Search results for: capillary electrophoresis-mass spectrometry

Authors: Ahmed Auda, Manjree Agarwala, Giles Hardya, Yonglin Rena

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Botrytis cinerea is a major pest for many plants. It can attack a wide range of plant parts. It can attack buds, flowers, and leaves, stems, and fruit. However, B. cinerea can be mixed with other diseases that cause the same damage. There are many species of botrytis and more than one different strains of each. Botrytis might infect the foliage of nursery stock stored through winter in damp conditions. There are no known resistant plants. Botrytis must have nutrients or food source before it infests the plant. Nutrients leaking from wounded plant parts or dying tissue like old flower petals give the required nutrients. From this food, the fungus becomes more attackers and invades healthy tissue. Dark to light brown rot forms in the ill tissue. High humidity conditions support the growth of this fungus. However, we suppose that selection pressure can act on the morphological and neurophysiologic filter properties of the receiver and on both the biochemical and the physiological regulation of the signal. Communication is implied when signal and receiver evolves toward more and more specific matching, culminating. In other hand, receivers respond to portions of a body odor bouquet which is released to the environment not as an (intentional) signal but as an unavoidable consequence of metabolic activity or tissue damage. Each year Botrytis species can cause considerable economic losses to plant crops. Even with the application of strict quarantine and control measures, these fungi can still find their way into crops and cause the imposition of onerous restrictions on exports. Blueberry fruit mould caused by a fungal infection usually results in major losses during post-harvest storage. Therefore, the management of infection in early stages of disease development is necessary to minimize losses. The overall purpose of this study will develop sensitive, cheap, quick and robust diagnostic techniques for the detection of B. cinerea in blueberry. The specific aim was designed to investigate the performance of volatile organic compounds (VOCs) in the detection and discrimination of blueberry fruits infected by fungal pathogens with an emphasis on Botrytis in the early storage stage of post-harvest.

Keywords: botrytis cinerea, blueberry, GC/MS, VOCs

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Authors: Anshuman Srivastava, Shiv Singh, Sheelendra Pratap Singh

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A simple, sensitive and effective gas chromatography tandem mass spectrometry (GC-MS/MS) method was developed for simultaneous analysis of multi pesticide residues (organophosphate, organochlorines, synthetic pyrethroids and herbicides) in food commodities using phenolic resin based activated carbon fibers (ACFs) as reversed-dispersive solid phase extraction (r-DSPE) sorbent in modified QuEChERS (Quick Easy Cheap Effective Rugged Safe) method. The acetonitrile-based QuEChERS technique was used for the extraction of the analytes from food matrices followed by sample cleanup with ACFs instead of traditionally used primary secondary amine (PSA). Different physico-chemical characterization techniques such as Fourier transform infrared spectroscopy, scanning electron microscopy, X-ray diffraction and Brunauer-Emmet-Teller surface area analysis were employed to investigate the engineering and structural properties of ACFs. The recovery of pesticides and herbicides was tested at concentration levels of 0.02 and 0.2 mg/kg in different commodities such as cauliflower, cucumber, banana, apple, wheat and black gram. The recoveries of all twenty-six pesticides and herbicides were found in acceptable limit (70-120%) according to SANCO guideline with relative standard deviation value < 15%. The limit of detection and limit of quantification of the method was in the range of 0.38-3.69 ng/mL and 1.26 -12.19 ng/mL, respectively. In traditional QuEChERS method, PSA used as r-DSPE sorbent plays a vital role in sample clean-up process and demonstrates good recoveries for multiclass pesticides. This study reports that ACFs are better in terms of removal of co-extractives in comparison of PSA without compromising the recoveries of multi pesticides from food matrices. Further, ACF replaces the need of charcoal in addition to the PSA from traditional QuEChERS method which is used to remove pigments. The developed method will be cost effective because the ACFs are significantly cheaper than the PSA. So the proposed modified QuEChERS method is more robust, effective and has better sample cleanup efficiency for multiclass multi pesticide residues analysis in different food matrices such as vegetables, grains and fruits.

Keywords: QuEChERS, activated carbon fibers, primary secondary amine, pesticides, sample preparation, carbon nanomaterials

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Authors: Alexander Chinyere Nwankpa, Nneka Ngozi Nwankpa

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In Nigeria, the Federal government is trying to make sure that everyone has access to enough food that is nutritiously adequate and safe. But in the southwest of Nigeria, notably in Ondo State, the most valuable minerals such as oil and gas, bitumen, kaolin, limestone talc, columbite, tin, gold, coal, and phosphate are abundant. Therefore, some regions of Ondo State are now linked to large quantities of natural radioactivity as a result of the mineral presence. In this work, the baseline radioactivity levels in some of the most important food crops in Ondo State were analyzed, allowing for the prediction of probable radiological health impacts. To this effect, maize (Zea mays), yam (Dioscorea alata) and cassava (Manihot esculenta) tubers were collected from the farmlands in the State because they make up the majority of food's nutritional needs. Ondo State was divided into eight zones in order to provide comprehensive coverage of the research region. At room temperature, the maize (Zea mays), yam (Dioscorea alata), and cassava (Manihot esculenta) samples were dried until they reached a consistent weight. They were pulverized, homogenized, and 250 g packed in a 1-liter Marinelli beaker and kept for 28 days to achieve secular equilibrium. The activity concentrations of Radium-226 (Ra-226), Thorium-232 (Th-232), and Potassium-40 (K-40) were determined in the food samples using Gamma-ray spectrometry. Firstly, the Hyper Pure Germanium detector was calibrated using standard radioactive sources. The gamma counting, which lasted for 36000s for each sample, was carried out in the Centre for Energy Research and Development, Obafemi Awolowo University, Ile-Ife, Nigeria. The mean activity concentration of Ra-226, Th-232 and K-40 for yam were 1.91 ± 0.10 Bq/kg, 2.34 ± 0.21 Bq/kg and 48.84 ± 3.14 Bq/kg, respectively. The content of the radionuclides in maize gave a mean value of 2.83 ± 0.21 Bq/kg for Ra-226, 2.19 ± 0.07 Bq/kg for Th-232 and 41.11 ± 2.16 Bq/kg for K-40. The mean activity concentrations in cassava were 2.52 ± 0.31 Bq/kg for Ra-226, 1.94 ± 0.21 Bq/kg for Th-232 and 45.12 ± 3.31 Bq/kg for K-40. The average committed effective doses in zones 6-8 were 0.55 µSv/y for the consumption of yam, 0.39 µSv/y for maize, and 0.49 µSv/y for cassava. These values are higher than the annual dose guideline of 0.35 µSv/y for the general public. Therefore, the values obtained in this work show that there is radiological contamination of some foodstuffs consumed in some parts of Ondo State. However, we recommend that systematic and appropriate methods also need to be established for the measurement of gamma-emitting radionuclides since these constitute important contributors to the internal exposure of man through ingestion, inhalation, or wound on the body.

Keywords: contamination, environment, radioactivity, radionuclides

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Authors: Mohamad Ammar Ayass, Natalya Griko, Victor Pashkov, Wanying Cao, Kevin Zhu, Jin Zhang, Lina Abi Mosleh

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Respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent for coronavirus disease 2019 (COVID-19). Early evidence pointed at the angiotensin-converting enzyme 2 (ACE-2) expressed on the epithelial cells of the lung as the main entry point of SARS-CoV-2 into the cells. The viral entry is mediated by the binding of the Receptor Binding Domain (RBD) of the spike protein that is expressed on the surface of the virus to the ACE-2 receptor. As the number of SARS-CoV-2 variants continues to increase, mutations arising in the RBD of SARS-CoV-2 may lead to the ineffectiveness of RBD targeted neutralizing antibodies. To address this limitation, the objective of this study is to develop a combination of aptamers that target different regions of the RBD, preventing the binding of the spike protein to ACE-2 receptor and subsequent viral entry and replication. A safe and innovative biomedical tool was developed to inhibit viral infection and reduce the harms of COVID-19. In the present study, DNA aptamers were developed against a recombinant trimer S protein using the Systematic Evolution of Ligands by Exponential enrichment (SELEX). Negative selection was introduced at round number 7 to select for aptamers that bind specifically to the RBD domain. A series of 9 aptamers (ADI2010, ADI2011, ADI201L, ADI203L, ADI205L, ADIR68, ADIR74, ADIR80, ADIR83) were selected and characterized with high binding affinity and specificity to the RBD of the spike protein. Aptamers (ADI25, ADI2009, ADI203L) were able to bind and pull down endogenous spike protein expressed on the surface of SARS-CoV-2 virus in COVID-19 positive patient samples and determined by liquid chromatography- tandem mass spectrometry analysis (LC-MS/MS). LC-MS/MS data confirmed that aptamers can bind to the RBD of the spike protein. Furthermore, results indicated that the combination of the 9 best aptamers inhibited the binding of the purified trimer spike protein to the ACE-2 receptor found on the surface of Vero E6 cells. In the same experiment, the combined aptamers displayed a better neutralizing effect than antibodies. The data suggests that the selected aptamers could be used in therapy to neutralize the effect of the SARS-CoV-2 virus by inhibiting the interaction between the RBD and ACE-2 receptor, preventing viral entry into target cells and therefore blocking viral replication.

