Search results for: lactate oxidase

Authors: Ayesha Imtiaz, Darlina Md. Naim

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Family Nemipteridae is among the most abundantly distributed family in Malaysian fish markets due to its high contribution to landing sites of Malaysia. Using an advanced molecular approach that used two mitochondrial (Cytochrome oxidase c I and Cytochrome oxidase b) and one nuclear gene (Recombination activating gene, RAGI) to expose cryptic diversity and phylogenetic relationships among commercially important species of family Nemipteridae. Our research covered all genera (including 31 species out total 45 species) of family Nemipteridae, distributed in Malaysia. We also found certain type of geographical barriers in the South China sea that reduces dispersal and stops a few species to intermix. Northside of the South China Sea (near Vietnam) does not allow genetic diversity to mix with the Southern side of the South China sea (Sarawak) and reduces dispersal. Straits of Malacca reduce the intermixing genetic diversity of South China Sea and the Indian Ocean.

Keywords: Nemipteridae, RAG I, south east Asia, Malaysia

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Authors: Maymona Al-Husari, Craig Murdoch, Steven Webb

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Some tumors are known to exhibit an extracellular pH that is more acidic than the intracellular, creating a 'reversed pH gradient' across the cell membrane and this has been shown to affect their invasive and metastatic potential. Tumour hypoxia also plays an important role in tumour development and has been directly linked to both tumour morphology and aggressiveness. In this paper, we present a hybrid mathematical model of intracellular pH regulation that examines the effect of oxygen and pH on tumour growth and morphology. In particular, we investigate the impact of pH regulatory mechanisms on the cellular pH gradient and tumour morphology. Analysis of the model shows that: low activity of the Na+/H+ exchanger or a high rate of anaerobic glycolysis can give rise to a 'fingering' tumour morphology; and a high activity of the lactate/H+ symporter can result in a reversed transmembrane pH gradient across a large portion of the tumour mass. Also, the reversed pH gradient is spatially heterogenous within the tumour, with a normal pH gradient observed within an intermediate growth layer, that is the layer between the proliferative inner and outermost layer of the tumour.

Keywords: acidic pH, cellular automaton, ebola, tumour growth

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Authors: Mehdi Ariana, Javad Hamedi

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Nisin is a commercial bacteriocin that is used as a food preservative and produced by Lactococcus lactis subsp. lactis. Nisin production through co-culture fermentation can be performed for increasing nisin quantities. Since lactate accumulation in the fermentation medium can prevent L. lactis growth and therefore reduce nisin production, the simultaneous culture of microorganisms can enhance L. lactis growth by a reduction in the amount of lactic acid. In this study, conducted coculture of L.lactis subsp. lactic and the yeast Yarrowia lipolytica. Both strains are cultured in a molasses-based medium that is mainly constructed of sucrose. Y. lipolytica is not able to use sucrose as a carbon source but is able to consume lactate and decrease lactic acid in the medium. So, Lactic acid consumption can increase pH value and stimulate L. lactis growth. The results showed the mixed culture increased L. lactis growth 6 times higher than that of pure culture and could enhance nisin activity by up to 40%.

Keywords: co-culture fermentation, lactococcus lactis subsp lactis, yarrowia lipolytica, nisin

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Authors: Kuei-Yu Chien, Nai-Wen Kan, Wan-Chun Wu, Guo-Dong Ma, Shu-Chen Chen

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Purpose: The purpose of this study was to investigate the responses of systolic blood pressure (SBP), diastolic blood pressure (DBP), heart rate (HR), rating of perceived exertion (RPE) and lactate following continued high-intensity interval exercise in water and on land. The results of studies can be an exercise program design reference for health care and fitness professionals. Method: A total of 20 volunteer postmenopausal women was included in this study. The inclusion criteria were: duration of menopause > 1 year; and sedentary lifestyle, defined as engaging in moderate-intensity exercise less than three times per week, or less than 20 minutes per day. Participants need to visit experimental place three times. The first time visiting, body composition was performed and participant filled out the questionnaire. Participants were assigned randomly to the exercise environment (water or land) in second and third time visiting. Water exercise testing was under water of trochanter level. In continuing jump testing, each movement consisted 10-second maximum volunteer jump for two sets. 50% heart rate reserve dynamic resting (walking or running) for one minute was within each set. SBP, DBP, HR, RPE of whole body/thigh (RPEW/RPET) and lactate were performed at pre and post testing. HR, RPEW, and RPET were monitored after 1, 2, and 10 min of exercise testing. SBP and DBP were performed after 10 and 30 min of exercise testing. Results: The responses of SBP and DBP after exercise testing in water were higher than those on land. Lactate levels after exercise testing in water were lower than those on land. The responses of RPET were lower than those on land post exercise 1 and 2 minutes. The heart rate recovery in water was faster than those on land at post exercise 5 minutes. Conclusion: This study showed water interval jump exercise induces higher cardiovascular responses with lower RPE responses and lactate levels than on-land jumps exercise in postmenopausal women. Fatigue is one of the major reasons to obstruct exercise behavior. Jump exercise could enhance cardiorespiratory fitness, the lower-extremity power, strength, and bone mass. There are several health benefits to the middle to older adults. This study showed that water interval jumping could be more relaxed and not tried to reach the same land-based cardiorespiratory exercise intensity.

Keywords: interval exercise, power, recovery, fatigue

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Authors: M. Omer Faruk, A. M. A. M. Zonaed Siddiki, M. Fazal Karim, Md. Masuduzzaman, S. Chowdhury, Md. Shafiqul Islam, M. Alamgir Hossain

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The dog tapeworms Echinococcus granulosus develop hydatid cysts in various organs in human and domestic animals worldwide including Bangladesh. The aim of this study was to identify and characterize the genotype of E. granulosus isolated from cattle using 12S rRNA and Cytochrome oxidase 1 (COX 1) genes. A total of 43 hydatid cyst samples were collected from 390 examined cattle samples derived from slaughterhouses. Among them, three cysts were fertile. Genomic DNA was extracted from germinal membrane and/or protoscoleces followed by PCR amplification of mitochondrial 12S rRNA and Cytochrome oxidase 1 gene fragments. The sequence data revealed existence of G1 (64.28%) and possible G3 (21.43%) genotypes for the first time in Bangladesh. The study indicates that common sheep strain G1 is the dominant subtype of E. granulosus in Chittagong region of Bangladesh. This will increase our understanding of the epidemiology of hydatidosis in the southern part of the country and will be useful to plan suitable control measures in the long run.

