Search results for: mitochondrial membrane

Authors: Nadine Wiesmann, Melanie Viel, Christoph Buhr, Rachel Tanner, Wolfgang Tremel, Juergen Brieger

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Recidivation of tumors and the development of resistances against the classical anti-tumor approaches represent a major challenge we face when treating cancer. In order to master this challenge, we are in desperate need of new treatment options beyond the beaten tracks. Zinc oxide nanoparticles (ZnO NPs) represent such an innovative approach. Zinc oxide is characterized by a high level of biocompatibility, concurrently ZnO NPs are able to exert anti-tumor effects. By concentration of the nanoparticles at the tumor site, tumor cells can specifically be exposed to the nanoparticles while low zinc concentrations at off-target sites are tolerated well and can be excreted easily. We evaluated the toxicity of ZnO NPs in vitro with the help of immortalized tumor cell lines and primary cells stemming from healthy tissue. Additionally, the Chorioallantoic Membrane Assay (CAM Assay) was employed to gain insights into the in vivo behavior of the nanoparticles. We could show that ZnO NPs interact with tumor cells as nanoparticulate matter. Furthermore, the extensive release of zinc ions from the nanoparticles nearby and within the tumor cells results in overload with zinc. Beyond that, ZnO NPs were found to further the generation of reactive oxygen species (ROS). We were able to show that tumor cells were more prone to the toxic effects of ZnO NPs at intermediate concentrations compared to fibroblasts. With the help of ZnO NPs covered by a silica shell in which FITC dye was incorporated, we were able to track ZnO NPs within tumor cells as well as within a whole organism in the CAM assay after injection into the bloodstream. Depending on the applied concentrations, selective tumor cell killing seems feasible. Furthermore, the combinational treatment of tumor cells with radiotherapy and ZnO NPs shows promising results. Still, further investigations are needed to gain a better understanding of the interaction between ZnO NPs and the human body to be able to pave the way for their application as an innovative anti-tumor agent in the clinics.

Keywords: metal oxide nanoparticles, nanomedicine, overcome resistances against classical treatment options, zinc oxide nanoparticles

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Authors: Hasan Türe, Kader Terzioglu, Evren Tunca

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Contamination of aqueous environment by heavy metal ions is a serious and complex problem, owing to their hazards to human being and ecological systems. The treatment methods utilized for removing metal ions from aqueous solution include membrane separation, ion exchange and chemical precipitation. However, these methods are limited by high operational cost. Recently, biobased beads are considered as promising biosorbent to remove heavy metal ions from water. The aim of present study was to characterize the alginate/perlite composite beads and to investigate the adsorption performance of obtained beads for removing Pb (II) from aqueous solution. Alginate beads were synthesized by ionic gelation methods and different amount of perlite (aljinate:perlite=1, 2, 3, 4, 5 wt./wt.) was incorporated into alginate beads. Samples were characterized by means of X-ray diffraction (XRD), thermogravimetric analysis (TGA), scanning electron microscopy (SEM). The effects of perlite level, the initial concentration of Pb (II), initial pH value of Pb(II) solution and effect of contact time on the adsorption capacity of beads were investigated by using batch method. XRD analysis indicated that perlite includes silicon or silicon and aluminum bearing crystalline phase. The diffraction pattern of perlite containing beads is similar to that of that perlite powder with reduced intensity. SEM analysis revealed that perlite was embedded into alginate polymer and SEM-EDX (Energy-Dispersive X-ray) showed that composite beads (aljinate:perlite=1) composed of C (41.93 wt.%,), O (43.64 wt.%), Na (10.20 wt.%), Al (0.74 wt.%), Si (2.72 wt.%) ve K (0.77 wt.%). According to TGA analysis, incorporation of perlite into beads significantly improved the thermal stability of the samples. Batch experiment indicated that optimum pH value for Pb (II) adsorption was found at pH=7 with 1 hour contact time. It was also found that the adsorption capacity of beads decreased with increases in perlite concentration. The results implied that alginate/perlite composite beads could be used as promising adsorbents for the removal of Pb (II) from wastewater. Acknowledgement: This study was supported by TUBITAK (Project No: 214Z146).

Keywords: alginate, adsorption, beads, perlite

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Authors: Syed Hadi Hasan, Devendra Kumar Singh, Viyaj Kumar

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Cr (VI) is a known toxic heavy metal and has been considered as a priority pollutant in water. The effluent of various industries including electroplating, anodizing baths, leather tanning, steel industries and chromium based catalyst are the major source of Cr (VI) contamination in the aquatic environment. Cr (VI) show high mobility in the environment and can easily penetrate cell membrane of the living tissues to exert noxious effects. The Cr (VI) contamination in drinking water causes various hazardous health effects to the human health such as cancer, skin and stomach irritation or ulceration, dermatitis, damage to liver, kidney circulation and nerve tissue damage. Herein, an attempt has been done to develop an efficient adsorbent for the removal of Cr (VI) from water. For this purpose nanosorbent composed of polyvinyl alcohol functionalized graphene oxide (GO/PVA) was prepared. Thus, obtained GO/PVA was characterized through FTIR, XRD, SEM, and Raman Spectroscopy. As prepared nanosorbent of GO/PVA was utilized for the removal Cr (VI) in batch mode experiment. The process variables such as contact time, initial Cr (VI) concentration, pH, and temperature were optimized. The maximum 99.8 % removal of Cr (VI) was achieved at initial Cr (VI) concentration 60 mg/L, pH 2, temperature 35 °C and equilibrium was achieved within 50 min. The two widely used isotherm models viz. Langmuir and Freundlich were analyzed using linear correlation coefficient (R2) and it was found that Langmuir model gives best fit with high value of R2 for the data of present adsorption system which indicate the monolayer adsorption of Cr (VI) on the GO/PVA. Kinetic studies were also conducted using pseudo-first order and pseudo-second order models and it was observed that chemosorptive pseudo-second order model described the kinetics of current adsorption system in better way with high value of correlation coefficient. Thermodynamic studies were also conducted and results showed that the adsorption was spontaneous and endothermic in nature.

Keywords: adsorption, GO/PVA, isotherm, kinetics, nanosorbent, thermodynamics

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Authors: Ananya Govindarajan

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Tetrahymena thermophila is a single-cell, eukaryotic organism that belongs to the Protozoa Kingdom. Tetrahymena thermophila is often used in signal transduction pathway studies because of its ability to model sensory input and the effects of environmental conditions such as chemicals and temperature. The recently discovered G37 chemorepellent receptor showed increased responsiveness to all chemorepellents. Investigating the mutant G37 Tetrahymena gene in various test solutions, including ferric chloride, ferrous sulfate, hydrogen peroxide, tetrazolium blue, potassium chloride, and dithiothreitol were performed to determine the role of oxidants and reducing agents with the mutant and wild-type cells (CU427) to assess the role of the receptor. Behavioral assays and recordings processed by ImageJ indicated that ferric chloride, hydrogen peroxide, and tetrazolium blue yielded little to no chemorepellent responses from G37 cells (<20% ARs). CU427 cells were over-responsive based on the mean percent of cells (>50% ARs). Reducing agents elicited chemorepellent responses from both G37 and CU427, in addition to potassium chloride. Cell responses were classified as over-responsive (>50% ARs). Dithiothreitol yielded unexpected results as G37 (37.0% ARs) and CU427 (38.1% ARs) had relatively similar responses and were only responsive and not over-responsive to the reducing agent test chemical solution. Ultimately, this indicates that the G37 receptor is more interactive with molecules that are reducing agents or non-oxidant compounds; G37 may be unable to sense and respond to oxidants effectively, further elucidating the pathways of the G37 strain and nature of this receptor. Results also indicate that the CSF most likely contained an oxidant, like ferric chloride. This research can be further applied to neuronal influences and how specific compounds may affect human neurons individually and their excitability as the responses model action potentials and membrane potential.

