Search results for: membrane resistance

Authors: Aw Adi Arryansyah

Abstract:

Using electronic stethoscope for detecting heartbeat sound, and breath sounds, are the effective way to investigate cardiovascular diseases. On the other side, technology is growing towards mobile. Almost everyone has a smartphone. Smartphone has many platforms. Creating mobile applications also became easier. We also can use HTML5 technology to creating mobile apps. Android is the most widely used type. This is the reason for us to make a wireless electronic stethoscope based on Android mobile. Android based Wireless Electronic Stethoscope designed by a simple system, uses sound sensors mounted membrane, then connected with Bluetooth module which will send the heart auscultation voice input data by Bluetooth signal to an android platform. On the software side, android will read the voice input then it will translate to beautiful visualization and release the voice output which can be regulated about how much of it is going to be released. We can change the heart beat sound into BPM data, and heart beat analysis, like normal beat, bradycardia or tachycardia.

Keywords: wireless, HTML 5, auscultation, bradycardia, tachycardia

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Authors: Cristiano Giordani, Marco Diociaiuti, Cecilia Bombelli, Laura Zanetti-Polzi, Marcello Belfiore, Raoul Fioravanti, Gianfranco Macchia

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We investigated induced functional effects by evaluating Ca2+-influx in liposomes and cell viability in HT22-DIFF neurons. Only solutions rich in unstructured Prefibrillar-Oligomers (PFOs) were able, in the presence of Monosialoganglioside-GM1 (GM1), to induce Ca2+-influx and were also neurotoxic, suggesting a correlation between the two phenomena. Thus, in the presence of GM1, we investigated the protein conformation and liposome modification due to the interaction. Circular Dichroism showed that GM1 fostered the formation of β-structures and Energy Filtered-Transmission Electron Microscopy that PFOs formed “amyloid-channels” as reported for Aβ. We speculate that electrostatic forces occurring between the positive PFOs and negative GM1 drive the initial binding, while the hydrophobic profile of the flexible PFO is responsible for the subsequent pore formation. Conversely, the rigid β-structured mature/fibers (MFs) and proto-fibers (PFs) were unable to induce membrane damage and Ca2+- influx.

Keywords: amyloid proteins, neurotoxicity, lipid-rafts, GM1

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Authors: Yeng Min Yi, Rosli Md Illias, Salehhuddin Hamdan

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Industrial biocatalysis is mainly based on the use of cell free or intracellular enzyme systems. However, the expensive cost and relatively lower operational stability of free enzymes limit practical use in industries. Cell surface display system can be used as a cost-efficient alternative to overcome the laborious purification and substrate transport limitation. In this research, TibA autotransporter from E. coli was used to display Aspergillus fumigatus xylanase (xyn). The amplified xyn was fused in between N-terminal signal peptide and C-terminal β-barrel of TibA. The cloned was transformed and expressed in E. coli BL21 (DE3). Outer membrane localization of TibA-xyn fusion protein was confirmed by SDS PAGE and western blot with expected size of 62.5 kDa. Functional display of xyn was examined by activity assay. Cell surface displayed xyn exhibited the highest activity at 37 °c, 0.3 mM IPTG. As a summary, TibA displaying system has the potential for further industrial applications. Moreover, this is the first report of the display of xylanase using TibA on the surface of E. coli.

Keywords: biocatalysis, cell surface display, Escherichia coli, TibA autotransporter

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Authors: S. B. Otieno1, F. Were, A. Afullo, K. Waza

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Background: Multi drug resistance HIV has emerged rendering the current conventional treatment of HIV ineffective. There is a need for new treatment regime which is cheap, effective and not prone to resistance development by HIV. Methods: In randomized clinical study of 68 HIV positive children 3 – 15 years to asses the efficacy of yeast selenium in HIV/AIDS patients, 50μ yeast selenium was administered to 34 children while in matched control of 34 were put on placebo. Blood samples and weight of the both groups which were taken every 3 months intervals up to 6 months, were analyzed by ELIZA for CD4T cells, the data was analyzed by SPSS version 16, WAZ scores were analyzed by Epi Info version 6. Results: No significant difference in age { χ2 (1, 62) =0.03, p =0.853}, cause of morbidity between test and controls {χ2 (1, 65) = 5.87, p= 0.015} and on condition of foster parents {χ2 ( 1,63) = 5.57, p= 0.0172} was observed. Children on selenium showed progressive improvement of WAZ and significant difference at six months {F (5,12) = =5.758, P=0.006}, and weight gain of up to 4.1 kilograms in six months, and significant CD4 T cell count increase t= -2.943, p<0.05 compared to matched controls t = -1.258 p> 0.05. CD4 T cell count increased among all age groups on test 3-5 years (+ 267.1),5-8 years (+200.3) 9-15 years (+71.2) cells/mm3 and in matched controls a decrease 3-5 years (-71), 5-8 years (-125) and 9-13 years (-10.1) cells/mm3 . No significant difference inCD4 T cell count between boys {F (2, 32) = 1.531 p= 0.232} and between boys {F (2, 49) = 1.040, p= 0.361} on test and between boys and girls {F (5, 81) = 1.379, p= 0.241} on test. Similarly no significant difference between boys and girls were observed {F (5, 86) = 1.168, p= 0.332}.In the test group there was significant positive correlation β =252.23 between weight for age (WAZ), and CD4 T Cell Count p=0.007, R2= 0.252, F< 0.05. In matched controls no significant correlation between weight gain and CD4 T cell count change was observed at six months p > 0.05. No positive correlation β =-138.23 was observed between CD4T Cell count, WAZ, p=0.934, R2 =0.0337 F >0.05. Majority (96.78%) of children on test either remained or progressed to WHO immunological stage I. Conclusion: From this study it can be concluded that yeast Selenium is effective in slowing the progress of HIV 1 in children from WHO clinical stage I by improving CD4 T cell count and hence the immunity.