Keywords: Echinococcus granulosus, Cox1, 12S rRNA, molecular characterization, Bangladesh

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Authors: Michal Kielbik, Izabela Szulc-Kielbik, Anna Brzostek, Jaroslaw Dziadek, Magdalena Klink

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The goal of many research groups in the world is to find new components that are important for survival of mycobacteria in the host cells. Mycobacterium tuberculosis (Mtb) possesses a number of enzymes degrading cholesterol that are considered to be an important factor for its survival and persistence in host macrophages. One of them - cholesterol oxidase (ChoD), although not being essential for cholesterol degradation, is discussed as a virulence compound, however its involvement in macrophages’ response to Mtb is still not sufficiently determined. The recognition of tubercle bacilli antigens by pathogen recognition receptors is crucial for the initiation of the host innate immune response. An important receptor that has been implicated in the recognition and/or uptake of Mtb is Toll-like receptor type 2 (TLR2). Engagement of TLR2 results in the activation and phosphorylation of intracellular signaling proteins including IRAK-1 and -4, TRAF-6, which in turn leads to the activation of target kinases and transcription factors responsible for bactericidal and pro-inflammatory response of macrophages. The aim of these studies was a detailed clarification of the role of Mtb cholesterol oxidase as a virulence factor affecting the TLR2 signaling pathway in human macrophages. As human macrophages the THP-1 differentiated cells were applied. The virulent wild-type Mtb strain (H37Rv), its mutant lacking a functional copy of gene encoding cholesterol oxidase (∆choD), as well as complimented strain (∆choD–choD) were used. We tested the impact of Mtb strains on the expression of TLR2-depended signaling proteins (mRNA level, cytosolic level and phosphorylation status). The cytokine and bactericidal response of THP-1 derived macrophages infected with Mtb strains in relation to TLR2 signaling pathway dependence was also determined. We found that during the 24-hours of infection process the wild-type and complemented Mtb significantly reduced the cytosolic level and phosphorylation status of IRAK-4 and TRAF-6 proteins in macrophages, that was not observed in the case of ΔchoD mutant. Decreasement of TLR2-dependent signaling proteins, induced by wild-type Mtb, was not dependent on the activity of proteasome. Blocking of TLR2 expression, before infection, effectively prevented the induced by wild-type strain reduction of cytosolic level and phosphorylation of IRAK-4. None of the strains affected the surface expression of TLR2. The mRNA level of IRAK-4 and TRAF-6 genes were significantly increased in macrophages 24 hours post-infection with either of tested strains. However, the impact of wild-type Mtb strain on both examined genes was significantly stronger than its ΔchoD mutant. We also found that wild-type strain stimulated macrophages to release high amount of immunosuppressive IL-10, accompanied by low amount of pro-inflammatory IL-8 and bactericidal nitric oxide in comparison to mutant lacking cholesterol oxidase. The influence of wild-type Mtb on this type of macrophages' response strongly dependent on fully active IRAK-1 and IRAK-4 signaling proteins. In conclusion, Mtb using cholesterol oxidase causes the over-activation of TLR2 signaling proteins leading to the reduction of their cytosolic level and activity resulting in the modulation of macrophages response to allow its intracellular survival. Supported by grant: 2014/15/B/NZ6/01565, National Science Center, Poland

Keywords: Mycobacterium tuberculosis, cholesterol oxidase, macrophages, TLR2-dependent signaling pathway

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Authors: Jyoti Kumari, Pavuluri Srinivasa Rao

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Jackfruit (ArtocarpusheterophyllusL.) is an underutilized yet highly nutritious fruit with unique flavour, known for its therapeutic and culinary properties. Fresh jackfruit bulb has a very short shelf life due to high moisture and sugar content leading to microbial and enzymatic browning, hindering its consumer acceptability and marketability. An attempt has been made for the preservation of the ripe jackfruit bulbs, by the application of high pressure (HP) over a range of 200-500 MPa at ambient temperature for dwell times ranging from 5 to 20 min. The physicochemical properties of jackfruit bulbs such as the pH, TSS, and titrable acidity were not affected by the pressurization process. The ripening index of the fruit bulb also decreased following HP treatment. While the ascorbic acid and antioxidant activity of jackfruit bulb were well retained by high pressure processing (HPP), the total phenols and carotenoids showed a slight increase. The HPP significantly affected the colour and textural properties of jackfruit bulb. High pressure processing was highly effective in reducing the browning index of jackfruit bulbs in comparison to untreated bulbs. The firmness of the bulbs improved upon the pressure treatment with longer dwelling time. The polyphenol oxidase has been identified as the most prominent oxidative enzyme in the jackfruit bulb. The enzymatic activity of polyphenol oxidase and peroxidase were significantly reduced by up to 40% following treatment at 400 MPa/15 min. HPP of jackfruit bulbs at ambient temperatures is shown to be highly beneficial in improving the shelf stability, retaining its nutrient profile, color, and appearance while ensuring the maximum inactivation of the spoilage enzymes.

Keywords: antioxidant capacity, ascorbic acid, carotenoids, color, HPP-high pressure processing, jackfruit bulbs, polyphenol oxidase, peroxidase, total phenolic content

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Authors: K. Raza, S. Gul, M. Ali

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Concrete is a fundamental building material, extensively utilized by the construction industry. Problems related to the strength of concrete is an immense issue for the sustainability of concrete structures. Concrete mostly loses its strength due to the cracks produced in it by shrinkage or hydration process. This study aims to enhance the strength and service life of the concrete structures by incorporating bio-based admixture in the concrete. By the injection of bio-based admixture (BBA) in concrete, it will self-heal the cracks by producing calcium carbonate. Minimization of cracks will compact the microstructure of the concrete, due to which strength will increase. For this study, Bacillus subtilis will be used as a bio-based admixture (BBA) in concrete. Calcium lactate up to 1.5% will be used as the food source for the Bacillus subtilis in concrete. Two formulations containing 0 and 5% of Bacillus subtilis by weight of cement, will be used for the casting of concrete specimens. Direct mixing method will be adopted for the usage of bio-based admixture in concrete. Compressive strength test will be carried out after 28 days of curing. Scanning electron microscopy (SEM) and X-ray diffraction analysis (XRD) will be performed for the examination of micro-structure of concrete. Results will be drawn by comparing the test results of 0 and 5% the formulations. It will be recommended to use to bio-based admixture (BBA) in concrete for columns because of the satisfactory increase in the compressive strength of concrete.