Keywords: aptamer, ACE-2 receptor, binding inhibitor, COVID-19, spike protein, SARS-CoV-2, treatment

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Authors: H. M. El-Rafie, A. H. Abou Zeid, R. S. Mohammed, A. A. Sleem

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Acrocarpus is a genus of flowering plants in the legume family Fabaceae which considered as a large and economically important family. This study aimed to investigate the phytoconstituents of the petroleum ether extract (PEE) of Acrocarpus fraxinofolius bark by Gas chromatography coupled with mass spectrometry (GC/MS) analysis of its fractions (fatty acid and unsaponifiable matter). Concerning this, identification of 52 compounds constituting 97.03 % of the total composition of the unsaponifiable matter fraction. Cycloeucalenol was found to be the major compound representing 32.52% followed by 4a, 14a-dimethyl-A8~24(28)-ergostadien (26.50%) and ß-sitosterol(13.74%), furthermore Gas liquid chromatography (GLC) analysis of the sterol fraction revealed the identification of cholesterol (7.22 %), campesterol (13.30 %), stigmasterol (10.00 %) and β - sitosterol (69.48 %). Meanwhile, the identification of 33 fatty acids representing 90.71% of the total fatty acid constituents. Methyl-9,12-octadecadienoate (40.39%) followed by methyl hexadecanoate (23.64%) were found to be the major compounds. On the other hand, column chromatography and Thin layer chromatography (TLC) fractionation of PEE separate the triterpenoid: 21β-hydroxylup-20(29)-en-3-one and β- amyrin which were structurally identified by spectroscopic analysis (NMR, MS and IR). PEE has been biologically evaluated for 1: management of diabetes in alloxan induced diabetic rats 2: cytotoxic activity against four human tumor cell lines (Cervix carcinoma cell line[HELA], Breast carcinoma cell line [MCF7], Liver carcinoma cell line[HEPG2] and Colon carcinoma cell line[HCT-116] 3: hepatoprotective activity against CCl4-induced hepatotoxicity in rats and the activity was studied by assaying the serum marker enzymes like AST, ALT, and ALP. Concerning this, the anti-diabetic activity exhibited by 100mg of PEE extract was 74.38% relative to metformin (100% potency). It also showed a significant anti-proliferative activity against MCF-7 (IC50= 2.35µg), Hela(IC50=3.85µg) and HEPG-2 (IC50= 9.54µg) compared with Doxorubicin as reference drug. The hepatoprotective activity was evidenced by significant decrease in liver function enzymes, i.e. AST, ALT and ALP by (29.18%, 28.26%, and 34.11%, respectively using silymarin as the reference drug, compared to their concentration levels in an untreated group with liver damage induced by CCl₄. This study was performed for the first time on the bark of this species.

Keywords: Acrocarpus fraxinofolius, antidiabetic, cytotoxic, hepatoprotective

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Authors: Lynda K. Eneh, Okal N. Mike, Anna-Karin Borg-Karlson, Ulrike Fillinger, Jenny M. Lindh

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The responses of individual gravid Anopheles gambiae sensu stricto (s.s.) to hay infusion volatiles were evaluated under laboratory conditions. Such infusions have long been known to be effective baits for monitoring mosquitoes that vector arboviral and filarial diseases but have previously not been tested for malaria vectors. Hay infusions were prepared by adding sun-dried Bermuda grass to lake water and leaving the mixture in a covered bucket for three days. The proportions of eggs laid by gravid An. gambiae s.s. in diluted (10%) and concentrated infusions ( ≥ 25%) was compared to that laid in lake water in two-choice egg-count bioassays. Furthermore, with the aim to develop a method that can be used to collect volatiles that influence the egg-laying behavior of malaria mosquitoes, different volatile trapping methods were investigated. Two different polymer-traps eluted using two different desorption methods and three parameters were investigated. Porapak®-Q traps and solvent desorption was compared to Tenax®-TA traps and thermal desorption. The parameters investigated were: collection time (1h vs. 20h), addition of salt (0.15 g/ml sodium chloride (NaCl) vs. no NaCl), and stirring the infusion (0 vs. 300 rpm). Sample analysis was with gas chromatography-mass spectrometry (GC-MS). An. gambiae s.s was ten times less likely to lay eggs in concentrated hay infusion than in lake water. The volatiles were best characterized by thermally desorbed Tenax traps, collected for 20 hours from infusion aliquots with sodium chloride added. Ten volatiles identified from headspace and previously indicated as putative oviposition semiochemicals for An. gambiae s.s. or confirmed semiochemicals for other mosquito species were tested in egg-count bioassays. Six of these (3-methylbutanol, phenol, 4-methylphenol, nonanal, indole and 3-methylindole), when added to lake water, were avoided for egg-laying when lake water was offered as the alternative in dual-choice egg count bioassays. These compounds likely contribute to the unfavorable oviposition responses towards hay infusions. This difference in oviposition response of different mosquito species should be considered when designing control measures.

Keywords: Anopheles gambiae, oviposition behaviour, egg-count cage bioassays, hay infusions, volatile detection, semiochemicals

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Authors: Jelka Pleadin, Nina Kudumija, Ana Vulic, Manuela Zadravec, Tina Lesic, Mario Skrivanko, Irena Perkovic, Nada Vahcic

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Products of animal origin, such as meat and meat products, can contribute to human mycotoxins’ intake coming as a result of either indirect transfer from farm animals exposed to naturally contaminated grains and feed (carry-over effects) or direct contamination with moulds or naturally contaminated spice mixtures used in meat production. Ochratoxin A (OTA) is mycotoxin considered to be of the outermost importance from the public health standpoint in connection with meat products. The aim of this study was to investigate the occurrence of OTA in different traditional meat products circulating on Croatian markets during 2018, produced by a large number of households situated in eastern and north Croatian regions using a variety of technologies. Concentrations of OTA were determined in traditional meat products (n = 70), including dry fermented sausages (Slavonian kulen, Slavonian sausage, Istrian sausage and domestic sausage; n = 28), dry-cured meat products (pancetta, pork rack and ham; n = 22) and cooked sausages (liver sausages, black pudding sausages and pate; n = 20). OTA was analyzed by use of quantitative screening immunoassay method (ELISA) and confirmed for positive samples (higher than the limit of detection) by liquid chromatography tandem mass spectrometry (LC-MS/MS) method. Whereas the bacon samples contaminated with OTA were not found, its level in dry fermented sausages ranged from 0.22 to 2.17 µg/kg and in dry-cured meat products from 0.47 to 5.35 µg/kg, with in total 9% of positive samples. Besides possible primary contamination of these products arising due to improper manufacturing or/and storage conditions, observed OTA contamination could also be the consequence of secondary contamination that comes as a result of contaminated feed the animals were fed on. OTA levels obtained in cooked sausages ranged from 0.32 to 4.12 µg/kg (5% of positives) and could probably be linked to the contaminated raw materials (liver, kidney and spices) used in the sausages production. The results showed an occasional OTA contamination of traditional meat products, pointing that to avoid such contamination on households these products should be produced and processed under standardized and well-controlled conditions. Further investigations should be performed in order to identify mycotoxin-producing moulds on the surface of the products and to define preventative measures that can reduce the contamination of traditional meat products during their production on households and period of storage.

Keywords: Croatian households, ochratoxin-A, traditional cooked sausages, traditional dry-cured meat products

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Authors: A. Abdi, A. Abbasi Daloee, A. Barari

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Aim: The adaptations that occur in human body after doing exercises training are a factor to help healthy people stay away from certain diseases. One of the main adaptations is a change in blood circulation, especially in vessels. The increase of capillary density is dependent on the balance between angiogenic and angiostatic factors. Most studies show that the changes made to angiogenic developmental factors resulted from physical exercises indicate the low level of stimulators compared with inhibitors. It is believed that the plasma level of VEGF-A, the important angiogenic factor, is reduced after physical exercise. Findings indicate that the extract of crataegus plant reduces the platelet-derived growth factor receptor (PDGFR) autophosphorylation in human's fibroblast. More importantly, crataegus (1 to 100 mg in liter) clearly leads to the inhibition of PDGFR autophosphorylation in vascular smooth muscle cells (VSMCs). Angiogenesis is a process that can be classified into physiological and pathophysiological forms. collagen XVIII is a part of extracellular protein and heparan sulfate proteoglycans in vascular epithelial and endothelial basement membrane cause the release of endostatin from noncollagenous collagen XVIII. Endostatin inhibits the growth of endothelial cells, inhibits angiogenesis, weakens different types of cancer, and the growth of tumors. The purpose of the current study was to investigate the effect of intensive aerobic running with or without aqueous extraction of black Crataegus elbursensis on Collagen XVIII in male rats. Design: Thirty-two Wistar male rats (4-6 weeks old, 125-135 gr weight) were acquired from the Pasteur's Institute (Amol, Mazandaran), and randomly assigned into control (n = 16) and training (n = 16) groups. Rats were further divided into saline-control (SC) (n=8), saline-training (ST) (n=8), crataegus pentaegyna extraction -control (CPEC) (n=8), and crataegus pentaegyna extraction - training (CPET) (n=8). The control (SC and CPEC) groups remained sedentary; whereas the training groups underwent a high running exercise program. plasma were excised and immediately frozen in liquid nitrogen. Statistical analysis was performed using a one way analysis of variance and Tukey test. Significance was accepted at P = 0.05. Results: The results show that aerobic exercise group had the highest concentration collagen XVIII compared to other groups and then respectively black crataegus, training-crataegus and control groups. Conclusion: In general, researchers in this study concluded that the increase of collagen XVIII (albeit insignificant) as a result of physical activity and consumption of black crataegus extract could possibly serve as a regional inhibitor of angiogenesis and another evidence for the anti-cancer effects of physical activities. Since the research has not managed in this study to measure the amount of plasma endostatin, it is suggested that both indices are measured with important angiogenic factors so that we can have a more accurate interpretation of changes to angiogenic and angiostatic factors resulted from physical exercises.