Keywords: selenium, HIV, AIDS, WAZ

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Authors: Zakia Hbellaq

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The Mediterranean region is considered a hotbed for climate change. Morocco is a semi-arid Mediterranean country facing water shortages and poor water quality. Its limited water resources limit the activities of various economic sectors. Most of Morocco's territory is in arid and desert areas. The potential water resources are estimated at 22 billion m3, which is equivalent to about 700 m3/inhabitant/year, and Morocco is in a state of structural water stress. Strictly speaking, the Kingdom of Morocco is one of the “very riskiest” countries, according to the World Resources Institute (WRI), which oversees the calculation of water stress risk in 167 countries. The surprising results of the Institute (WRI) rank Morocco as one of the riskiest countries in terms of water scarcity, ranking 3.89 out of 5, thus occupying the 23rd place out of a total of 167 countries, which indicates that the demand for water exceeds the available resources. Agriculture with a score of 3.89 is most affected by water stress from irrigation and places a heavy burden on the water table. Irrigation is an unavoidable technical need and has undeniable economic and social benefits given the available resources and climatic conditions. Irrigation, and therefore the agricultural sector, currently uses 86% of its water resources, while industry uses 5.5%. Although its development has undeniable economic and social benefits, it also contributes to the overfishing of most groundwater resources and the surprising decline in levels and deterioration of water quality in some aquifers. In this context, REUSE is one of the proposed solutions to reduce the water footprint of the agricultural sector and alleviate the shortage of water resources. Indeed, wastewater reuse, also known as REUSE (reuse of treated wastewater), is a step forward not only for the circular economy but also for the future, especially in the context of climate change. In particular, water reuse provides an alternative to existing water supplies and can be used to improve water security, sustainability, and resilience. However, given the introduction of organic trace pollutants or, organic micro-pollutants, the absorption of emerging contaminants, and decreasing salinity, it is possible to tackle innovative capabilities to overcome these problems and ensure food and health safety. To this end, attention will be paid to the adoption of an integrated and attractive approach, based on the reinforcement and optimization of the treatments proposed for the elimination of the organic load with particular attention to the elimination of emerging pollutants, to achieve this goal. , membrane bioreactors (MBR) as stand-alone technologies are not able to meet the requirements of WHO guidelines. They will be combined with heterogeneous Fenton processes using persulfate or hydrogen peroxide oxidants. Similarly, adsorption and filtration are applied as tertiary treatment In addition, the evaluation of crop performance in terms of yield, productivity, quality, and safety, through the optimization of Trichoderma sp strains that will be used to increase crop resistance to abiotic stresses, as well as the use of modern omics tools such as transcriptomic analysis using RNA sequencing and methylation to identify adaptive traits and associated genetic diversity that is tolerant/resistant/resilient to biotic and abiotic stresses. Hence, ensuring this approach will undoubtedly alleviate water scarcity and, likewise, increase the negative and harmful impact of wastewater irrigation on the condition of crops and the health of their consumers.

Keywords: water scarcity, food security, irrigation, agricultural water footprint, reuse, emerging contaminants

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Authors: F. Madani, M. Doudi, L. Rahimzadeh Torabi

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The irregular consumption of current antibiotics contributes to an escalation in antibiotic resistance among urinary pathogens on a global scale. The objective of this research was to investigate the process of biologically synthesized silver nanoparticles through the utilization of Zataria multiflora extract. Additionally, the study aimed to evaluate the efficacy of these synthesized nanoparticles in inhibiting the growth of multi-drug resistant negative bacillus bacteria, which commonly contribute to urinary tract infections. The botanical specimen utilized in the current research investigation was Z. multiflora, and its extract was produced employing the Soxhlet extraction technique. The study examined the green synthesis conditions of silver nanoparticles by considering three key parameters: the quantity of extract used, the concentration of silver nitrate salt, and the temperature. The particle dimensions were ascertained using the Zetasizer technique. In order to identify synthesized Silver nanoparticles TEM, XRD, and FTIR methods were used. For evaluating the antibacterial effects of nanoparticles synthesized through a biological method, different concentrations of silver nanoparticles were studied on 140 cases of Multiple drug resistance (MDR) bacteria strains Escherichia coli, Klebsiella pneumoniae, Enterobacter aerogenes, Proteus vulgaris,Citrobacter freundii, Acinetobacter bumanii and Pseudomonas aeruginosa, (each genus of bacteria, 20 samples), which all were MDR and cause urinary tract infections, for identification of bacteria were used of PCR test and laboratory methods (Agar well diffusion and Microdilution methods) to assess their sensitivity to Nanoparticles. The data were subjected to analysis using the statistical software SPSS, specifically employing nonparametric Kruskal-Wallis and Mann-Whitney tests. This study yielded noteworthy findings regarding the impacts of varying concentrations of silver nitrate, different quantities of Z. multiflora extract, and levels of temperature on nanoparticles. Specifically, it was observed that an increase in the concentration of silver nitrate, extract amount, and temperature resulted in a reduction in the size of the nanoparticles synthesized. However, the impact of the aforementioned factors on the index of particle diffusion was found to be statistically non-significant. According to the transmission electron microscopy (TEM) findings, the particles exhibited predominantly spherical morphology, with a diameter spanning from 25 to 50 nanometers. Nanoparticles in the examined sample. Nanocrystals of silver. FTIR method illustrated that the spectrums of Z. multiflora and synthesized nanoparticles had clear peaks in the ranges of 1500-2000, and 3500 - 4000. The obtained results of antibacterial effects of different concentrations of silver nanoparticles on according to agar well diffusion and microdilution method, biologically synthesized nanoparticles showed 1000 mg /ml highest and lowest mean inhibition zone diameter in E. coli, A. bumanii 23 and 15mm, respectively. MIC was observed for all of bacteria 125 mg/ml and for A. bumanii 250 mg/ml. Comparing the growth inhibitory effect of chemically synthesized the results obtained from the experiment indicated that both nanoparticles and biologically synthesized nanoparticles exhibit a notable growth inhibition effect. Specifically, the chemical method of synthesizing nanoparticles demonstrated the highest level of growth inhibition at a concentration of 62.5 mg/mL The present study demonstrated an inhibitory effect on bacterial growth, facilitating the causative factors of urine infection and multidrug resistance (MDR).

Keywords: multiple drug resistance, negative bacillus bacteria, urine infection, Zataria multiflora

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Authors: Sandeep Vasaikar, Lary Obi

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Rapid and discriminative genotyping methods are useful for determining the clonality of the isolates in nosocomial or household outbreaks. Multilocus sequence typing (MLST) is a nucleotide sequence-based approach for characterising bacterial isolates. The genetic diversity and the clinical relevance of the drug-resistant Klebsiella isolates from Mthatha are largely unknown. For this reason, prospective, experimental study of the molecular epidemiology of Klebsiella isolates from patients being treated in Mthatha over a three-year period was analysed. Methodology: PCR amplification and sequencing of the drug-resistance-associated genes, and multilocus sequence typing (MLST) using 7 housekeeping genes mdh, pgi, infB, FusAR, phoE, gapA and rpoB were conducted. A total of 32 isolates were analysed. Results: The percentages of multidrug-resistant (MDR), extensively drug-resistance (XDR) and pandrug-resistant (PDR) isolates were; MDR 65.6 % (21) and XDR and PDR with 0 % each. In this study, K. pneumoniae was 19/32 (59.4 %). MLST results showed 22 sequence types (STs) were identified, which were further separated by Maximum Parsimony into 10 clonal complexes and 12 singletons. The most dominant group was Klebsiella pneumoniae with 23/32 (71.8 %) isolates, Klebsiella oxytoca as a second group with 2/32 (6.25 %) isolates, and a single (3.1 %) K. varricola as a third group while 6 isolates were of unknown sequences. Conclusions/significance: A phylogenetic analysis of the concatenated sequences of the 7 housekeeping genes showed that strains of K. pneumoniae form a distinct lineage within the genus Klebsiella, with K. oxytoca and K. varricola its nearest phylogenetic neighbours. With the analysis of 7 genes were determined 1 K. variicola, which was mistakenly identified as K. pneumoniae by phenotypic methods. Two misidentifications of K. oxytoca were found when phenotypic methods were used. No significant differences were observed between ESBL blaCTX-M, blaTEM and blaSHV groups in the distribution of Sequence types (STs) or Clonal complexes (CCs).

