Search results for: plasma protein binding

Authors: Mohammed Sabbah, Prospero Di Pierro, Raffaele Porta

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Protein-based edible films and coatings have attracted an increasing interest in recent years since they might be used to protect pharmaceuticals or improve the shelf life of different food products. Among them, several plant proteins represent an abundant, inexpensive and renewable raw source. These natural biopolymers are used as film forming agents, being able to form intermolecular linkages by various interactions. However, without the addition of a plasticizing agent, many biomaterials are brittle and, consequently, very difficult to be manipulated. Plasticizers are generally small and non-volatile organic additives used to increase film extensibility and reduce its crystallinity, brittleness and water vapor permeability. Plasticizers normally act by decreasing the intermolecular forces along the polymer chains, thus reducing the relative number of polymer-polymer contacts, producing a decrease in cohesion and tensile strength and thereby increasing film flexibility allowing its deformation without rupture. The most commonly studied plasticizers are polyols, like glycerol (GLY) and some mono or oligosaccharides. In particular, GLY not only increases film extensibility but also migrates inside the film network often causing the loss of desirable mechanical properties of the material. Therefore, replacing GLY with a different plasticizer might help to improve film characteristics allowing potential industrial applications. To improve film properties, it seemed of interest to test as plasticizers some cationic small molecules like polyamines (PAs). Putrescine, spermidine (SPD), and spermine are PAs widely distributed in nature and of particular interest for their biological activities that may have some beneficial health effects. Since PAs contains amino instead of hydroxyl functional groups, they are able to trigger ionic interactions with negatively charged proteins. Bitter vetch (Vicia ervilia; BV) is an ancient grain legume crop, originated in the Mediterranean region, which can be found today in many countries around the world. This annual Vicia genus shows several favorable features, being their seeds a cheap and abundant protein source. The main objectives of this study were to investigate the effect of different concentrations of SPD on the mechanical and permeability properties of films prepared with native or heat denatured BV proteins in the presence of different concentrations of SPD and/or GLY. Therefore, a BV seed protein concentrate (BVPC), containing about 77% proteins, was used to prepare film forming solutions (FFSs), whereas GLY and SPD were added as film plasticizers, either singly or in combination, at various concentrations. Since a primary plasticizer is generally defined as a molecule that when added to a material makes it softer, more flexible and easier to be processed, our findings lead to consider SPD as a possible primary plasticizer of protein-based films. In fact, the addition of millimolar concentrations of SPD to BVPC FFS allowed obtaining handleable biomaterials with improved properties. Moreover, SPD can be also considered as a secondary plasticizer, namely an 'extender', because of its ability even to enhance the plasticizing performance of GLY. In conclusion, our studies indicate that innovative edible protein-based films and coatings can be obtained by using PAs as new plasticizers.

Keywords: edible films, glycerol, plasticizers, polyamines, spermidine

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Authors: Shenghu Feng, Jun Cheng

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The molecular mechanism underlying nonalcoholic fatty liver disease (NAFLD) progression to hepatocellular carcinoma (HCC) remains unknown. In this study, immunohistochemistry staining result showed that NS5ABP37 protein expression decreased as with increasing degree of HCC malignancy. In agreement, NS5ABP37 protein overexpression significantly suppressed cell proliferation, caused G1/S cell cycle arrest, and induced apoptosis by increasing caspase-3/7 activity and cleaved caspase-3 levels. In addition, NS5ABP37 overexpression resulted in decreased intracellular TG and TC contents, with level reduction in SREBPs and downstream effectors. Furthermore, NS5ABP37 overexpression decreased SREBP1c and SREBP2 levels by inducing their respective promoters. Finally, ROS levels and ER-stress were both induced by NS5ABP37 overexpression. These findings together demonstrate that NS5ABP37 inhibits cancer cell proliferation and promotes apoptosis, by altering SREBP-dependent lipogenesis and cholesterogenesis in HepG2 cells and inducing oxidative stress and ER stress.

Keywords: NS5ABP37, liver cancer, lipid metabolism, oxidative stress, ER stress

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Authors: Amira E. Derbalah, Doaa M. Zaghloul

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10 clinically healthy hemal nodes were collected from male bulls aged 2-3 years. Light microscopy revealed a capsule of connective tissue consisted mainly of collagen fiber surrounding hemal node, numerous erythrocytes were found in wide subcapsular sinus under the capsule. The parenchyma of the hemal node was divided into cortex and medulla. Diffused lymphocytes, and lymphoid follicles, having germinal centers were the main components of the cortex, while in the medulla there was wide medullary sinus, diffused lymphocytes and few lymphoid nodules. The area occupied with lymph nodules was larger than that occupied with non-nodular structure of lymphoid cords and blood sinusoids. Electron microscopy revealed the cellular components of hemal node including elements of circulating erythrocytes intermingled with lymphocytes, plasma cells, mast cells, reticular cells, macrophages, megakaryocytes and endothelial cells lining the blood sinuses. The lymphocytes were somewhat triangular in shape with cytoplasmic processes extending between adjacent erythrocytes. Nuclei were triangular to oval in shape, lightly stained with clear nuclear membrane indentation and clear nucleoli. The reticular cells were elongated in shape with cytoplasmic processes extending between adjacent lymphocytes, rough endoplasmic reticulum, ribosomes and few lysosomes were seen in their cytoplasm. Nucleus was elongated in shape with less condensed chromatin. Plasma cells were oval to irregular in shape with numerous dilated rough endoplasmic reticulum containing electron lucent material occupying the whole cytoplasm and few mitochondria were found. Nuclei were centrally located and oval in shape with heterochromatin emarginated and often clumped near the nuclear membrane. Occasionally megakaryocytes and mast cells were seen among lymphocytes. Megakaryocytes had multilobulated nucleus and free ribosomes often appearing as small aggregates in their cytoplasm, while mast cell had their characteristic electron dense granule in the cytoplasm, few electron lucent granules were found also, we conclude that, the main function of the hemal node of cattle is proliferation of lymphocytes. No role for plasma cell in erythrophagocytosis could be suggested.

Keywords: cattle, electron microscopy, hemal node, histology, immune system

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Authors: Thomais Tsoulia, Arvind Y. M. Sundaram, Stine Braaen, Øyvind Haugland, Espen Rimstad, Øystein Wessel, Maria K. Dahle

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Fish red blood cells (RBC) are nucleated, and in addition to their function in gas exchange, they have been characterized as mediators of immune responses. Salmonid RBC are the major target cells of Piscineorthoreovirus (PRV), a virus associated with heart and skeletal muscle inflammation (HSMI) in farmed Atlantic salmon. The activation of antiviral response genesin RBChas previously been described in ex vivo and in vivo PRV-infection models, but not explored in the initial virus encounter phase. In the present study, mRNA transcriptome responses were explored in erythrocytes from individual fish, kept ex vivo, and exposed to purified PRV for 24 hours. The responses were compared to responses in macrophage-like salmon head kidney (SHK-1) and endothelial-like Atlantic salmon kidney (ASK) cells, none of which support PRV replication. The comparative analysis showed that the antiviral response to PRV was strongest in the SHK-1 cells, with a set of 80 significantly induced genes (≥ 2-fold upregulation). In RBC, 46 genes were significantly upregulated, while ASK cells were not significantly responsive. In particular, the transcriptome analysis of RBC revealed that PRV significantly induced interferon regulatory factor 1 (IRF1) and interferon-induced protein with tetratricopeptide repeats 5-like (IFIT9). However, several interferon-regulated antiviral genes which have previously been reported upregulated in PRV infected RBC in vivo (myxovirus resistance (Mx), interferon-stimulated gene 15 (ISG15), toll-like receptor 3 (TLR3)), were not significantly induced after 24h of virus stimulation. In contrast to RBC, these antiviral response genes were significantly upregulated in SHK-1. These results confirm that RBC are involved in the innate immune response to viruses, but with a delayed antiviral response compared to SHK-1. A notable difference is that interferon regulatory factor 1 (IRF-1) is the most strongly induced gene in RBC, but not among the significantly induced genes in SHK-1. Putative differences in the binding, recognition, and response to PRV, and any link to effects on the ability of PRV to replicate remains to be explored.

