Search results for: cellular uptake

Authors: D. H. Tejavathi

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Medicinal plants are globally valuable sources of herbal products. Wild populations of many medicinal plants are facing threat of extinction because of their narrow distribution, endemicity, and degradation of specific habitats. Micropropagation is an established in vitro technique by which large number of clones can be obtained from a small bit of explants in a short span of time within a limited space. Mycorrhization can minimize the transient transplantation shock, experienced by the micropropagated plants when they are transferred from lab to land. AM fungal association improves the physiological status of the host plants through better uptake of water and nutrients, particularly phosphorus. Consequently, the growth performance and biosynthesis of active principles are significantly enhanced in AM fungal treated plants. Bacopa monnieri, Andrographis paniculata, Agave vera-curz, Drymaria cordata and Majorana hortensis, important medicinal plants used in various indigenous systems of medicines, are selected for the present study. They form the main constituents of many herbal formulations. Standard in vitro techniques were followed to obtain the micropropagated plants. Shoot tips and nodal segments were used as explants. Explants were cultured on Murashige and Skoog, and Phillips and Collins media supplemented with various combinations of growth regulators. Multiple shoots were obtained on a media containing both auxins and cytokinins at various concentrations and combinations. Multiple shoots were then transferred to rooting media containing auxins for root induction. Thus, obtained in vitro regenerated plants were subjected to brief acclimatization before transferring them to land. One-month-old in vitro plants were treated with AM fungi, and the symbiotic effect on the overall growth parameters was analyzed. It was found that micropropagation coupled with mycorrhization has significant effect on the enhancement of biomass and biosynthesis of active principles in these selected medicinal plants. In vitro techniques coupled with mycorrhization have opened a possibility of obtaining better clones in respect of enhancement of biomass and biosynthesis of active principles. Beneficial effects of AM fungal association with medicinal plants are discussed.

Keywords: cultivation, medicinal plants, micropropagation, mycorrhization

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Authors: Dileep Maarisetty, Janaki Komandur, Saroj S. Baral

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In the current work, photocatalytic degradation of phenol was carried both in UV and visible light to find the slowest step that is limiting the rate of photo-degradation process. Characterization such as XRD, SEM, FT-IR, TEM, XPS, UV-DRS, PL, BET, UPS, ESR and zeta potential experiments were conducted to assess the credibility of catalysts in boosting the photocatalytic activity. To explore the synergy, TiO₂ was doped with graphene and alumina. The orbital hybridization with alumina doping (mediated by graphene) resulted in higher electron transfer from the conduction band of TiO₂ to alumina surface where oxygen reduction reactions (ORR) occur. Besides, the doping of alumina and graphene introduced defects into Ti lattice and helped in improving the adsorptive properties of modified photo-catalyst. Results showed that these defects promoted the oxygen reduction reactions (ORR) on the catalyst’s surface. ORR activity aims at producing reactive oxygen species (ROS). These ROS species oxidizes the phenol molecules which is adsorbed on the surface of photo-catalysts, thereby driving the photocatalytic reactions. Since mass transfer is considered as rate limiting step, various mathematical models were applied to the experimental data to probe the best fit. By varying the parameters, it was found that intra-particle diffusion was the slowest step in the degradation process. Lagergren model gave the best R² values indicating the nature of rate kinetics. Similarly, different adsorption isotherms were employed and realized that Langmuir isotherm suits the best with tremendous increase in uptake capacity (mg/g) of TiO₂-rGO-Al₂O₃ as compared undoped TiO₂. This further assisted in higher adsorption of phenol molecules. The results obtained from experimental, kinetic modelling and adsorption isotherms; it is concluded that apart from changes in surface, optoelectronic and morphological properties that enhanced the photocatalytic activity, the intra-particle diffusion within the catalyst’s pores serve as rate-limiting step in deciding the fate of photo-catalytic degradation of phenol.