Keywords: phylogenetic analysis, phylogeny, klebsiella phylogenetic, klebsiella

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Authors: Fabio T. Costa, Sergio E. Moya, Marcelo H. Sousa

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Nanocomposites designed by embedding magnetic nanoparticles into a polymeric matrix stand out as ideal magnetic-hybrid and magneto-responsive materials as sorbents for removal of pollutants in environmental applications. Covalent coupling is often desired for the immobilization of species on these nanocomposites, in order to keep them permanently bounded, not desorbing or leaching over time. Moreover, unwanted adsorbates can be separated by successive washes/magnetic separations, and it is also possible to recover the adsorbate covalently bound to the nanocomposite surface through detaching/cleavage protocols. Thus, in this work, we describe the preparation and characterization of highly-magnetizable chloromethylated polystyrene-based nanocomposite beads for selective covalent coupling in environmental applications. For synthesis optimization, acid resistant core-shelled maghemite (γ-Fe₂O₃) nanoparticles were coated with oleate molecules and directly incorporated into the organic medium during a suspension polymerization process. Moreover, the cross-linking agent ethylene glycol dimethacrylate (EGDMA) was utilized for co-polymerization with the 4-vinyl benzyl chloride (VBC) to increase the resistance of microbeads against leaching. After characterizing samples with XRD, ICP-OES, TGA, optical, SEM and TEM microscopes, a magnetic composite consisting of ~500 nm-sized cross-linked polymeric microspheres embedding ~8 nm γ-Fe₂O₃ nanoparticles was verified. This nanocomposite showed large room temperature magnetization (~24 emu/g) due to the high content in maghemite (~45 wt%) and resistance against leaching even in acidic media. Moreover, the presence of superficial chloromethyl groups, probed by FTIR and XPS spectroscopies and confirmed by an amination test can selectively adsorb molecules through the covalent coupling and be used in molecular separations as shown for the selective removal of 4-aminobenzoic acid from a mixture with benzoic acid.

Keywords: nanocomposite, magnetic nanoparticle, covalent separation, pollutant removal

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Authors: Ahmet Aslan, Safiye Meric Acikel, Raziye Hilal Senay, Sinan Akgol

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Different chemical substances and too much water are used during leather production. Therefore, the waste water load of the leather industry is harmful to the environment. One of the pollution sources is the production of leather coloring process is a further need to focus on the removal of dye waste waters subject. These water-soluble dyes have a small organic molecular size. Besides the environmental hazards, these dyes cannot be underestimated, they also have harmful effects on human health. In this study, poly(hydroxyethyl methacrylate-glycidyl methacrylate) p(HEMA-GMA) hydrogel membranes were synthesized by UV polymerization method. The hydrogel synthesized is modified with imino diacetic acid (IDA) and then chelated with Cr (III) ions. The chelating capacity of the membranes was determined according to the time, pH and concentration parameters. Dynamic swelling test, elemental analysis, ninhydrin analysis and adsorption, desorption and reusability performances of membranes were also determined.

Keywords: adsorption, dye, leather, p(HEMA-GMA)-IDA

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Authors: K. Suresh, R. Arunkumar

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The objective of this study is to investigate the preparation of gramine nano suspension and protective effect of Gramine on the apoptosis of laryngeal cancer cells cell line (HEp-2 and KB). The growth inhibition rate of Hep-2 and KB cells in vitro were measured by MTT assay and apoptosis by, levels of reactive oxygen species, mitochondrial membrane potential, morphological changes and flowcytometry. Based on the results, we determined the effective doses of gramine as 127.23µm/ml for 24 hr and 119.81 µm/ml for 48hr in hep-2 cell line and 147.58 µm ml for 24 hr and 123.74µm µm/ml for 48hr in KB cell line. cytotoxicity effects of gramine were confirmed by treatment of HEp-2 cell and KB cell with IC50 concentration of gramine resulted in sequences of events marked by the enhance the apoptosis accompanied by loss of cell viability, modulation of reactive oxygen species and cell cycle arrest through the induction of G0/G1 phase arrest on HEp-2 cells. Our study suggests that the nanosuspension of gramine possesses the more cytotoxic effect of cancer cells and a novel candidate for cancer chemoprevention.

Keywords: apoptosis, HEp-2 cell line, KB cell line mitochondria, gramine, nanosuspension

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Authors: Chia-Chih Wei, Yun-Qi Li, Ssu-Ying Chen, Hsuan-Jung Chen, Hsi-Wen Yang, Chih-Yuan Chen, Chien-Chon Chen

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In this paper we used high quality anodization technique to make colored wind chime with a nano-tube structure anodic film, which controls the length to diameter ratio of an aluminum rod and controls the oxide film structure on the surface of the aluminum rod by anodizing method. The research experiment used hard anodization to grow a controllable thickness of anodic film on aluminum alloy surface. The hard anodization film has high hardness, high insulation, high temperature resistance, good corrosion resistance, colors, and mass production properties can be further applied to transportation, electronic products, biomedical fields, or energy industry applications. This study also in-depth research and detailed discussion in the related process of aluminum alloy surface hard anodizing including pre-anodization, anodization, and post-anodization. The experiment parameters of anodization including using a mixed acid solution of sulfuric acid and oxalic acid as an anodization electrolyte, and control the temperature, time, current density, and final voltage to obtain the anodic film. In the experiments results, the properties of anodic film including thickness, hardness, insulation, and corrosion characteristics, microstructure of the anode film were measured and the hard anodization efficiency was calculated. Thereby obtaining different transmission speeds of sound in the aluminum rod and different audio sounds can be presented on the aluminum rod. Another feature of the present invention is the use of anodizing method dyeing method, laser engraving patterning and electrophoresis method to make colored aluminum wind chimes.