Keywords: aerobic running, Crataegus elbursensis, Collagen XVIII

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Authors: A. Shaygani, R. Saifi, M. S. Saidi, M. Sani

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Particle size distribution, the most important characteristics of aerosols, is obtained through electrical characterization techniques. The dynamics of charged nano-particles under the influence of electric field in electrical mobility spectrometer (EMS) reveals the size distribution of these particles. The accuracy of this measurement is influenced by flow conditions, geometry, electric field and particle charging process, therefore by the transfer function (transfer matrix) of the instrument. In this work, a wire-cylinder corona charger was designed and the combined field-diffusion charging process of injected poly-disperse aerosol particles was numerically simulated as a prerequisite for the study of a multi-channel EMS. The result, a cloud of particles with non-uniform charge distribution, was introduced to the EMS. The flow pattern and electric field in the EMS were simulated using computational fluid dynamics (CFD) to obtain particle trajectories in the device and therefore to calculate the reported signal by each electrometer. According to the output signals (resulted from bombardment of particles and transferring their charges as currents), we proposed a modification to the size of detecting rings (which are connected to electrometers) in order to evaluate particle size distributions more accurately. Based on the capability of the system to transfer information contents about size distribution of the injected particles, we proposed a benchmark for the assessment of optimality of the design. This method applies the concept of Von Neumann entropy and borrows the definition of entropy from information theory (Shannon entropy) to measure optimality. Entropy, according to the Shannon entropy, is the ''average amount of information contained in an event, sample or character extracted from a data stream''. Evaluating the responses (signals) which were obtained via various configurations of detecting rings, the best configuration which gave the best predictions about the size distributions of injected particles, was the modified configuration. It was also the one that had the maximum amount of entropy. A reasonable consistency was also observed between the accuracy of the predictions and the entropy content of each configuration. In this method, entropy is extracted from the transfer matrix of the instrument for each configuration. Ultimately, various clouds of particles were introduced to the simulations and predicted size distributions were compared to the exact size distributions.

Keywords: aerosol nano-particle, CFD, electrical mobility spectrometer, von neumann entropy

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Authors: Gordana Branković, Dejan Dodig, Vesna Pajić, Vesna Kandić, Desimir Knežević, Nenad Đurić

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The wheat grain quality properties are influenced by genotype, environmental conditions and genotype × environment interaction (GEI). The increasing request of more nutritious wheat products will direct future breeding programmes. Therefore, the aim of investigation was to determine: i) variability of the protein content (PC), wet gluten content (WG) and Zeleny sedimentation volume (ZS); ii) components of variance, heritability in a broad sense (hb2), and expected genetic advance as percent of mean (GAM) for PC, WG, and ZS; iii) correlations between PC, WG, ZS, and most important agronomic traits; in order to assess expected breeding success versus environmental impact for these quality traits. The plant material consisted of 30 genotypes of bread wheat (Triticum aestivum L. ssp. aestivum) and durum wheat (Triticum durum Desf.). The trials were sown at the three test locations in Serbia: Rimski Šančevi, Zemun Polje and Padinska Skela during 2010-2011 and 2011-2012. The experiments were set as randomized complete block design with four replications. The plot consisted of five rows of 1 m2 (5 × 0.2 m × 1 m). PC, WG and ZS were determined by the use of Near infrared spectrometry (NIRS) with the Infraneo analyser (Chopin Technologies, France). PC, WG and ZS, in bread wheat, were in the range 13.4-16.4%, 22.8-30.3%, and 39.4-67.1 mL, respectively, and in durum wheat, in the range 15.3-18.1%, 28.9-36.3%, 37.4-48.3 mL, respectively. The dominant component of variance for PC, WG, and ZS, in bread wheat, was genotype with the genetic variance/GEI variance (VG/VG × E) relation of 3.2, 2.9 and 1.0, respectively, and in durum wheat was GEI with the VG/VG × E relation of 0.70, 0.69 and 0.49, respectively. hb2 and GAM values for PC, WG and ZS, in bread wheat, were 94.9% and 12.6%, 93.7% and 18.4%, and 86.2% and 28.1%, respectively, and in durum wheat, 80.7% and 7.6%, 79.7% and 10.2%, and 74% and 11.2%, respectively. The most consistent through six environments, statistically significant correlations, for bread wheat, were between PC and spike length (-0.312 to -0.637); PC, WG, ZS and grain number per spike (-0.320 to -0.620; -0.369 to -0.567; -0.301 to -0.378, respectively); PC and grain thickness (0.338 to 0.566), and for durum wheat, were between PC, WG, ZS and yield (-0.290 to -0.690; -0.433 to -0.753; -0.297 to -0.660, respectively); PC and plant height (-0.314 to -0.521); PC, WG and spike length (-0.298 to -0.597; -0.293 to -0.627, respectively); PC, WG and grain thickness (0.260 to 0.575; 0.269 to 0.498, respectively); PC, WG and grain vitreousness (0.278 to 0.665; 0.357 to 0.690, respectively). Breeding success can be anticipated for ZS in bread wheat due to coupled high values for hb2 and GAM, suggesting existence of additive genetic effects, and also for WG in bread wheat, due to very high hb2 and medium high GAM. The small, and medium, negative correlations between PC, WG, ZS, and yield or yield components, indicate difficulties to select simultaneously for high quality and yield, depending on linkage for particular genetic arrangements to be broken by recombination.

Keywords: bread and durum wheat, genetic advance, protein and wet gluten content, Zeleny sedimentation volume

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Authors: Arezoo Rahmani, Rinez Thapa, Juha-Matti Aalto, Petri Turhanen, Jouko Vepsalainen, Vesa-PekkaLehto, Joakim Riikonen

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Production of Scandium (Sc) is a complicated process because Sc is found only in low concentrations in ores and the concentration of Sc is very low compared with other metals. Therefore, utilization of typical extraction processes such as solvent extraction is problematic in scandium extraction. The Adsorption/desorption method can be used, but it is challenging to prepare materials, which have good selectivity, high adsorption capacity, and high stability. Therefore, efficient and environmentally friendly methods for Sc extraction are needed. In this study, the nanoporous composite material was developed for extracting Sc from an Sc ore. The nanoporous composite material offers several advantageous properties such as large surface area, high chemical and mechanical stability, fast diffusion of the metals in the material and possibility to construct a filter out of the material with good flow-through properties. The nanoporous silicon material was produced by first stabilizing the surfaces with a silicon carbide layer and then functionalizing the surface with bisphosphonates that act as metal chelators. The surface area and porosity of the material were characterized by N₂ adsorption and the morphology was studied by scanning electron microscopy (SEM). The bisphosphonate content of the material was studied by thermogravimetric analysis (TGA). The concentration of metal ions in the adsorption/desorption experiments was measured with inductively coupled plasma mass spectrometry (ICP-MS). The maximum capacity of the material was 25 µmol/g Sc at pH=1 and 45 µmol/g Sc at pH=3, obtained from adsorption isotherm. The selectivity of the material towards Sc in artificial solutions containing several metal ions was studied at pH one and pH 3. The result shows good selectivity of the nanoporous composite towards adsorption of Sc. Scandium was less efficiently adsorbed from solution leached from the ore of Sc because of excessive amounts of iron (Fe), aluminum (Al) and titanium (Ti) which disturbed the adsorption process. For example, the concentration of Fe was more than 4500 ppm, while the concentration of Sc was only three ppm, approximately 1500 times lower. Precipitation methods were developed to lower the concentration of the metals other than Sc. Optimal pH for precipitation was found to be pH 4. The concentration of Fe, Al and Ti were decreased by 99, 70, 99.6%, respectively, while the concentration of Sc decreased only 22%. Despite the large reduction in the concentration of other metals, more work is needed to further increase the relative concentration of Sc compared with other metals to efficiently extract it using the developed nanoporous composite material. Nevertheless, the developed material may provide an affordable, efficient and environmentally friendly method to extract Sc on a large scale.