Keywords: ORR, phenol degradation, photo-catalyst, rate kinetics

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Authors: Igor Vishnevskyi

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Introduction: The urinary bladder undergoes repeated strain during its working cycle, suggesting the presence of an efficient support system, force transmission, and mechanical amplification. The concept of a "hydrostatic skeleton" (HS) could contribute to our understanding of the functional relationships among bladder constituents. Methods: A multidisciplinary literature review was conducted to identify key features of the HS and to gather evidence supporting its applicability in urinary bladder biomechanics. The collected evidence was synthesized to propose a framework for understanding the potential hydrostatic properties of the urinary bladder based on existing knowledge and HS principles. Results: Our analysis revealed similarities in biomechanical features between living fluid-filled structures and the urinary bladder. These similarities include the geodesic arrangement of fibres, the role of enclosed fluid (urine) in force transmission, prestress as a determinant of stiffness, and the ability to maintain shape integrity during various activities. From a biomechanical perspective, urine may be considered an essential component of the bladder. The hydrostatic skeleton, with its autonomy and flexibility, may provide insights for researchers involved in bladder engineering. Discussion: The concept of a hydrostatic skeleton offers a holistic perspective for understanding bladder function by considering multiple mechanical factors as a single structure with emergent properties. Incorporating viewpoints from various fields on HS can help identify how this concept applies to live fluid-filled structures or organs and reveal its broader relevance to biological systems, both natural and artificial. Conclusion: The hydrostatic skeleton (HS) design principle can be applied to the urinary bladder. Understanding the bladder as a structure with HS can be instrumental in biomechanical modelling and engineering. Further research is required to fully elucidate the cellular and molecular mechanisms underlying HS in the bladder.

Keywords: hydrostatic skeleton, urinary bladder morphology, shape integrity, prestress, biomechanical modelling

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Authors: Sarra K. Ikhlef, Onita Basu

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Natural organic matter (NOM) removal efficiency using drinking water treatment biological filters can be highly influenced by backwashing conditions. Backwashing has the ability to remove the accumulated biomass and particles in order to regenerate the biological filters' removal capacity and prevent excessive headloss buildup. A lab scale system consisting of 3 biological filters was used in this study to examine the implications of different backwash strategies on biological filtration performance. The backwash procedures were evaluated based on their impacts on dissolved organic carbon (DOC) removals, biological filters’ biomass, backwash water volume usage, and particle removal. Results showed that under nutrient limited conditions, the simultaneous use of air and water under collapse pulsing conditions lead to a DOC removal of 22% which was significantly higher (p>0.05) than the 12% removal observed under water only backwash conditions. Employing a bed expansion of 20% under nutrient supplemented conditions compared to a 30% reference bed expansion while using the same amount of water volume lead to similar DOC removals. On the other hand, utilizing a higher bed expansion (40%) lead to significantly lower DOC removals (23%). Also, a backwash strategy that reduced the backwash water volume usage by about 20% resulted in similar DOC removals observed with the reference backwash. The backwash procedures investigated in this study showed no consistent impact on biological filters' biomass concentrations as measured by the phospholipids and the adenosine tri-phosphate (ATP) methods. Moreover, none of these two analyses showed a direct correlation with DOC removal. On the other hand, dissolved oxygen (DO) uptake showed a direct correlation with DOC removals. The addition of the extended terminal subfluidization wash (ETSW) demonstrated no apparent impact on DOC removals. ETSW also successfully eliminated the filter ripening sequence (FRS). As a result, the additional water usage resulting from implementing ETSW was compensated by water savings after restart. Results from this study provide insight to researchers and water treatment utilities on how to better optimize the backwashing procedure for the goal of optimizing the overall biological filtration process.