Keywords: antiviral responses, atlantic salmon, piscine orthoreovirus-1, red blood cells, RNA-seq

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Authors: Asif Arsalan

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The mortality rate of type 2 diabetes increasing day by day at an alarming rate. Changing lifestyle and environment have contributory effect in increase rate of type 2 diabetes mellitus. This study introduces a new method in physiotherapy field of treating a disease like diabetes, and gives the new way to control the diabetes without medicines.50 patients were selected on the basis of inclusion and exclusion criteria and were assigned to receive either TENS (group A) on the bilateral ST36 acupoints at a frequency of 25 Hz with intensity of 9 mA or placebo (group B) treatment for 5 minutes for 7 days. The blood glucose level was measured at both pre and post stimulation. Stimulation was given after 3 hours of food on every day regularly on stipulated time.There was significant improvement (P<0.05) in random blood sugar level of type 2 diabetes mellitus. It has been found TENS on bilateral ST36 acupoints have an effect to control plasma glucose level for type 2 diabetic mellitus patients and can be used without having any side effect. This study gives new idea to treat the type 2 diabetes conservatively with the TENS. As there are some study that TENS had been used to treat nausea, spasticity etc. condition by stimulating the acupoint but it is the very first time that TENS has been used to treat diabetes like disease. This study help the physiotherapy community to spread the physiotherapy treatment in other branches of the medical field and this gives a new identity for the physiotherapy. This also gives the benefit to patients to take a safe and cost effective treatment for the diabetes, and make the new use of TENS to treat other condition rather than pain.

Keywords: acupoint, plasma glucose level, type 2 diabetic mellitus, TENS

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Authors: Muhammed Saeed Rasheed

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Knowledge of the zinc and phytic acid (PA) concentrations of staple cereal crops are essential when evaluating the nutritional health of national and regional populations. In the present study, a total of 120 farmers’ fields in Iraqi Kurdistan were surveyed for zinc status in soil and wheat grain samples; wheat is the staple carbohydrate source in the region. Soils were analysed for total concentrations of phosphorus (PT) and zinc (ZnT), available P (POlsen) and Zn (ZnDTPA) and for pH. Average values (mg kg-1) ranged between 403-3740 (PT), 42.0-203 (ZnT), 2.13-28.1 (POlsen) and 0.14-5.23 (ZnDTPA); pH was in the range 7.46-8.67. The concentrations of Zn, PA/Zn molar ratio and estimated Zn bioavailability were also determined in wheat grain. The ranges of Zn and PA concentrations (mg kg⁻¹) were 12.3-63.2 and 5400 – 9300, respectively, giving a PA/Zn molar ratio of 15.7-30.6. A trivariate model was used to estimate intake of bioaccessible Zn, employing the following parameter values: (i) maximum Zn absorption = 0.09 (AMAX), (ii) equilibrium dissociation constant of zinc-receptor binding reaction = 0.680 (KP), and (iii) equilibrium dissociation constant of Zn-PA binding reaction = 0.033 (KR). In the model, total daily absorbed Zn (TAZ) (mg d⁻¹) as a function of total daily nutritional PA (mmole d⁻¹) and total daily nutritional Zn (mmole Zn d⁻¹) was estimated assuming an average wheat flour consumption of 300 g day⁻¹ in the region. Consideration of the PA and Zn intake suggest only 21.5±2.9% of grain Zn is bioavailable so that the effective Zn intake from wheat is only 1.84-2.63 mg d-1 for the local population. Overall results suggest available dietary Zn is below recommended levels (11 mg d⁻¹), partly due to low uptake by wheat but also due to the presence of large concentrations of PA in wheat grains. A crop breeding program combined with enhanced agronomic management methods is needed to enhance both Zn uptake and bioavailability in grains of cultivated wheat types.

Keywords: phosphorus, zinc, phytic acid, phytic acid to zinc molar ratio, zinc bioavailability

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Authors: Nour S. Abdelrahman, Seunghyun Hong, Hassan A. Arafat, Daniel Choi, Faisal Al Marzooqi

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Abstract—The advancement of lithium extraction methods from water sources, particularly saltwater brine, is gaining prominence in the lithium recovery industry due to its cost-effectiveness. Traditional techniques like recrystallization, chemical precipitation, and solvent extraction for metal recovery from seawater or brine are energy-intensive and exhibit low efficiency. Moreover, the extensive use of organic solvents poses environmental concerns. As a result, there's a growing demand for environmentally friendly lithium recovery methods. Membrane-based separation technology has emerged as a promising alternative, offering high energy efficiency and ease of continuous operation. In our study, we explored the potential of lithium-selective sieve channels constructed from layers of 2D graphene oxide and MXene (transition metal carbides and nitrides), integrated with surface – SO₃₋ groups. The arrangement of these 2D sheets creates interplanar spacing ranging from 0.3 to 0.8 nm, which forms a barrier against multivalent ions while facilitating lithium-ion movement through nano capillaries. The introduction of the sulfonate group provides an effective pathway for Li⁺ ions, with a calculated binding energy of Li⁺ – SO³⁻ at – 0.77 eV, the lowest among monovalent species. These modified membranes demonstrated remarkably rapid transport of Li⁺ ions, efficiently distinguishing them from other monovalent and divalent species. This selectivity is achieved through a combination of size exclusion and varying binding affinities. The graphene oxide channels in these membranes showed exceptional inter-cation selectivity, with a Li⁺/Mg²⁺ selectivity ratio exceeding 104, surpassing commercial membranes. Additionally, these membranes achieved over 94% rejection of MgCl₂.