Keywords: adsorption, nanoporous silicon, ore solution, scandium

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Authors: Luofu Liu, Fei Xiao Jr., Fei Xiao

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Through the analysis technology of gas chromatography-mass spectrometry (GC-MS), the composition and distribution characteristics of aromatic hydrocarbons in the Chepaizi area of the Junggar Basin were analyzed in detail. Based on that, the biological input, maturity of crude oils and sedimentary environment of the corresponding source rocks were determined and the origin types of crude oils were divided. The results show that there are three types of crude oils in the study area including Type I, Type II and Type III oils. The crude oils from the 1st member of the Neogene Shawan Formation are the Type I oils; the crude oils from the 2nd member of the Neogene Shawan Formation are the Type II oils; the crude oils from the Cretaceous Qingshuihe and Jurassic Badaowan Formations are the Type III oils. For the Type I oils, they show a single model in the late retention time of the chromatogram of total aromatic hydrocarbons. The content of triaromatic steroid series is high, and the content of dibenzofuran is low. Maturity parameters related to alkyl naphthalene, methylphenanthrene and alkyl dibenzothiophene all indicate low maturity for the Type I oils. For the Type II oils, they have also a single model in the early retention time of the chromatogram of total aromatic hydrocarbons. The content of naphthalene and phenanthrene series is high, and the content of dibenzofuran is medium. The content of polycyclic aromatic hydrocarbon representing the terrestrial organic matter is high. The aromatic maturity parameters indicate high maturity for the Type II oils. For the Type III oils, they have a bi-model in the chromatogram of total aromatic hydrocarbons. The contents of naphthalene series, phenanthrene series, and dibenzofuran series are high. The aromatic maturity parameters indicate medium maturity for the Type III oils. The correlation results of triaromatic steroid series fingerprint show that the Type I and Type III oils have similar source and are both from the Permian Wuerhe source rocks. Because of the strong biodegradation and mixing from other source, the Type I oils are very different from the Type III oils in aromatic hydrocarbon characteristics. The Type II oils have the typical characteristics of terrestrial organic matter input under oxidative environment, and are the coal oil mainly generated by the mature Jurassic coal measure source rocks. However, the overprinting effect from the low maturity Cretaceous source rocks changed the original distribution characteristics of aromatic hydrocarbons to some degree.

Keywords: oil source, geochemistry, aromatic hydrocarbons, crude oils, chepaizi area, Junggar Basin

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Authors: Mónica P. Cala, Juan S. Carreño, Roland J.W. Meesters

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Recently, collection of biological fluids on special filter papers has become a popular micro-sampling technique. Especially, the dried blood spot (DBS) micro-sampling technique has gained much attention and is momently applied in various life sciences reserach areas. As a result of this popularity, DBS are not only intensively competing with the venous blood sampling method but are at this moment widely applied in numerous bioanalytical assays. In particular, in the screening of inherited metabolic diseases, pharmacokinetic modeling and in therapeutic drug monitoring. Recently, microsampling techniques were also introduced in “omics” areas, whereunder metabolomics. For a metabolic profiling study we applied micro-sampling of biological fluids (blood and plasma) from healthy controls and from women with breast cancer. From blood samples, dried blood and plasma samples were prepared by spotting 8uL sample onto pre-cutted 5-mm paper disks followed by drying of the disks for 100 minutes. Dried disks were then extracted by 100 uL of methanol. From liquid blood and plasma samples 40 uL were deproteinized with methanol followed by centrifugation and collection of supernatants. Supernatants and extracts were evaporated until dryness by nitrogen gas and residues derivated by O-methyxyamine and MSTFA. As internal standard C17:0-methylester in heptane (10 ppm) was used. Deconvolution and alignment of and full scan (m/z 50-500) MS data were done by AMDIS and SpectConnect (http://spectconnect.mit.edu) software, respectively. Statistical Data analysis was done by Principal Component Analysis (PCA) using R software. The results obtained from our preliminary study indicate that the use of dried blood/plasma on paper disks could be a powerful new tool in metabolic profiling. Many of the metabolites observed in plasma (liquid/dried) were also positively identified in whole blood samples (liquid/dried). Whole blood could be a potential substitute matrix for plasma in Metabolomic profiling studies as well also micro-sampling techniques for the collection of samples in clinical studies. It was concluded that the separation of the different sample methodologies (liquid vs. dried) as observed by PCA was due to different sample treatment protocols applied. More experiments need to be done to confirm obtained observations as well also a more rigorous validation .of these micro-sampling techniques is needed. The novelty of our approach can be found in the application of different biological fluid micro-sampling techniques for metabolic profiling.

Keywords: biofluids, breast cancer, metabolic profiling, micro-sampling

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Authors: Laura Rueda-Gensini, Jader Rodríguez, Juan C. Cruz, Carolina Munoz-Camargo

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There is an emerging interest in botanicals and plant extracts for medicinal practices due to their widely reported health benefits. A large variety of phytochemicals found in plants have been correlated with antioxidant, immunomodulatory, and analgesic properties, which makes plant-derived products promising candidates for modulating the progression and treatment of numerous diseases. Non-centrifugal cane sugar (NCS), in particular, has been known for its high antioxidant and nutritional value, but composition-wise variability due to changing environmental and processing conditions have considerably limited its use in the nutraceutical and biomedical fields. This work is therefore aimed at assessing the effect of thermal exposure during NCS production over its bioactive composition and, in turn, its therapeutic value. Accordingly, two modified dehydration methods are proposed that employ: (i) vacuum-aided evaporation, which reduces the necessary temperatures to dehydrate the sample, and (ii) window refractance evaporation, which reduces thermal exposure time. The biochemical composition of NCS produced under these two methods was compared to traditionally-produced NCS by estimating their total polyphenolic and protein content with Folin-Ciocalteu and Bradford assays, as well as identifying the major phenolic compounds in each sample via HPLC-coupled mass spectrometry. Their antioxidant activities were also compared as measured by their scavenging potential of ABTS and DPPH radicals. Results show that the two modified production methods enhance polyphenolic and protein yield in resulting NCS samples when compared to traditional production methods. In particular, reducing employed temperatures with vacuum-aided evaporation demonstrated to be superior at preserving polyphenolic compounds, as evidenced both in the total and individual polyphenol concentrations. However, antioxidant activities were not significantly different between these. Although additional studies should be performed to determine if the observed compositional differences affect other therapeutic activities (e.g., anti-inflammatory, analgesic, and immunoprotective), these results suggest that reducing thermal exposure holds great promise for the production of natural products with enhanced nutritional value.

Keywords: non-centrifugal cane sugar, polyphenolic compounds, thermal processing, antioxidant activity

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Authors: Rose Vanessa Bandeira Reidel, Bernardo Melai, Pier Luigi Cioni, Luisa Pistelli

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Ziziphus jujuba Miller is a common species of the Ziziphus genus (Rhamnaceae family) native to the tropics and subtropics known for its edible fruits, fresh consumed or used in healthy food, as flavoring and sweetener. Many phytochemicals and biological activities are described for this species. In this work, the aroma profiles emitted in vivo by whole fresh organs (leaf, bud flower, flower, green and red fruits) were analyzed separately by mean of solid phase micro-extraction (SPME) coupled with gas chromatography mass spectrometry (GC-MS). The emitted volatiles from different plant parts were analysed using Supelco SPME device coated with polydimethylsiloxane (PDMS, 100µm). Fresh plant material was introduced separately into a glass conical flask and allowed to equilibrate for 20 min. After the equilibration time, the fibre was exposed to the headspace for 15 min at room temperature, the fibre was re-inserted into the needle and transferred to the injector of the CG and CG-MS system, where the fibre was desorbed. All the data were submitted to multivariate statistical analysis, evidencing many differences amongst the selected plant parts and their developmental stages. A total of 144 compounds were identified corresponding to 94.6-99.4% of the whole aroma profile of jujube samples. Sesquiterpene hydrocarbons were the main chemical class of compounds in leaves also present in similar percentage in flowers and bud flowers where (E, E)-α-farnesene was the main constituent in all cited plant parts. This behavior can be due to a protection mechanism against pathogens and herbivores as well as resistance to abiotic factors. The aroma of green fruits was characterized by high amount of perillene while the red fruits release a volatile blend mainly constituted by different monoterpenes. The terpenoid emission of flesh fruits has important function in the interaction with animals including attraction of seed dispersers and it is related to a good quality of fruits. This study provides for the first time the chemical composition of the volatile emission from different Ziziphus jujuba organs. The SPME analyses of the collected samples showed different patterns of emission and can contribute to understand their ecological interactions and fruit production management.