Keywords: bio-based admixture, Bacillus subtilis, calcium lactate, compressive strength

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Authors: Abdolrasoul Daneshjoo

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The aim of this study was studying the effect of acute creatine supplementation on physiological variables of continuous and intermittent soccer activities of men soccer players. 32 soccer players from Tarbiat Moalem University aged (22/3+-1/6) volunteered for this research and were divided into two groups randomly. Both experimental and control groups after 6 days taking supplementation were tested. For measuring height and weight meter and balance were used. Questionnaire for health background, lactate electro, heart beat measuring polar electro, continuous and intermittent training program and time recorder were used for data collection. For data analysis descriptive statistical techniques, two-way ANOVA and F test were used. The result of this study showed increased significantly in heart rate in control group. For control group heart beat was (71/6 +- 3/5) and for experimental group it was (75/3 +- 4/9). No significant differences were observed in players weight after taking creatine.

Keywords: heartbeat, lactate Blood, creatine, soccer players of Tarbiat Moalem University

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Authors: Sangeeta Palekar, Neeraj Rangwani, Akash Poddar, Jayu Kalambe

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The bio-medical analysis is an indispensable procedure for identifying health-related diseases like diabetes. Monitoring the glucose level in our body regularly helps us identify hyperglycemia and hypoglycemia, which can cause severe medical problems like nerve damage or kidney diseases. This paper presents a method for predicting the glucose concentration in blood samples using image processing and machine learning algorithms. The glucose solution is prepared by the glucose oxidase (GOD) and peroxidase (POD) method. An experimental database is generated based on the colorimetric technique. The image of the glucose solution is captured by the raspberry pi camera and analyzed using image processing by extracting the RGB, HSV, LUX color space values. Regression algorithms like multiple linear regression, decision tree, RandomForest, and XGBoost were used to predict the unknown glucose concentration. The multiple linear regression algorithm predicts the results with 97% accuracy. The image processing and machine learning-based approach reduce the hardware complexities of existing platforms.

Keywords: artificial intelligence glucose detection, glucose oxidase, peroxidase, image processing, machine learning

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Authors: Noor-ul-Ain, Muhammad Tahir Khan, Kashif Khan, Adeela Ajmal, Hamid Mustafa

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The objective of this study was to evaluate the preservative and upgrading potential of urea (70g/kg DM) added to high moisture oat silage at laboratory scale trial and urea was hydrolysed 95%. Microbial activity measured by pH and volatile fatty acids (VFA) and lactate production was reduced (p<0.001) by the urea addition. The pH of oat silage (without treated) was measured 5.7 and increased up to 8.00 on average while; volatile fatty acids (VFA) concentration was decreased. Relative proportions of fermentation acids changed after urea addition, increasing the acetate and butyrate and decreasing the propionate and lactate proportions. The addition of urea to oat silages increased (P<0.001) water soluble and ammonium nitrogen of the forage. These nitrogen fractions represented more than 40% of total nitrogen. After urea addition, total nitrogen content of oat silages increased from 21.0 g/kg DM to 28 g/kg DM. Application of urea at a rate of 70 g/kg DM significantly increased (P<0.001) the in situ degradation of neutral-detergent fibre after 48h of rumen incubation (NDF-situ). The NDF-situ was 200 g/kg NDF higher on oat forages ensiled with urea than on oat forages ensiled without urea. Oat silages can be effectively preserved and upgraded by ensiling with 70 g urea/kg dry matter. Further studies are required to evaluate voluntary intake of this forage.

Keywords: oat, silage, urea, pH, forage

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Authors: Mohammad Java Esmaeily

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Background: We evaluate the effects of tissue and organ perfusion during and after coronary artery bypass graft surgery with either colloid (Voluven) or crystalloid (Lactated ringers) as a prime solution. Materials and Methods: In this prospective randomized-controlled trial study, 70 patients undergoing on-pump coronary artery bypass graft surgery were randomly assigned to receive either colloid (Voluven) or crystalloid (Lactated ringer's) as a prime solution for initiation of cardiopulmonary bypass machine procedure. Tissue and organ perfusion markers, including lactate, troponin I, liver and renal function tests and electrolytes, were measured sequentially before induction (T1) to the second days after surgery (T5). Results: With the exception of chloride and potassium levels, no significant differences were detected in other measurements, and laboratory results were identical entirely in the two groups. Conclusion: Voluven® (hydroxyethyl starch, HES 130/0.4) has a not significant difference in comparison with crystalloid (Lactated ringer's) as priming solution on the basis of organ and tissue perfusion tests assessment.

Keywords: prime, colloid, crystalloid, lactate, troponin, hydroxyethyl starch

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Authors: Khaled S. Al Salhen

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Aldehyde oxidase (AO) and xanthine oxidoreductase (XOR) catalyze the oxidation of many different N-heterocyclic compounds as well as aliphatic and aromatic aldehydes to their corresponding lactam and carboxylic acids respectively. The present study examines the oxidation of dimethylamino-cinnamaldehyde (DMAC), vanillin and phenanthridine by AO and xanthine by XOR from Drosophila cytosol. Therefore, the results obtained in the present study showed the DMAC, vanillin and phenanthridine substrates used were found to be good substrates of Drosophila AO and xanthine is the preferred substrate for Drosophila XOR. Km values of AO substrates were observed with DMAC (50±5.4 µM), phenanthridine (80±9.1 µM) and vanillin (303±11.7 µM) respectively for Drosophila cytosol. The Km values for DMAC and phenanthridine were ~6 and ~4 fold lower than that for vanillin as a substrate. The Km for XOR with xanthine using NAD+ as an electron acceptor was 27±4.1 µM. Relatively low Vmax values were obtained with phenanthridine (1.78±0.38 nmol/min/mg protein) and DMAC (1.80±0.35 nmol/min/mg protein). The highest Vmax was obtained from Drosophila cytosol with vanillin (7.58±2.11 nmol/min/mg protein). It is concluded these results that AO and XOR in Drosophila were able to catalyse the biotransformation of numerous substrates of the well-characterised mammalian AO and XOR. The kinetic parameters have indicated that the activity of AO of Drosophila may be a significant factor the oxidation of aromatic aldehyde compounds.