Keywords: tetrahymena thermophila, signal transduction, chemosensory, oxidant, reducing agent

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Authors: Anca Dinischiotu, Sorina Nicoleta Voicu

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Silica nanoparticles (SiO2-NPs) are widely used in consumer products such as paints, plastics, insulation materials, tires, concrete production, as well as in gene delivery systems and imaging procedures. Environmental human exposure to them occurs during utilization of these products, in a time-dependent manner, the uptake being by topic and inhalation route especially. SiO2-NPs enter cells and induce membrane damage, oxidative stress and inflammatory reactions in a concentration-dependent manner. In this study, MRC-5 cells (human fetal lung fibroblasts) were exposed to amorphous SiO2-NPs at a dose of 62.5 μg/ml for 24, 48 and 72 hours. The size distribution of NPs was a lognormal function, in the range 3-14 nm. A time-dependent decrease of total reduced glutathione concentration by 36%, 50%, and 78% and an increase of NO level by 62%, 32%, respectively 24% compared to control were noticed. An up-regulation of NF-kB expression by 20%, 50% respectively 10% and of Nrf-2 by 139%, 58%, and 16% compared to control after 24, 48 and 72 hours was noticed also. The expression of IL-1β, IL-6, IL-8, and COX-2 was up-regulated in a time-dependent manner. Also, the expression of MMP-2 and MMP-9 were down-regulated after 48 and 72 hours, whereas their activities raised in a time-dependent manner. Exposure of cells to NPs up-regulated the expression of inducible NO synthase, as previously was shown, and probably this is the reason for the increased level of NO, that can react with the thiol groups of reduced glutathione molecules, diminishing its concentration Nrf2 is a transcription factor translocated in nucleus, under oxidative stress, where downstream gene expression activates in order to modulate the adaptive intracellular response against oxidative stress. The cross-talk between Nrf2 and NF-kB activities regulates the inflammatory processes. The activation of NF-kB could activate up-regulation of IL-1β, IL-6, and IL-8. The increase of COX-2 expression could be correlated with IL-1β one. Also, probably in response to the pro-inflammatory cytokines, MMP-2 and MMP-9 were induced and activated. In conclusion, the exposure of MRC-5 cells to SiO2-NPs generated inflammation in a time-dependent manner.

Keywords: inflammation, MRC-5 cells, oxidative stress, silica nanoparticles

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Authors: Juan Alfredo Guevara Carrio, Swamy Toolahalli Thipperudra, Riddhi Naik Dharmeshbhai, Sergio Graniero Echeverrigaray, Jose Vitorio Emiliano, Antonio Helio Castro

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In order to advance the hydrogen economy, several industrial sectors can potentially benefit from the trillions of stimulus spending for post-coronavirus. Blending hydrogen into natural gas pipeline networks has been proposed as a means of delivering it during the early market development phase, using separation and purification technologies downstream to extract the pure H₂ close to the point of end-use. This first step has been mentioned around the world as an opportunity to use existing infrastructures for immediate decarbonisation pathways. Among current technologies used to extract hydrogen from mixtures in pipelines or liquid carriers, membrane separation can achieve the highest selectivity. The most efficient approach for the separation of H₂ from other substances by membranes is offered from the research of 2D layered materials due to their exceptional physical and chemical properties. Graphene-based membranes, with their distribution of pore sizes in nanometers and angstrom range, have shown fundamental and economic advantages over other materials. Their combination with the structure of ceramic and geopolymeric materials enabled the synthesis of nanocomposites and the fabrication of membranes with long-term stability and robustness in a relevant range of physical and chemical conditions. Versatile separation modules have been developed for hydrogen separation, which adaptability allows their integration in industrial prototypes for applications in heavy transport, steel, and cement production, as well as small installations at end-user stations of pipeline networks. The developed membranes and prototypes are a practical contribution to the technological challenge of supply pure H₂ for the mentioned industries as well as hydrogen energy-based fuel cells.

Keywords: graphene nano-composite membranes, hydrogen separation and purification, separation modules, indsutrial prototype

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Authors: Silvia Di Giacomo, Marcello Locatelli, Simone Carradori, Francesco Cacciagrano, Chiara Toniolo, Gabriela Mazzanti, Luisa Mannina, Stefania Cesa, Antonella Di Sotto

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Hyperglycemia represents the main pathogenic factor in the development of diabetes complications and has been found associated with mitochondrial dysfunction and oxidative stress, which in turn increase cell dysfunction. Therefore, counteract oxidative species appears to be a suitable strategy for preventing the hyperglycemia-induce cell damage and support the pharmacotherapy of diabetes and metabolic diseases. Antidiabetic potential of many food sources has been linked to the presence of polyphenolic metabolites, particularly flavonoids such as quercetin and its glycosylated form rutin. In line with this evidence, in the present study, we assayed the potential anti-hyperglycemic activity of an ethyl acetate extract from the peel of Punica granatum L. var. Dente di Cavallo (PGE), a fruit well known to traditional medicine for the beneficial properties of its edible juice. The effect of the extract on the glucidic metabolism has been evaluated by assessing its ability to inhibit α-amylase and α-glucosidase, two digestive enzymes responsible for the hydrolysis of dietary carbohydrates: their inhibition can delay the carbohydrate digestion and reduce glucose absorption, thus representing an important strategy for the management of hyperglycemia. Also, the PGE ability to block the release of advanced glycated end-products (AGEs), whose accumulation is known to be responsible for diabetic vascular complications, was studied. The iron-reducing and chelating activities, which are the primary mechanisms by which AGE inhibitors stop their metal-catalyzed formation, were evaluated as possible antioxidant mechanisms. At last, the phenolic content of PGE was characterized by chromatographic and spectrophotometric methods. Our results displayed the ability of PGE to inhibit α-amylase enzyme with a similar potency to the positive control: the IC₅₀ values were 52.2 (CL 27.7 - 101.2) µg/ml and 35.6 (CL 22.8 - 55.5) µg/ml for acarbose and PGE, respectively. PGE also inhibited the α-glucosidase enzyme with about a 25 higher potency than the positive controls of acarbose and quercetin. Furthermore, the extract exhibited ferrous and ferric ion chelating ability, with a maximum effect of 82.1% and 80.6% at a concentration of 250 µg/ml respectively, and reducing properties, reaching the maximum effect of 80.5% at a concentration of 10 µg/ml. At last, PGE was found able to inhibit the AGE production (maximum inhibition of 82.2% at the concentration of 1000 µg/ml), although with lower potency with respect to the positive control rutin. The phytochemical analysis of PGE displayed the presence of high levels of total polyphenols, tannins, and flavonoids, among which ellagic acid, gallic acid and catechin were identified. Altogether these data highlight the ability of PGE to control the carbohydrate metabolism at different levels, both by inhibiting the metabolic enzymes and by affecting the AGE formation likely by chelating mechanisms. It is also noteworthy that peel from pomegranate, although being a waste of juice production, can be reviewed as a nutraceutical source. In conclusion, present results suggest the possible role of PGE as a remedy for preventing hyperglycemia complications and encourage further in vivo studies.

