be an independent predictive factor in ICU mortality. Also, a rising compared to a falling blood lactate concentration over the first 24 hours can be associated with significant increase in the risk of mortality.

Keywords: critically ill patients, lactate indices, mortality in intensive care, anaerobic metabolism

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Authors: Rahile Ozturk, Esra Maltas

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This study aimed to determine the effect of vinclozolin on some biochemical characteristics of Galleria mellonella (Lepidoptera: Pyralidae) which is an economically harmful species damaging the honeycomb in beekeeping. For experimental groups, the eggs obtained from stock were dropped into the mixed feed of vinclozolin at different doses (20, 40 and 60 ppm) and had the larvae fed with this feed. As result of the addition of vinclozolin at concentrations of 20, 40 and 60 ppm, glycogen contents of G. mellonella were determined and a significant reduction in the amount of glycogen was observed with increasing concentration of vinclozolin. In this study, activity of catalase enzyme, particularly effective in defense mechanism, activity of xanthine oxidase involved in nucleotide metabolism and activity of glucose oxidase in the metabolism of carbohydrates were measured. When compared with the results from control groups, the enzyme activities of the larvaes fed with the feed including 20, 40 and 60 ppm of vinclozolin were observed to vary or remain constant. Accordingly, glucose oxidase and catalase activities increased with the increase in amount of vinclozolin in the feed and the activity of xanthine oxidase remained stable.

Keywords: Catalase, Galleria mellonella, glucose oxidase, vinclozolin, xanthine oxidase.

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Authors: M. S. Kilic, S. Korkut, B. Hazer

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Newly synthesized Polypropylene-g-Polyethylene glycol polymer was first time used for a compartment-less enzymatic fuel cell. Working electrodes based on Polypropylene-g-Polyethylene glycol were operated as unmediated and mediated system (with ferrocene and gold/cobalt oxide nanoparticles). Glucose oxidase and bilirubin oxidase was selected as anodic and cathodic enzyme, respectively. Glucose was used as fuel in a single-compartment and membrane-less cell. Maximum power density was obtained as 0.65 nW cm-2, 65 nW cm-2, and 23500 nW cm-2 from the unmediated, ferrocene and gold/cobalt oxide modified polymeric film, respectively. Power density was calculated to be ~16000 nW cm-2 for undiluted wastewater sample with gold/cobalt oxide nanoparticles including system.

Keywords: bilirubin oxidase, enzymatic fuel cell, glucose oxidase, nanoparticles

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Authors: Mohamed Khalifa Zayet, Salma Belal Eiid, Mushira Mohamed Dahaba

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Introduction: Malignant tumors may not be easily detected by traditional radiographic techniques especially in an anatomically complex area like maxillofacial region. At the same time, the advent of biological functional MRI was a significant footstep in the diagnostic imaging field. Objective: The purpose of this study was to define the malignant metabolic profile of maxillofacial lesions using diffusion MRI and magnetic resonance spectroscopy, as adjunctive aids for diagnosing of such lesions. Subjects and Methods: Twenty-one patients with twenty-two lesions were enrolled in this study. Both morphological and functional MRI scans were performed, where T1, T2 weighted images, diffusion-weighted MRI with four apparent diffusion coefficient (ADC) maps were constructed for analysis, and magnetic resonance spectroscopy with qualitative and semi-quantitative analyses of choline and lactate peaks were applied. Then, all patients underwent incisional or excisional biopsies within two weeks from MR scans. Results: Statistical analysis revealed that not all the parameters had the same diagnostic performance, where lactate had the highest areas under the curve (AUC) of 0.9 and choline was the lowest with insignificant diagnostic value. The best cut-off value suggested for lactate was 0.125, where any lesion above this value is supposed to be malignant with 90 % sensitivity and 83.3 % specificity. Despite that ADC maps had comparable AUCs still, the statistical measure that had the final say was the interpretation of likelihood ratio. As expected, lactate again showed the best combination of positive and negative likelihood ratios, whereas for the maps, ADC map with 500 and 1000 b-values showed the best realistic combination of likelihood ratios, however, with lower sensitivity and specificity than lactate. Conclusion: Diffusion weighted imaging and magnetic resonance spectroscopy are state-of-art in the diagnostic arena and they manifested themselves as key players in the differentiation process of orofacial tumors. The complete biological profile of malignancy can be decoded as low ADC values, high choline and/or high lactate, whereas that of benign entities can be translated as high ADC values, low choline and no lactate.

Keywords: diffusion magnetic resonance imaging, magnetic resonance spectroscopy, malignant tumors, maxillofacial

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Authors: Kyung-Soon Choi, Ji-Young Hwang, Young-Hee Pyo

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Vinegars have been used as an alternative remedy for treating gout, but the scientific basis remains to be elucidated. In this study, acetic acid fermentation was applied for the first time to Monascus-fermented soybeans to examine its effect on the bioactive components together with the xanthine oxidase inhibitory (XOI) activity of the soy vinegar. The content of total phenols (0.47~0.97 mg gallic acid equivalents/mL) and flavonoids (0.18~0.39 mg quercetin equivallents/mL) were spectrophotometrically determined, and the content of organic acid (10.22~59.76 mg/mL) and isoflavones (6.79~7.46 mg/mL) were determined using HPLC-UV. The analytical method for ubiquinones (0.079~0.276 μg/mL) employed saponification before solvent extraction and quantification using LC-MS. Soy vinegar also showed significant XOI (95.3%) after 20 days of acetic acid fermentation at 30 °C. The results suggest that soy vinegar has potential as a novel medicinal food.