Keywords: anodization, colored, high quality, wind chime, nano-tube

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Authors: Kwan-Ho Kim, Jin-Hong Park

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The multi-valued logic (MVL) circuits, which can handle more than two logic states, are one of the promising solutions to overcome the bit density limitations of conventional binary logic systems. Recently, tunneling devices such as Esaki diode and resonant tunneling diode (RTD) have been extensively explored to construct the MVL circuits. These tunneling devices present a negative differential resistance (NDR) phenomenon in which a current decreases as a voltage increases in a specific applied voltage region. Due to this non-monotonic current behavior, the tunneling devices have more than two threshold voltages, consequently enabling construction of MVL circuits. Recently, the emergence of two dimensional (2D) van der Waals (vdW) crystals has opened up the possibility to fabricate such tunneling devices easily. Owing to the defect-free surface of the 2D crystals, a very abrupt junction interface could be formed through a simple stacking process, which subsequently allowed the implementation of a high-performance tunneling device. Here, we report a vdW heterostructure based tunneling device with multiple threshold voltages, which was fabricated with black phosphorus (BP) and hafnium diselenide (HfSe₂). First, we exfoliated BP on the SiO₂ substrate and then transferred HfSe₂ on BP using dry transfer method. The BP and HfSe₂ form type-Ⅲ heterojunction so that the highly doped n+/p+ interface can be easily implemented without additional electrical or chemical doping process. Owing to high natural doping at the junction, record high peak to valley ratio (PVCR) of 16 was observed to the best our knowledge in 2D materials based NDR device. Furthermore, based on this, we first demonstrate the feasibility of the ternary latch by connecting two multi-threshold voltage devices in series.

Keywords: two dimensional van der Waals crystal, multi-valued logic, negative differential resistnace, tunneling device

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Authors: T. Büyüksırıt, H. Kuleaşan

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Natural antimicrobials are used to preserve foods that can be found in plants, animals, and microorganisms. Antimicrobial substances are natural or artificial agents that produced by microorganisms or obtained semi/total chemical synthesis are used at low concentrations to inhibit the growth of other microorganisms. Food borne pathogens and spoilage microorganisms are inactivated by the use of antagonistic microorganisms and their metabolites. Yeasts can produce toxic proteins or glycoproteins (toxins) that cause inhibition of sensitive bacteria and yeast species. Antimicrobial substance producing phenotypes belonging different yeast genus were isolated from different sources. Toxins secreted by many yeast strains inhibiting the growth of other yeast strains. These strains show antimicrobial activity, inhibiting the growth of mold and bacteria. The effect of antimicrobial agents produced by yeasts can be extremely fast, and therefore may be used in various treatment procedures. Rapid inhibition of microorganisms is possibly caused by microbial cell membrane lipopolysaccharide binding and in activation (neutralization) effect. Antimicrobial agents inhibit the target cells via different mechanisms of action.

Keywords: antimicrobial agents, yeast, toxic protein, glycoprotein

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Authors: Salma H. Abu Hafsa, A. Mendonca, B. Brehm-Stecher, A. A. Hassan, S. A. Ibrahim

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Enterococci are important inhabitants of the animal intestine and are widely used in probiotic products. A probiotic strain is expected to possess several desirable properties in order to exert beneficial effects. Therefore, the objective of this study was to isolate and characterize strains of Enterococcus sp. from chicken cecal and fecal samples to determine potential probiotic properties. Enterococci were isolated from thirty one chicken cecal and fecal samples collected from a local farm. In vitro studies were performed to assess antibacterial activity (using agar well diffusion and cell free supernatant broth technique against Salmonella enterica serotype Enteritidis), susceptibility to antibiotics (amoxycillin, cotrimoxazole, chloramphenicol, cefuroxime, ceftriaxone, ciprofloxacin, and nalidixic acid), survival in acidic conditions, resistance to bile salts, and their survival during simulated gastric juice conditions at pH 2.5. Isolates were identified using biochemical and molecular assays (API 50 CHL, and API ZYM kits followed by 16S rDNA gene sequence analysis). Two strains were identified, of which, Enteroccocus faecium was capable of inhibiting the growth of S. enteritidis and was susceptible to a wide range of antibiotics. In addition, the isolated strain exhibited significant resistance under highly acidic conditions (pH=2.5) for 8 hours and survived well in bile salt at 0.2% for 24 hours and showing ability to survive in the presence of simulated gastric juice at pH 2.5. Based on these results, the E. faecium isolate fulfills some of the criteria to be considered as a probiotic strain and therefore, could be used as a feed additive with good potential for controlling S. enteritidis in chickens. However, in vivo studies are needed to determine the safety of the strain.

Keywords: acid tolerance, antimicrobial activity, Enterococcus faecium, probiotic

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Authors: A. Chiker, A. Benamor, A. Haddad, Y. Hadji, M. Hadji

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Metal matrix composites (MMCs) have been of big interest for a few decades. Their major drawback lies in their enhanced mechanical performance over unreinforced alloys. They found ground in many engineering fields, such as aeronautics, aerospace, automotive, and other structural applications. One of the most used alloys as a matrix is nickel alloys, which meet the need for high-temperature mechanical properties; some attempts have been made to develop nickel base composites reinforced by high melt point and high modulus particulates. Among the carbides used as reinforcing particulates, chromium carbide is interesting for wear applications; it is widely used as a tribological coating material in high-temperature applications requiring high wear resistance and hardness. Moreover, a set of properties make it suitable for use in MMCs, such as toughness, the good corrosion and oxidation resistance of its three polymorphs -the cubic (Cr23C6), the hexagonal (Cr7C3), and the orthorhombic (Cr3C2)-, and it’s coefficient of thermal expansion that is almost equal to that of metals. The in-situ synthesis of CrC-reinforced Ni matrix composites could be achieved by the powder metallurgy route. To ensure the in-situ reactions during the sintering process, the use of phase precursors is necessary. Recently, new precursor materials have been proposed; these materials are called MAX phases. The MAX phases are thermodynamically stable nano-laminated materials displaying unusual and sometimes unique properties. These novel phases possess Mn+1AXn chemistry, where n is 1, 2, or 3, M is an early transition metal element, A is an A-group element, and X is C or N. Herein, the pressureless sintering method is used to elaborate Ni/Cr2AlC composites. Four composites were elaborated from 5, 10, 15 and 20 wt% of Cr2AlC MAX phase precursor which fully reacted with Ni-matrix at 1100 °C sintering temperature for 4 h in argon atmosphere. XRD results showed that Cr2AlC MAX phase was totally decomposed forming chromium carbide Cr7C3, and the released Al and Cr atoms diffused in Ni matrix giving rise to γ-Ni(Al,Cr) solid solution and γ’-Ni3(Al,Cr) intermetallic. Scanning Electron Microscopy (SEM) of the elaborated samples showed the presence of nanosized Cr7C3 reinforcing particles embedded in the Ni metal matrix, which have a direct impact on the tribological properties of the composites and their hardness. All the composites exhibited higher hardness than pure Ni; whereas adding 15 wt% of Cr2AlC gives the highest hardness (1.85 GPa). Using a ball-on-disc tribometer, dry sliding tests for the elaborated composites against 100Cr6 steel ball were studied under different applied loads. The microstructures and worn surface characteristics were then analyzed using SEM and Raman spectroscopy. The results show that all the composites exhibited better wear resistance compared to pure Ni, which could be explained by the formation of a lubricious tribo-layer during sliding and the good bonding between the Ni matrix and the reinforcing phases.