Keywords: anti-hyperglycemic activity, antioxidant properties, nutraceuticals, polyphenols, pomegranate

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Authors: Suneeta Gireesh Panicker

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Aim: Aminopeptidase P (APP) is an enzyme that has specificity for proline, can specifically cleave Xaa-Proline peptides and is a metallo-aminopeptidase. The bonds nearby to the imino acid proline are tough to cleave by many peptidases, but APP can specifically break peptide bonds engaged with proline. Membrane-bound form and a cytosolic form are the two forms in which this enzyme exists. The exact physiological function of APP remains unclear and hence the present work attempts to determine it. Methods: In the present study, the expression pattern of cytosolic Aminopeptidase P (DAP) was determined in all the embryonic stages and larval stages of wild-type Drosophila by using polyclonal monospecific antibodies. To show the presence of DAP RNA in embryonic and larval stages, RNA in situ hybridization was performed. DAP promoter-LacZ fusion reporter gene vector was used to construct transgenic embryos to study the regulation pattern of DAP. To study the DAP expression profile, a transgenic fly consisting of a DAP promoter with β-gal and GFP reporter genes in front of it was constructed. Results: DAP protein expression was observed in neuroectodermal cells, posterior midgut primordium, proctodeum, ventral neuroblast and primordial stomatogastric nervous system. It was observed in the ventral cord and midgut in stage 12. The completely developed embryos showed the intense occurrence of it in the ventral cord and gut region. The eye-antennal disc, wing disc and leg disc also showed the presence of DAP protein. LacZ expression in transgenic embryos also showed the same pattern. Conclusion: Similar to various known multiple-functional proteins, DAP could be one with different functions at different stages and in different cells. Data presented here designates DAP functions in the early embryonic and imaginal dics differentiation and development, suggesting that it may be required for the metabolism of proteins like neuropeptides and tachykinins.

Keywords: aminopeptidase P, in situ hybridization, transgenic fly, embryonic stages

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Authors: Stephanie Hernández-Carreón, Judith Virginia Ríos-Arana

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Microplastics (MPs) are plastic particles smaller than 5 mm that has been detected in soil, air, organisms, and mostly water around the world. Most studies have focused on MPs detection in marine waters, and less so in freshwater, such is the case of Mexico, where studies about MPs in freshwaters are limited. One of the most important rivers in the country is The Rio Grande/Río Bravo, a natural border between Mexico and the United States. Its waters serve different purposes, such as fishing, habitat to endemic species, electricity generation, agriculture, and drinking water sources, among others. Despite its importance, the river’s waters have not been analyzed to determine the presence of MPs; therefore, the purpose of this research is to determine if the Rio Bravo/Rio Grande is polluted with microplastics. For doing so, three sites (Borderland, Casa de Adobe, and Guadalupe) along the El Paso-Juárez metroplex have been sampled: 30 L of water were filtered through a plankton net (64 µm) in each site and sediments-composed samples were collected. Water samples and sediments were 1) digested with a hydrogen peroxide solution (30%), 2) resuspended in a calcium chloride solution (1.5 g/cm3) to separate MPs, and 3) filtered through a 0.45 µm nitrocellulose membrane. Processed water samples were dyed with Nile Red (1 mg/ml ethanol) and analyzed by fluorescence microscopy. Two water samples have been analyzed until January 2023: Casa de Adobe and Borderland finding a concentration of 5.67 particles/L and 5.93 particles/L, respectively. Three types of particles were observed: fibers, fragments, and films, fibers being the most abundant. These data, as well as the data obtained from the rest of the samples, will be analyzed by an ANOVA (α=0.05). The concentrations and types of particles found in the Río Bravo correspond with other studies on rivers associated with urban environments and agricultural activities in China, where a range of 3.67—10.7 particles/L was reported in the Wei River. Even though we are in the early stages of the study, and three new sites will be sampled and analyzed in 2023 to provide more data about this issue in the river, this presents the first evidence of microplastic pollution in the Rio Grande.

Keywords: microplastics, fresh water, Rio Bravo, fluorescence microscopy

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Authors: Lanlan Xiao, Yang Liu, Shuo Chen, Bingmei Fu

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Most cancer-related deaths are due to metastasis. Metastasis is a complex, multistep processes including the detachment of cancer cells from the primary tumor and the migration to distant targeted organs through blood and/or lymphatic circulations. During hematogenous metastasis, the emigration of tumor cells from the blood stream through the vascular wall into the tissue involves arrest in the microvasculature, adhesion to the endothelial cells forming the microvessel wall and transmigration to the tissue through the endothelial barrier termed as extravasation. The narrow slit between endothelial cells that line the microvessel wall is the principal pathway for tumor cell extravasation to the surrounding tissue. To understand this crucial step for tumor hematogenous metastasis, we used Dissipative Particle Dynamics method to investigate an individual cell passing through a narrow slit numerically. The cell membrane was simulated by a spring-based network model which can separate the internal cytoplasm and surrounding fluid. The effects of the cell elasticity, cell shape and cell surface area increase, and slit size on the cell transmigration through the slit were investigated. Under a fixed driven force, the cell with higher elasticity can be elongated more and pass faster through the slit. When the slit width decreases to 2/3 of the cell diameter, the spherical cell becomes jammed despite reducing its elasticity modulus by 10 times. However, transforming the cell from a spherical to ellipsoidal shape and increasing the cell surface area only by 3% can enable the cell to pass the narrow slit. Therefore the cell shape and surface area increase play a more important role than the cell elasticity in cell passing through the narrow slit. In addition, the simulation results indicate that the cell migration velocity decreases during entry but increases during exit of the slit, which is qualitatively in agreement with the experimental observation.

Keywords: dissipative particle dynamics, deformability, surface area increase, cell migration

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Authors: Abhishek Soti, Aditya Sharma, Akhilendra Bhushan Gupta

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Fouled reverse osmosis membranes are primarily characterized by Scanning Electron Microscopy (SEM) and Energy Dispersive X-ray Spectrometer (EDS) for a detailed investigation of foulants; however, this has severe limitations on several accounts. Apart from inaccuracy in spectral properties and inevitable interferences and interactions between sample and instrument, misidentification of elements due to overlapping peaks is a significant drawback of EDS. This paper discusses this limitation by analyzing fouled polyamide RO membranes derived from community RO plants of Rajasthan treating brackish water via a combination of results obtained from EDS and Raman spectroscopy and cross corroborating with ICP-MS analysis of water samples prepared by dissolving the deposited salts. The anomalous behavior of different morphic forms of CaCO₃ in aqueous suspensions tends to introduce false reporting of the presence of certain heavy metals and rare earth metals in the scales of the fouled RO membranes used for treating brackish groundwater when analyzed using the commonly adopted techniques like SEM-EDS or Raman spectrometry. Peaks of CaCO₃ reflected in EDS spectra of the membrane were found to be misinterpreted as Scandium due to the automatic assignment of elements by the software. Similarly, the morphic forms merged with the dominant peak of CaCO₃ might be reflected as a single peak of Molybdenum in the Raman spectrum. A subsequent ICP-MS analysis of the deposited salts showed that both Sc and Mo were below detectable levels. It is always essential to cross-confirm the results through a destructive analysis method to avoid such interferences. It is further recommended to study different morphic forms of CaCO₃ scales, as they exhibit anomalous properties like reverse solubility with temperature and hence altered precipitation tendencies, for an accurate description of the composition of scales, which is vital for the smooth functioning of RO systems.