Keywords: aldehyde oxidase, xanthine oxidoreductase, dimethylamino-cinnamaldehyde, vanillin, phenanthridine, Drosophila melanogaster

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Authors: Yijun Lai, Saber Khederzadeh, Lingshaung Han

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Species in Thunnus are organized due to the similarity between them. The closeness between T. maccoyii, T. thynnus, T. Tonggol, T. atlanticus, T. albacares, T. obsesus, T. alalunga, and T. orientails are in different degrees. However, the genetic pattern of differentiation has not been presented based on individuals yet, to the author’s best knowledge. Hence, we aimed to analyze the difference in individuals level of tuna species to identify the factors that contribute to the maternal lineage variety using Cytochrome c oxidase subunit I (COXI) gene sequences. Our analyses provided evidence of sharing lineages in the Thunnus. A phylogenetic analysis revealed that these lineages are basal to the other sequences. We also showed a close connection between the T. tonggol, T. thynnus, and T. albacares populations. Also, the majority of the T. orientalis samples were clustered with the T. alalunga and, then, T. atlanticus populations. Phylogenetic trees and migration modeling revealed high proximity of T. thynnus sequences to a few T. orientalis and suggested possible gene flow with T. tonggol and T. albacares lineages, while all T. obsesus samples indicated unique clustering with each other. Our results support the presence of old maternal lineages in Thunnus, as a legacy of an ancient wave of colonization or migration.

Keywords: Thunnus Tuna, phylogeny, maternal lineage, COXI gene

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Authors: Thouraya Majoul

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Obesity is now a real public health issue throughout the world, and it is well-established that obesity leads to cardiovascular diseases. The prevention and treatment of obesity using nutritional supplements has become a realistic and effective approach. This study was carried out to analyze the incidence of a high-fat diet on rat heart metabolism as well as on fatty acids composition, then to investigate the eventual protective effects of a grape seed extract (GSE). The experimental design consisted of three rat groups subjected to three different conditions; standard (SD), high-fat diet (HFD) and HFD+GSE (HG). We showed that GSE counteracted the effect of HFD on fatty acid composition, namely, docosapentaenoic acid, docosahexaenoic acid, arachidonic acid (ARA), palmitic acid (PA) and palmitoleic acid. Besides, GSE treatment restored HFD-altered metabolic pathways through the recovery of some cardiac enzyme activities such as lipase, glucose 6 phosphate dehydrogenase and pyruvate dehydrogenase. The cardiac lactate level and lactate dehydrogenase activity were also analyzed in relation to HFD and GSE administration. To our knowledge, this is the first study showing the anti-obesity and cardioprotective effects of GSE in relation to fatty acid composition and some cardiac enzymes, supporting its role as a therapeutic agent of obesity.

Keywords: Grape seed extract, phenolic, obesity, cardioprotective, lipotoxicity, energy metabolism

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Authors: V. Surena, B. Nazemisalman, F. Noghrehkar

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Introduction: Gingival overgrowth (GO) is a common side effect involving users of antiepileptic, immunosuppressive and calcium channel blocker drugs. Cyclosporine and phenytoin are amongst the most widely used drugs associated with GO. Gingival fibroblasts seem to have a significant role in the production of certain enzymes after administration of the drugs contributing to GO. Previous studies have shown a higher prevalence of GO in children and adolescents. The aim of this study was to compare normal human gingival fibroblasts with those exposed to Cyclosporine or phenytoin in measuring the production levels of certain enzymes that could have a possible role in GO. Methods: samples were obtained from the gingival biopsies of seven adult and seven children and were cultured into plates. With the growth of fibroblast cells, they were treated with or without either Cyclosporine or phenytoin. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to determine the expressed levels of R-EGF, cathepsin B,L, Lysyl oxidase, COL1, TGF β1, MMP-1,2, and TIMP1. Results: according to RT-PCR analyses, the expressed levels of R-EGF, cathepsin B, L, Lysyl oxidase, COL1, TGF β1, MMP-1, 2 and TIMP1 were affected by Cyclosporine and phenytoin. TGF-β1, TIMP, Cathepsin B and EGF showed comparable values in the adult and pediatric groups. Conclusions: Different expressed levels of enzymes after treatment of the gingival fibroblasts of adults and pediatrics with phenytoin or Cyclosporine could be the reason for the higher severity of GO in children. More studies need to be performed on the pathogenesis of GO at different age groups.

Keywords: cyclosporine, fibroblasts, phenytoin, gingivae

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Authors: Mattea Carmen Castrovilli, Antonella Cartoni

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Electrospray ionisation (ESI), a well-established technique widely used to produce ion beams of biomolecules in mass spectrometry (ESI-MS), can be used for ambient soft landing of enzymes on a specific substrate. In this work, we show how the ambient electrospray deposition (ESD) technique can be successfully exploited for manufacturing a promising, green-friendly electrochemical amperometric laccase-based biosensor with unprecedented reuse and storage performance. These biosensors have been manufactured by spraying a laccase solution of 2μg/μL at 20% of methanol on a commercial carbon screen printed electrode (C-SPE) using a custom ESD set-up. The laccase-based ESD biosensor has been tested against catechol compounds in the linear range 2-100 μM, with a limit of detection of 1.7 μM, without interference from cadmium, chrome, arsenic, and zinc and without any memory effects, but showing a matrix effect in lake and well water. The ESD biosensor shows enhanced performances compared to the ones fabricated with other immobilization methods, like drop-casting. Indeed, it retains 100% activity up to two months of storage at ambient conditions without any special care and working stability up to 63 measurements on the same electrode just prepared and 20 on a one-year-old electrode subjected to redeposition together with a 100% resistance to use of the same electrode in subsequent days. The ESD method is a one-step, environmentally friendly method that allows the deposition of the bio-recognition layer without using any additional chemicals. The promising results in terms of storage and working stability also obtained with the more fragile lactate oxidase enzyme suggest these improvements should be attributed to the ESD technique rather than to the bioreceptor, highlighting how the ESD could be useful in reducing pollution from disposable devices. Acknowledgment: The understanding at the molecular level of this promising biosensor by using different spectroscopies, microscopies and analytical techniques is the subject of our PRIN 2022 project ESILARANTE.