Keywords: acetic acid fermentation, bioactive component, soy vinegar, xanthine oxidase inhibitory activity

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Authors: Seyhan Sahan-Firat, Meryem Temiz-Resitoglu, Demet Sinem Guden, Sefika Pinar Kucukkavruk, Bahar Tunctan, Ayse Nihal Sari, Zumrut Kocak

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We hypothesized that mTOR inhibition may prevent the multiple organ failures following severe multiple tissue injury associated with increased NADPH oxidase system activity occur in zymosan-induced systemic inflammation. Therefore, we investigated the role of mTOR in oxidative/nitrosative stress associated with increase in NADPH oxidase activity in zymosan-induced systemic inflammation model in rats. Male Wistar rats received saline (4 ml/kg, i.p.) and zymosan (500 mg/kg, i.p.) at time 0. Saline, or zymosan-treated rats were given rapamycin (1 mg/kg, i.p.) 1 h after saline or zymosan injections. Rats were sacrified 4 h after zymosan challenge and kidney, heart, thoracic aorta, and superior mesenteric artery were collected. NADPH oxidase activity, p22phox, gp91phox, and p47phox protein expression and nitrotyrosine levels were measured in tissue samples. Zymosan administration caused an increase in NADPH oxidase activity, p22phox, gp91phox, and p47phox protein expression and nitrotyrosine levels in kidney, heart, thoracic aorta, and superior mesenteric artery. These changes caused by zymosan reversed by rapamycin, a selective mTOR inhibitor. Rapamycin alone had no effect on the parameters measured. Our results demonstrated that zymosan-induced oxidative/nitrosative stress presumably due to enhanced activity of NADPH oxidase, expression of p22phox, gp91phox, and p47phox and production of peroxynitrite were mediated by mTOR. [This work was financially supported by Research Foundation of Mersin University (2016-2-AP3-1900)].

Keywords: oxidative stress, mTOR, nitrosative stress, zymosan

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Authors: Jung-Feng Hsieh, Chu Ze Chen

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Curcumin is the main active compound of turmeric that can inhibit the activities of α-glucosidase and xanthine oxidase (XO). α-Glucosidase and XO inhibitors are widely used to treat patients with diabetes mellitus and gout, respectively; therefore, the objective of this research was to evaluate the inhibitory activities of curcumin and its analogs against α-glucosidase and XO. Our results demonstrated that CM-F had the strongest antioxidant activity with a half-maximal effective concentration (EC50) of 9.39 ± 0.16 μM, which was superior to vitamin E (EC50=17.03 ± 0.09 μM). CM-F also exhibited potent inhibitory activity against XO with an IC50 value of 6.14 ± 0.38 μM and enzyme kinetic results revealed competitive inhibition of XO. We also found that CM-1 and CM-2 inhibited α-glucosidase with IC50 values of 21.06 ± 0.92 μM and 5.95 ± 0.09 μM, respectively, and kinetic studies indicated that both CM-1 and CM-2 are mixed competitive inhibitors of α-glucosidase. Furthermore, docking simulation identified five hydrogen bonds between XO and CM-F; however, only one and two hydrogen bonds are involved in CM-1 and CM-2 binding to α-glucosidase, respectively. Accordingly, curcumin and its analogs have the potential to be used in the treatment of patients with diabetes mellitus and gout.

Keywords: curcumin, α-glucosidase, inhibitor, xanthine oxidase

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Authors: Timothy Quinn, Ronald Croce, Aliaksandr Leuchanka, Justin Walker

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Many endurance athletes train at or just below an intensity associated with their lactate threshold (LT) and often the heart rate (HR) that these athletes use for their LT are above their true LT-HR measured in a laboratory. Training above their true LT-HR may lead to overtraining and injury. Few athletes have the capability of measuring their LT in a laboratory and rely on perception to guide them, as accurate field tests to determine LT are limited. Therefore, the purpose of this study was to determine if an 8-minute field test could accurately define the HR associated with LT as measured in the laboratory. On Day 1, fifteen male runners (mean±SD; age, 27.8±4.1 years; height, 177.9±7.1 cm; body mass, 72.3±6.2 kg; body fat, 8.3±3.1%) performed a discontinuous treadmill LT/maximal oxygen consumption (LT/VO2max) test using a portable metabolic gas analyzer (Cosmed K4b2) and a lactate analyzer (Analox GL5). The LT (and associated HR) was determined using the 1/+1 method, where blood lactate increased by 1 mMol•L-1 over baseline followed by an additional 1 mMol•L-1 increase. Days 2 and 3 were randomized, and the athletes performed either an 8-minute run on the treadmill (TM) or on a 160-m indoor track (TR) in an effort to cover as much distance as possible while maintaining a high intensity throughout the entire 8 minutes. VO2, HR, ventilation (VE), and respiratory exchange ratio (RER) were measured using the Cosmed system, and rating of perceived exertion (RPE; 6-20 scale) was recorded every minute. All variables were averaged over the 8 minutes. The total distance covered over the 8 minutes was measured in both conditions. At the completion of the 8-minute runs, blood lactate was measured. Paired sample t-tests and pairwise Pearson correlations were computed to determine the relationship between variables measured in the field tests versus those obtained in the laboratory at LT. An alpha level of <0.05 was required for statistical significance. The HR (mean +SD) during the TM (167+9 bpm) and TR (172+9 bpm) tests were strongly correlated to the HR measured during the laboratory LT (169+11 bpm) test (r=0.68; p<0.03 and r=0.88; p<0.001, respectively). Blood lactate values during the TM and TR tests were not different from each other but were strongly correlated with the laboratory LT (r=0.73; p<0.04 and r=0.66; p<0.05, respectively). VE (Lmin-1) was significantly greater during the TR (134.8+11.4 Lmin-1) as compared to the TM (123.3+16.2 Lmin-1) with moderately strong correlations to the laboratory threshold values (r=0.38; p=0.27 and r=0.58; p=0.06, respectively). VO2 was higher during TR (51.4 mlkg-1min-1) compared to TM (47.4 mlkg-1min-1) with correlations of 0.33 (p=0.35) and 0.48 (p=0.13), respectively to threshold values. Total distance run was significantly greater during the TR (2331.6+180.9 m) as compared to the TM (2177.0+232.6 m), but they were strongly correlated with each other (r=0.82; p<0.002). These results suggest that an 8-minute running field test can accurately predict the HR associated with the LT and may be a simple test that athletes and coaches could implement to aid in training techniques.