Keywords: reverse osmosis, foulant analysis, groundwater, EDS, artifacts

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Authors: Azri Yamina Mounia, Zitouni Dalila, Aziza Majda, Tou Insaf, Sadi Meriem

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To confront the fossil fuel crisis and the consequences of global warning, many efforts were devoted to develop alternative electricity generation and attracted numerous researchers, especially in the microbial fuel cell field, because it allows generating electric energy and degrading multiple organics compounds at the same time. However, one of the main constraints on power generation is the slow rate of oxygen reduction at the cathode electrode. This paper describes the potential of algal biomass (Chlorella vulgaris) as photosynthetic cathodes, eliminating the need for a mechanical air supply and the use of often expensive noble metal cathode catalysts, thus improving the sustainability and cost-effectiveness of the MFC system. During polarizations, MFC power density using algal biomass was 0.4mW/m², whereas the MFC with mechanic aeration showed a value of 0.2mW/m². Chlorella vulgaris was chosen due to its fastest growing. C. vulgaris grown in BG11 medium in sterilized Erlenmeyer flask. C. vulgaris was used as a bio‐cathode. Anaerobic activated sludge from the plant of Beni‐Messous WWTP(Algiers) was used in an anodic compartment. A dual‐chamber reactor MFC was used as a reactor. The reactor has been fabricated in the laboratory using plastic jars. The cylindrical and rectangular jars were used as the anode and cathode chambers, respectively. The volume of anode and cathode chambers was 0.8 and 2L, respectively. The two chambers were connected with a proton exchange membrane (PEM). The plain graphite plates (5 x 2cm) were used as electrodes for both anode and cathode. The cyclic voltammetry analysis of oxygen reduction revealed that the cathode potential was proportional to the amount of oxygen available in the cathode surface electrode. In the case of algal aeration, the peak reduction value of -2.18A/m² was two times higher than in mechanical aeration -1.85A/m². The electricity production reached 70 mA/m² and was stimulated immediately by the oxygen produced by algae up to the value of 20 mg/L.

Keywords: Chlorella vulgaris, cyclic voltammetry, microbial fuel cell, oxygen reduction

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Authors: Ankush M. Dewle, Suditi Bhattacharya, Prachi R. Abhang, Savita Datar, Ajay J. Jog, Rupesh K. Srivastava, Geetanjali Tomar

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Objectives: The aim of this study was to investigate the quality of mesenchymal stem cells (MSCs) obtained from ageing gingival tissues, in order to suggest their potential role in autologous stem cell therapy for old individuals. Methods: MSCs were isolated from gingival tissues of young (18-45 years) and old (above 45 years) donors by enzymatic digestion. MSCs were analysed for cfu-f, surface marker expression by flow-cytometry and multilineage differentiation potential. The angiogenic potential was compared in a chick embryo yolk sac membrane model. The aging and differentiation markers including SA-β-galactosidase and p21 respectively were analysed by staining and flow-cytometry analysis. Additionally, osteogenic markers such as glucocorticoid receptor (GR), vitamin D receptor (VDR) were measured by flow-cytometry and RT-qPCR was performed for quantification of osteogenic gene expression. Alizarin Red S and alkaline phosphatase (ALP) activity were also quantitated. Results: Gingival MSCs (GMSCs) from both the age groups were similar in their morphology and displayed cfu-f. They had similar expression of MSC surface markers and p21, comparable rate of proliferation and differentiated to all the four lineages. GMSCs from young donors had a higher adipogenic differentiation potential as compared to the old GMSCs. Moreover, these cells did not display a significant difference in ALP activity probably due to comparable expression of GR, VDR, and osteogenic genes. Conclusions: Ageing of GMSCs occurs at a much slower rate than stem cells from other sources. Thus we suggest GMSCs as an excellent candidate for autologous stem cell therapy in degenerative diseases of elderly individuals. Clinical Significance: GMSCs could help overcome the setbacks in clinical implementation of autologous stem cell therapy for regenerative medicine in all age group of patient.

Keywords: bone regeneration, cell therapy, senescence, stem cell

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Authors: Lubna Azmi, Ila Shukla, Shyam Sundar Gupta, Padam Kant, Ch. V. Rao

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Gastric ulcer is a major global health threat, and it is the leading cause of stomach cancer death worldwide. Helicobacter pylori bacteriumis the most important etiologic factor for gastric ulcer. This infection is highly pervasive in South Asian developing countries, especially in India, Nepal, Srilanka etc. due to diversification in geographic area. Pathophysiology of gastric mucosal damage associated with non-invasive bacterium has not justified in detail, but it leads to change in histopathology, immunochemistry of the gastric and duodenal reason of host. The mechanism responsible for bacteria tissue tropism and mucosal damage in stomach variance during the disease is not clearly described and understood scientifically in treatment and control of pathogenic organisms. Polyphenols are secondary metabolites of plants and are generally involved in defense against aggression by pathogens. 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxychromen-4-one and 1-hydroxy-5,7-dimethoxy-2-naphthalene-carboxaldehyde are polyphenolic compound obtained from popular Indian medicinal plants ghavpatta (ArgeriaspeciosaLinn.f) andBael (Aeglemarmelos) have long been used in traditional Ayurvedic Indian medicine for various diseases. They have promising effects on ulcer, as detailed investigation has made in our laboratory. Therefore, the aim of present study is to explore membrane –dependent morphogenesis of H. pylori and associated apoptosis-mediated cell death. Based on this we analyzed immune gene expression in stomach of experimental animals with H. pylori, using quantitative reverse transcription polymerase chain reaction(q RT-PCR). This revealed rapid induction of prostaglandin, interferon I (INF-I), interferon II (INF-II) and INF-I associated genes in the infected animal. Ultrastructural changes associated with H. pylori will be taken for advanced studies. This investigation shows that the biomarkers eradicate H. pylori bacterium caused gastric ulcer which is a major risk factor for gastric cancer.