Keywords: reuse, storage performance, immobilization, electrospray deposition, biosensor, laccase, catechol detection, green chemistry

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Authors: Ofonime U. M. John, Justina I. R. Udotong, Victor O. Nwaugo, Ime R. Udotong

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Oily wastes from oil and gas exploration and production (O&G E&P) activities were remediated for twelve weeks using Soil-Poultry dropping amendment. Culture-dependent microbiological, chemical and enzymatic techniques were employed to assess the efficacy of remediation process. Microbiological activities of the remediated wastes showed increased hydrocarbonoclastic microbial populations with increased remediation time; 2.7±0.1 x 105cfu/g to 8.3 ± 0.04 x106cfu/g for hydrocarbon utilizing bacteria, 1.7 ± 0.2 x103cfu/g to 6.0 ± 0.01 x 104cfu/g for hydrocarbon utilizing fungi and 2.2 ± 0.1 x 102cfu/g to 6.7 ± 0.1 x 103cfu/g for hydrocarbon utilizing actinomycetes. Bacteria associated with the remediated wastes after the remediation period included the genera Bacillus, Psuedomonas, Beijerinckia, Acinetobacter, Alcaligenes and Serratia. Fungal isolates included species of Penicillium, Aspergillus and Cladosporium, while the Actinomycetes included species of Rhodococcus, Nocardia and Streptomyces. Slight fluctuations in pH values between 6.5± 0.2 and 7.1 ± 0.08 were recorded throughout the process, while total petroleum hydrocarbon (TPH) content decreased from 89, 900 ± 0.03mg/kg to 425 ± 0.1 mg/kg after twelve weeks of remediation. The polycyclic aromatic hydrocarbon (PAH) levels decreased with increased remediation time; naphthalene, flourene, pheneanthrene, anthracene, pyrene, chrysene and benzo(b)flouranthene showed decreased values < 0.01 after twelve weeks of remediation. Enzyme activities revealed increased dehydrogenase and urease activities with increased remediation time and decreased phenol oxidase activity with increased remediation period. There was a positive linear correlation between densities of hydrocarbonoclastic microbes and dehydrogenase activity. On the contrary, phenol oxidase and urease activities showed negative correlation with microbial population. Results of this study confirmed that remediation of oily wastes using soil-poultry dropping amendment can result in eco-friendly O&G E&P wastes. It also indicates that urease and phenol oxidase activities can be reliable indices/tools to monitor PAH levels and rates of petroleum hydrocarbon degradation.

Keywords: dehydrogenase activity, oily wastes, remediation, soil-poultry dropping amendment

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Authors: Uc-Cayetano E. G., Ake-Uh O. E., Villanueva-Mena I. E., Ordonez L. C.

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Carbon nanotubes (CNTs) and other graphitic nanostructures are materials with extraordinary physical, physicochemical and electrochemical properties which are being aggressively investigated for a variety of sensing applications. Thus, sensing of biological molecules such as proteins, DNA, glucose and other enzymes using either single wall or multiwall carbon nanotubes (MWCNTs) has been widely reported. Despite the current progress in this area, the electrochemical response of CNTs used in a variety of sensing arrangements still needs to be improved. An alternative towards the enhancement of this CNTs' electrochemical response is to chemically (or physically) modify its surface. The influence of the decoration with iron oxide nanoparticles in different types of MWCNTs on the amperometric sensing of glucose, urea, and cholesterol in solution is investigated. Commercial MWCNTs were oxidized in acid media and subsequently decorated with iron oxide nanoparticles; finally, the enzymes glucose oxidase, urease, and cholesterol oxidase are chemically immobilized to oxidized and decorated MWCNTs for glucose, urease, and cholesterol electrochemical sensing. The results of the electrochemical characterizations consistently show that the presence of iron oxide nanoparticles decorating the surface of MWCNTs enhance the amperometric response and the sensitivity to increments in glucose, urease, and cholesterol concentration when compared to non-decorated MWCNTs.

Keywords: WCNTs, enzymes, oxidation, decoration

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Authors: Gourav Jain, Sameer Dixit, Surjeet Kumar Arya, Praveen C. Verma

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Plant cell wall plays a major role in defence mechanism against biotic and abiotic stress as it constitutes the physical barrier between the microenvironment and internal component of the cell. It is a complex structure composed of mostly carbohydrates among which cellulose and hemicelluloses are most abundant that is embedded in a matrix of pectins and proteins. Multiple enzymes have been reported which plays a vital role in cell wall modification, Pectin Methylesterase (PME) is one of them which catalyses the demethylesterification of homogalacturonans component of pectin which releases acidic pectin and methanol. As emitted methanol is toxic to the insect pest, we use PME gene for the better methanol production. In the current study we showed overexpression of PME gene isolated from Withania somnifera under the insect inducible promoter causes enhancement of methanol production at the time of insect feeds to plants, and that provides better insect resistance property. We found that the 85-90% mortality causes by transgenic tobacco in both chewing (Spodoptera litura larvae and Helicoverpa armigera) and sap-sucking (Aphid, mealybug, and whitefly) pest. The methanol content and emission level were also enhanced by 10-15 folds at different inducible time point interval (15min, 30min, 45min, 60min) which would be analysed by Purpald/Alcohol Oxidase method.