Keywords: blood lactate, heart rate, running, training

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Authors: Osama Moustafa Zayed

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Aim of the work: To evaluate the effectiveness of venous glucose, levels of serum lactate and base deficit in polytraumatized patients as simple parameters to predict the mortality in these patients. Compared to the predictive value of Trauma and injury severity (TRISS) and Acute Physiology And Chronic Health Evaluation IV (APACHE IV). Introduction: Trauma is a serious global health problem, accounting for approximately one in 10 deaths worldwide. Trauma accounts for 5 million deaths per year. Prediction of mortality in trauma patients is an important part of trauma care. Several trauma scores have been devised to predict injury severity and risk of mortality. The trauma and injury severity score (TRISS) was most common used. Regardless of the accuracy of trauma scores, is based on an anatomical description of every injury and cannot be assigned to the patients until a full diagnostic procedure has been performed. So we hypothesized that alterations in admission glucose, lactate levels and base deficit would be an early and easy rapid predictor of mortality. Patient and Method: a comparative cross-sectional study. 282 Polytraumatized patients attended to the Emergency Department(ED) of the Suez Canal university Hospital constituted. The period from 1/1/2012 to 1/4/2013 was included. Results: We found that the best cut off value of TRISS probability of survival score for prediction of mortality among poly-traumatized patients is = 90, with 77% sensitivity and 89% specificity using area under the ROC curve (0.89) at (95%CI). APACHE IV demonstrated 67% sensitivity and 95% specificity at 95% CI at cut off point 99. The best cutoff value of Random Blood Sugar (RBS) for prediction of mortality was>140 mg/dl, with 89%, sensitivity, 49% specificity. The best cut off value of base deficit for prediction of mortality was less than -5.6 with 64% sensitivity, 93% specificity. The best cutoff point of lactate for prediction of mortality was > 2.6 mmol/L with 92%, sensitivity, 42% specificity. Conclusion: According to our results from all evaluated predictors of mortality (laboratory and scores) and mortality based on the estimated cutoff values using ROC curves analysis, the highest risk of mortality was found using a cutoff value of 90 in TRISS score while with laboratory parameters the highest risk of mortality was with serum lactate > 2.6 . Although that all of the three parameter are accurate in predicting mortality in poly-traumatized patients and near with each other, as in serum lactate the area under the curve 0.82, in BD 0.79 and 0.77 in RBS.

Keywords: APACHE IV, emergency department, polytraumatized patients, serum lactate

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Authors: R. D. Esposito, V. M. Barberá, P. García Morales, P. Dorado Rodríguez, J. Sanz, M. Fuentes, D. Planes Meseguer, M. Saceda, L. Fernández Fornos, M. P. Ventero

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Results obtained by our group on glioblastoma multiforme (GBM) primary cultures , show a dramatic potentiation of radiation effects when 2 units/ml of D-amino acid oxidase (DAO) enzyme are added, free or immobilized in magnetic nanoparticles, to irradiated samples just after the irradiation. Cell cultures were exposed to radiation doses of 7Gy and 15Gy of 6 MV photons from a clinical linear accelerator. At both doses, we observed a clear enhancing effect of radiation-induced damages due to the addition of DAO.

Keywords: D-amino Acid Oxidase (DAO) enzyme, magnetic particles, nanotechnology, radiation therapy enhancement

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Authors: Nafisa Komilova, Plamena Angelova, Andrey Abramov, Ulugbek Mirkhodjaev

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Parkinson’s disease (PD) is a progressive neurodegenerative disorder which is induced by the loss of dopaminergic neurons in the midbrain. The mechanism of neurodegeneration is associated with the aggregation of misfolded proteins, oxidative stress, and mitochondrial disfunction. Considering this, the process of removal of unwanted organelles or proteins by autophagy is vitally important in neurons, and activation of these processes could be protective in PD. Short-time acidification of cytosol can activate mitophagy and autophagy, and here we used sodium pyruvate and sodium lactate in human fibroblasts with PD mutations (Pink1, Pink1/Park2, α-syn triplication, A53T) to induce changes in intracellular pH. We have found that both lactate and pyruvate in millimolar concentrations can induce short-time acidification of cytosol in these cells. It induced activation of mitophagy and autophagy in control and PD fibroblasts and protected against cell death. Importantly, the application of lactate to acute brain slices of control and Pink1 knockout mice also induced a reduction of pH in neurons and astrocytes that increase the level of mitophagy. Thus, acidification of cytosol by compounds which play important role in cell metabolism also can activate mitophagy and autophagy and protect cells in the familial form of PD.