Keywords: gastric ulcer, Helicobacter pylori, immunochemistry, polyphenols

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Authors: Tony Jehi, Sujatha Rajaram, Nader majzoub, Joan Sabate

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Aging significantly increases the incidence of various cardiometabolic diseases, including cardiovascular disease (CVD). To combat CVD and its associated risk factors, it is imperative to adopt a healthy dietary pattern that is rife with beneficial nutrient and non-nutrient compounds. Unsaturated fats, specifically n-3 polyunsaturated fatty acids (n-3 PUFA), have cardio-protective effects; the opposite is true for saturated fatty acids. What role, if any, does the biomarker of fatty acid intake (specific fatty acids in the erythrocyte) play in predicting cardiometabolic risk among the elderly, a population highly susceptible to increased mortality and morbidity from CVD risk factors, remains unclear. This was a secondary analysis of the Walnuts and Healthy Aging Study. Briefly, elderly (n=192, mean age 69 y) participants followed their usual diet and were randomized into two groups to either eat walnuts daily or abstain from eating walnuts for a period of 2 years. The purpose was to identify potential associations between erythrocyte membrane fatty acids and cardiometabolic risk factors (body weight, blood pressure, blood lipids, and fasting glucose). Erythrocyte n-3 PUFA were inversely associated with total cholesterol (ß = -3.83; p= 0.02), triglycerides (ß = -7.66; p= <0.01), and fasting glucose (ß = -0.19; p=0.03). Specifically, erythrocyte ALA (ß= -1.59; P = 0.04) and DPA (ß= -0.62; P=0.04) were inversely associated with diastolic blood pressure and fasting glucose, respectively. N-6 PUFAs were positively associated with systolic blood pressure (ß=1.10; P=0.02). Mono-unsaturated fatty acids were positively associated with TAG (ß = 4.16; P=0.03). Total saturated fatty acids were not associated with any cardiometabolic risk factors. No association was found between any erythrocyte fatty acid and body weight. In conclusion, erythrocyte n-3 PUFA may be used as a biomarker to predict the cardiometabolic risk among healthy elders, providing support for the American Heart Association guidelines for including n-3 PUFA for preventing CVD.

Keywords: cardiometabolic diseases, erythrocyte fatty acids, elderly, n-3 PUFA

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Authors: Luz del Carmen Rubí Félix Peña, Jose Adan Felix-Ortiz, Ely Sara Lopez-Alvarez, Wenceslao Valenzuela-Quiñonez

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On a global scale, Mexico is the sixth largest producer of farmed white shrimp (Penaeus vannamei). The activity suffered significant economic losses due to acute hepatopancreatic necrosis (AHPND) caused by a strain of Vibrio parahaemolyticus. For control, the first option is the application of antibiotics in food, causing changes in the environment and bacterial communities, which has produced greater virulence and resistance of pathogenic bacteria. An alternative treatment is silver nanoparticles (AgNPs) generated by green synthesis, which have shown an antibacterial capacity by destroying the cell membrane or denaturing the cell. However, the doses at which these are effective are still unknown. The aim is to calculate the minimum inhibitory concentration (MIC) using the Gompertz, Richard, and Logistic model of biosynthesized AgNPs against a strain of V. parahaemolyticus. Through the testing of different formulations of AgNPs synthesized from Euphorbia prostrate (Ep) extracts against V. parahaemolyticus causing AHPND in white shrimp. Aqueous and ethanol extracts were obtained, and the concentration of phenols and flavonoids was quantified. In the antibiograms, AgNPs were formulated in ethanol extracts of Ep (20 and 30%). The inhibition halo at well dilution test were 18±1.7 and 17.67±2.1 mm against V. parahaemolyticus. A broth microdilution was performed with the inhibitory agents (aqueous and ethanolic extracts and AgNPs) and 20 μL of the inoculum of V. parahaemolyticus. The MIC for AgNPs was 6.2-9.3 μg/mL and for ethanol extract of 49-73 mg/mL. The Akaike index (AIC) was used to choose the Gompertz model for ethanol extracts of Ep as the best data descriptor (AIC=204.8, 10%; 45.5, 20%, and 204.8, 30%). The Richards model was at AgNPs ethanol extract with AIC=-9.3 (10%), -17.5 (20 and 30%). The MIC calculated for EP extracts with the modified Gompertz model were 20 mg/mL (10% and 20% extract) and 40 mg/mL at 30%, while Richard was winner for AgNPs-synthesized it was 5 μg/mL (10% and 20%) and 8 μg/mL (30%). The solver tool Excel was used for the calculations of the models and inhibition curves against V.parahaemolyticus.

Keywords: green synthesis, euphorbia prostata, phenols, flavonoids, bactericide

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Authors: Saman Khan, Imrana Naseem

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Copper is an essential trace element required for pro-angiogenic co-factors including vascular endothelial growth factor (VEGF). Elevated levels of copper are found in various types of cancer including prostrate, colon, breast, lung and liver for angiogensis and metastasis. Therefore, targeting copper via copper-specific chelators in cancer cells can be developed as effective anticancer treatment strategy. In continuation of our pursuit to design and synthesize copper chelators, herein we opted for a reaction to incorporate di-(2-picolyl) amine in 3-(bromoacetyl) coumarin (parent backbone) for the synthesis of complex 1. We evaluated lipid peroxidation, protein carbonylation, ROS generation, DNA damage and consequent apoptosis by complex 1 in exogenously added Cu(II) in human peripheral lymphocytes (simulate malignancy condition). Results showed that Cu(II)-complex 1 interaction leads to cell proliferation inhibition, apoptosis, ROS generation and DNA damage in human lymphocytes, and these effects were abrogated by cuprous chelator neocuproine and ROS scavengers (thiourea, catalase, SOD). This indicates that complex 1 cytotoxicity is due to redox cycling of copper to generate ROS which leads to pro-oxidant cell death in cancer cells. To further confirm our hypothesis, using the rat model of diethylnitrosamine (DEN) induced hepatocellular carcinoma; we showed that complex 1 mediates DNA breakage and cell death in isolated carcinoma cells. Membrane permeant copper chelator, neocuproine, and ROS scavengers inhibited the complex 1-mediated cellular DNA degradation and apoptosis. In summary, complex 1 anticancer activity is due to its copper chelation capability. These results will provide copper chelation as an effective targeted cancer treatment strategy for selective cytotoxic action against malignant cells without affecting normal cells.

Keywords: cancer treatment, copper chelation, ROS generation, DNA damage, redox cycling, apoptosis

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Authors: Aurora Gitto, Philipp Proksch

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The presence of various microorganisms (bacteria, fungi) in surface water and groundwater represents an important issue for human health worldwide. The matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS) has emerged as a promising and reliable tool for bacteria identification in clinical diagnostic microbiology and environmental strains thanks to an ionization technique that uses a laser energy absorbing matrix to create ions from large molecules with minimal fragmentation. The study aims first to conceptualise and set up library information and create a comprehensive database of MALDI-TOF-MS spectra from environmental water samples. The samples were analysed over a year (2021-2022) using membrane filtration methodology (0.45 μm and 0.22 μm) and then isolated on R2A agar for a period of 5 days and Yeast extract agar growing at 22 °C up to 4 days and 37 °C for 48 hours. The undetected organisms by MALDI-TOF-MS were analysed by PCR and then sequenced. The information obtained by the sequencing was further implemented in the MALDI-TOF-MS library. Among the culturable bacteria, the results show how the incubator temperature affects the growth of some genera instead of others, as demonstrated by Pseudomonas sp., which grows at 22 °C, compared to Bacillus sp., which is abundant at 37 °C. The bacteria community shows a variation in composition also between the media used, as demonstrated with R2A agar which has been defined by a higher presence of organisms not detected compared to YEA. Interesting is the variability of the Genus over one year of sampling and how the seasonality impacts the bacteria community; in fact, in some sampling locations, we observed how the composition changed, moving from winter to spring and summer. In conclusion, the bacteria community in groundwater and river bank filtration represents important information that needs to be added to the library to simplify future water quality analysis but mainly to prevent potential risks to human health.