Keywords: biological filtration, backwashing, collapse pulsing, ETSW

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Authors: Polina Prokopovich

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Despite the well-established dependence of cartilage mechanical properties on the frequency of the applied load, most research in the field is carried out in either load-free or constant load conditions because of the complexity of the equipment required for the determination of time-dependent properties. These simpler analyses provide a limited representation of cartilage properties thus greatly reducing the impact of the information gathered hindering the understanding of the mechanisms involved in this tissue replacement, development and pathology. More complex techniques could represent better investigative methods, but their uptake in cartilage research is limited by the highly specialised training required and cost of the equipment. There is, therefore, a clear need for alternative experimental approaches to cartilage testing to be deployed in research and clinical settings using more user-friendly and financial accessible devices. Frequency dependent material properties can be determined through rheometry that is an easy to use requiring a relatively inexpensive device; we present how a commercial rheometer can be adapted to determine the viscoelastic properties of articular cartilage. Frequency-sweep tests were run at various applied normal loads on immature, mature and trypsinased (as model of osteoarthritis) cartilage samples to determine the dynamic shear moduli (G*, G′ G″) of the tissues. Moduli increased with increasing frequency and applied load; mature cartilage had generally the highest moduli and GAG depleted samples the lowest. Hydraulic permeability (KH) was estimated from the rheological data and decreased with applied load; GAG depleted cartilage exhibited higher hydraulic permeability than either immature or mature tissues. The rheometer-based methodology developed was validated by the close comparison of the rheometer-obtained cartilage characteristics (G*, G′, G″, KH) with results obtained with more complex testing techniques available in literature. Rheometry is relatively simpler and does not require highly capital intensive machinery and staff training is more accessible; thus the use of a rheometer would represent a cost-effective approach for the determination of frequency-dependent properties of cartilage for more comprehensive and impactful results for both healthcare professional and R&D.

Keywords: tissue, rheometer, biomaterial, cartilage

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Authors: Rita Emília Szabó, Róbert Polanek, Tünde Tőkés, Zoltán Szabó, Szabolcs Czifrus, Katalin Hideghéty

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Purpose: Several feature of zebrafish are making them amenable for investigation on therapeutic approaches such as ionizing radiation. The establishment of zebrafish model for comprehensive radiobiological research stands in the focus of our investigation, comparing the radiation effect curves of neutron and photon irradiation. Our final aim is to develop an appropriate vertebrate model in order to investigate the relative biological effectiveness of laser driven ionizing radiation. Methods and Materials: After careful dosimetry series of viable zebrafish embryos were exposed to a single fraction whole-body neutron-irradiation (1,25; 1,875; 2; 2,5 Gy) at the research reactor of the Technical University of Budapest and to conventional 6 MeV photon beam at 24 hour post-fertilization (hpf). The survival and morphologic abnormalities (pericardial edema, spine curvature) of each embryo were assessed for each experiment at 24-hour intervals from the point of fertilization up to 168 hpf (defining the dose lethal for 50% (LD50)). Results: In the zebrafish embryo model LD50 at 20 Gy dose level was defined and the same lethality were found at 2 Gy dose from the reactor neutron beam resulting RBE of 10. Dose-dependent organ perturbations were detected on macroscopic (shortening of the body length, spine curvature, microcephaly, micro-ophthalmia, micrognathia, pericardial edema, and inhibition of yolk sac resorption) and microscopic (marked cellular changes in skin, cardiac, gastrointestinal system) with the same magnitude of dose difference. Conclusion: In our observations, we found that zebrafish embryo model can be used for investigating the effects of different type of ionizing radiation and this system proved to be highly efficient vertebrate model for preclinical examinations.

Keywords: ionizing radiation, LD50, relative biological effectiveness, zebrafish embryo

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Authors: Rahma Goussi, Walid Derbali, Arafet Manaa, Simone Cantamessa, Graziella Berta, Chedly Abdelly, Roberto Barbato