Keywords: Rhamnaceae, aroma profile, jujube organs, HS-SPME, GC-MS

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Authors: K. Thiel, P. Patrikainen, C. Nagy, D. Fitzpatrick, E.-M. Aro, P. Kallio

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Photosynthetic cyanobacteria have been recognized as potential future biotechnological hosts for the direct conversion of CO₂ into chemicals of interest using sunlight as the solar energy source. However, in order to develop commercially viable systems, the flux of electrons from the photosynthetic light reactions towards specified target chemicals must be significantly improved. The objective of the study was to investigate whether the autotrophic production efficiency of specified end-metabolites can be improved in engineered cyanobacterial cells by rescuing excited electrons that are normally lost to molecular oxygen due to the cyanobacterial flavodiiron protein Flv1/3. Natively Flv1/3 dissipates excess electrons in the photosynthetic electron transfer chain by directing them to molecular oxygen in Mehler-like reaction to protect photosystem I. To evaluate the effect of flavodiiron inactivation on autotrophic production efficiency in the cyanobacterial host Synechocystis sp. PCC 6803 (Synechocystis), sucrose was selected as the quantitative reporter and a representative of a potential end-product of interest. The concept is based on the native property of Synechocystis to produce sucrose as an intracellular osmoprotectant when exposed to high external ion concentrations, in combination with the introduction of a heterologous sucrose permease (CscB from Escherichia coli), which transports the sucrose out from the cell. In addition, cell growth, photosynthetic gas fluxes using membrane inlet mass spectrometry and endogenous storage compounds were analysed to illustrate the consequent effects of flv deletion on pathway flux distributions. The results indicate that a significant proportion of the electrons can be lost to molecular oxygen via Flv1/3 even when the cells are grown under high CO₂ and that the inactivation of flavodiiron activity can enhance the photosynthetic electron flux towards optionally available sinks. The flux distribution is dependent on the light conditions and the genetic context of the Δflv mutants, and favors the production of either sucrose or one of the two storage compounds, glycogen or polyhydroxybutyrate. As a conclusion, elimination of the native Flv1/3 reaction and concomitant introduction of an engineered product pathway as an alternative sink for excited electrons could enhance the photosynthetic electron flux towards the target endproduct without compromising the fitness of the host.

Keywords: cyanobacterial engineering, flavodiiron proteins, redirecting electron flux, sucrose

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Authors: I. Ahumada, L. Ascar, C. Pedraza, J. Montecino

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It has been found that the addition of biosolids to soil is beneficial to soil health, enriching soil with essential nutrient elements. Although this sludge has properties that allow for the improvement of the physical features and productivity of agricultural and forest soils and the recovery of degraded soils, they also contain trace elements, organic trace and pathogens that can cause damage to the environment. The application of these biosolids to land without the total reclamation and the treated wastewater can transfer these compounds into terrestrial and aquatic environments, giving rise to potential accumulation in plants. The general aim of this study was to evaluate the bioavailability of bisphenol A (BPA), and 17 α-ethynyl estradiol (EE2) in a soil-biosolid system using wheat (Triticum aestivum) plant assays and a predictive extraction method using a solution of hydroxypropyl-β-cyclodextrin (HPCD) to determine if it is a reliable surrogate for this bioassay. Two soils were obtained from the central region of Chile (Lo Prado and Chicauma). Biosolids were obtained from a regional wastewater treatment plant. The soils were amended with biosolids at 90 Mg ha-1. Soils treated with biosolids, spiked with 10 mgkg-1 of the EE2 and 15 mgkg-1 and 30 mgkg-1of BPA were also included. The BPA, and EE2 concentration were determined in biosolids, soils and plant samples through ultrasound assisted extraction, solid phase extraction (SPE) and gas chromatography coupled to mass spectrometry determination (GC/MS). The bioavailable fraction found of each one of soils cultivated with wheat plants was compared with results obtained through a cyclodextrin biosimulator method. The total concentration found in biosolid from a treatment plant was 0.150 ± 0.064 mgkg-1 and 12.8±2.9 mgkg-1 of EE2 and BPA respectively. BPA and EE2 bioavailability is affected by the organic matter content and the physical and chemical properties of the soil. The bioavailability response of both compounds in the two soils varied with the EE2 and BPA concentration. It was observed in the case of EE2, the bioavailability in wheat plant crops contained higher concentrations in the roots than in the shoots. The concentration of EE2 increased with increasing biosolids rate. On the other hand, for BPA, a higher concentration was found in the shoot than the roots of the plants. The predictive capability the HPCD extraction was assessed using a simple linear correlation test, for both compounds in wheat plants. The correlation coefficients for the EE2 obtained from the HPCD extraction with those obtained from the wheat plants were r= 0.99 and p-value ≤ 0.05. On the other hand, in the case of BPA a correlation was not found. Therefore, the methodology was validated with respect to wheat plants bioassays, only in the EE2 case. Acknowledgments: The authors thank FONDECYT 1150502.

Keywords: emerging compounds, bioavailability, biosolids, endocrine disruptors

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Authors: Carmela Monterroso, Marc Romero-Estonllo, Carlos Pascual, Beatriz Rodríguez-Garrido

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The mobilization of heavy metals from polluted soils causes their transfer to natural waters, with consequences for ecosystems and human health. Phytostabilization techniques are applied to reduce this mobility, through the establishment of a vegetal cover and the application of soil amendments. In this work, the capacity of different organic amendments to improve water quality and reduce the mobility of metals in mine-tailings was evaluated. A field pilot test was carried out with leaching columns installed on an old Cu mine ore (NW of Spain) which forms part of the PhytoSUDOE network of phytomanaged contaminated field sites (PhytoSUDOE/ Phy2SUDOE Projects (SOE1/P5/E0189 and SOE4/P5/E1021)). Ten columns (1 meter high by 25 cm in diameter) were packed with untreated mine tailings (control) or those treated with organic amendments. Applied amendments were based on different combinations of municipal wastes, bark chippings, biomass fly ash, and nanoparticles like aluminum oxides or ferrihydrite-type iron oxides. During the packing of the columns, rhizon-samplers were installed at different heights (10, 20, and 50 cm) from the top, and pore water samples were obtained by suction. Additionally, in each column, a bottom leachate sample was collected through a valve installed at the bottom of the column. After packing, the columns were sown with grasses. Water samples were analyzed for: pH and redox potential, using combined electrodes; salinity by conductivity meter: bicarbonate by titration, sulfate, nitrate, and chloride, by ion chromatography (Dionex 2000); phosphate by colorimetry with ammonium molybdate/ascorbic acid; Ca, Mg, Fe, Al, Mn, Zn, Cu, Cd, and Pb by flame atomic absorption/emission spectrometry (Perkin Elmer). Porewater and leachate from the control columns (packed with unamended mine tailings) were extremely acidic and had a high concentration of Al, Fe, and Cu. In these columns, no plant development was observed. The application of organic amendments improved soil conditions, which allowed the establishment of a dense cover of grasses in the rest of the columns. The combined effect of soil amendment and plant growth had a positive impact on water quality and reduced mobility of aluminum and heavy metals.

Keywords: leaching, organic amendments, phytostabilization, polluted soils

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Authors: Pompilia Buzatu, Hazim Qiblawey, Albert Odai, Jana Jamaleddin, Mustafa Nasser, Simon J. Judd

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Membrane bioreactor (MBR) technology is challenged by the tendency for the membrane permeability to decrease due to ‘clogging’. Clogging includes ‘sludging’, the filling of the membrane channels with sludge solids, and ‘ragging’, the aggregation of short filaments to form long rag-like particles. Both sludging and ragging demand manual intervention to clear out the solids, which is time-consuming, labour-intensive and potentially damaging to the membranes. These factors impact on costs more significantly than membrane surface fouling which, unlike clogging, is largely mitigated by the chemical clean. However, practical evaluation of MBR clogging has thus far been limited. This paper presents the results of recent work attempting to quantify sludging and clogging based on simple bench-scale tests. Results from a novel ragging simulation trial indicated that rags can be formed within 24-36 hours from dispersed < 5 mm-long filaments at concentrations of 5-10 mg/L under gently agitated conditions. Rag formation occurred for both a cotton wool standard and samples taken from an operating municipal MBR, with between 15% and 75% of the added fibrous material forming a single rag. The extent of rag formation depended both on the material type or origin – lint from laundering operations forming zero rags – and the filament length. Sludging rates were quantified using a bespoke parallel-channel test cell representing the membrane channels of an immersed flat sheet MBR. Sludge samples were provided from two local MBRs, one treating municipal and the other industrial effluent. Bulk sludge properties measured comprised mixed liquor suspended solids (MLSS) concentration, capillary suction time (CST), particle size, soluble COD (sCOD) and rheology (apparent viscosity μₐ vs shear rate γ). The fouling and sludging propensity of the sludge was determined using the test cell, ‘fouling’ being quantified as the pressure incline rate against flux via the flux step test (for which clogging was absent) and sludging by photographing the channel and processing the image to determine the ratio of the clogged to unclogged regions. A substantial difference in rheological and fouling behaviour was evident between the two sludge sources, the industrial sludge having a higher viscosity but less shear-thinning than the municipal. Fouling, as manifested by the pressure increase Δp/Δt, as a function of flux from classic flux-step experiments (where no clogging was evident), was more rapid for the industrial sludge. Across all samples of both sludge origins the expected trend of increased fouling propensity with increased CST and sCOD was demonstrated, whereas no correlation was observed between clogging rate and these parameters. The relative contribution of fouling and clogging was appraised by adjusting the clogging propensity via increasing the MLSS both with and without a commensurate increase in the COD. Results indicated that whereas for the municipal sludge the fouling propensity was affected by the increased sCOD, there was no associated increased in the sludging propensity (or cake formation). The clogging rate actually decreased on increasing the MLSS. Against this, for the industrial sludge the clogging rate dramatically increased with solids concentration despite a decrease in the soluble COD. From this was surmised that sludging did not relate to fouling.