Keywords: water quality, MALDI-TOF-MS, sequencing, library

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Authors: Matheus Müller, Delma Vidal

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The present paper aims to expose two techniques of dewatering for sludge, analyzing its operations and dewatering processes, aiming at improving the conditions of disposal of residues with high liquid content. It describes the field tests performed on two geotextile systems, a closed geotextile tube and an open geotextile drying bed, both of which are submitted to two filling cycles. The sludge used in the filling cycles for the field trials is from the water treatment plant of the Technological Center of Aeronautics – CTA, in São José dos Campos, Brazil. Data about volume and height abatement due to the dewatering and consolidation were collected per time, until it was observed constancy. With the laboratory analysis of the sludge allied to the data collected in the field, it was possible to perform a critical comparative study between the observed and the scientific literature, in this way, this paper expresses the data obtained and compares them with the bibliography. The tests were carried out on three fronts: field tests, including the filling cycles of the systems with the sludge from CTA, taking measurements of filling time per cycle and maximum filling height per cycle, heights against the abatement by dewatering of the systems over time; tests carried out in the laboratory, including the characterization of the sludge and removal of material samples from the systems to ascertain the solids content within the systems per time and; comparing the data obtained in the field and laboratory tests with the scientific literature. Through the study, it was possible to perceive that the process of densification of the material inside a closed system, such as the geotextile tube, occurs faster than the observed in the drying bed system. This process of accelerated densification can be brought about by the pumping pressure of the sludge in its filling and by the confinement of the residue through the permeable geotextile membrane (allowing water to pass through), accelerating the process of densification and dewatering by its own weight after the filling with sludge.

Keywords: consolidation, dewatering, geotextile drying bed, geotextile tube

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Authors: Madiha El Awamie, Catherine Rees

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Natural preservatives have been used as alternatives to traditional chemical preservatives; however, a limited number have been commercially developed and many remain to be investigated as sources of safer and effective antimicrobials. In this study, we have been investigating the antimicrobial activity of an extract of Glycyrrhiza glabra (liquorice) that was provided as a waste material from the production of liquorice flavourings for the food industry, and to investigate if this retained the expected antimicrobial activity so it could be used as a natural preservative. Antibacterial activity of liquorice extract was screened for evidence of growth inhibition against eight species of Gram-negative and Gram-positive bacteria, including Listeria monocytogenes, Listeria innocua, Staphylococcus aureus, Enterococcus faecalis and Bacillus subtilis. The Gram-negative bacteria tested include Pseudomonas aeruginosa, Escherichia coli and Salmonella typhimurium but none of these were affected by the extract. In contrast, for all of the Gram-positive bacteria tested, growth was inhibited as monitored using optical density. However parallel studies using viable count indicated that the cells were not killed meaning that the extract was bacteriostatic rather than bacteriocidal. The Minimum Inhibitory Concentration [MIC] and Minimum Bactericidal Concentration [MBC] of the extract was also determined and a concentration of 50 µg ml^-1 was found to have a strong bacteriostatic effect on Gram-positive bacteria. Microscopic analysis indicated that there were changes in cell shape suggesting the cell wall was affected. In addition, the use of a reporter strain of Listeria transformed with the bioluminescence genes luxABCDE indicated that cell energy levels were reduced when treated with either 12.5 or 50 µg ml^-1 of the extract, with the reduction in light output being proportional to the concentration of the extract used. Together these results suggest that the extract is inhibiting the growth of Gram-positive bacteria only by damaging the cell wall and/or membrane.

Keywords: antibacterial activity, bioluminescence, Glycyrrhiza glabra, natural preservative

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Authors: Mitiku Tefera

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Background: Meconium-stained amniotic fluid is one of the primary causes of birth asphyxia. Each year, over five million neonatal deaths occur worldwide due to meconium-stained amniotic fluid, with 90% of these deaths due to birth asphyxia. In Ethiopia meconium-stained amniotic fluid is under investigated, specifically in North Shoa Zone Amhara region Ethiopia. Objective: The aim of this study was to assess the magnitude of meconium-stained amniotic fluid and associated factors among women who gave birth in the North Shoa Zone Hospital’s Amhara Region, Ethiopia, in 2022. Methods: An institutional-based, cross-sectional study was conducted among 628 women who gave birth at North Shoa Zone Hospitals, Amhara, Ethiopia. The study was conducted from 08/June-08/August 2022. Two-stage cluster sampling was used to recruit study participants. The data was collected by using a structured interview-administered questionnaire and chart review. The collected data was entered into Epi-Data Version 4.6 and exported to SPSS Version 25. Logistics regression was employed, and a p-value <0.05 was considered significant. Result: The magnitude of meconium-stained amniotic fluid was 30.3%. Women presented with normal hematocrit level 83% less likely develop meconium-stained amniotic fluid. Women had mid-upper arm circumference value was less than 22.9cm(AOR=1.9; 95% CI;1.18-3.20), obstructed labor(AOR=3.6; 95% CI;1.48-8.83), prolonged labor ≥ 15hr (AOR=7.5; 95% CI ;7.68-13.3), the premature rapture of the membrane (AOR=1.7; 95% CI; 3.22-7.40), fetal tachycardia(AOR=6.2; 95% CI; 2.41-16.3) and Bradycardia (AOR=3.1; 95% CI;1.93-5.28) were significant association with meconium stained amniotic fluid. Conclusion: The magnitude of meconium-stained amniotic fluid, which was high. In this study, MUAC value <22.9 cm, obstructed and prolonged labor, PROM, bradycardia, and tachycardia were factors associated with meconium-stained amniotic fluid. A follow-up study and pooled similar articles will be mentioned for better evidence, enhancing intrapartum services and strengthening early detection of meconium-stained amniotic fluid for the health of the mother and baby.

Keywords: women, meconium-staned amniotic fluid, magnitude, Ethiopia

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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

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Authors: Huyen T. Pham, Nhue P. Nguyen, Tien Q. Phi, Phuong T. Dang, Hy G. Le

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Since the marine environmental conditions are extremely different from the other ones, so that marine actinomycetes might produce novel bioactive compounds. Therefore, actinomycete strains were screened from marine water and sediment samples collected from the coastal areas of Northern Vietnam. Ninety-nine actinomycete strains were obtained on starch-casein agar media by dilution technique, only seven strains, named HP112, HP12, HP411, HPN11, HP 11, HPT13 and HPX12, showed significant antibacterial activity against both gram-positive and gram-negative bacteria (Bacillus subtilis ATCC 6633, Staphylococcus epidemidis ATCC 12228, Escherichia coli ATCC 11105). Further studies were carried out with the most active HP411strain against Candida albicans ATCC 10231. This strain could grow rapidly on starch casein agar and other media with high salt containing 7-10% NaCl at 28-30oC. Spore-chain of HP411 showed an elongated and circular shape with 10 to 30 spores/chain. Identification of the strain was carried out by employing the taxonomical studies including the 16S rRNA sequence. Based on phylogenetic and phenotypic evidence it is proposed that HP411 to be belongs to species Streptomyces variabilis. The potent of the crude extract of fermentation broth of HP411that are effective against wide range of pathogens: both gram-positive, gram-negative and fungi. Further studies revealed that the crude extract HP411 could obtain the anticancer activity for cancer cell lines: Hep-G2 (liver cancer cell line); RD (cardiac and skeletal muscle letters cell line); FL (membrane of the uterus cancer cell line). However, the actinomycetes from marine ecosystem will be useful for the discovery of new drugs in the furture.