Keywords: composites, microscopy, sintering, wear

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Authors: Antonio Marco Miotti, Antonella Ruffatto, Giampaola Basso, Antonio Madia, Giulia Zavatta, Emanuela Salvatico, Emanuela Zilli

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A National Action Plan to fight antimicrobial resistance was launched in Italy in 2017. In order to reduce inappropriate exposure to antibiotics and infections from multi-drug resistant bacteria, it is essential to set up a structured system of surveillance and monitoring of the implementation of National Action Plan standards, including antimicrobial consumption, with a special focus on quinolones, third generation cephalosporins and carbapenems. A quantitative estimate of antibiotic consumption (defined daily dose - DDD - consumption per 100 days of hospitalization) has been provided by the Pharmaceutical Service to the Hospital of Cittadella, ULSS 6 Euganea – Health Trust (District of Padua) for the years 2019 (before the pandemic), 2020 and 2021 for all classes of antibiotics. Multidisciplinary meetings have been organized monthly by the local Antimicrobial Stewardship Group. Between 2019 and 2021, an increase in the consumption of carbapenems in the Intensive Care Unit (from 12.2 to 18.2 DDD, + 49.2%) and a decrease in Medical wards (from 5.3 to 2.6 DDD, - 50.9%) was reported; a decrease in the consumption of quinolones in Intensive Care Unit (from 17.2 to 10.8 DDD, - 37.2%), Medical wards (from 10.5 to 6.6 DDD, - 37.1%) and Surgical wards (from 10.2 to 9.3 DDD, - 8.8%) was highlighted; an increase in the consumption of third generation cephalosporins in Medical wards (from 18.1 to 22.6 DDD, + 24,1%) was reported. Finally, after an increase in the consumption of macrolides between 2020 and 2019, in 2021, a decrease was reported in the Intensive Care Unit (DDD: 8.0 in 2019, 18.0 in 2020, 6.4 in 2021) and Medical wards (DDD: 9.0 in 2019, 13.7 in 2020, 10.9 in 2021). Constant monitoring of antimicrobial consumption and timely identifying of warning situations that may need a specific intervention are the cornerstone of Antimicrobial Stewardship programs, together with analysing data on bacterial resistance rates and infections from multi-drug resistant bacteria.

Keywords: carbapenems, quinolones, antimicrobial, stewardship

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Authors: A. T. Adetunji, F. B. Lewu, R. Mundembe

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Plants require nitrogen (N) to support desired production levels. Nitrogen is available to plants in the form of nitrate or ammonium, which are transported into the cell with the aid of various transport proteins. Ammonium transporters (AMTs) play a role in the uptake of ammonium, the form in which nitrogen is preferentially absorbed by plants. Solanum tuberosum AMT1 (StAMT1) was characterized using molecular biology and bioinformatics methods. Nucleotide database sequences were used to design AMT1-specific primers which were used to amplify the AMT1 internal regions. Nucleotide sequencing, alignment and phylogenetic analysis assigned StAMT1 to the AMT1 family. The deduced amino acid sequences showed that StAMT1 is 92%, 83% and 76% similar to Solanum lycopersicum LeAMT1.1, Lotus japonicus LjAMT1.1 and Solanum lycopersicum LeAMT1.2 respectively. StAMT1 fragments were shown to correspond to the 5th - 10th trans-membrane domains. Residue StAMT1 D15 is predicted to be essential for ammonium transport, while mutations of StAMT1 S76A may further enhance ammonium transport.

Keywords: ammonium transporter, bioinformatics, nitrogen, primers, Solanum tuberosum

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Authors: Mario Alejandro Grisales, M. Daniela Chimá, Gilberto Bejarano Gaitán

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High entropy alloys (HEA) and nitride (HEN) are currently very attractive to the automotive, aerospace, metalworking and materials forming manufacturing industry, among others, for exhibiting higher mechanical properties, wear resistance, and thermal stability than binary and ternary alloys. In this work medium-entropy coatings of TiTaZrNb and the nitrides of (TiTaZrNb)Nx were synthesized on to AISI 420 and M2 steel samples by the direct current magnetron sputtering technique. The influence of the bias voltage supplied to the substrate on the microstructure, chemical- and phase composition of the matrix coating was evaluated, and the effect of nitrogen flow on the microstructural, mechanical and tribological properties of the corresponding nitrides was studied. A change in the crystalline structure from BCC for TiTaZrNb coatings to FCC for (TiTaZrNb)Nx was observed, that is associated with the incorporation of nitrogen into the matrix and the consequent formation of a solid solution of (TiTaZrNb)Nx. An increase in hardness and residual stresses was observed with increasing bias voltage for TiTaZrNb, reaching 12.8 GPa for the coating deposited with a bias of -130V. In the case of (TiTaZrNb)Nx nitride, a greater hardness of 23 GPa is achieved for the coating deposited with a N2 flow of 12 sccm, which slightly drops to 21.7 GPa for that deposited with N2 flow of 15 sccm. The slight reduction in hardness could be associated with the precipitation of the TiN and ZrN phases that are formed at higher nitrogen flows. The specific wear rate of the deposited coatings ranged between 0.5xexp13 and 0.6xexp13 N/m2. The steel substrate exhibited an average hardness of 2.0 GPa and a specific wear rate of 203.2exp13 N/m2. Both the hardness and the specific wear rate of the synthesized nitride coatings were higher than that of the steel substrate, showing a protective effect of the steel against wear.

Keywords: medium entropy coatings, hard coatings, magnetron sputtering, tribology, wear resistance

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Authors: Faramarz Khoshnoudian

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A seismic stability assessment of the concrete gravity dam was performed. Initially (Phase 1), a linear response spectrum analysis was performed to verify the potential for crack formation. The result shows the possibility of developing cracks in the upstream face of the dam close to the lowest gallery, which were sufficiently long that the dam would not be stable following the earthquake. The results show the dam has potentially inadequate seismic and post-earthquake resistance and recommended an update of the stability analysis.