Keywords: clogging, membrane bioreactors, ragging, sludge

Procedia PDF Downloads 154

Authors: Mir Md. Abdus Salam, Muhammad Mohsin, Pertti Pulkkinen, Paavo Pelkonen, Ari Pappinen

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Soil and water pollution caused by extensive mining practices can adversely affect environmental components, such as humans, animals, and plants. Despite a generally positive contribution to society, mining practices have become a serious threat to biological systems. As metals do not degrade completely, they require immobilization, toxicity reduction, or removal. A greenhouse experiment was conducted to evaluate the effects of lime and N100 (11-amino-1-hydroxyundecylidene) chelate amendment on the growth and phytoextraction potential of the willow variety Klara (S. viminalis × S. schwerinii × S. dasyclados) grown in soils heavily contaminated with copper (Cu). The plants were irrigated with tap or processed water (mine wastewater). The sequential extraction technique and inductively coupled plasma-mass spectrometry (ICP-MS) tool were used to determine the extractable metals and evaluate the fraction of metals in the soil that could be potentially available for plant uptake. The results suggest that the combined effects of the contaminated soil and processed water inhibited growth parameter values. In contrast, the accumulation of Cu in the plant tissues was increased compared to the control. When the soil was supplemented with lime and N100; growth parameter and resistance capacity were significantly higher compared to unamended soil treatments, especially in the contaminated soil treatments. The combined lime- and N100-amended soil treatment produced higher growth rate of biomass, resistance capacity and phytoextraction efficiency levels relative to either the lime-amended or the N100-amended soil treatments. This study provides practical evidence of the efficient chelate-assisted phytoextraction capability of Klara and highlights its potential as a viable and inexpensive novel approach for in-situ remediation of Cu-contaminated soils and mine wastewaters. Abandoned agricultural, industrial and mining sites can also be utilized by a Salix afforestation program without conflict with the production of food crops. This kind of program may create opportunities for bioenergy production and economic development, but contamination levels should be examined before bioenergy products are used.

Keywords: copper, Klara, lime, N100, phytoextraction

Procedia PDF Downloads 125

Authors: Meng-Ying Chiu, Yu-Fang Huang, Pei-Wei Wang, Yi-Ru Wang, Yi-Shuan Shao, Mei-Lien Chen

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Background: Organophosphate pesticides (OPs) are the most heavily used pesticides in agriculture in Taiwan. Therefore, they are commonly detected in general public including pregnant women and children. These compounds are proven endocrine disrupters that may affect the neural development in humans. The aim of this study is to assess the OPs exposure of children in 2 years of age and to examine the association between the exposure concentrations and neurodevelopmental effects in children. Methods: In a prospective cohort of 280 mother-child pairs, urine samples of prenatal and postnatal were collected from each participant and analyzed for metabolites of OPs by using gas chromatography-mass spectrometry. Six analytes were measured including dimethylphosphate (DMP), dimethylthiophosphate (DMTP), dimethyldithiophosphate (DMDTP), diethylphosphate (DEP), diethylthiophosphate (DETP), and diethyldithiophosphate (DEDTP). This study created a combined concentration measure for dimethyl compounds (DMs) consisting of the three dimethyl metabolites (DMP, DMTP, and DMDTP), for diethyl compounds (DEs) consisting of the three diethyl metabolites (DEP, DETP, and DEDTP) and six dialkyl phosphate (DAPs). The Bayley Scales of Infant and Toddler Development (Bayley-III) was used to assess children's cognitive behavior at 2 years old. The association between OPs exposure and Bayley-III scale score was determined by using the Mann-Whitney U test. Results: The measurements of urine samples are still on-going. This preliminary data are the report of 56 children aged 2 from the cohort. The detection rates for DMP, DMTP, DMDTP, DEP, DETP, and DEDTP are 80.4%, 69.6%, 64.3%, 64.3%, 62.5%, and 75%, respectively. After adjusting the creatinine concentrations of urine, the median (nmol/g creatinine) of urinary DMP, DMTP, DMDTP, DEP, DETP, DEDTP, DMs, DEs, and DAPs are 153.14, 53.32, 52.13, 19.24, 141.65, 192.17, 308.8, 311.6, and 702.11, respectively. The concentrations of urine are considerably higher than that in other countries. Children’s cognitive behavior was used three scales for Bayley-III, including cognitive, language and motor. In Mann-Whitney U test, the higher levels of DEs had significantly lower motor score (p=0.037), but no significant association was found between the OPs exposure levels and the score of either cognitive or language. Conclusion: The limited sample size suggests that Taipei children are commonly exposed to OPs and OPs exposure might affect the cognitive behavior of young children. This report will present more data to verify the results. The predictors of OPs concentrations, such as dietary pattern will also be included.

Keywords: biomonitoring, children, neurodevelopment, organophosphate pesticides exposure

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Authors: Begum Rokeya, Rahelee Zinnat, Fatema Jebunnesa, Israt Ara Hossain, A. Rahman

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Background and Aims: Arsenic in drinking water is a global environmental health problem, and the exposure may increase dyslipidemia and cerebrovascular diseases mortalities, most likely through causing atherosclerosis. However, the mechanism of lipid metabolism, atherosclerosis formation, arsenic exposure and impact in pregnancy is still unclear. Recent epidemiological evidences indicate close association between inorganic arsenic exposure via drinking water and Dyslipidemia. However, the exact mechanism of this arsenic-mediated increase in atherosclerosis risk factors remains enigmatic. We explore the association of the effect of arsenic on serum lipid profile in pregnant subjects. Methods: A total 200 pregnant mother screened in this study from arsenic exposed area. Our study group included 100 exposed subjects were cases and 100 Non exposed healthy pregnant were controls requited by a cross-sectional study. Clinical and anthropometric measurements were done by standard techniques. Lipidemic status was assessed by enzymatic endpoint method. Urinary As was measured by inductively coupled plasma-mass spectrometry and adjusted with specific gravity and Arsenic exposure was assessed by the level of urinary arsenic level > 100 μg/L was categorized as arsenic exposed and < 100 μg/L were categorized as non-exposed. Multivariate logistic regression and Student’s t - test was used for statistical analysis. Results: Systolic and diastolic blood pressure both were significantly higher in the Arsenic exposed pregnant subjects compared to the Non-exposed group (p<0.001). Arsenic exposed subjects had 2 times higher chance of developing hypertensive pregnancy (Odds Ratio 2.2). In parallel to the findings in Ar exposed subjects showed significantly higher proportion of triglyceride and total cholesterol and low density of lipo protein when compare to non- arsenic exposed pregnant subjects. Significant correlation of urinary arsenic level was also found with SBP, DBP, TG, T chol and serum LDL-Cholesterol. On multivariate logistic regression showed urinary arsenic had a positive association with DBP, SBP, Triglyceride and LDL-c. Conclusion: In conclusion, arsenic exposure may induce dyslipidemia like atherosclerosis through modifying reverse cholesterol transport in cholesterol metabolism. For decreasing atherosclerosis related mortality associated with arsenic, preventing exposure from environmental sources in early life is an important element.

Keywords: Arsenic Exposure, Dyslipidemia, Gestational Diabetes Mellitus, Serum lipid profile

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Authors: Tim Bolle, Franziska Kreimendahl, Thomas Gries, Stefan Jockenhoevel

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The therapeutic treatment of extensive, deep wounds is limited. Autologous split-skin grafts are used as a so-called ‘gold standard’. Most common deficits are the defects at the donor site, the risk of scarring as well as the limited availability and quality of the autologous grafts. The aim of this project is a tissue engineered dermal-epidermal skin replacement to overcome the limitations of the gold standard. A key requirement for the development of such a three-dimensional implant is the formation of a functional capillary-like network inside the implant to ensure a sufficient nutrient and gas supply. Tailored three-dimensional warp knitted spacer fabrics are used to reinforce the mechanically week fibrin gel-based scaffold and further to create a directed in vitro pre-vascularization along the parallel-oriented pile yarns within a co-culture. In this study various three-dimensional warp knitted spacer fabrics were developed in a factorial design to analyze the influence of the machine parameters such as the stitch density and the pattern of the fabric on the scaffold performance and further to determine suitable parameters for a successful fibrin gel-incorporation and a physiological performance of the scaffold. The fabrics were manufactured on a Karl Mayer double-bar raschel machine DR 16 EEC/EAC. A fine machine gauge of E30 was used to ensure a high pile yarn density for sufficient nutrient, gas and waste exchange. In order to ensure a high mechanical stability of the graft, the fabrics were made of biocompatible PVDF yarns. Key parameters such as the pore size, porosity and stress/strain behavior were investigated under standardized, controlled climate conditions. The influence of the input parameters on the mechanical and morphological properties as well as the ability of fibrin gel incorporation into the spacer fabric was analyzed. Subsequently, the pile yarns of the spacer fabrics were colonized with Human Umbilical Vein Endothelial Cells (HUVEC) to analyze the ability of the fabric to further function as a guiding structure for a directed vascularization. The cells were stained with DAPI and investigated using fluorescence microscopy. The analysis revealed that the stitch density and the binding pattern have a strong influence on both the mechanical and morphological properties of the fabric. As expected, the incorporation of the fibrin gel was significantly improved with higher pore sizes and porosities, whereas the mechanical strength decreases. Furthermore, the colonization trials revealed a high cell distribution and density on the pile yarns of the spacer fabrics. For a tailored reinforcing structure, the minimum porosity and pore size needs to be evaluated which still ensures a complete incorporation of the reinforcing structure into the fibrin gel matrix. That will enable a mechanically stable dermal graft with a dense vascular network for a sufficient nutrient and oxygen supply of the cells. The results are promising for subsequent research in the field of reinforcing mechanically weak biological scaffolds and develop functional three-dimensional scaffolds with an oriented pre-vascularization.