Keywords: marine actinomycetes, antibacterial, anticancer, Streptomyces variabilis

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Authors: Joana Antunes, Thomas Levée, Barbara Radovani, Anu Suonpää, Paulina Saloranta, Liji Sobhana, Petri Ihalainen

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Lignin is an important component in the exploitation of lignocellulosic biomass. It has been shown that within the next years, the yield of added-value lignin-based chemicals and materials will generate renewable alternatives to oil-based products (e.g. polymeric composites, resins and adhesives) and enhance the economic feasibility of biorefineries. In this paper, a novel technology for lignin valorisation (METNIN™) is presented. METNIN™ is based on the oxidative action of an alkaliphilic enzyme in aqueous alkaline conditions (pH 10-11) at mild temperature (40-50 °C) combined with a cascading membrane operation, yielding a collection of lignin fractions (from oligomeric down to mixture of tri-, di- and monomeric units) with distinct molecular weight distribution, low polydispersity and favourable physicochemical properties. The alkaline process conditions ensure the high processibility of crude lignin in an aqueous environment and the efficiency of the enzyme, yielding better compatibility of lignin towards targeted applications. The application of a selected lignin fraction produced by METNIN™ as a suitable lignopolyol to completely replace a commercial polyol in polyurethane rigid foam formulations is presented as a prototype. Liquid lignopolyols with a high lignin content were prepared by oxypropylation and their full utilization in the polyurethane rigid foam formulation was successfully demonstrated. Moreover, selected technical specifications of different foam demonstrators were determined, including closed cell count, water uptake and compression characteristics. These specifications are within industrial standards for rigid foam applications. The lignin loading in the lignopolyol was a major factor determining the properties of the foam. In addition to polyurethane foam demonstrators, other examples of lignin-based products related to resins and sizing applications will be presented.

Keywords: enzyme, lignin valorisation, polyol, polyurethane foam

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Authors: Negin Bayat, Nahid HasanZadeh

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Effluents of different industries such as metalworking, battery industry, mining, including heavy metal are considered problematic issues for both humans and the environment. These heavy metals include cadmium, copper, zinc, nickel, chromium, cyanide, lead, etc. Different physicochemical and biological methods are used to remove heavy metals, such as sedimentation, coagulation, flotation, chemical precipitation, filtration, membrane processes (reverse osmosis and nanofiltration), ion exchange, biological methods, adsorption with activated carbon, etc. These methods are generally either expensive or ineffective. In recent years, considerable attention has been given to the removal of heavy metal ions from solution by absorption using discarded and low-cost materials. In this study, nickel removal using an adsorption process by eucalyptus powdered leaves and poplar ash was investigated. This is an applied study. The effect of various parameters on metal removal, such as pH, amount of adsorbent, contact time, and stirring speed, was studied using a discontinuous method. This research was conducted in aqueous solutions on the laboratory scale. Then, optimum absorption conditions were obtained. Then, the study was conducted on real wastewater samples. In addition, the nickel concentration in the wastewater before and after the absorption process was measured. In all experiments, the remaining nickel was measured using an atomic absorption spectrometry device at 382 nm wavelength after an appropriate time and filtration. The results showed that increasing both adsorbent and pH parameters increase the metal removal rate. Nickel removal increased at the first 60 minutes. Then, the absorption rate remained constant and reached equilibrium. A desired removal rate with 40 mg in 100 ml adsorbent solution at pH = 9.5 was observed. According to the obtained results, the best absorption rate was observed at 40 mg dose using a combination of eucalyptus leaves and poplar ash in this study, which was equal to 99.76%. Thus, this combined method can be used as an inexpensive and effective absorbent for the removal of nickel from aqueous solutions.

Keywords: absorption, wastewater, nickel, poplar ash, eucalyptus leaf, treatment

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Authors: Axel Mancebo, Ana M. Bada, Angel Casacó, Bárbara González, Avelina León, María E. Arteaga, Consuelo González, Belinda Sánchez, Adriana Carr, Nuris Ledón, Arianna Iglesias

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Despite the many preventive and therapeutic modalities aimed at curing cancer, it remains as a serious world health problem. Promising recent developments suggest that cancer immunotherapy may be the next great hope for cancer treatment. EGFRs are receptor tyrosine kinases and it is considered an important therapeutic target related with tumor progression, and several types of molecular therapies, including monoclonal antibodies, small molecules, and vaccines, have been developed to target the HER family of receptors. On the other hand, gangliosides are membrane glycosphingolipids that contain two variants of sialic acid, the N-acetylated (NeuAc) and the N-glycolylated (NeuGc) variant. The high expression of this antigen-specific molecule has been associated with malignant tumor progression and immunosuppressive mechanisms, so ganglioside could be considered as the target for cancer immunotherapy. We have been working for several years in the safety evaluation of cancer vaccines targeting these two systems, the EGF receptor and ganglioside. We presented in this work results of repeated dose toxicity studies performed in Sprague Dawley rats and Cynomolgus monkeys, including clinical observations, body weight and rectal temperature measuring, clinical pathology analysis, gross necropsy and histological examination in rodent studies, and immunological evaluation. Immunizations were capable of inducing mainly inflammatory effects at the injection site, with findings largely attributable to the adjuvants used and probably enhanced by the immunological properties of the antigens. In general, these vaccines were shown to be well tolerated, and these studies in relevant species allow treating cancer patients with tumors during long periods with relative weight safety margin.

Keywords: cancer vaccines, safety, toxicology, rats, non human primates

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Authors: Ahmed Mohamed El-Shamy, Akram Abbas El-Awady, Mahmoud Taha Eldestawy

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Background: The present study was to evaluate clinically and radiographically the combined effect of split crest technique by ultrasonic bone surgery and platelet concentrates in implant site development. Methods: Forty patients with narrow ridge were participated in this study. Patients were assigned randomly into one of the following four groups according to treatment: Group 1: Patients received split-crest technique by ultrasonic bone surgery with implant placement. Group 2: Patients received split-crest technique by ultrasonic bone surgery with implant placement and PRF. Group 3: Patients received split-crest technique by ultrasonic bone surgery with implant placement and PRP. Group 4: Patients received split-crest technique by ultrasonic bone surgery with implant placement and collagen membrane. Modified plaque index, modified sulcus bleeding index, and implant stability were recorded as a baseline and measured again at 3 and 6 months. CBCT scans were taken immediately after surgery completion and at 9 months to evaluate bone density at the bone-implant interface. Results after 6 months; collagen group showed statistically significantly lower mean modified bleeding index than the other groups. After 3 months, the PRF group showed statistically significantly higher mean implant stability with ostell ISQ units' than the other groups. After 6 months, the PRF group showed statistically significantly higher mean implant stability with ostell ISQ units' than the other groups. After 6 months, the PRF group showed statistically significantly higher mean bone density than the collagen group. Conclusion: Ultrasonic bone surgery in split-crest technique can be a successful option for increasing implant stability values throughout the healing period. The use of a combined technique of ultrasonic bone surgery with PRF and simultaneous implant placement potentially improves osseointegration (bone density). PRF membranes represent advanced technology for the stimulation and acceleration of bone regeneration.