Keywords: methanol, Pectin methylesterase, inducible promoters, Purpald/Alcohol oxidase

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Authors: Thanura Randula Abeywardena

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This study recognized the effectiveness of cold-water immersion recovery post exhaustive short-term exercise. The purpose of this study was to understand if 16- 20°C of cold-water immersion would be beneficial in a tropical environment to achieve optimal recovery in sprint swim performance in comparison to 10-15°C of water immersion. Two 100m-sprint swim performance times were measured along with blood lactate (BLa), heart rate (HR) and rate of perceived exertion (RPE) in a 25m swimming pool with full body head out horizontal water immersions of 10-15°C, 16-20°C and 29-32°C (pool temperature) for 10 minutes followed by 5 minutes of seated passive rest outside; in between the two swim performances. Twelve well-trained adult swimmers (5 male and 5 female) within the top twenty in the Sri Lankan national swimming championships in 100m Butterfly and Freestyle in the years 2020 & 2021 volunteered for this study. One-way ANOVA analysis (p<0.05) suggested performance time, Bla and HR had no significant differences between the 3 conditions after the second sprint; however, RPE was significantly different with p=0.034 between 10-15°C and 16-20°C immersion conditions. The study suggested that the recovery post the two cold-water immersion conditions were similar in terms of performance and physiological factors; however, the 16-20°C temperature had a better “feel good” factor post sprint 2. Further study is recommended as there was participant bias with the swimmers not reaching optimal levels in sprint 1. Therefore, they might have possibly fully recovered before sprint 2, invalidating the physiological effect of recovery.

Keywords: hydrotherapy, blood lactate, fatigue, recovery, sprint-performance, sprint-swimming

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Authors: R. M. Srivastava, Subbanna A.R.N.S, Md Abbas Ahmad, S. P.More, Shivashankar, B. Kalyanbabu

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The similar morphology associated with high genetic variability poses problems in phylogenetic studies of Helicoverpa armigera (Hubner). To identify genetic variation of North Western Himalayan population’s, partial (Mid to terminal region) cytochrome c oxidase subunit I (COX-1) gene was amplified and sequenced for three populations collected from Pantnagar, Almora, and Chinyalisaur. The alignment of sequences with other two populations, Nagpur representing central India population and Anhui, China representing complete COX-1 sequence revealed unanimity in middle region with eleven single nucleotide polymorphisms (SNPs) in Nagpur populations. However, the consensus is missing when approaching towards terminal region, which is associated with 15 each SNPs and pair base substitutions in Chinyalisaur populations. In minimum evolution tree, all the five populations were majorly separated into two clades, one comprising of only Nagpur population and the other with rest. Amongst, North Western populations, Chinyalisaur one is promising by farming a separate clade. The pairwise genetic distance ranges from 0.025 to 0.192 with the maximum between H. armigera populations of Nagpur and Chinyalisaur. This genetic isolation of populations can be attributed to a key role of topological barriers of weather and mountain ranges and temporal barriers due to cropping patterns.

Keywords: cytochrome c oxidase subunit I, northwestern Himalayan population, Helicoverpa armigera (Noctuidae: Lepidoptera), phylogenetic relationship, genetic variation

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Authors: Boris Nemzer, Tania Reyes-Izquierdo, Zbigniew Pietrzkowski

Abstract:

Glucosinolate is a family of glucosides that can be found in a family of brassica vegetables. Upon the damage of the plant, glucosinolate breakdown by an internal enzyme myrosinase (thioglucosidase; EC 3.2.3.1) into isothiocyanates, such as sulforaphane. Sulforaphane is formed by glucoraphanin cleaving the sugar off by myrosinase and rearranged. Sulforaphane nitrile is formed in the same reaction as sulforaphane with the active of epithiospecifier protein (ESP). Most common food processing procedure would break the plant and mix the glucoraphanin and myrosinase together, and the formed sulforaphane would be further degraded. The purpose of this study is to understand the glucoraphanin/sulforaphane and the myrosinase activity of broccoli seeds germinated at a different time and technological processing conditions that keep the activity of the enzyme to form sulforaphane. Broccoli seeds were germinated in the house. Myrosinase activities were tested as the glucose content using glucose assay kit and measured UV-Vis spectrophotometer. Glucosinolates were measured by HPLC/DAD. Sulforaphane was measured using HPLC-DAD and GC/MS. The 6 hr germinated sprouts have a myrosinase activity 32.2 mg glucose/g, which is comparable with 12 and 24 hour germinated seeds and higher than dry seeds. The glucoraphanin content in 6 hour germinated sprouts is 13935 µg/g which is comparable to 24 hour germinated seeds and lower than the dry seeds. GC/MS results show that the amount of sulforaphane is higher than the amount of sulforaphane nitrile in seeds, 6 hour and 24 hour germinated seeds. The ratio of sulforaphane and sulforaphane nitrile is high in 6 hour germinated seeds, which indicates the inactivated ESP in the reaction. After evaluating the results, the short time germinated seeds can be used as the source of glucoraphanin and myrosinase supply to form potential higher sulforaphane content. Broccoli contains glucosinolates, glucoraphanin (4-methylsulfinylbutyl glucosinolate), which is an important metabolite with health-promoting effects. In the pilot clinical study, we observed the effects of a glucosinolates/glucoraphanin-rich extract from short time germinated broccoli seeds on blood adenosine triphosphate (ATP), reactive oxygen species (ROS) and lactate levels. A single dose of 50 mg of broccoli sprouts extract increased blood levels of ATP up to 61% (p=0.0092) during the first 2 hours after the ingestion. Interestingly, this effect was not associated with an increase in blood ROS or lactate. When compared to the placebo group, levels of lactate were reduced by 10% (p=0.006). These results indicate that broccoli germinated seed extract may positively affect the generation of ATP in humans. Due to the preliminary nature of this work and promising results, larger clinical trials are justified.