Keywords: ion permeation, lithium extraction, membrane-based separation, nanotechnology

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Authors: Meskure Pak, Nihal Buyukuslu

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During Ramadan Muslims refrain from eating and drinking from dawn to sunset. Transformation of the eating habits cause profound changes in body composition. This study was performed during Ramadan of June-July 2015 with the regular fasting healthy adults (15 women and 15 men). The participants who were not fasting the whole month, have chronic diseases, pregnant and lactated were excluded. All attendances were informed about study. Written informed consent was taken from the voluntary participants. The work was approved by the Ethics and Research Committee of Istanbul Medipol University, Turkey. A questionnaire was conducted to determine the nutritional status, demographic and anthropometric data at the beginning, in the middle and at the end of Ramadan. Statistical Package for Social Sciences version 18.0 (SPSS, Chicago, IL, USA) was used for analyses. The mean ages of women and men were 34.4±9.45 and 28.9±10.55 years respectively. The BMI values (kg/m2) were slightly increased in men (26.5±3.9 to 26.2±3.7) and decreased in women (22.5±3.5 to 23.3±4.5). However the differences in BMI values between genders were not significant. Waist circumferences (WC) (cm) decreased in both women (80.2±14.6 to 79.4±17.7) and men (98.9±8.4 to 95.2±11.0) through the Ramadan. Fat percentages of women (27.0±71) increased in the middle of Ramadan (28.4±7.8) and decreased at the end of Ramadan (27.8±8.3). The fat percentages of men (21.5±6.3) were not affected in the middle of Ramadan (21.5±6.4) however decreased at the end of Ramadan (20.8±6.2). The total change in fat mass from beginning to end of Ramadan was higher in women than in men. The daily energy intake was higher in men than in women during Ramadan. In the middle of Ramadan energy intake (kcal) was reached to the highest level (2057.8±693.1) and at the end of Ramadan it decreased to the beginning level (1656.7±553.2) for men. However, daily energy intake of women slightly decreased from the beginning (1410.0±359.7) to the end (1409.2±366.7) of Ramadan. The comparison of energy intake between men and women was significant in the middle of Ramadan (p < 0.05). Water consumptions for both groups were increased in Ramadan fasting period. In comparison with the beginning of Ramadan, daily carbohydrate and fat consumptions increased and the consumption of protein decreased for men and for women at the end of Ramadan. The gender comparison resulted in a significant increase for protein and carbohydrate consumption of men in the middle of Ramadan (p < 0.05). In the first two weeks, the daily energy intake, the consumption of carbohydrates and fats seemed to increase for both men and women. However the later days of Ramadan daily fat consumption decreased to the level of beginning consumption levels which may indicate the nutritional adaptation period. In spite of the consumption of protein sources such as meat, poultry and egg increased, the decrease in the total amount of protein consumed in Ramadan may be due to a decrease in the consumption of milk and dairy products. In conclusion, the nutritional habits and preferred foods changed during Ramadan as a result affected the body composition.

Keywords: body composition, fasting, nutritional status, Ramadan

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Authors: Mohamed Moussaoui

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A series of thirty-one potential inhibitors targeting the epidermal growth factor receptor kinase (EGFR), derived from quinazoline, underwent 3D-QSAR analysis using CoMFA and CoMSIA methodologies. The training and test sets of quinazoline derivatives were utilized to construct and validate the QSAR models, respectively, with dataset alignment performed using the lowest energy conformer of the most active compound. The best-performing CoMFA and CoMSIA models demonstrated impressive determination coefficients, with R² values of 0.981 and 0.978, respectively, and Leave One Out cross-validation determination coefficients, Q², of 0.645 and 0.729, respectively. Furthermore, external validation using a test set of five compounds yielded predicted determination coefficients, R² test, of 0.929 and 0.909 for CoMFA and CoMSIA, respectively. Building upon these promising results, eighteen new compounds were designed and assessed for drug likeness and ADMET properties through in silico methods. Additionally, molecular docking studies were conducted to elucidate the binding interactions between the selected compounds and the enzyme. Detailed molecular dynamics simulations were performed to analyze the stability, conformational changes, and binding interactions of the quinazoline derivatives with the EGFR kinase. These simulations provided deeper insights into the dynamic behavior of the compounds within the active site. This comprehensive analysis enhances the understanding of quinazoline derivatives as potential anti-cancer agents and provides valuable insights for lead optimization in the early stages of drug discovery, particularly for developing highly potent anticancer therapeutics

Keywords: 3D-QSAR, CoMFA, CoMSIA, ADMET, molecular docking, quinazoline, molecular dynamic, egfr inhibitors, lung cancer, anticancer

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Authors: Zohreh Bayat, Dariush Minai-Tehrani

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Pseudomonas aeruginosa is a Gram-negative, rode shape and aerobic bacterium that has shown to be resistance to many antibiotics. This resistance makes the bacterium very harmful in some diseases. It can also generate diseases in any part of the gastrointestinal tract from oropharynx to rectum. P. aeruginosa has become an important cause of infection, especially in patients with compromised host defense mechanisms. One of the most important reasons that make P. aeruginosa an emerging opportunistic pathogen in patients is its ability to use various compounds as carbon sources. Lipase is an enzyme that catalyzes the hydrolysis of lipids. Most lipases act at a specific position on the glycerol backbone of lipid substrate. Some lipases are expressed and secreted by pathogenic organisms during the infection. Muscarinic antagonist used as an antispasmodic and in urinary incontinence. The drug has little effect on glandular secretion or the cardiovascular system. It does have some local anesthetic properties and is used in gastrointestinal, biliary, and urinary tract spasms. Aim: In this study the inhibitory effect of a muscarinic antagonist on lipase of P. aeruginosa was investigated. Methods: P. aeruginosa was cultured in minimal salt medium with 1% olive oil as carbon source. The cells were harvested and the supernatant, which contained lipase, was used for enzyme assay. Results: Our results showed that the drug can inhibit P. aeruginosa lipase by competitive manner. In the presence of different concentrations of the drug, the Vmax (2 mmol/min/mg protein) of enzyme did not change, while the Km raised by increasing the drug concentration. The Ki (inhibition constant) and IC50 (the half maximal inhibitory concentration) value of drug was estimated to be about 30 uM and 60 uM which determined that the drug binds to enzyme with high affinity. Maximum activity of the enzyme was observed at pH 8 in the absence and presence of muscarinic antagonist, respectively. The maximum activity of lipase was observed at 600C and the enzyme became inactive at 900C. Conclusion: The muscarinic antagonist drug could inhibit lipase of P. aeruginosa and changed the kinetic parameters of the enzyme. The drug binded to enzyme with high affinity and did not chang the optimum pH of the enzyme. Temperature did not affect the binding of drug to musmuscarinic antagonist.

Keywords: Pseudomonas aeruginosa, drug, enzyme, inhibition

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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: Sundaram Arulmozhiraja, Kanade Shimizu, Yuta Yamamoto, Satoshi Ichikawa, Maenaka Katsumi, Hiroaki Tokiwa