Keywords: fibrin-gel, skin replacement, spacer fabric, pre-vascularization

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Authors: Andrea M. Rivas-Castillo, Marlenne Gómez-Ramirez, Isela Rodríguez-Pozos, Norma G. Rojas-Avelizapa

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Spent catalysts are considered as hazardous residues of major concern, mainly due to the simultaneous presence of several metals in elevated concentrations. Although hydrometallurgical, pyrometallurgical and chelating agent methods are available to remove and recover some metals contained in spent catalysts; these procedures generate potentially hazardous wastes and the emission of harmful gases. Thus, biotechnological treatments are currently gaining importance to avoid the negative impacts of chemical technologies. To this end, diverse microorganisms have been used to assess the removal of metals from spent catalysts, comprising bacteria, archaea and fungi, whose resistance and metal uptake capabilities differ depending on the microorganism tested. Acidophilic sulfur oxidizing bacteria have been used to investigate the biotreatment and extraction of valuable metals from spent catalysts, namely Acidithiobacillus thiooxidans and Acidithiobacillus ferroxidans, as they present the ability to produce leaching agents such as sulfuric acid and sulfur oxidation intermediates. In the present work, the ability of A. thiooxidans DSM 26636 for the bioleaching of metals contained in five different spent catalysts was assessed by growing the culture in modified Starkey mineral medium (with elemental sulfur at 1%, w/v), and 1% (w/v) pulp density of each residue for up to 21 days at 30 °C and 150 rpm. Sulfur-oxidizing activity was periodically evaluated by determining sulfate concentration in the supernatants according to the NMX-k-436-1977 method. The production of sulfuric acid was assessed in the supernatants as well, by a titration procedure using NaOH 0.5 M with bromothymol blue as acid-base indicator, and by measuring pH using a digital potentiometer. On the other hand, Inductively Coupled Plasma - Optical Emission Spectrometry was used to analyze metal removal from the five different spent catalysts by A. thiooxidans DSM 26636. Results obtained show that, as could be expected, sulfuric acid production is directly related to the diminish of pH, and also to highest metal removal efficiencies. It was observed that Al and Fe are recurrently removed from refinery spent catalysts regardless of their origin and previous usage, although these removals may vary from 9.5 ± 2.2 to 439 ± 3.9 mg/kg for Al, and from 7.13 ± 0.31 to 368.4 ± 47.8 mg/kg for Fe, depending on the spent catalyst proven. Besides, bioleaching of metals like Mg, Ni, and Si was also obtained from automotive spent catalysts, which removals were of up to 66 ± 2.2, 6.2±0.07, and 100±2.4, respectively. Hence, the data presented here exhibit the potential of A. thiooxidans DSM 26636 for the simultaneous bioleaching of metals contained in spent catalysts from diverse provenance.

Keywords: bioleaching, metal removal, spent catalysts, Acidithiobacillus thiooxidans

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Authors: Mingjiang Dai, Qian Shi, Fang Hu, Songsheng Lin, Huijun Hou, Chunbei Wei

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A diamond-like carbon (DLC) is considered as a promising protective film since its high hardness and excellent tribological properties. However, DLC films are very sensitive to the environmental condition, its friction coefficient could dramatic change in high humidity, therefore, limited their further application in aerospace, the watch industry, and micro/nano-electromechanical systems. Therefore, most studies focus on the low friction coefficient of DLC films at a high humid environment. However, this is out of satisfied in practical application. An important thing was ignored is that the DLC coated components are usually used in the diversed environment, which means its friction coefficient may evidently change in different humid condition. As a result, the invalidation of DLC coated components or even sometimes disaster occurred. For example, DLC coated minisize gears were used in the watch industry, and the customer may frequently transform their locations with different weather and humidity even in one day. If friction coefficient is not stable in dry and high moisture conditions, the watch will be inaccurate. Thus, it is necessary to investigate the stable tribological behavior of DLC films in various environments. In this study, a-C:H:Si films were deposited by multi-function magnetron sputtering system, containing one ion source device and a pair of SiC dual mid-frequent targets and two direct current Ti/C targets. Hydrogenated carbon layers were manufactured by sputtering the graphite target in argon and methane gasses. The silicon was doped in DLC coatings by sputtering silicon carbide targets and the doping content were adjusted by mid-frequent sputtering current. The microstructure of the film was characterized by Raman spectrometry, X-ray photoelectron spectroscopy, and transmission electron microscopy while its friction behavior under different humidity conditions was studied using a ball-on-disc tribometer. The a-C:H films with Si content from 0 to 17at.% were obtained and the influence of Si content on the structure and tribological properties under the relative humidity of 50% and 85% were investigated. Results show that the a-C:H:Si film has typical diamond-like characteristics, in which Si mainly existed in the form of Si, SiC, and SiO2. As expected, the friction coefficient of a-C:H films can be effectively changed after Si doping, from 0.302 to 0.176 in RH 50%. The further test shows that the friction coefficient value of a-C:H:Si film in RH 85% is first increase and then decrease as a function of Si content. We found that the a-C:H:Si films with a Si content of 3.75 at.% show a stable friction coefficient of 0.13 in different humidity environment. It is suggestion that the sp3/sp2 ratio of a-C:H films with 3.75 at.% Si was higher than others, which tend to form the silica-gel-like sacrificial layers during friction tests. Therefore, the films deliver stable low friction coefficient under controlled RH value of 50 and 85%.

Keywords: diamond-like carbon, Si doping, moisture environment, table low friction coefficient

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Authors: Sarah A. Alotaibi, Mohammed A. Almutairi, Abdullah A. Alsayari, Adibah M. Almutairi, Somaiah K. Almubayedh

Abstract:

Concerns over the presence of acrylamide in food date back to 2002, when Swedish scientists stated that, in carbohydrate-rich foods, amounts of acrylamide were formed when cooked at high temperatures. Similar findings were reported by other researchers which, consequently, caused major international efforts to investigate dietary exposure and the subsequent health complications in order to properly manage this issue. Due to this issue, in this work, we aim to determine the acrylamide level in different foods (coffee, potato chips, biscuits, and baby food) commonly consumed by the Saudi population. In a total of forty-three samples, acrylamide was detected in twenty-three samples at levels of 12.3 to 2850 µg/kg. In reference to the food groups, the highest concentration of acrylamide was found in coffee samples (<12.3-2850 μg/kg), followed by potato chips (655-1310 μg/kg), then biscuits (23.5-449 μg/kg), whereas the lowest acrylamide level was observed in baby food (<14.75 – 126 μg/kg). Most coffee, biscuits and potato chips products contain high amount of acrylamide content and also the most commonly consumed product. Saudi adults had a mean exposure of acrylamide for coffee, potato, biscuit, and cereal (0.07439, 0.04794, 0.01125, 0.003371 µg/kg-b.w/day), respectively. On the other hand, exposure to acrylamide in Saudi infants and children to the same types of food was (0.1701, 0.1096, 0.02572, 0.00771 µg/kg-b.w/day), respectively. Most groups have a percentile that exceeds the tolerable daily intake (TDI) cancer value (2.6 µg/kg-b.w/day). Overall, the MOE results show that the Saudi population is at high risk of acrylamide-related disease in all food types, and there is a chance of cancer risk in all age groups (all values ˂10,000). Furthermore, it was found that in non-cancer risks, the acrylamide in all tested foods was within the safe limit (˃125), except for potato chips, in which there is a risk for diseases in the population. With potato and coffee as raw materials, additional studies were conducted to assess different factors, including temperature, cocking time, and additives affecting the acrylamide formation in fried potato and roasted coffee, by systematically varying processing temperatures and time values, a mitigation of acrylamide content was achieved when lowering the temperature and decreasing the cooking time. Furthermore, it was shown that the combination of the addition of chitosan and NaCl had a large impact on the formation.