Keywords: broccoli glucosinolates, glucoraphanin, germinated seeds, myrosinase, adenosine triphosphate

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Authors: B. Nakhostin-Roohi, F. Khoshkhahesh, KH. Parandak, R. Ramazanzadeh

Abstract:

Introduction: The protective effect of antioxidants in diminishing the post-exercise rise of serum CK and LDH in individuals trained for competitive sports has come to light in recent years. This study was conducted to assess the effect of Two-week L-carnitine supplementation on exercise-induced muscle damage, as well as antioxidant capacity after a bout of strenuous exercise in active healthy young men. Methodology: Twenty active healthy men volunteered for this study. Participants were randomized in a double-blind placebo-controlled fashion into two groups: L-carnitine (C group; n = 10) and placebo group (P group; n = 10). The participants took supplementation (2000 mg L-carnitine) or placebo (2000 mg lactose) daily for 2weeks before the main trial. Then, participants ran 14 km. Blood samples were taken before supplementation, before exercise, immediately, 2h and 24h after exercise. Creatine kinase (CK), and lactate dehydrogenase (LDH), and total antioxidant capacity (TAC) were measured. Results: Serum CK and LDH significantly increased after exercise in both groups (p < 0.05). Serum LDH was significantly lower in C group than P group 2h and 24h after exercise (p < 0.05). Furthermore, CK was significantly lower in C group compared with P group just 24h after exercise (p < 0.05). Plasma TAC increased significantly 14 days after supplementation and 24h after exercise in C group compared with P group (p < 0.05). Discussion and conclusion: These results suggest two-week daily oral supplementation of L-carnitine has been able to promote antioxidant capacity before and after exercise and decrease muscle damage markers through possibly inhibition of exercise-induced oxidative stress.

Keywords: L-carnitine, muscle damage, creatine kinase, Lactate dehydrogenase

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Authors: Zain Ul Abdin, Mirza Aizaz Asim, Rao Sohail Ahmad Khan, Luqman Amrao, Fiaz Hussain, Hasooba Hira, Saqi Kosar Abbas

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The success of Integrated vector management programmes mainly depends on the correct identification of insect vector species involved in vector borne diseases. Based on molecular data the most important insect species involved as vectors for Zoonotic diseases in Pakistan were identified. The precise and accurate identification of such type of organism is only possible through molecular based techniques like “DNA barcoding”. Morphological species identification in insects at any life stage, is very challenging, therefore, DNA barcoding was used as a tool for rapid and accurate species identification in a wide variety of taxa across the globe and parallel studies revealed that DNA barcoding data can be effectively used in resolving taxonomic ambiguities, detection of cryptic diversity, invasion biology, description of new species etc. A comprehensive survey was carried out for the collection of insects (both adult and immature stages) in district Faisalabad, Pakistan and their DNA was extracted and mitochondrial cytochrome oxidase subunit I (COI-59) barcode sequences was used for molecular identification of immature and adult life stage.This preliminary research work opens new frontiers for developing sustainable insect vectors management programmes for saving lives of mankind from fatal diseases.

Keywords: zoonotic diseases, cytochrome oxidase, and insect vectors, CO1

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Authors: H. C. Ozdamar , O. Kilic-Erkek, H. E. Akkaya, E. Kilic-Toprak, M. Bor-Kucukatay

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Nordic hamstring exercise (NHE) is used to increase hamstring muscle strength, prevent injuries. The aim of this study was to reveal the acute, long-term effects of 10-week NHE, followed by 5, 10-week detraining on anthropometric measurements, flexibility, anaerobic power, muscle architecture, damage, fatigue, oxidative stress, plasma viscosity (PV), blood lactate levels. 40 sedentary, healthy male volunteers underwent 10 weeks of progressive NHE followed by 5, 10 weeks of detraining. Muscle architecture was determined by ultrasonography, stiffness by strain elastography. Anaerobic power was assessed by double-foot standing, long jump, vertical jump, flexibility by sit-lie, hamstring flexibility tests. Creatine kinase activity, oxidant/antioxidant parameters were measured from venous blood by a commercial kit, whereas PV was determined using a cone-plate viscometer. The blood lactate level was measured from the fingertip. NHE allowed subjects to lose weight, this effect was reversed by detraining for 5 weeks. Exercise caused an increase in knee angles measured by a goniometer, which wasn’t affected by detraining. 10-week NHE caused a partially reversed increase in anaerobic performance upon detraining. NHE resulted in increment of biceps femoris long head (BFub) area, pennation angle, which was reversed by detraining of 10-weeks. Blood lactate levels, muscle pain, fatigue were increased after each exercise session. NHE didn’t change oxidant/antioxidant parameters; 5-week detraining resulted in an increase in total oxidant capacity (TOC) and oxidative stress index (OSI). Detraining of 10 weeks caused a reduction of these parameters. Acute exercise caused a reduction in PV at 1 to 10 weeks. Pre-exercise PV measured on the 10th week was lower than the basal value. Detraining caused the increment of PV. The results may guide the selection of the exercise type to increase performance and muscle strength. Knowing how much of the gains will be lost after a period of detraining can contribute to raising awareness of the continuity of the exercise. This work was supported by PAU Scientific Research Projects Coordination Unit (Project number: 2018SABE034)

Keywords: anaerobic power, detraining, Nordic hamstring exercise, oxidative stress, plasma viscosity

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Authors: Samet Aktaş, Nurtekin Erkmen, Faruk Guven, Halil Taskin

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This study aimed to determine the effects of 3 versus 3 small-sided games (SSG) with different recovery times on soma physiological parameters in soccer players. Twelve soccer players from Regional Amateur League volunteered for this study (mean±SD age, 20.50±2.43 years; height, 177.73±4.13 cm; weight, 70.83±8.38 kg). Subjects were performing soccer training for five days per week. The protocol of the study was approved by the local ethic committee in School of Physical Education and Sport, Selcuk University. The subjects were divided into teams with 3 players according to Yo-Yo Intermittent Recovery Test. The field dimension was 26 m wide and 34 m in length. Subjects performed two times in a random order a series of 3 bouts of 3-a-side SSGs with 3 min and 5 min recovery durations. In SSGs, each set were performed with 6 min duration. The percent of maximal heart rate (% HRmax), blood lactate concentration (LA) and Rated Perceived Exertion (RPE) scale points were collected before the SSGs and at the end of each set. Data were analyzed by analysis of variance (ANOVA) with repeated measures. Significant differences were found between %HRmax in before SSG and 1st set, 2nd set, and 3rd set in both SSG with 3 min recovery duration and SSG with 5 min recovery duration (p<0.05). Means of %HRmax in SSG with 3 min recovery duration at both 1st and 2nd sets were significantly higher than SSG with 5 min recovery duration (p<0.05). No significant difference was found between sets of either SSGs in terms of LA (p>0.05). LA in SSG with 3 min recovery duration was higher than SSG with 5 min recovery duration at 2nd sets (p<0.05). RPE in soccer players was not different between SSGs (p>0.05).In conclusion, this study demonstrates that exercise intensity in SSG with 3 min recovery durations is higher than SSG with 5 min recovery durations.