Keywords: dental implants, split-crest, PRF, PRP

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Authors: P. Saha, R. Ganguly, N. K .Singha, A. Pich

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Microgel, a smart class of material, has drawn attention in the past few years due to its response to external stimuli like temperature, pH, and ionic strength of the solution. Among them, one type of polymer becomes soluble, and the other becomes insoluble in water upon heating displaying upper critical solution temperature (UCST) (e.g., polysulfobetaine, PSB) and lower critical solution temperature (LCST) (e.g., poly(N-vinylcaprolactam, PVCL)) respectively. Polyzwitterions, electrically neutral polymers are biocompatible, biodegradable, and non-cytotoxic in nature, and presence of zwitterionic pendant group in the main backbone makes them stable against temperature and pH variations and strong hydration capability in salt solution promotes them to be used as interfacial bio-adhesion resistance material. Majority of zwitterionic microgels have been synthesized in mini- emulsion technique using free radical polymerization approach. Here, a new route to synthesize dual thermo-responsive PVCL microgels decorated with appreciable amount of zwitterionic PSB chains was developed by a purely water-based surfactant-free reversible addition–fragmentation chain transfer (RAFT) precipitation polymerization. PSB macro-RAFTs having different molecular weights were synthesized and utilized for surface-grafting with PVCL microgels varying the macro-RAFT concentration using N,N′-methylenebis(acrylamide) (BIS) as cross-linker. Increasing the PSB concentration in the PVCL microgels resulted in a linear increase in UCST but decrease in hydrodynamic radius due to strong intrachain coulombic attraction forces acting between the opposite charges present in the zwitterionic groups. Anti- fouling effect was observed on addition of BSA protein solution on the microgel-coated membrane surfaces as studied by fluorescence spectrophotoscopy.

Keywords: microgels, polyzwitterions, upper critical solution temperature-lower critical solution temperature, UCST-LCST, ionic crosslinking

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Authors: Ntokozo Dambuza, Maryna Van De Venter, Trevor Koekemoer

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Oxidative stress contributes to the pathogenesis of many diseases and consequently antioxidant therapy has attracted much attention as a potential therapeutic strategy. Regardless of the quantities ingested, antioxidants need to reach the diseased tissues at concentrations sufficient to combat oxidative stress. Bioavailability is thus a defining criterion for the therapeutic efficacy of antioxidants. In addition, therapeutic antioxidants must possess biologically relevant characteristics which can target the specific molecular mechanisms responsible for disease related oxidative stress. While many chemical antioxidant assays are available to quantify antioxidant capacity, they relate poorly to the biological environment and provide no information as to the bioavailability. The present comparative study thus aims to characterise green and fermented rooibos extracts, well recognized for their exceptional antioxidant capacity, in terms of antioxidant bioavailability and efficacy in a disease relevant cellular setting. Chinese green tea antioxidant activity was also evaluated. Chemical antioxidant assays (FRAP, DPPH and ORAC) confirmed the potent antioxidant capacity of both green and fermented rooibos, with green rooibos possessing antioxidant activity superior to that of fermented rooibos and Chinese green tea. Bioavailability was assessed using the PAMPA assay and the results indicate that green and fermented rooibos have a permeation coefficient of 5.7 x 10-6 and 6.9 x 10-6 cm/s, respectively. Chinese green tea permeability coefficient was 8.5 x 10-6 cm/s. These values were comparable to those of rifampicin, which is known to have a high permeability across intestinal epithelium with a permeability coefficient of 5 x 10 -6 cm/s. To assess the antioxidant efficacy in a cellular context, U937 and red blood cells were pre-treated with rooibos and Chinese green tea extracts in the presence of a dye DCFH-DA and then exposed to oxidative stress. Green rooibos exhibited highest activity with an IC50 value of 29 μg/ml and 70 μg/ml, when U937 and red blood cells were exposed oxidative stress, respectively. Fermented rooibos and Chinese green tea had IC50 values of 61 μg/ml and 57 μg/ml for U937, respectively, and 221 μg/ml and 405 μg/ml for red blood cells, respectively. These results indicate that fermented and green rooibos extracts were able to permeate the U937 cells and red blood cell membrane and inhibited oxidation of DCFH-DA to a fluorescent DCF within the cells.

Keywords: rooibos, antioxidants, permeability, bioavailability

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Authors: Adriano Losso, Birgit Dämon, Stefan Mayr

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In vascular plants, water flows from roots to leaves in a metastable state, and even a small perturbation of the system can lead a sudden transition from the liquid to the vapor phase, resulting in xylem embolism (cavitation). Xylem embolism, induced by drought stress and/or freezing stress is caused by the aspiration of gaseous bubbles into xylem conduits from adjacent gas-filled compartments through pit membrane pores (‘air seeding’). At water potentials less negative than the threshold for air seeding, the surface tension (γ) stabilizes the air-water interface and thus prevents air from passing the pit pores. This hold is probably also true for conifers, where this effect occurs at the edge of the sealed torus. Accordingly, it was experimentally demonstrated that γ influences air seeding, but information on the relevance of this effect under field conditions is missing. In this study, we analyzed seasonal changes in γ of the xylem sap in two conifers growing at the alpine timberline (Picea abies and Pinus mugo). In addition, cut branches were perfused (40 min perfusion at 0.004 MPa) with different γ solutions (i.e. distilled and degassed water, 2, 5 and 15% (v/v) ethanol-water solution corresponding to a γ of 74, 65, 55 and 45 mN m-1, respectively) and their vulnerability to drought-induced embolism analyzed via the centrifuge technique (Cavitron). In both species, xylem sap γ changed considerably (ca. 53-67 and ca. 50-68 mN m-1 in P. abies and P. cembra, respectively) over the season. Branches perfused with low γ solutions showed reduced resistance against drought-induced embolism in both species. A significant linear relationship (P < 0.001) between P12, P50 and P88 (i.e. water potential at 12, 50 and 88% of the loss of conductivity) and xylem sap γ was found. Based on this correlation, a variation in P50 between -3.10 and -3.83 MPa (P. abies) and between -3.21 and -4.11 MPa (P. mugo) over the season could be estimated. Results demonstrate that changes in γ of the xylem sap can considerably influence a tree´s resistance to drought-induced embolism. They indicate that vulnerability analyses, normally conducted at a γ near that of pure water, might often underestimate vulnerabilities under field conditions. For studied timberline conifers, seasonal changes in γ might be especially relevant in winter, when frost drought and freezing stress can lead to an excessive embolism.

Keywords: conifers, Picea abies, Pinus mugo, timberline

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