Keywords: crack propgation, concrete gravity dam, seismic, assesment

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Authors: Daisy Biswas, Samar Kanti Bose, M. Mozaffar Hossain

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The development of value added composite products from bamboo with the application of gluing technology can play a vital role in economic development and also in forest resource conservation of any country. In this study, the gluability of Bambusa balcooa and Bambusa vulgaris, two locally grown bamboo species of Bangladesh was assessed. As the culm wall thickness of bamboos decreases from bottom to top, a culm portion of up to 5.4 m and 3.6 m were used from the base of B. balcooa and B. vulgaris, respectively, to get rectangular strips of uniform thickness. The color of the B. vulgaris strips was yellowish brown and that of B. balcooa was reddish brown. The strips were treated in borax-boric, bleaching and carbonization for extending the service life of the laminates. The preservative treatments changed the color of the strips. Borax–boric acid treated strips were reddish brown. When bleached with hydrogen peroxide, the color of the strips turned into whitish yellow. Carbonization produced dark brownish strips having coffee flavor. Chemical constituents for untreated and treated strips were determined. B. vulgaris was more acidic than B. balcooa. Then the treated strips were used to develop three-layered bamboo laminated panel. Urea formaldehyde (UF) and polyvinyl acetate (PVA) were used as binder. The shear strength and abrasive resistance of the panel were evaluated. It was found that the shear strength of the UF-panel was higher than the PVA-panel for all treatments. Between the species, gluability of B. vulgaris was better and in some cases better than hardwood species. The abrasive resistance of B. balcooa is slightly higher than B. vulgaris; however, the latter was preferred as it showed well gluability. The panels could be used as structural panel, floor tiles, flat pack furniture component, and wall panel etc. However, further research on durability and creep behavior of the product in service condition is warranted.

Keywords: Bambusa balcooa, Bambusa vulgaris, polyvinyl acetate, urea formaldehyde

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Authors: Shanawaz Patil, Mohamed Haneef, K. S. Narayanaswamy

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In the recent years, aluminum metal matrix composites were most widely used, which are finding wide applications in various field such as automobile, aerospace defense etc., due to their outstanding mechanical properties like low density, light weight, exceptional high levels of strength, stiffness, wear resistance, high temperature resistance, low coefficient of thermal expansion and good formability. In the present work, an effort is made to study the effect of heat treatment on mechanical properties of aluminum 7075 alloy reinforced with constant weight percentage of naturally occurring mineral beryl and varying weight percentage of graphene. The hybrid composites are developed with 0.5 wt. %, 1wt.%, 1.5 wt.% and 2 wt.% of graphene and 6 wt.% of beryl  by stir casting liquid metallurgy route. The cast specimens of unreinforced aluminum alloy and hybrid composite samples were prepared for heat treatment process and subjected to solutionizing treatment (T6) at a temperature of 490±5 ^oC for 8 hours in a muffle furnace followed by quenching in boiling water. The microstructure analysis of as cast and heat treated hybrid composite specimens are examined by scanning electron microscope (SEM). The tensile test and hardness test of unreinforced aluminum alloy and hybrid composites are examined. The wear behavior is examined by pin-on disc apparatus. The results of as cast specimens and heat treated specimens were compared. The heat treated Al7075-Beryl-Graphene hybrid composite had better properties and significantly improved the ultimate tensile strength, hardness and reduced wear loss when compared to aluminum alloy and  as cast hybrid composites.

Keywords: beryl, graphene, heat treatment, mechanical properties

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Authors: Ali Naeemy, Mir Ghasem Hoseini

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In this study, for the first time, an analytical method developed and validated by combining electrochemical impedance spectroscopy and electromembrane extraction (EIS-EME) by Vulcan/poly pyrrole nanocomposite modified screen-printed electrode (PPY–VU/SPE) for accurately quantifying zolpidem. EME parameters optimized, including solvent composition, voltage, pH adjustments and extraction time. Zolpidem was transferred from a donor solution (pH 5) to an acceptor solution (pH 13) using a hollow fiber in 1-octanol as a membrane, driven by a 60 V voltage for 25 minutes, ensuring precise and selective extraction. In comparison with SPE, VU/SPE and PPY/SPE, the PPY–VU/SPE was much more efficient for ZP oxidation. Calibration curves with good linearity were obtained in the concentration range of 2-75 µmol L-1 using the EIS-EME with the detection limit of 0.5 µmol L-1 . Finally, the EIS-EME by using the PPY– VU/SPE was successfully used to determine ZP in tablet dosage form, urine and plasma samples. Keywords: Electrochemical impedance spectroscopy, Electromembrane extraction, Zolpidem, Vulcan, poly pyrrole, Screen printed electrode

Keywords: electrochemical impedance spectroscopy, electromembrane extraction, screen printed electrode, zolpidem

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Authors: Piyaporn Buaklang, Narisa Kengtrong Bordeerat

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The use of nanoparticles is increasing worldwide and there are many nanotech-based daily products available in the market. The toxicity of nanoparticles results from their extremely small size which can be transported easily into the blood stream and other organs. We aimed to study the genotoxicity of two nanoparticles, Titanium dioxide (TiO2-NPs) and Zinc oxide (ZnO-NPs), in TK6 cells by micronucleus assay. The cells were tested at 8, 24, and 48 hours after exposed to 0.10, 0.25, 0.50 and 1.00 µg/mL of TiO2-NPs particles size < 25 nm and < 100 nm and to ZnO-NPs at 1, 10, 50, and 100 µg/mL, particles size < 50 nm and < 100 nm. At 24 hours of incubation transmission electron microscope (TEM) revealed that the nanoparticles TiO2-NPs at 1.00 µg/mL and ZnO-NPs at 10 µg/mL were able to be taken into the cells and induced the production of increasing amount of micronucleus in dose-dependent manner. The effect of the two nanoparticles on chromosome aberration indicated that TiO2-NPs and ZnO-NPs are genotoxic. In addition, the toxicity of TiO2-NPs was found to be 10 times more toxic than ZnO-NPs after 24 hours exposure. Analysis showed that the TiO2-NPs induced formation of micronucleus was both time and dose dependent, whereas the genotoxicity of ZnO-NPs was only dose dependent. In conclusion, TiO2-NPs and ZnO-NPs were able to transport through the cells membrane and directly genotoxic to TK6 cells in dose-dependent manner.