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Drug discovery is shifting from small molecule based drugs targeting local active site to middle molecules (MM) targeting large, flat, and groove-shaped binding sites, for example, protein-protein interface because at least half of all targets assumed to be involved in human disease have been classified as “difficult to drug” with traditional small molecules. Hence, MMs such as peptides, natural products, glycans, nucleic acids with various high potent bioactivities become important targets for drug discovery programs in the recent years as they could be used for ‘undruggable” intracellular targets. Cell membrane permeability is one of the key properties of pharmacodynamically active MM drug compounds and so evaluating this property for the potential MMs is crucial. Computational prediction for cell membrane permeability of molecules is very challenging; however, recent advancement in the molecular dynamics simulations help to solve this issue partially. It is expected that MMs with high membrane permeability will enable drug discovery research to expand its borders towards intracellular targets. Further to understand the chemistry behind the permeability of MMs, it is necessary to investigate their conformational changes during the permeation through membrane and for that their interactions with the membrane field should be studied reliably because these interactions involve various non-bonding interactions such as hydrogen bonding, -stacking, charge-transfer, polarization dispersion, and non-classical weak hydrogen bonding. Therefore, parameters-based classical mechanics calculations are hardly sufficient to investigate these interactions rather, quantum mechanical (QM) calculations are essential. Fragment molecular orbital (FMO) method could be used for such purpose as it performs ab initio QM calculations by dividing the system into fragments. The present work is aimed to study the cell permeability of middle molecules using molecular dynamics simulations and FMO-QM calculations. For this purpose, a natural compound syringolin and its analogues were considered in this study. Molecular simulations were performed using NAMD and Gromacs programs with CHARMM force field. FMO calculations were performed using the PAICS program at the correlated Resolution-of-Identity second-order Moller Plesset (RI-MP2) level with the cc-pVDZ basis set. The simulations clearly show that while syringolin could not permeate the membrane, its selected analogues go through the medium in nano second scale. These correlates well with the existing experimental evidences that these syringolin analogues are membrane-permeable compounds. Further analyses indicate that intramolecular -stacking interactions in the syringolin analogues influenced their permeability positively. These intramolecular interactions reduce the polarity of these analogues so that they could permeate the lipophilic cell membrane. Conclusively, the cell membrane permeability of various middle molecules with potent bioactivities is efficiently studied using molecular dynamics simulations. Insight of this behavior is thoroughly investigated using FMO-QM calculations. Results obtained in the present study indicate that non-bonding intramolecular interactions such as hydrogen-bonding and -stacking along with the conformational flexibility of MMs are essential for amicable membrane permeation. These results are interesting and are nice example for this theoretical calculation approach that could be used to study the permeability of other middle molecules. This work was supported by Japan Agency for Medical Research and Development (AMED) under Grant Number 18ae0101047.

Keywords: fragment molecular orbital theory, membrane permeability, middle molecules, molecular dynamics simulation

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Authors: V. Karuppaiah, J. C. Padaria, C. Srivastava

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The tobacco caterpillar, Spodoptera litura Fab (Lepidoptera: Noctuidae) is a polyphagous pest various field and horticulture crops in India. Pest had virtually developed resistance to all commonly used insecticides. Enhanced detoxification is the prime mechanism that is dictated by detoxification different enzymes and carboxylesterase is one of the major enzyme responsible development of resistance. In India, insecticide resistance studies on S. litura are mainly deployed on detoxification enzymes activity and investigation at gene level alteration i.e. at nucleotide level is very merger. In the present study, we collected the S. litura larvae from three different cauliflower growing belt viz., IARI, New Delhi (Delhi), Palari, Sonepat (Haryana) and Varanasi (Uttar Pradesh) to study the role of carboxylesterase activity and its gene level variation The CarE activity was measured using UV-VIS spectrophotometer with 3rd instar larvae of S. litura. The elevated activity of CarE was observed in Sonepat strain (28.09 ± 0.09 µmol/min/mg of protein) followed by Delhi (26.72 ± 0.04 µmol/min/mg of protein) and Varanasi strain (10.00 ± 0.44 µmol/min/mg of protein) of S. litura. The genomic DNA was isolated from 3rd instar larvae and CarE gene was amplified using a primer sequence, F:5’tccagagttccttgtcaggcac3’; R:5’ctgcatcaagcatgtctc3. CarE gene, about 500bp was partially amplified, sequenced and submitted to NCBI (Accession No. KF835886, KF835887 and KF835888). The sequence data revealed polymorphism at nucleotide level in all the three strains and gene found to have 88 to 97% similarity with previous available nucleotide sequences of S. litura, S. littoralis and S. exiqua. The polymorphism at the nucleotide level could be a reason for differential activity of carboxylesterase enzymes among the strains. However, investigation at gene expression level would be useful to analyze the overproduction of carboxylesterase enzyme.

Keywords: carboxylesterase, CarE gene, nucleotide polymorphism, insecticide resistance, spodoptera litura

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Authors: Gadanya, A.M., Ponfa, S., Jibril, M.M., Abubakar, S. M.

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Background: Edible insects such as grasshopper are important sources of food for humans, and have been consumed as traditional foods by many indigenous communities especially in Africa, Asia, and Latin America. These communities have developed their skills and techniques in harvesting, preparing, consuming, and preserving edible insects, widely contributing to the role played by the use of insects in human nutrition. Aim/ objective: This study was aimed at determining the effect of insect enriched sprouted soyabean /millet based food on some gut health markers in albino rats. Methods. Four different formulations of Complementary foods (i.e Complementary Food B (CFB): sprouted millet (SM), Complementary Food C (CFC): sprouted soyabean (SSB), Complementary Food D (CFD): sprouted soybean and millet (SSBM) in a ratio of (50:50) and Complementary Food E (CFE): insect (grasshopper) enriched sprouted soybean and millet (SSBMI) in a ratio of (50:25:25)) were prepared. Proximate composition and short chain fatty acid contents were determined. Thirty albino rats were divided into5 groups of six rats each. Group 1(CDA) were fed with basal diet and served as a control group, while groups 2,3,4 and 5 were fed with the corresponding complimentary foods CFB, CFC, CFD and CFE respectively daily for four weeks. Concentrations of fecal protein, serum total carotenoids and nitric oxide were determined. DNA extraction for molecular isolation and characterization were carried out followed by PCR, the use of mega 11 software and NCBI blast for construction of the phylogenetic tree and organism identification respectively. Results: Significant increase (P<0.05) in percentage ash, fat, protein and moisture contents, as well as short chain fatty acid (acetate, butyrate and propionate) concentrations were recorded in the insect enriched sprouted composite food (CFE) when compared with the CFA, CFB, CFC and CFD composite food. Faecal protein, carotenoid and nitric oxide concentrations were significantly lower (P>0.05) in group 5 in comparison to groups 1to 4. Ruminococcus bromii and Bacteroidetes were molecularly isolated and characterized by 16s rRNA from the sprouted millet/sprouted soybean and the insect enriched sprouted soybean/sprouted millet based food respectively. The presence of these bacterial strains in the feaces of the treated rats is an indication that the gut of the treated rats is colonized by good gut bacteria, hence, an improved gut health. Conclusion: Insect enriched sprouted soya bean/sprouted millet based complementary diet showed a high composition of ash, fat, protein and fiber. Thus, could increase the availability of short chain fatty acids whose role to the host organism cannot be overemphasized. It was also found to have decrease the level of faecal protein, carotenoid and nitric oxide in the serum which is an indication of an improvement in the immune system function.