Keywords: Parkinson's disease, mutations, mitophagy, autophagy

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Authors: Pawasuth Saengdee, Chamras Promptmas, Ting Zeng, Silke Leimkühler, Ulla Wollenberger

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Sulfite has been used as a versatile preservative to limit the microbial growth and to control the taste in some food and beverage. However, it has been reported to cause a wide spectrum of severe adverse reactions. Therefore, it is important to determine the amount of sulfite in food and beverage to ensure consumer safety. An efficient electrocatalytic biosensor for sulfite detection was developed by immobilizing of human sulfite oxidase (hSO) on 3-aminoproplytriethoxysilane (APTES) modified indium tin oxide (ITO) electrode. Cyclic voltammetry was employed to investigate the electrochemical characteristics of the hSO modified ITO electrode for various pretreatment and binding conditions. Amperometry was also utilized to demonstrate the current responses of the sulfite sensor toward sodium sulfite in an aqueous solution at a potential of 0 V (vs. Ag/AgCl 1 M KCl). The proposed sulfite sensor has a linear range between 0.5 to 2 mM with a correlation coefficient 0.972. Then, the additional polymer layer of PVA was introduced to extend the linear range of sulfite sensor and protect the enzyme. The linear range of sulfite sensor with 5% coverage increases from 2.8 to 20 mM at a correlation coefficient of 0.983. In addition, the stability of sulfite sensor with 5% PVA coverage increases until 14 days when kept in 0.5 mM Tris-buffer, pH 7.0 at 4 8C. Therefore, this sensor could be applied for the detection of sulfite in the real sample, especially in food and beverage.

Keywords: sulfite oxidase, bioelectrocatalytsis, indium tin oxide, direct electrochemistry, sulfite sensor

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Authors: Bhavini Patel, Aslihan Ugun-Klusek, Ellen Billet

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The two main contributing factors to familial and idiopathic form of Parkinson’s disease (PD) are oxidative stress and altered proteolysis. Monoamine oxidase-A (MAO-A) plays a significant role in redox homeostasis by producing reactive oxygen species (ROS) via deamination of for example, dopamine. The ROS generated induces chemical modification of proteins resulting in altered biological function. The ubiquitin-proteasome system, which consists of three different types or proteolytic activity, namely “chymotrypsin-like” activity (CLA), “trypsin-like” activity (TLA) and “post acidic-like” activity (PLA), is responsible for the degradation of ubiquitinated proteins. Defects in UPS are known to be strongly correlated to PD. Herein, the effect of ROS generated by MAO-A on proteasome activity and the effects of proteasome inhibition on MAO-A protein levels in WT, mock and MAO-A overexpressed (MAO-A+) SHSY5Y neuroblastoma cell lines were investigated. The data in this study report increased proteolytic activity when MAO-A protein levels are significantly increased, in particular CLA and PLA. Additionally, 20S proteasome inhibition induced a decrease in MAO-A levels in WT and mock cells in comparison to MAO-A+ cells in which 20S proteasome inhibition induced increased MAO-A levels to be further increased at 48 hours of inhibition. This study supports the fact that MAO-A could be a potential pharmaceutical target for neuronal protection as data suggests that endogenous MAO-A levels may be essential for modulating cell death and survival.

Keywords: monoamine oxidase, neurodegeneration, Parkinson's disease, proteasome

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Authors: Diana Puigserver Cuerda, Jofre Herrero Ferran, José M. Carmona Perez

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In the transition zone between aquifers and basal aquitards, DNAPL-pools of chlorinated solvents are more recalcitrant than at other depths in the aquifer. Although degradation of carbon tetrachloride (CT) and chloroform (CF) occurs in this zone, this is a slow process, which is why an adequate remediation strategy is necessary. The working hypothesis of this study is that the biostimulation of the transition zone of an aquifer contaminated by CT and CF can be an effective remediation strategy. This hypothesis has been tested in a site on an unconfined aquifer in which the major contaminants were CT and CF of industrial origin and where the hydrochemical background was rich in other compounds that can hinder natural attenuation of chloromethanes. Field studies and five laboratory microcosm experiments were carried out at the level of groundwater and sediments to identify: i) the degradation processes of CT and CF; ii) the structure of microbial communities; and iii) the microorganisms implicated on this degradation. For this, concentration of contaminants and co-contaminants (nitrate and sulfate), Compound Specific Isotope Analysis, molecular techniques (Denaturing Gradient Gel Electrophoresis) and clone library analysis were used. The main results were: i) degradation processes of CT and CF occurred in groundwater and in the lesser conductive sediments; ii) sulfate-reducing conditions in the transition zone were high and similar to those in the source of contamination; iii) two microorganisms (Azospira suillum and a bacterium of the Clostridiales order) were identified in the transition zone at the field and lab experiments that were compatible with the role of carrying out the reductive dechlorination of CT, CF and their degradation products (dichloromethane and chloromethane); iv) these two microorganisms were present at the high starting concentrations of the microcosm experiments (similar to those in the source of DNAPL) and continued being present until the last day of the lactate biostimulation; and v) the lactate biostimulation gave rise to the fastest and highest degradation rates and promoted the elimination of other electron acceptors (e.g. nitrate and sulfate). All these results are evidence that lactate biostimulation can be effective in remediating the source and plume, especially in the transition zone, and highlight the environmental relevance of the treatment of contaminated transition zones in industrial contexts similar to that studied.