Keywords: nanoparticles, genotoxicity, human lymphoblast cells (TK6), micronucleus

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Authors: Min-Hwa Lim, Mi-Jeong Park, Ho-Young Jung

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Imidazolium-brominated polyphenylene oxide (Im-bPPO) is based on the functionalization of bromomethylated poly(2,6-dimethyl-1,4-phenylene oxide) (BPPO) using 1-Methylimdazole. For the purpose of long cycle life of vanadium redox battery (VRB), the chemical stability of Im-bPPO, sPPO (sulfonated 2,6-dimethyl-1,4-phenylene oxide) and Fumatech membranes were evaluated firstly in the 0.1M vanadium (V) solution dissolved in 3M sulfuric acid (H2SO4) for 72h, and UV analyses of the degradation products proved that ether bond in PPO backbone was vulnerable to be attacked by vanadium (V) ion. It was found that the membranes had slightly weight loss after soaking in 2 ml distilled water included in STS pressure vessel for 1 day at 200◦C. ATR-FT-IR data indicated before and after the degradation of the membranes. Further evaluation on the degradation mechanism of the menbranes were carried out in Fenton’s reagent solution for 72 h at 50 ◦C and analyses of the membranes before and after degradation confirmed the weight loss of the membranes. The Fumatech membranes exhibited better performance than AEM and CEM, but Nafion 212 still suffers chemical degradation.

Keywords: vanadium redox flow battery, ion exchange membrane, permeability, degradation, chemical stability

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Authors: Xiping Guo, Fanglin Ge, Ping Guan

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Refractory high entropy alloys are alternative materials expected to be employed at high temperatures. The comprehensive changes of microstructure and properties of NbTiZrCrAl refractory high entropy alloys are systematically studied by adjusting Al content. Five kinds of button alloy ingots with different contents of Al in NbTiZrCrAlX (X=0, 0.2, 0.5, 0.75, 1.0) were prepared by vacuum non-consumable arc melting technology. The microstructure analysis results show that the five alloys are composed of BCC solid solution phase rich in Nb and Ti and Laves phase rich in Cr, Zr, and Al. The addition of Al changes the structure from hypoeutectic to hypereutectic, increases the proportion of Laves phase, and changes the structure from cubic C15 to hexagonal C14. The hardness and fracture toughness of the five alloys were tested at room temperature, and the compressive mechanical properties were tested at 1000℃. The results showed that the addition of Al increased the proportion of Laves phase and decreased the proportion of the BCC phase, thus increasing the hardness and decreasing the fracture toughness at room temperature. However, at 1000℃, the strength of 0.5Al and 0.75Al alloys whose composition is close to the eutectic point is the best, which indicates that the eutectic structure is of great significance for the improvement of high temperature strength of NbTiZrCrAl refractory high entropy alloys. The five alloys were oxidized for 1 h and 20 h in static air at 1000℃. The results show that only the oxide film of 0Al alloy falls off after oxidizing for 1 h at 1000℃. After 20h, the oxide film of all the alloys fell off, but the oxide film of alloys containing Al was more dense and complete. By producing protective oxide Al₂O₃, inhibiting the preferential oxidation of Zr, promoting the preferential oxidation of Ti, and combination of Cr₂O₃ and Nb₂O₅ to form CrNbO₄, Al significantly improves the high temperature oxidation resistance of NbTiZrCrAl refractory high entropy alloys.

Keywords: NbTiZrCrAl, refractory high entropy alloy, al content, microstructural evolution, room temperature mechanical properties, high temperature compressive strength, oxidation resistance

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Authors: Konstantine Eristavi

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The paper will contribute to the discussion on the pitfalls, limits, and possibilities of legal and rights discourse in opposing large infrastructural projects in the context of neoliberal globalisation. To this end, the paper will analyse the struggle against the Namakhvani HPP project in Georgia. The latter has been hailed by the government as one of the largest energy projects in the history of the country, with an enormous potential impact on energy security, energy independence, economic growth, and development. This takes place against the backdrop of decades of market-led -or neoliberal- model of development in Georgia, characterised by structural adjustments, deregulation, privatisation, and Laissez-Fair approach to foreign investment. In this context, the Georgian state vies with other low and middle-income countries for foreign capital by offering to potential investors, on the one hand, exemptions from social and environmental regulations and, on the other hand, huge legal concessions and safeguards, thereby participating in what is often called a “race to the bottom.” The Namakhvani project is a good example of this. At every stage, the project has been marred with violations of laws and regulations concerning transparency, participation, social and environmental regulations, and so on. Moreover, the leaked contract between the state and the developer reveals the contractual safeguards which effectively insulate the investment throughout the duration of the contract from the changes in the national law that might adversely affect investors’ rights and returns. These clauses, aimed at preserving investors' economic position, place the contract above national law in many respects and even conflict with fundamental constitutional rights. In response to the perceived deficiencies of the project, one of the largest and most diverse social movements in the history of post-soviet Georgia has been assembled, consisting of the local population, conservative and leftist groups, human rights and environmental NGOs, etc. Crucially, the resistance movement is actively using legal tools. In order to analyse both the limitations and possibilities of legal discourse, the paper will distinguish between internal and immanent critiques. Law as internal critique, in the context of the struggles around the Namakhvani project, while potentially fruitful in hindering the project, risks neglecting and reproducing those factors -e.g., the particular model of development- that made such contractual concessions and safeguards and concomitant rights violations possible in the first place. On the other hand, the use of rights and law as part of immanent critique articulates a certain incapacity on the part of the addressee government to uphold existing laws and rights due to structural factors, hence, pointing to a need for a fundamental change. This 'ruptural' form of legal discourse that the movement employs makes it possible to go beyond the discussion around the breaches of law and enables a critical deliberation on the development model within which these violations and extraordinary contractual safeguards become necessary. It will be argued that it is this form of immanent critique that expresses the emancipatory potential of legal discourse.