Keywords: gut-health, insect, millet, soybean, sprouted

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Authors: Sahana Selladurai, Christine DeWolf

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Lung surfactant comprises a lipid-protein film that coats the alveolar surface and serves to prevent alveolar collapse upon repeated breathing cycles. Exposure of lung surfactant to high concentrations of airborne pollutants, for example tropospheric ozone in smog, can chemically modify the lipid and protein components. These chemical changes can impact the film functionality by decreasing the film’s collapse pressure (minimum surface tension attainable), altering it is mechanical and flow properties and modifying lipid reservoir formation essential for re-spreading of the film during the inhalation process. In this study, we use Langmuir monolayers spread at the air-water interface as model membranes where the compression and expansion of the film mimics the breathing cycle. The impact of ozone exposure on model lung surfactant films is measured using a Langmuir film balance, Brewster angle microscopy and a pendant drop tensiometer as a function of film and sub-phase composition. The oxidized films are analyzed using mass spectrometry where lipid and protein oxidation products are observed. Oxidation is shown to reduce surface activity, alter line tension (and film morphology) and in some cases visibly reduce the viscoelastic properties of the film when compared to controls. These reductions in functionality of the films are highly dependent on film and sub-phase composition, where for example, the effect of oxidation is more pronounced when using a physiologically relevant buffer as opposed to water as the sub-phase. These findings can lead to a better understanding on the impact of continuous exposure to high levels of ozone on the mechanical process of breathing, as well as understanding the roles of certain lung surfactant components in this process.

Keywords: lung surfactant, oxidation, ozone, viscoelasticity

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Authors: K. Nazan Turhan, Tuğçe Ersan

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The population of the world is approximately 8 billion, and it increases uncontrollably and irrepressibly, leading to an increase in consumption. This situation causes crucial problems, and food waste is one of these. The Food and Agriculture Organization of the United Nations (FAO) defines food waste as the discarding or alternative utilization of food that is safe and nutritious for the consumption of humans along the entire food supply chain, from primary production to end household consumer level. In addition, according to the estimation of FAO, one-third of all food produced for human consumption is lost or wasted worldwide every year. Wasting food endangers natural resources and causes hunger. For instance, excessive amounts of food waste cause greenhouse gas emissions, contributing to global warming. Therefore, waste management has been gaining significance in the last few decades at both local and global levels due to the expected scarcity of resources for the increasing population of the world. There are several ways to recover food waste. According to the United States Environmental Protection Agency’s Food Recovery Hierarchy, food waste recovery ways are source reduction, feeding hungry people, feeding animals, industrial uses, composting, and landfill/incineration from the most preferred to the least preferred, respectively. Bioethanol, biodiesel, biogas, agricultural fertilizer and animal feed can be obtained from food waste that is generated by different food industries. In this project, feeding animals was selected as a food waste recovery method and food waste of a plant was used to provide ingredient uniformity. Grasshoppers were used as a protein source. In other words, the project was performed to develop a dog food product by recovery of the plant’s food waste after following some steps. The collected food waste and purchased grasshoppers were sterilized, dried and pulverized. Then, they were all mixed with 60 g agar-agar solution (4%w/v). 3 different aromas were added, separately to the samples to enhance flavour quality. Since there are differences in the required amounts of different species of dogs, fulfilling all nutritional needs is one of the problems. In other words, there is a wide range of nutritional needs in terms of carbohydrates, protein, fat, sodium, calcium, and so on. Furthermore, the requirements differ depending on age, gender, weight, height, and species. Therefore, the product that was developed contains average amounts of each substance so as not to cause any deficiency or surplus. On the other hand, it contains more protein than similar products in the market. The product was evaluated in terms of contamination and nutritional content. For contamination risk, detection of E. coli and Salmonella experiments were performed, and the results were negative. For the nutritional value test, protein content analysis was done. The protein contents of different samples vary between 33.68% and 26.07%. In addition, water activity analysis was performed, and the water activity (aw) values of different samples ranged between 0.2456 and 0.4145.

Keywords: food waste, dog food, animal nutrition, food waste recovery

Procedia PDF Downloads 63

Authors: Loredana E. Nita, Aurica P. Chiriac, Elena Stoleru, Alina Diaconu, Tudorachi Nita

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Self-assembly processes are an attractive method to form new and complex structures between macromolecular compounds to be used for specific applications. In this context, intramolecular and intermolecular bonds play a key role during self-assembling processes in preparation of carrier systems of bioactive substances. Polyelectrolyte complexes (PECs) are formed through electrostatic interactions, and though they are significantly below of the covalent linkages in their strength, these complexes are sufficiently stable owing to the association processes. The relative ease way of PECs formation makes from them a versatile tool for preparation of various materials, with properties that can be tuned by adjusting several parameters, such as the chemical composition and structure of polyelectrolytes, pH and ionic strength of solutions, temperature and post-treatment procedures. For example, protein-polyelectrolyte complexes (PPCs) are playing an important role in various chemical and biological processes, such as protein separation, enzyme stabilization and polymer drug delivery systems. The present investigation is focused on evaluation of the PPC formation between a synthetic polypeptide (poly(aspartic acid) – PAS) and a natural protein (bovine serum albumin - BSA). The PPC obtained from PAS and BSA in different ratio was investigated by corroboration of various techniques of characterization as: spectroscopy, microscopy, thermo-gravimetric analysis, DLS and zeta potential determination, measurements which were performed in static and/or dynamic conditions. The static contact angle of the sample films was also determined in order to evaluate the changes brought upon surface free energy of the prepared PPCs in interdependence with the complexes composition. The evolution of hydrodynamic diameter and zeta potential of the PPC, recorded in situ, confirm changes of both co-partners conformation, a 1/1 ratio between protein and polyelectrolyte being benefit for the preparation of a stable PPC. Also, the study evidenced the dependence of PPC formation on the temperature of preparation. Thus, at low temperatures the PPC is formed with compact structure, small dimension and hydrodynamic diameter, close to those of BSA. The behavior at thermal treatment of the prepared PPCs is in agreement with the composition of the complexes. From the contact angle determination results the increase of the PPC films cohesion, which is higher than that of BSA films. Also, a higher hydrophobicity corresponds to the new PPC films denoting a good adhesion of the red blood cells onto the surface of PSA/BSA interpenetrated systems. The SEM investigation evidenced as well the specific internal structure of PPC concretized in phases with different size and shape in interdependence with the interpolymer mixture composition.

Keywords: polyelectrolyte – protein complex, bovine serum albumin, poly(aspartic acid), self-assembly

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Authors: Imene Soufli, Abdelkrim Hablal, Manel Amri, Moussa Labsi, Rania Sihem Boussa, Nassim Sid Idris, Chafia Touil-Boukoffa

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Crohn’s Disease (CD) is a relapsing–remitting inflammatory bowel disease with a progressive course. The aim of our study was to evaluate the relationship between the immunomarkers: Nitric Oxide (NO), pro-inflammatory cytokines, and blood count-based ratios and the outcome of corticosteroid or anti-TNF-α therapy in patients with complicated Crohn’s Disease. In this context, we evaluated the NLR as the ratio of neutrophil count to lymphocyte count, PLR as the ratio of platelet counts to lymphocyte count, and MLR as the ratio of monocyte count to lymphocyte count in patients and controls. Furthermore, we assessed NO production by the Griess method in plasma along with iNOS and NF-κB expression by immunofluorescence method in intestinal tissues of patients and controls. In the same way, we evaluated plasma TNF-α, IL-17A, and IL-10 levels using ELISA. Our results indicate that blood count-based ratios NLR, PLR, and MLR were significantly higher in patients compared to controls. In addition, increased systemic levels of NO, TNF-α, and IL-17A and colonic expression of iNOS and NF-κB were observed in the same patients. Interestingly, the high ratio of NLR and MLR, as well as NO production, was significantly decreased in treated patients. Collectively, our findings suggest that Nitric Oxide, as well as the blood count-based ratios (NLR, PLR, MLR), could constitute useful immuno-markers in complicated Crohn’s Disease, predicting the response to treatment