Keywords: Azospira suillum, lactate biostimulation of carbon tetrachloride and chloroform, reductive dechlorination, transition zone between aquifer and aquitard

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Authors: A. Fordham, A. Hudson

Abstract:

Background: Studies have suggested 16-25% of normotensive trauma patients with no clinical signs of shock have abnormal lactate and BD readings evidencing shock; a phenomenon known as occult hypoperfusion (OH). In light of the scarce quantity of evidence currently documenting OH, this study aimed to identify the prevalence of OH in the pre-hospital setting and explore ways to improve its identification and management. Methods: A quantitative retrospective data analysis was carried out on 75 sets of patient records for trauma patients treated by Kent Surrey Sussex Air Ambulance Trust between November 2013 and October 2014. The KSS HEMS notes and subsequent ED notes were collected. Trends between patients’ SBP on the scene, whether or not they received PRBCs on the scene as well as lactate and BD readings in the ED were assessed. Patients’ KSS HEMS notes written by the HEMS crew were also reviewed and recorded. Results: -Suspected OH was identified in 7% of the patients who did not receive PRBCs in the pre-hospital phase. -SBP heavily influences the physicians’ decision of whether or not to transfuse PRBCs in the pre-hospital phase. Preliminary conclusions: OH is an under-studied and underestimated phenomenon. We suggest a prospective trial is carried out to evaluate whether detecting trauma patients’ tissue perfusion status in the pre-hospital phase using portable devices capable of measuring serum BD and/or lactate could aid more accurate detection and management of all haemorrhaging trauma patients, including patients with OH.

Keywords: occult hypoperfusion, PRBC transfusion, point of care testing, pre-hospital emergency medicine, trauma

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Authors: Naouel Boussoualim, Hayat Trabsa, Imane Krache, Seddik Khennouf, Noureddine Charef, Lekhmici Arrar, Abderrahmane Baghiani

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Purpose: To evaluate the in-vitro inhibition of xanthine oxidase (purified from bovine milk) by extracts of Lycium arabicum, as well as it is in vivo hypouricemic and renal protective effects. Methods: Four extracts of Lycium arabicum, methanol (CrE), chloroform (ChE), ethyl acetate (EaE) and aqueous (AqE) extracts, were screened for their total phenolics and potential inhibitory effects on purified bovine milk xanthine oxidase (XO) activity by measuring the formation of uric acid or superoxide radical. The mode of inhibition was investigated and compared with the standard drugs, allopurinol, quercitin, and catechin. To evaluate their hypouricemic effect, the extracts were administered to potassium oxonate-induced hyperuricemic mice at a dose of 50 mg/kg body weight. Results: The results showed that EaE had the highest content of phenolic compounds and was the most potent inhibitor of uric acid formation (IC50 = 0.017 ± 0.001 mg/mL) and formation of superoxide (IC50 = 0.035 ± 0.001 mg/ml). Lineweaver-Burk analysis showed that CrE and EaE inhibited XO competitively, whereas the inhibitory activities exerted by ChE and AqE were of a mixed type. Intraperetoneal injection of L. arabicum extracts (50 mg/kg) elicited hypouricemic actions in hyperuricemic mice. Hyperuricemic mice presented a serum uric acid concentration of 4.71 ± 0.29 mg/L but this was reduced to 1.78 ± 0.11 mg/L by EaE, which was the most potent hyporuricemic extract. Conclusion: L. arabicum fractions have a strong inhibitory effect on xanthine oxidase and and also have a significantly lowering effect on serum and liver creatinine and urea levels in hyperuricemic mice.

Keywords: lycium arabicum, uric acid, creatinine, superoxide, phenolic compounds, flavonoids, hyperuricemia

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Authors: Aida Kalantari, Boyang Ji, Tao Chen, Ivan Mijakovic

Abstract:

3-hydroxypropanoic acid (3-HP) is one of the most important biomass-derivable platform chemicals that can be converted into a number of industrially important compounds. There have been several attempts at production of 3-HP from renewable sources in cell factories, focusing mainly on Escherichia coli, Klebsiella pneumoniae, and Saccharomyces cerevisiae. Despite the significant progress made in this field, commercially exploitable large-scale production of 3-HP in microbial strains has still not been achieved. In this study, we investigated the potential of Bacillus subtilis to be used as a microbial platform for bioconversion of glycerol into 3-HP. Our recombinant B. subtilis strains overexpress the two-step heterologous pathway containing glycerol dehydratase and aldehyde dehydrogenase from various backgrounds. The recombinant strains harboring the codon-optimized synthetic pathway from K. pneumoniae produced low levels of 3-HP. Since the enzymes in the heterologous pathway are sensitive to oxygen, we had to perform our experiments in micro-aerobic conditions. Under these conditions, the cell produces lactate in order to regenerate NAD+, and we found the lactate production to be in competition with the production of 3-HP. Therefore, based on the in silico predictions, we knocked out the glycerol kinase (glpk), which in combination with growth on glucose, resulted in improving the 3-HP titer to 1 g/L and the removal of lactate. Cultivation of the same strain in an enriched medium improved the 3-HP titer up to 7.6 g/L. Our findings provide the first report of successful introduction of the biosynthetic pathway for conversion of glycerol into 3-HP in B. subtilis.