Keywords: small-sided games, soccer, heart rate, lactate

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Authors: Anguo Chen, Huajie Chen, Caimei Yang, Qihua Hong, Jun Feng

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The experiment was conducted with 360 one-day-old Avian commercial broilers to study the effects of dietary cinnamon extract (CE), garlic extract (GE) and yucca extract (YE) on growth performance and serum biochemical parameters in broilers. The chickens were randomly divided equally into 4 treatment groups, each group with 3 replications, and received the same basal corn-bean diets included a starter from 1 d to 21 d and then a grower until 42 d, added with recommended dose 250 mg/kg CE, 25 mg/kg GE and 10 mg/kg YE to relevant group, respectively. The birds were kept in a stainless steel net coop each replication with 24 h light and were fed and drunk ad libitum. At 21 d and 42 d of age, 6 chicks were respectively picked out from every group and were bled to collect serum samples and intestinal samples for laboratory analysis. The results showed that the average daily gain (ADG) of CE, GE and YE group were increased by 7.20% (P<0.05), 3.43% (P>0.05) and 4.89% (P>0.05), feed gain ratio (F/G) was improved by 9.71% (P<0.05), 3.40% (P>0.05) and 3.40% (P>0.05) compared with the control, respectively. At 21 d of age, the content of serum urea nitrogen (SUN) and serum uric acid (SUA) and the activity of serum xanthine oxidase (SXO) in CE group were reduced by 35.17% (P<0.01), 13.73% (P<0.01) and 16.33% (P<0.05) compared with the control, respectively. At 42 d of age, SUN and SUA level and SXO activity were lowered by 24.35% (P<0.01), 15.49% (P<0.05) and 23.09% (P<0.01), respectively. The SXO activity in CE group was decreased by 14.86% (P<0.01) and 15.34%(P<0.01) compare with GE and YE group, respectively. Also, adding CE, GE and YE into broiler diets resulted in lower UN and UA level of intestinal contents. It is clear that CE was more significantly decreased the SXO activity and SUA levels than GE and YE, especially at the latter period, thereby it may play a more important role in improving the growth performance of broilers.

Keywords: cinnamon extract, broiler, growth performance, serum uric acid, serum xanthine oxidase

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Authors: Gowri Balaji, Muthusamy Nachiyappan, Ramalingam Venugopal

Abstract:

Amongst the wide spectrum of pesticides, pyrethroids are preferably used rather than organochlorine, organophosphorous and carbamates pesticides due to their high effectiveness. Synthetic pyrethroids which are the chemicals used for the pest control in agriculture are now being excessively used in India. The aim of the present study was to evaluate the adverse effect of cypermethrin on the fresh water fish Cyprinus carpio, the common carp. The effect was assessed by comparing the biochemical parameters in the blood and liver tissues of control fishes with three experimental group of fishes exposed with cypermethrin for 7 days 1/15 Lc50 (E1) 1/10 Lc50 (E2) and 1/5 Lc50 values (E3). After 7 days of exposure, blood was collected and liver and gills was dissected out. The activities of acid phosphatase, alkaline phosphatase, lactate dehydrogenase, aspartate aminotransferase and alanine aminotransferase were estimated by standard spectrophotometric techniques in the blood, liver and gills tissue homogenate. Lactate dehydrogenase was significantly decreased in E2 and E3 experimental groups. The activities of acid phosphatase, alkaline phosphatase, aspartate aminotransferase and alanine aminotransferase were significantly altered in the experimental groups. All the biochemical parameters studied were adversely affected in the liver and gills of cypermethrin exposed fish. The results obtained from the present study of cypermethrin exposed fishes indicate a marked toxic effect of cypermethrin and also its dose dependent impact on different organs of the fish.

Keywords: cypermethrin, Cyprinus carpio, ALT, AST, LDH, liver, gills

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Authors: Anahita Izadyar, My Ni Van, Kayleigh Amber Rodriguez, Ilwoo Seok, Elizabeth E. Hood

Abstract:

Using a recombinant enzyme derived from corn and a simple modification, we fabricated a facile, fast, and cost-beneficial biosensor to measure glucose. The Nafion/ Plant Produced Mn Peroxidase (PPMP)– glucose oxidase (GOx)- Bovine serum albumin (BSA) /Au electrode showed an excellent amperometric response to detect glucose. This biosensor is capable of responding to a wide range of glucose—20.0 µM−15.0 mM and has a lower detection limit (LOD) of 2.90µM. The reproducibility response using six electrodes is also very substantial and indicates the high capability of this biosensor to detect a wide range of 3.10±0.19µM to 13.2±1.8 mM glucose concentration. Selectivity of this electrode was investigated in an optimized experimental solution contains 10% diet green tea with citrus containing ascorbic acid (AA), and citric acid (CA) in a wide concentration of glucose at 0.02 to 14.0mM with an LOD of 3.10µM. Reproducibility was also investigated using 4 electrodes in this sample and shows notable results in the wide concentration range of 3.35±0.45µM to of 13.0 ± 0.81 mM. We also used other voltammetry methods to evaluate this biosensor. We applied linear sweep voltammetry (LSV) and this technique shows a wide range of 0.10−15.0 mM to detect glucose with a lower detection limit of 19.5µM. The performance and strength of this enzyme biosensor were the simplicity, wide linear ranges, sensitivities, selectivity, and low limits of detection. We expect that the modified biosensor has the potential for monitoring various biofluids.

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

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