Keywords: complicated crohn’s disease, nitric oxide, blood count-based ratios, treatments, pro-inflammatory cytokines

Procedia PDF Downloads 79

Authors: Rabia Göçmen, Gülşah Kanbur, Sinan Sefa Parlat

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This study was conducted to determine that carrot powder obtain by different drying methods (oven and vacuum-freeze dryer) of carrot unfit for human consumption that whether feed additives in animal nutrition or not. Carrots randomly divided 2 groups. First group was dried by using oven, second group was by using vacuum freeze dryer methods. Dried carrot prepared from fresh carrot was analysed nutrient matter (energy, crude protein, crude oil, crude ash, beta carotene, mineral concentration and colour). The differences between groups in terms of energy, crude protein, ash, Ca and Mg was not significant (P> 0,05). Crude oil, P, beta carotene content and colour values (L, a, b) with vacuum-freeze dryer group was greater than oven group (P<0,05). Consequently, carrot powder obtained by drying the vacuum-freeze dryer method can be used as a source of carotene.

Keywords: carrot, vacuum freeze dryer, oven, beta carotene

Procedia PDF Downloads 324

Authors: Fatima Arrutia, Francisco Amador Riera

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The meat industry generates large volumes of blood as a result of meat processing. Several industrial procedures have been implemented in order to treat this by-product, but are focused on the production of low-value products, and in many cases, blood is simply discarded as waste. Besides, in addition to economic interests, there is an environmental concern due to bloodborne pathogens and other chemical contaminants found in blood. Consequently, there is a dire need to find extensive uses for blood that can be both applicable to industrial scale and able to yield high value-added products. Blood has been recognized as an important source of protein. The main blood serum protein in mammals is serum albumin. One of the top trends in food market is functional foods. Among them, bioactive peptides can be obtained from protein sources by microbiological fermentation or enzymatic and chemical hydrolysis. Bioactive peptides are short amino acid sequences that can have a positive impact on health when administered. The main drawback for bioactive peptide production is the high cost of the isolation, purification and characterization techniques (such as chromatography and mass spectrometry) that make unaffordable the scale-up. On the other hand, membrane technologies are very suitable to apply to the industry because they offer a very easy scale-up and are low-cost technologies, compared to other traditional separation methods. In this work, the possibility of obtaining bioactive peptide fractions from serum albumin by means of a simple procedure of only 2 steps (hydrolysis and membrane filtration) was evaluated, as an exploratory study for possible industrial application. The methodology used in this work was, firstly, a tryptic hydrolysis of serum albumin in order to release the peptides from the protein. The protein was previously subjected to a thermal treatment in order to enhance the enzyme cleavage and thus the peptide yield. Then, the obtained hydrolysate was filtered through a nanofiltration/ultrafiltration flat rig at three different pH values with two different membrane materials, so as to compare membrane performance. The corresponding permeates were analyzed by liquid chromatography-tandem mass spectrometry technology in order to obtain the peptide sequences present in each permeate. Finally, different concentrations of every permeate were evaluated for their in vitro antihypertensive and antioxidant activities though ACE-inhibition and DPPH radical scavenging tests. The hydrolysis process with the previous thermal treatment allowed achieving a degree of hydrolysis of the 49.66% of the maximum possible. It was found that peptides were best transmitted to the permeate stream at pH values that corresponded to their isoelectric points. Best selectivity between peptide groups was achieved at basic pH values. Differences in peptide content were found between membranes and also between pH values for the same membrane. The antioxidant activity of all permeates was high compared with the control only for the highest dose. However, antihypertensive activity was best for intermediate concentrations, rather than higher or lower doses. Therefore, although differences between them, all permeates were promising regarding antihypertensive and antioxidant properties.

Keywords: bioactive peptides, bovine serum albumin, hydrolysis, membrane filtration

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Authors: Hermalina Sinay, Estri L. Arumingtyas

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The research objective was to determine the expression level of dehydration responsive element binding/DREB gene of local corn cultivars from Kisar Island Maluku. The study design was a randomized block design with single factor consist of six local corn cultivars obtained from farmers in Kisar Island and one reference varieties wich has been released by the government as a drought-tolerant varieties and obtained from Cereal Crops Research Institute (ICERI) Maros South Sulawesi. Leaf samples were taken is the second leaf after the flag leaf at the 65 days after planting. Isolation of total RNA from leaf samples was carried out according to the protocols of the R & A-BlueTM Total RNA Extraction Kit and was used as a template for cDNA synthesis. The making of cDNA from total RNA was carried out according to the protocol of One-Step Reverse Transcriptase PCR Premix Kit. Real Time-PCR was performed on cDNA from reverse transcription followed the procedures of Real MODTM Green Real-Time PCR Master Mix Kit. Data obtained from the real time-PCR results were analyzed using relative quantification method based on the critical point / Cycle Threshold (CP / CT). The results of gene expression analysis of DREB gene showed that the expression level of the gene was highest obtained at Deep Yellow local corn cultivar, and the lowest one was obtained at the Rubby Brown Cob cultivar. It can be concluded that the expression level of DREB gene of Deep Yellow local corn cultivar was highest than other local corn cultivars and Srikandi variety as a reference variety.

Keywords: expression, level, DREB gene, local corn cultivars, Kisar Island, Maluku

Procedia PDF Downloads 299

Authors: Kang-Hyun Leem, Myung-Gyou Kim, Hye Kyung Kim

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Deer antler, traditionally used as a tonic and valuable drug in oriental medicine, has been considered to possess bone-strengthening activity. The upper section, mid section, and base of the antler has been known to exhibit different biological properties. Present study was performed to examine the effects of different parts of deer antler extract (DH) on osteogenic gene expressions in MG-63 cells and longitudinal bone growth in adolescent male rats. The expressions of osteogenic genes, collagen, alkaline phosphatase, osteocalcin, and osteopontin, were measured by quantitative real-time PCR. Longitudinal bone growth was measured in 3-week-old male Sprague-Dawley rats using fluorescence microscopy. To examine the effects on the growth plate metabolism, the total height of growth plate and bone morphogenetic protein-2 (BMP-2) were measured. Collagen and osteocalcin mRNA expressions were increased by all three parts of the DH treatment while osteopontin gene expression was not affected by any of the DH treatment. Alkaline phosphatase gene expression was increased by upper and mid part of DH while base part of DH fails to affect alkaline phosphatase gene expression. The upper and mid parts of the DH treatment enhanced longitudinal bone growth and total height of growth plate. The induction of BMP-2 protein expression in growth plate assessed by immunostaining was also promoted by upper and mid parts of the DH treatment. These results suggest that DH, especially upper and mid parts, stimulate osteogenic gene expressions and have the effect on bone growth in adolescent rats and might be used for the growth delayed adolescent and inherent growth failure patient.