Keywords: bacillus subtilis, glycerol, 3-hydroxypropanoic acid, metabolic engineering

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Authors: Angkrit Phitchayangkoon, Ar-Aishah Dadeh

Abstract:

Background: A key aim of triage is to identify the patients with high risk of cardiac arrest because they require intensive monitoring, resuscitation facilities, and early intervention. We aimed to identify the predicting factors such as initial vital signs, serum pH, serum lactate level, initial capillary blood glucose, and Modified Early Warning Score (MEWS) which affect the occurrence of cardiac arrest in an emergency department (ED). Methods: We conducted a retrospective data review of ED patients in an emergency department (ED) from 1 August 2014 to 31 July 2016. Significant variables in univariate analysis were used to create a multivariate analysis. Differentiation of predicting factors between cardiac arrest patient and non-cardiac arrest patients for occurrence of cardiac arrest in an emergency department (ED) was the primary outcome. Results: The data of 527 non-trauma patients with Emergency Severity Index (ESI) 1-3 were collected. The factors found to have a significant association (P < 0.05) in the non-cardiac arrest group versus the cardiac arrest group at the ED were systolic BP (mean [IQR] 135 [114,158] vs 120 [90,140] mmHg), oxygen saturation (mean [IQR] 97 [89,98] vs 82.5 [78,95]%), GCS (mean [IQR] 15 [15,15] vs 11.5 [8.815]), normal sinus rhythm (mean 59.8 vs 30%), sinus tachycardia (mean 46.7 vs 21.7%), pH (mean [IQR] 7.4 [7.3,7.4] vs 7.2 [7,7.3]), serum lactate (mean [IQR] 2 [1.1,4.2] vs 7 [5,10.8]), and MEWS score (mean [IQR] 3 [2,5] vs 5 [3,6]). A multivariate analysis was then performed. After adjusting for multiple factors, ESI level 2 patients were more likely to have cardiac arrest in the ER compared with ESI 1 (odds ratio [OR], 1.66; P < 0.001). Furthermore, ESI 2 patients were more likely than ESI 1 patients to have cardiovascular disease (OR, 1.89; P = 0.01), heart rate < 55 (OR, 6.83; P = 0.18), SBP < 90 (OR, 3.41; P = 0.006), SpO2 < 94 (OR, 4.76; P = 0.012), sinus tachycardia (OR, 4.32; P = 0.002), lactate > 4 (OR, 10.66; P = < 0.001), and MEWS > 4 (OR, 4.86; P = 0.028). These factors remained predictive of cardiac arrest at the ED. Conclusion: The factors related to cardiac arrest in the ED are ESI 1 patients, ESI 2 patients, patients diagnosed with cardiovascular disease, SpO2 < 94, lactate > 4, and a MEWS > 4. These factors can be used as markers in the event of simultaneous arrival of many patients and can help as a pre-state for patients who have a tendency to develop cardiac arrest. The hemodynamic status and vital signs of these patients should be closely monitored. Early detection of potentially critical conditions to prevent critical medical intervention is mandatory.

Keywords: cardiac arrest, predicting factor, emergency department, emergency patient

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Authors: Arleta Rifati Nixha, Mustafa Arslan, Adem Ergün, Nahit Gencer

Abstract:

Polyphenol oxidase (PPO), sometimes referred to as phenol oxidase, catecholase, phenolase, catechol oxidase, or even tyrosinase, is considered to be an o-dipenol. PPO (EC 1.14.18.1), a multifunctional copper containing enzyme, is widely distributed in nature. It catalyzes two distinct reactions of melanin synthesis: a hydroxylation of monophenols to o-diphenols (monophenolase activity) and an oxidation of o-diphenols to o-quinones (diphenolase activity), both using molecular oxygen. Additionaly, investigation demonstrated that various dermatological disorders, such as age spots and freckle, were caused by the accumulation of an excessive level of epidermal pigmentation. Tyrosinase has also been linked to Parkinson’s and other neurodegenerative diseases. Nitrogen heterocycles have received a great deal of attention in the literature because of biological properties. Especially, among these heterocyclic systems, pyridine containing compounds have been the subject of expanding research efforts in heteroaromatic and biological chemistry. The pyrido [2,3-d] pyrimidine heterocycles, which are those annelated to a pyrimidine ring, are important because of their wide range of biological and pharmaceutical applications (i.e., bronchodilators, vasodilators) and their anti-allergic, cardiotonic, antihypertensive, and hepatoprotective activities. In this study series of 12 new carbazole substituted pyridopyrimidine urea(thiourea) derivatives were synthesized and evaluated effect on PPO. Additionally, we presented structure-activity relationship analyses and theoretical calculations of the compounds.