Keywords: risk assessment, dietary exposure, MOA, acrylamide, hazard

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Authors: Nadine khadraoui, Rym Essid, Olfa Tabbene, Imen Chniba, Safa Boujemaa, Selim Jallouli, Nadia Fares, Behija Mlik, Boutheina Ben Abdelmoumen Mardassi

Abstract:

Background and aim: Mycoplasma hominis is an opportunistic pathogen that can cause various gynecological infections such cervicitis, infertility, and, less frequently, extra-genital infections. Previous studies on the antimicrobial susceptibility of Mycoplasma hominis Tunisian strains have highlighted a significant resistance, even multi-resistance, to the most used antibiotic in the therapy of consequential infections. To address this concern, the present study aimed for the alternative of phytotherapy. Peganum harmala seed extract was tested as an antibacterial agent against multidrug-resistant M.hominis clinical strains. Material and Methods: Peganum harmala plant was collected from Ain Sebaa, Tabarka, North West region of Tunisia in April 2018, air-dried, grounded and extracted by different solvents.The crude methanolic extract was further partitioned with n-HEX, DCM, EtOAC and n-BuOl. Antibacterial activity was evaluated against M. hominis ATCC 23114 and 20 M. hominis clinical strains.The antimycoplasmal activity was tested by the microdilution method, and MIC values were determined. Phytochemical analysis and hemolytic activity on human erythrocytes were also performed. The active fraction was then subjected to purification, and the chemical identification of the active compound was investigated. Results: Among the tested fractions, the n-BuOH extract was the most active fraction since it exhibited an inhibitory effect against M. hominis ATCC 23114 and 80% of the tested clinical strains with MIC between 125 and 1000 µg/ml. The phytochemical analysis of the n-BuOH revealed its metabolic abundance in polyphenols, flavonoids and condensed tannin with levels of 257.37 mg AGE/g, 172.27 mg EC/g and 58.27 mg EC/g, respectively. In addition, P. harmala n-BuOH extract exhibited potent bactericidal activity against all M. hominis isolates with CMB values ranging between 125 and 4000 µg/ml. Further, the active fraction exhibited weak cytotoxicity effect at active concentrations when tested on human erythrocytes. The active compound was identified by gas chromatography–mass spectrometry as an indole alkaloid harmaline. Conclusion: In summary, Peganum harmala extract demonstrated an interesting anti-mycoplasmal activity against M. hominis Tunisian strains. Therefore, it could be considered as a potential candidate for the treatment of consequential infections. However, further studies are necessary to evaluate its mechanism of action in mycoplasmas.

Keywords: mycoplasma hominis, peganum harmala, antibioresistance, phytotherapy, phytochemical analysis

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Authors: Jaco Oosthuizen, Nerina C. Van Der Merwe

Abstract:

The National Health Laboratory Services in Bloemfontien has been the diagnostic testing facility for 1830 patients for familial breast cancer since 1997. From the cohort, 540 were comprehensively screened using High-Resolution Melting Analysis or Next Generation Sequencing for the presence of point mutations and/or indels. Approximately 90% of these patients stil remain undiagnosed as they are BRCA1/2 negative. Multiplex ligation-dependent probe amplification was initially added to screen for copy number variation detection, but with the introduction of next generation sequencing in 2017, was substituted and is currently used as a confirmation assay. The aim was to investigate the viability of utilizing internationally designed copy number variation detection assays based on mostly European/Caucasian genomic data for use within a South African context. The multiplex ligation-dependent probe amplification technique is based on the hybridization and subsequent ligation of multiple probes to a targeted exon. The ligated probes are amplified using conventional polymerase chain reaction, followed by fragment analysis by means of capillary electrophoresis. The experimental design of the assay was performed according to the guidelines of MRC-Holland. For BRCA1 (P002-D1) and BRCA2 (P045-B3), both multiplex assays were validated, and results were confirmed using a secondary probe set for each gene. The next generation sequencing technique is based on target amplification via multiplex polymerase chain reaction, where after the amplicons are sequenced parallel on a semiconductor chip. Amplified read counts are visualized as relative copy numbers to determine the median of the absolute values of all pairwise differences. Various experimental parameters such as DNA quality, quantity, and signal intensity or read depth were verified using positive and negative patients previously tested internationally. DNA quality and quantity proved to be the critical factors during the verification of both assays. The quantity influenced the relative copy number frequency directly whereas the quality of the DNA and its salt concentration influenced denaturation consistency in both assays. Multiplex ligation-dependent probe amplification produced false positives due to ligation failure when ligation was inhibited due to a variant present within the ligation site. Next generation sequencing produced false positives due to read dropout when primer sequences did not meet optimal multiplex binding kinetics due to population variants in the primer binding site. The analytical sensitivity and specificity for the South African population have been proven. Verification resulted in repeatable reactions with regards to the detection of relative copy number differences. Both multiplex ligation-dependent probe amplification and next generation sequencing multiplex panels need to be optimized to accommodate South African polymorphisms present within the genetically diverse ethnic groups to reduce the false copy number variation positive rate and increase performance efficiency.

Keywords: familial breast cancer, multiplex ligation-dependent probe amplification, next generation sequencing, South Africa

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Authors: Daniel R. McDougall, Martin D. de Jonge, Gordon M. Miskelly, Duncan J. McGillivray, Andrew G. Jeffs

Abstract:

The chelating agent ethylenediaminetetraacetic acid (EDTA) is commonly added as a cure-all to seawater in aquaculture hatcheries around the world to reduce heavy metal toxicity, significantly improve the survival of larval shellfish, and to therefore improve the overall production efficiency of the aquaculture industry. However, EDTA is not a biodegradable chemical and is considered to be a persistent organic pollutant, which will accumulate in the environment over time. This makes the use of EDTA unsustainable environmentally, and therefore alternatives should be considered. Ethylenediaminedisuccinic acid (EDDS) is a biodegradable alternative to EDTA with very similar metal chelation properties. This study investigates the effect of EDTA and EDDS at two different concentrations, on metal concentrations found within developing New Zealand green-lipped mussel (Perna canaliculus) larvae. P. canaliculus is New Zealand’s main shellfish aquaculture species, providing a major export for New Zealand’s economy, with excellent potential for increased production in the near future. It is well known that the early stages of bivalve development are the most vulnerable to metal toxicity and P. canaliculus is no exception. The commercially used concentration (12 µmol L⁻¹) of EDTA added to P. canaliculus larval rearing tanks often increases the yield of D-larvae by over 80%. This concentration of EDTA and EDDS will be tested in this study, along with a lower concentration (3 µmol L⁻¹). After 48 hours of larval development, the D-larvae will be analyzed for heavy metal content with Inductively Coupled Plasma Mass Spectrometry (ICP-MS) and heavy metal distribution with synchrotron X-ray Fluorescence Microscopy (XFM). In this study, we found that EDDS also improves the yield of P. canaliculus larvae and could be a viable alternative to EDTA in aquaculture. Furthermore, results suggest a higher concentration of chelating agent is more effective for improving the yield of developing P. canaliculus larvae. Metals with significant differences in concentration with the addition of EDTA were Cr, Cu, Zn, Cd and Pb (P < 0.05). We observed for the first time to the author’s best knowledge, metal distribution within 100 µm P. canaliculus D-larvae using synchrotron XFM and found changes in the distribution of metals with the addition of EDTA. XFM also has the potential to provide information about the chemical state of the metals within mussel larvae. This research provides greater insight into the reasons for the effectiveness of adding the chelating agent to aquaculture culture water, and a more environmentally conscious alternative to the currently used EDTA, which could be extremely valuable for the aquaculture industry.

Keywords: EDDS, EDTA, heavy metals, P. canaliculus, toxicity, water treatment

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Authors: Emmanuel Ikechuckwu Onwubuya, Afees Adebayo Oladejo

Abstract:

Background: The use of medicinal plants in the treatment of chronic diseases especially myocardial infarction, is gaining wide acceptance globally. Andrographis paniculata (Acanthaceae) is a medicinal plant commonly known as the king of bitters in Nigeria and has been acclaimed for several therapeutic activities. Materials and methods: This study investigated the cardio-protective effect of the leaf extract of A. paniculata in isoproterenol-induced myocardial infarction. Fresh green leaves of A paniculata were harvested from the Faculty of Agriculture farmland, Nnamdi Azikiwe University, Awka, Nigeria. Identification and authentication of the plant were carried out at the Department of Botany, Nnamdi Azikiwe University and a voucher specimen was deposited at the herbarium. The plant material was then shredded, air-dried under shade and pulverized. The fine powders obtained were weighed and extraction was done via a solvent combination of water and ethanol (3:7) for 72 hr via maceration. The filtrate gotten was evaporated to dryness to obtain the ethanol extract, which was used for further bioassay study. The bioactive constituents of the plant extract were quantitatively analyzed by Gas chromatography-mass spectrometry (GC-MS). The animals were administered the extract of A. paniculata orally for seven days at a divided dose of 100 mg/kg, 200 mg/kg and 400 mg/kg body weights. On the eighth day, myocardial infarction was induced through subcutaneous administration of isoproterenol at a dose of 150 mg/kg/day diluted in 2 ml of saline on two consecutive days. Subsequently, the blood pressures were monitored and blood was collected for bioassay studies. Results: The results of the study showed that the leaf extract of A. paniculata was rich in Dodecanoic acid (8.261%), 4-Dibenzofuranamine (6.03%), Cyclotrisiloxane (4.679 %). The findings also showed a significant decrease (p>0.05) in the Mean arterial blood pressure, heart rate, aspartate transaminase, alanine transaminase, creatinine kinase and lactate dehydrogenase activities of the treatment group compared with the untreated control group while the antioxidant (superoxide dismutase, catalase and glutathione) activities were significantly increased in the treatment group, compared with the untreated control group. Conclusion: The findings of this work have shown that the leaf of A. paniculata was rich in bioactive compounds, which could be synthesized to produce plant-based products to fight cardiovascular diseases, especially myocardial infarction.

Keywords: cardiovascular disease, myocardial infarction, medicinal plant, andrographis paniculata, isoproterenol

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