Keywords: bone morphogenetic protein-2, deer antler, longitudinal bone growth, osteogenic genes

Procedia PDF Downloads 379

Authors: Naveen Kumar B. T., Anuj Tyagi, Niraj Kumar Singh, Visanu Boonyawiwat, Shanthanagouda A. H., Orawan Boodde, Shankar K. M., Prakash Patil, Shubhkaramjeet Kaur

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Since 2009, Acute Hepatopancreatic Necrosis Disease (AHPND) outbreaks have increased rapidly, and these have led to the major economic losses to the global shrimp industry. In comparison to other treatments, passive immunity and monoclonal antibody (MAb) based farmer level kit have proved their importance in controlling and treating the diseases in the shrimp industry. In the present study, MAbs were produced against the recombinant PirB protein Vibrio parahaemolyticus strain causing AHPND. Briefly, Balb/C mice were immunized with rPirB at 15 days interval, and antibody titer was determined by ELISA. Spleen cells from mice showing high antibody titer were fused with SP2O myeloma cells for hybridoma production. Among 130 hybridomas, four showed high antibody titer and positive reactivity in an immunoblot assay. In Western blot assay, three out of four MAbs (4C4, 2C2 and 4G3) showed reactivity to rPirB protein. However, in the natural host, only Mab clone 4G3 show strong reactivity (with a strain of V. parahemolyticus causing EMS/AHPND). These clones also showed reactivity with less than 20 kDa proteins in AHPND free V. parahaemolyticus (Thailand stain). Further, on from MAb 4G3 clone, four panels of single cell MAbs clones (G3F5, G3B8, G3H2, and G3D6) were produced of which three showed strong positive reactivity to rPirB protein in the Western blot. These MAbs have potential for controlling and prevention of the AHPND through passive immunity and development of filed level rapid diagnostic kits.

Keywords: shrimp, economic loss, AHPND, MAb

Procedia PDF Downloads 253

Authors: Panadda Rojpibulstit, Suthathip Kittisenachai, Songchan Puthong, Sirikul Manochantr, Pornpen Gamnarai, Sasichai Kangsadalampai, Sittiruk Roytrakul

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Hepatocellular carcinoma (HCC) is the most primary hepatic cancer worldwide. Nowadays a targeted therapy via monoclonal antibodies (mAbs) specific to tumor-associated antigen is continually developed in HCC treatment. In this regard, after establishing and consequently exploring Hep88 mAb’s tumoricidal effect on hepatocellular carcinoma cell line (HepG2 cell line), the Hep88 mAb’s specific Ag from both membrane and cytoplasmic fractions of HepG2 cell line was identified by 2-D gel electrophoresis and western blot analysis. After in-gel digestion and subsequent analysis by liquid chromatography-mass spectrometry (LC-MS), mortalin (HSPA9) and alpha-enolase were identified. The recombinant proteins specific to Hep88 mAb were cloned and expressed in E.coli BL21 (DE3). Moreover, alteration of HepG2 and Chang liver cell line after being induced by Hep88 mAb for 1-3 days was investigated using a transmission electron microscope. The result demonstrated that Hep88 mAb can bind to the recombinant mortalin (HSPA9) andalpha-enolase. In addition, gradual appearance of mitochondria vacuolization and endoplasmic reticulum dilatation were observed. Taken together, paraptosis-like programmed cell death (PCD) of HepG2 is induced by binding of mortalin (HSPA9) and alpha-enolase to Hep88 mAb. Mortalin depletion by formation of Hep88 mAb-mortalin (HSPA9) complex might initiate transcription-independent of p53-mediated apoptosis. Additionally, Hep88 mAb-alpha-enolase complex might initiate HepG2 cells energy exhaustion by glycolysis pathway obstruction. These results imply that Hep88 mAb might be a promising tool for development of an effective treatment of HCC in the next decade.

Keywords: Hepatocellular carcinoma, Monoclonal antibody, Paraptosis-like program cell death, Transmission electron microscopy, mortalin (HSPA9), alpha-enolase

Procedia PDF Downloads 361

Authors: Masataka Inada, Masanao Kinoshita, Nobuaki Matsumori

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It has been demonstrated that lipid molecules in biological membranes are responsible for the functionalization and structuration of membrane proteins. However, it is still unclear how the interaction of lipid molecules with membrane proteins is correlated with the function of the membrane proteins. Here we first developed an evaluation method for the interaction between membrane proteins and lipid molecules via surface plasmon resonance (SPR) analysis. Bacteriorhodopsin (bR), which was obtained by the culture of halobacteria, was used as a membrane protein. We prepared SPR sensor chips covered with self-assembled monolayer containing mercaptocarboxylic acids, and immobilized bR onto them. Then, we evaluated the interactions with various lipids that have different structures. As a result, the halobacterium-specific glycolipid S-TGA-1 was found to have much higher affinity with bRs than other lipids. This is probably due to not only hydrophobic and electrostatic interactions but also hydrogen bonds with sugar moieties in the glycolipid. Next, we analyzed the roles of the lipid in the structuration and functionalization of bR. CD analysis showed that S-TGA-1 could promote trimerization of bR monomers more efficiently than any other lipids. Flash photolysis further indicated that bR trimers formed by S-TGA-1 reproduced the photocyclic activity of bR in purple membrane, halobacterium-membrane. These results suggest that S-TGA-1 promotes trimerization of bR through strong interactions and consequently fulfills the bR’s function efficiently.

Keywords: membrane protein, lipid, interaction, bacteriorhodopsin, glycolipid

Procedia PDF Downloads 253

Authors: Sidra Shaw, Sarenna Shaw, Chae Joon Lee, Irimpan Mathews, Eric Koehn

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One of the biggest public health concerns of our time is increasing antimicrobial resistance. As of 2019, the CDC has documented more than 2.8 million serious antibiotic resistant infections in the United States. Currently, antibiotic resistant infections are directly implicated in over 750,000 deaths per year globally. On our current trajectory, British economist Jim O’Neill predicts that by 2050, an additional 10 million people (about half the population of New York) will die annually due to drug resistant infections. As a result, new biochemical pathways must be targeted to generate next generation antibiotic drugs that will be effective against drug resistant bacteria. One enticing target is the biosynthesis of DNA within bacteria, as few drugs interrupt this essential life process. Thymidylate synthase enzymes are essential for life as they catalyze the synthesis of a DNA building block, 2′-deoxythymidine-5′-monophosphate (dTMP). In humans, the thymidylate synthase enzyme (TSase) has been shown to be distinct from the flavin-dependent thymidylate synthase (FDTS) produced by many pathogenic bacteria. TSase and FDTS have distinct structures and mechanisms of catalysis, which should allow selective inhibition of FDTS over human TSase. Currently, C. diff is one of the most antibiotic resistant bacteria, and no drugs that target thymine biosynthesis exist for C. diff. Here we present the initial biochemical characterization of FDTS from C. diff. Specifically, we examine enzyme kinetics and binding features of this enzyme to determine the nature of interaction with ligands/inhibitors and understand the molecular mechanism of catalysis. This research will provide more insight into the targetability of the C. diff FDTS enzyme for novel antibiotic drugs.