Keywords: carbazole, pyridopyrimidine, urea, thiourea, tyrosinase inhibitors

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Authors: Gulnur Arabaci

Abstract:

Heavy metals are one of the major groups of contaminants in the environment and many of them are toxic even at very low concentration in plants and animals. However, some metals play important roles in the biological function of many enzymes in living organisms. Metals such as zinc, iron, and cooper are important for survival and activity of enzymes in plants, however heavy metals can inhibit enzyme which is responsible for defense system of plants. Polyphenol oxidase (PPO) is a copper-containing metalloenzyme which is responsible for enzymatic browning reaction of plants. Enzymatic browning is a major problem for the handling of vegetables and fruits in food industry. It can be increased and effected with many different futures such as metals in the nature and ground. In the present work, PPO was isolated and characterized from green leaves of red poppy plant (Papaver rhoeas). Then, the effect of some known antibrowning agents which can form complexes with metals and metals were investigated on the red poppy PPO activity. The results showed that glutathione was the most potent inhibitory effect on PPO activity. Cu(II) and Fe(II) metals increased the enzyme activities however, Sn(II) had the maximum inhibitory effect and Zn(II) and Pb(II) had no significant effect on the enzyme activity. In order to reduce the effect of heavy metals, the effects of metal-antibrowning agent complexes on the PPO activity were determined. EDTA and metal complexes had no significant effect on the enzyme. L-ascorbic acid and metal complexes decreased but L-ascorbic acid-Cu(II)-complex had no effect. Glutathione–metal complexes had the best inhibitory effect on Red poppy leaf PPO activity.

Keywords: inhibition, metal, red poppy, poly phenol oxidase (PPO)

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Authors: Yasser M. Saad, Heba El-Sebaie Abd El-Sadek

Abstract:

Some Metapenaeus monoceros cox1 gene fragments were isolated, purified, sequenced, and comparatively analyzed with some other Crustacean Cox1 gene sequences (obtained from National Center for Biotechnology Information). This work was designed for testing the efficiency of this system in reconstruction of phylogenetic relations among some Crustacean species belonging to four genera (Metapenaeus, Artemia, Daphnia and Calanus). The single nucleotide polymorphism and haplotype diversity were calculated for all estimated mt-DNA fragments. The genetic distance values were 0.292, 0.015, 0.151, and 0.09 within Metapenaeus species, Calanus species, Artemia species, and Daphnia species, respectively. The reconstructed phylogenetic tree is clustered into some unique clades. Cytochrome oxidase subunit 1 gene (cox1) was a powerful system in reconstruction of phylogenetic relations among evaluated crustacean species.

Keywords: crustaceans, genetics, Cox1, phylogeny

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Authors: Inkyong Yi

Abstract:

Acute kidney injury (AKI) in trauma patients is known to be associated with multiple factors, especially shock and consequent inadequate renal perfusion, yet its clinical presentation is little known in severe trauma patients. Our aim was to investigate the clinical presentation of acute kidney injury and its outcome in severe trauma patients at a level I trauma center. A total of 93 consecutive adult trauma patients with an injury severity score (ISS) of more than 15 were analyzed retrospectively from our Level I trauma center data base. Patients with direct renal injury were excluded. Patients were dichotomized into two groups, according to the presence of AKI. Various clinical parameters were compared between two groups, with Student’s T test and Mann-Whitney’s U test. The AKI group was further dichotomized into patients who recovered within seven days, and those who required more than 7days for recovery or those who did not recover at all. Various clinical parameters associated with outcome were further analyzed. Patients with AKI (n=33, 35%) presented with significantly higher age (61.4±17.3 vs. 45.4±17.3, p < 0.0001), incidence of comorbidities (hypertension; 51.5% vs. 13.3%, OR 6.906 95%CI 2.515-18.967, diabetes; 27.3% vs. 6.7%, OR 5.250, 95%CI 1.472-18.722), odds of head and neck trauma (69.7% vs. 41.7%, OR 3.220, 95%CI 1.306-7.942) and presence of shock during emergency room care (66.7% vs 21.7% OR 7.231, 95%CI, 2.798-18.687). Among AKI patients, patients who recovered within 1 week showed lower peak lactate (4.7mmol/L, 95%CI 2.9-6.5 vs 7.3mmol/L, 95%CI 5.0-9.6, p < 0.0287), lesser units of transfusion during first 24 hours (pRBC; 20.4unit, 95%CI 12.5-28.3 vs. 58.9unit, 95%CI 39.4-78.5, p=0.0003, FFP; 16.6unit, 95%CI 6.8-26.4 vs. 56.1unit, 95%CI 26.9-85.2, p=0.0027). In severe trauma patients, patients with AKI showed different clinical presentations and worse outcomes. Initial presence of shock and higher DIC profiles may be important risk factors for AKI in severe trauma patients. In patients with AKI, peak lactate level and amounts of transfusion are related to recovery.

Keywords: acute kidney injury, lactate, transfusion, trauma

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Authors: Ehsan Motamedian

Abstract:

Finding all optimal flux distributions of a metabolic model is an important challenge in systems biology. In this paper, a new algorithm is introduced to identify all alternate optimal solutions of a large scale metabolic network. The algorithm reduces the model to decrease computations for finding optimal solutions. The algorithm was implemented on the Escherichia coli metabolic model to find all optimal solutions for lactate and acetate production. There were more optimal flux distributions when acetate production was optimized. The model was reduced from 1076 to 80 variable fluxes for lactate while it was reduced to 91 variable fluxes for acetate. These 11 more variable fluxes resulted in about three times more optimal flux distributions. Variable fluxes were from 12 various metabolic pathways and most of them belonged to nucleotide salvage and extra cellular transport pathways.