Keywords: law, resistance, development, rights

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

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

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

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Authors: Shashikant Iyengar, Jasmeet Kaur, Anup Singh, Arun Kumar, Ira Sahay

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Insulin resistance (IR) is a condition that occurs when cells in the body become less responsive to insulin, leading to higher levels of both insulin and glucose in the blood. This condition is linked to metabolic syndromes, including diabetes. It is crucial to address IR promptly after diagnosis to prevent long-term complications associated with high insulin and high blood glucose. This four-month case study highlights the importance of treating the underlying condition to manage diabetes effectively. Insulin is essential for regulating blood sugar levels by facilitating the uptake of glucose into cells for energy or storage. In IR individuals, cells are less efficient at taking up glucose from the blood resulting in elevated blood glucose levels. As a result of IR, beta cells produce more insulin to make up for the body's inability to use insulin effectively. This leads to high insulin levels, a condition known as hyperinsulinemia, which further impairs glucose metabolism and can contribute to various chronic diseases. In addition to regulating blood glucose, insulin has anti-catabolic effects, preventing the breakdown of molecules in the body, such as inhibiting glycogen breakdown in the liver, inhibiting gluconeogenesis, and inhibiting lipolysis. If a person is insulin-sensitive or metabolically healthy, an optimal level of insulin prevents fat cells from releasing fat and promotes the storage of glucose and fat in the body. Thus optimal insulin levels are crucial for maintaining energy balance and plays a key role in metabolic processes. During the four-month study, researchers looked at the impact of a low-carb dietary (LCD) intervention on two male individuals (A & B) who had Type-2 diabetes. Althoughvneither of these individuals were obese, they were both slightly overweight and had abdominal fat deposits. Before the trial began, important markers such as fasting blood glucose (FBG), triglycerides (TG), high-density lipoprotein (HDL) cholesterol, and Hba1c were measured. These markers are essential in defining metabolic health, their individual values and variability are integral in deciphering metabolic health. The ratio of TG to HDL is used as a surrogate marker for IR. This ratio has a high correlation with the prevalence of metabolic syndrome and with IR itself. It is a convenient measure because it can be calculated from a standard lipid profile and does not require more complex tests. In this four-month trial, an improvement in insulin sensitivity was observed through the ratio of TG/HDL, which, in turn, improves fasting blood glucose levels and HbA1c. For subject A, HbA1c dropped from 13 to 6.28, and for subject B, it dropped from 9.4 to 5.7. During the trial, neither of the subjects were taking any diabetic medications. The significant improvements in their health markers, such as better glucose control, along with an increase in energy levels, demonstrate that incorporating LCD interventions can effectively manage diabetes.

Keywords: metabolic disorder, insulin resistance, type-2 diabetes, low-carb nutrition

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Authors: J. J. Mathew

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Pulmonary tuberculosis is highly prevalent in the Indian subcontinent. Most cases of pulmonary tuberculosis are diagnosed with sputum examinations and the vast majority of these are undertaken by the government run establishments. However, mycobacterial cultures are not routinely done, unless drug resistance is detected based on clinical response. Modern diagnostic tests like bronchoscopy and Genexpert are not routinely employed in the government institutions for the diagnosis of pulmonary tuberculosis, but have been accepted widely by good private institutions. The utility of these investigations in the private sector is not yet well recognized. This retrospective study aims to assess the usefulness of bronchoscopy and Genexpert in the diagnosis of pulmonary tuberculosis in quaternary care private hospital in India. 30 patients with respiratory symptoms raising the possibility of tuberculosis based on clinical and radiological features, but without any significant sputum production, were subject to bronchoscopy and BAL samples taken for microbiological studies, including Genexpert. 6 out of the 30 patients were found to be Genexpert positive and none of them showed Rifampicin resistance. All the 6 cases had upper zone predominant disease. One of the 6 cases of tuberculosis had another co-existent bacterial infection according to the routine culture studies. 6 other cases were proven to be due to other bacterial infections alone, 2 had a malignant diagnosis and the remaining cases were thought to be non-infective pathologies. The Genexpert results were made available within 48 hours in the 6 positive cases. All of them were commenced on standard anti-tuberculous regimen with excellent clinical response. The other infective cases were also managed successfully based on the drug susceptibilities. The study has shown the usefulness of these investigations as early intervention enabled diagnosis facilitating treatment and prevention of any clinical deterioration. The study lends support to early bronchoscopy and Genexpert testing in suspected cases of pulmonary tuberculosis without significant sputum production, in a high prevalence country which normally relies on sputum examination for the diagnosis of pulmonary tuberculosis.

Keywords: pulmonary, tuberculosis, bronchoscopy, genexpert

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Authors: Jan Masak, Eva Kvasnickova, Vladimir Scholtz, Olga Matatkova, Marketa Valkova, Alena Cejkova

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Microbial colonization of medical instruments, catheters, implants, etc. is a serious problem in the spread of nosocomial infections. Biofilms exhibit enormous resistance to environment. The resistance of biofilm populations to antibiotic or biocides often increases by two to three orders of magnitude in comparison with suspension populations. Subjects of interests are substances or physical processes that primarily cause the destruction of biofilm, while the released cells can be killed by existing antibiotics. In addition, agents that do not have a strong lethal effect do not cause such a significant selection pressure to further enhance resistance. Non-thermal plasma (NTP) is defined as neutral, ionized gas composed of particles (photons, electrons, positive and negative ions, free radicals and excited or non-excited molecules) which are in permanent interaction. In this work, the effect of NTP generated by the cometary corona with a metallic grid on the formation and stability of biofilm and metabolic activity of cells in biofilm was studied. NTP was applied on biofilm populations of Staphylococcus epidermidis DBM 3179, Pseudomonas aeruginosa DBM 3081, DBM 3777, ATCC 15442 and ATCC 10145, Escherichia coli DBM 3125 and Candida albicans DBM 2164 grown on solid media on Petri dishes and on the titanium alloy (Ti6Al4V) surface used for the production joint replacements. Erythromycin (for S. epidermidis), polymyxin B (for E. coli and P. aeruginosa), amphotericin B (for C. albicans) and ceftazidime (for P. aeruginosa) were used to study the combined effect of NTP and antibiotics. Biofilms were quantified by crystal violet assay. Metabolic activity of the cells in biofilm was measured using MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) colorimetric test based on the reduction of MTT into formazan by the dehydrogenase system of living cells. Fluorescence microscopy was applied to visualize the biofilm on the surface of the titanium alloy; SYTO 13 was used as a fluorescence probe to stain cells in the biofilm. It has been shown that biofilm populations of all studied microorganisms are very sensitive to the type of used NTP. The inhibition zone of biofilm recorded after 60 minutes exposure to NTP exceeded 20 cm², except P. aeruginosa DBM 3777 and ATCC 10145, where it was about 9 cm². Also metabolic activity of cells in biofilm differed for individual microbial strains. High sensitivity to NTP was observed in S. epidermidis, in which the metabolic activity of biofilm decreased after 30 minutes of NTP exposure to 15% and after 60 minutes to 1%. Conversely, the metabolic activity of cells of C. albicans decreased to 53% after 30 minutes of NTP exposure. Nevertheless, this result can be considered very good. Suitable combinations of exposure time of NTP and the concentration of antibiotic achieved in most cases a remarkable synergic effect on the reduction of the metabolic activity of the cells of the biofilm. For example, in the case of P. aeruginosa DBM 3777, a combination of 30 minutes of NTP with 1 mg/l of ceftazidime resulted in a decrease metabolic activity below 4%.

Keywords: anti-biofilm activity, antibiotic, non-thermal plasma, opportunistic pathogens

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