Keywords: flavin-dependent thymidylate synthase, FDTS, clostridioides difficile, C. diff, antibiotic resistance, DNA synthesis, enzyme kinetics, binding features

Procedia PDF Downloads 104

Authors: Issam Lakdhar, Akram Alhussein, Juan Creus

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With the huge increase in world energy consumption, researchers are working to find other alternative sources of energy instead of fossil fuel one causing many environmental problems as the production of greenhouse effect gases. Hydrogen is considered a green energy source, which its combustion does not cause environmental pollution. The transport and the storage of the gas molecules or the other products containing this smallest chemical element in metallic structures (pipelines, tanks) are crucial issues. The dissolve and the permeation of hydrogen into the metal lattice lead to the formation of hydride phases and the embrittlement of structures. To protect the metallic structures, a surface treatment could be a good solution. Among the different techniques, magnetron sputtering is used to elaborate micrometric coatings capable of slowing down or stop hydrogen permeation. In the plasma environment, the deposition parameters of new thin-film metallic glasses Al-Ti-W were optimized and controlled in order to obtain, hydrogen barrier. Many characterizations were carried out (SEM, XRD and Nano-indentation…) to control the composition and understand the influence of film microstructure and chemical composition on the hydrogen permeation through the coatings. The coating performance was evaluated under two hydrogen production methods: chemical and electrochemical (cathodic protection) techniques. The hydrogen quantity absorbed was experimentally determined using the Thermal-Desorption Spectroscopy method (TDS)). An ideal ATW thin film was developed and showed excellent behavior against the diffusion of hydrogen.

Keywords: thin films, hydrogen, PVD, plasma technology, electrochemical properties

Procedia PDF Downloads 184

Authors: Aditya Arya, Hairin Taha, Ataul Karim Khan, Nayiar Shahid, Hapipah Mohd Ali, Mustafa Ali Mohd

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This study has investigated the antidiabetic and antioxidant potential of Pseudovaria macrophylla bark extract on streptozotocin–nicotinamide induced type 2 diabetic rats. LCMS-QTOF and NMR experiments were done to determine the chemical composition in the methanolic bark extract. For in vivo experiments, the STZ (60 mg/kg/b.w, 15 min after 120 mg/kg/1 nicotinamide, i.p.) induced diabetic rats were treated with methanolic extract of Pseuduvaria macrophylla (200 and 400 mg/kg∙bw) and glibenclamide (2.5 mg/kg) as positive control respectively. Biochemical parameters were assayed in the blood samples of all groups of rats. The pro-inflammatory cytokines, antioxidant status and plasma transforming growth factor βeta-1 (TGF-β1) were evaluated. The histological study of the pancreas was examined and its expression level of insulin was observed by immunohistochemistry. In addition, the expression of glucose transporters (GLUT 1, 2 and 4) were assessed in pancreas tissue by western blot analysis. The outcomes of the study displayed that the bark methanol extract of Pseuduvaria macrophylla has potentially normalized the elevated blood glucose levels and improved serum insulin and C-peptide levels with significant increase in the antioxidant enzyme, reduced glutathione (GSH) and decrease in the level of lipid peroxidation (LPO). Additionally, the extract has markedly decreased the levels of serum pro-inflammatory cytokines and transforming growth factor beta-1 (TGF-β1). Histopathology analysis demonstrated that Pseuduvaria macrophylla has the potential to protect the pancreas of diabetic rats against peroxidation damage by downregulating oxidative stress and elevated hyperglycaemia. Furthermore, the expression of insulin protein, GLUT-1, GLUT-2 and GLUT-4 in pancreatic cells was enhanced. The findings of this study support the anti-diabetic claims of Pseudovaria macrophylla bark.

Keywords: diabetes mellitus, Pseuduvaria macrophylla, alkaloids, caffeic acid

Procedia PDF Downloads 357

Authors: H. Lahlali F. El Haouzi, A.M.Al-Baradi, I. El Aboudi, M. El Azhari, A. Mdarhri

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Polytetrafluoroethylene (PTFE) is a high performance thermoplastic polymer with exceptional physical and chemical properties, such as a high melting temperature, high thermal stability, and very good chemical resistance. Nevertheless, manufacturing PTFE is problematic due to its high melt viscosity (10 12 Pa.s). In practice, it is by now well established that this property presents a serious problem when the classical methods are used to synthesized the dense PTFE materials in particularly hot pressing, high temperature extrusion. In this framework, we use here a new process namely spark plasma sintering (SPS) to elaborate PTFE samples from the micro metric particles powder. It consists in applying simultaneous electric current and pressure directly on the sample powder. By controlling the processing parameters of this technique, a series of PTFE samples are easy obtained and associated to remarkably short time as is reported in an early work. Our central goal in the present study is to understand how the non conventional SPS affects the mechanical properties at room temperature. For this end, a second commercially series of PTFE synthesized by using the extrusion method is investigated. The first data according to the tensile mechanical properties are found to be superior for the first set samples (SPS). However, this trend is not observed for the results obtained from the compression testing. The observed macro-behaviors are correlated to some physical properties of the two series of samples such as their crystallinity or density. Upon a close examination of these properties, we believe the SPS technique can be seen as a promising way to elaborate the polymer having high molecular mass without compromising their mechanical properties.

Keywords: PTFE, extrusion, Spark Plasma Sintering, physical properties, mechanical behavior

Procedia PDF Downloads 308

Authors: Yun-Chin Chung, Nileema Divate, Gen-Hung Chen, Pei-Ru Huang, Rupesh Divate

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Many environmental factors, such as glucose concentration, ethanol, temperature, osmotic pressure and pH, decrease the production rate of ethanol using yeast as a starter. Fermentation starters with high tolerance to various stresses are always demanded for brewing industry. Trehalose, a storage carbohydrate in cell wall of yeast, plays an important role in tolerance of environmental stress by preserving integrity of plasma membrane and stabilizing proteins. Furan aldehydes are toxic to yeast and the growth rate of yeast is significantly reduced if furan aldehydes were present in the fermentation medium. In yeast, aldehyde reductase is involved in the detoxification of reactive aldehydes and consequently the growth of yeast is improved. The aims of this study were to construct a genetic recombinant Saccharomyces cerevisiae or Pichia pastoris with furfural and HMF degrading and high ethanol tolerance capacities. Yeast strains were engineered by genetic recombination for overexpression of trehalose-6-phosphate synthase gene (tps1) and aldehyde reductase gene (ari1). TPS1 gene was cloned from S. cerevisiae by reverse transcription-polymerase chain reaction (RT-PCR) and then ligated with pGAPZαC vector. The constructed vector, pGAPZC-tps1, was transformed to recombinant yeasts strain with overexpression of ari1. The transformants with pGAPZC-tps1-ari1 were generated called STA (S. cerevisiae) and PTA (P. pastoris) with overexpression of tps1, ari1. PCR with tps1-specific primers and western blot with his-tag confirmed the gene insertion and protein expression of tps1 in the transformants, respectively. The neutral trehalase gene (nth1) of STA was successfully deleted and the novel strain STAΔN will be used for further study, including the measurement of trehalose concentration and ethanol, furfural tolerance assay.

Keywords: genetic recombinant, yeast, ethanol tolerance, trehalase, aldehyde reductase

Procedia PDF Downloads 422