Keywords: flux variability, metabolic network, mixed-integer linear programming, multiple optimal solutions

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Authors: Violina Angelova, Stefan Krustev

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The aim of this work was to evaluate the effectiveness of different soil test extractants for the determination of available soil phosphorus in five internationally certified standard soils, sludge and clay (NCS DC 85104, NCS DC 85106, ISE 859, ISE 952, ISE 998). The certified samples were extracted with the following methods/extractants: CaCl₂, CaCl₂ and DTPA (CAT), double lactate (DL), ammonium lactate (AL), calcium acetate lactate (CAL), Olsen, Mehlich 3, Bray and Kurtz I, and Morgan, which are commonly used in soil testing laboratories. The phosphorus in soil extracts was measured colorimetrically using Spectroquant Pharo 100 spectrometer. The methods used in the study were evaluated according to the recovery of available phosphorus, facility of application and rapidity of performance. The relationships between methods are examined statistically. A good agreement of the results from different soil test was established for all certified samples. In general, the P values extracted by the nine extraction methods significantly correlated with each other. When grouping the soils according to pH, organic carbon content and clay content, weaker extraction methods showed analogous trends; also among the stronger extraction methods, common tendencies were found. Other factors influencing the extraction force of the different methods include soil: solution ratio, as well as the duration and power of shaking the samples. The mean extractable P in certified samples was found to be in the order of CaCl₂ < CAT < Morgan < Bray and Kurtz I < Olsen < CAL < DL < Mehlich 3 < AL. Although the nine methods extracted different amounts of P from the certified samples, values of P extracted by the different methods were strongly correlated among themselves. Acknowledgment: The financial support by the Bulgarian National Science Fund Projects DFNI Н04/9 and DFNI Н06/21 are greatly appreciated.

Keywords: available soil phosphorus, certified samples, determination, soil test extractants

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Authors: Navodit Goel, Prabir K. Paul

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Tomato (Solanum lycopersicum) is attacked by Pseudomonas syringae pv. tomato causing speck lesions on the leaves leading to severe economic casualty. In the present study, aqueous fruit extracts of Azadirachta indica (neem) were sprayed on a single node of tomato plants grown under controlled contamination-free conditions. The treatment of plants was performed with neem fruit extract either alone or along with the pathogen. The parameters of observation were activities of polyphenol oxidase (PPO) and lysozyme, and isoform analysis of PPO; both at the treated leaves as well as untreated leaves away from the site of extract application. Polyphenol oxidase initiates phenylpropanoid pathway resulting in the synthesis of quinines from cytoplasmic phenols and production of reactive oxygen species toxic to broad spectrum microbes. Lysozyme is responsible for the breakdown of bacterial cell wall. The results indicate the upregulation of PPO and lysozyme activities in both the treated and untreated leaves along with de novo expression of newer PPO isoenzymes (which were absent in control samples). The appearance of additional PPO isoenzymes in bioelicitor-treated plants indicates that either the isoenzymes were expressed after bioelicitor application or the already expressed but inactive isoenzymes were activated by it. Lysozyme activity was significantly increased in the plants when treated with the bioelicitor or the pathogen alone. However, no new isoenzymes of lysozyme were expressed upon application of the extract. Induction of resistance by neem fruit extract could be a potent weapon in eco-friendly plant protection strategies.

Keywords: Azadirachta indica, lysozyme, polyphenol oxidase, Solanum lycopersicum

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Authors: Menan M. Elsayed, Hussein A. Heshmat

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The aim of the study is to determine the effect of 100 mg/d Pregnenolone on biological variables after plyometric training for volleyball players. Methods: 15 male volleyball players participated in this study. Serum levels of testosterone, creatine phosphokinase (CPK), lactate, and glucose were measured before and post-exercise. Results: Testosterone was not altered, while creatine phosphokinase (CPK), lactate, and glucose levels significantly decreased. It is recommended to use Pregnenolone administration to decreased muscle damage and delayed fatigue for volleyball players after plyometric training. In conclusion, this study demonstrated that oral Pregnenolone administration of 100 mg/d might decrease muscle damage and delayed fatigue which may affect positively the volleyball players after a plyometric training bout.

Keywords: biological variables, plyometric exercise program, pregnenolone, volleyball player

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Authors: Anahita Izadyar

Abstract:

Using a recombinant enzyme derived from corn and a simple modification, we are fabricating a facile, fast, and cost-beneficial novel biosensor to measure glucose. We are applying Plant Produced Mn Peroxidase (PPMP), glucose oxidase (GOx), polyaniline (PANI) as conductive polymer and gold nanoparticles (AuNPs) on Au electrode using electrochemical response to detect glucose. We applied the entrapment method of enzyme composition, which is generally used to immobilize conductive polymer and facilitate electron transfer from the enzyme oxidation-reduction center to the sample solution. In this work, the oxidation of glucose on the modified gold electrode was quantified with Linear Sweep Voltammetry(LSV). We expect that the modified biosensor has the potential for monitoring various biofluids.

Keywords: plant-produced manganese peroxidase, enzyme-based biosensors, glucose, modified gold nanoparticles electrode, polyaniline

Procedia PDF Downloads 163