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Salinity is one of the most important abiotic affecting plant growth and productivity worldwide. Photosynthesis, together with cell growth, is among the primary processes to be affected by salinity. Here, we report the effects of salinity stress on the primary processes of photosynthesis in a model halophyte Thellungiella Salsuginea. Plants were cultivated in hydroponic system with different NaCl concentrations (0, 100, 200 and 400 mM) during 2 weeks. The obtained results showed an obvious change in the photosynthetic efficiency of photosystem I (PSI) and phostosytem II (PSII), related to NaCl concentration supplemented to the medium and the stress duration considered. With moderate salinity (100 and 200 mM NaCl), no significant variation was observed in photosynthetic parameters of PSI and PSII and Chl fluorescence whatever the time of stress application. Also, the photosynthesis apparatus Fo, Fm and Fv fluorescence, as well as Fv/Fm were not affected by salt stress. While a significant decrease was observed on quantum yields Y(I), Y(II) and electron transport rate ETR(I), ETR(II) under high salt treatment (400 mM NaCl) with prolonged period (15 days). This reduction is quantitatively compensated by a corresponding increase of energy dissipation Y(NPQ) and a progressive decrease in Fv/Fm under salt treatment. The intensity of the OJIP fluorescence transient decreased with increase in NaCl concentration, with a major effect observed during prolonged period of salt stress. Ultrastructural analysis with Light Microscopy and Transmission Electron Microscopy of T. salsuginea chloroplasts showed some cellular changes, such as the shape of the mesophyll cells and number of chloroplast/cell only under higher NaCl concentration. Salt-stress caused the swelling of thylakoids in T. Salsuginea mesophyll with more accumulation of starch as compared to control plant.

Keywords: fluorescence, halophyte, photosynthesis, salt stress

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Authors: Sebastien Relave, Christophe Desrayaud, Aurelien Vilani, Alexey Sova

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Laser beam melting (LBM) is an additive manufacturing process that enables complex 3D parts to be designed. This process is now commonly employed for various applications such as chemistry or energy, requiring the use of stainless steel grades. LBM can offer comparable and sometimes superior mechanical properties to those of wrought materials. However, we observed an anisotropic microstructure which results from the process, caused by the very high thermal gradients along the building axis. This microstructure can be harmful depending on the application. For this reason, control and prediction of the microstructure are important to ensure the improvement and reproducibility of the mechanical properties. This study is focused on the 316L SS grade and aims at understanding the solidification and transformation mechanisms during process. Experiments to analyse the nucleation and growth of the microstructure obtained by the LBM process according to several conditions. These samples have been designed on different type of support bulk and lattice. Samples are produced on ProX DMP 200 LBM device. For the two conditions the analysis of microstructures, thanks to SEM and EBSD, revealed a single phase Austenite with preferential crystallite growth along the (100) plane. The microstructure was presented a hierarchical structure consisting columnar grains sizes in the range of 20-100 µm and sub grains structure of size 0.5 μm. These sub-grains were found in different shapes (columnar and cellular). This difference can be explained by a variation of the thermal gradient and cooling rate or element segregation while no sign of element segregation was found at the sub-grain boundaries. A high dislocation concentration was observed at sub-grain boundaries. These sub-grains are separated by very low misorientation walls ( < 2°) this causes a lattice of curvature inside large grain. A discussion is proposed on the occurrence of these microstructures formation, in regard of the LBM process conditions.

Keywords: selective laser melting, stainless steel, microstructure

Procedia PDF Downloads 157

Authors: Wajiha Javed, Arsalan Jabbar, Nelofer Mehboob, Muhammad Tafseer, Zahid Memon

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Introduction: Researchers globally have strived to explore diverse factors that augment the continuation and uptake of family planning methods. Clients’ satisfaction is one of the core determinants facilitating continuation of family planning methods. There is a major debate yet scanty evidence to contrast public and private sectors with respect to client satisfaction. The objective of this study is to compare quality-of-care provided by public and private sectors of Pakistan through a client satisfaction lens. Methods: We used Pakistan Demographic Heath Survey 2012-13 dataset (Sindh province) on a total of 3133 Married Women of Reproductive Age (MWRA) aged 15-49 years. Source of family planning (public/private sector) was the main exposure variable. Outcome variable was client satisfaction judged by ten different dimensions of client satisfaction. Means and standard deviations were calculated for continuous variable while for categorical variable frequencies and percentages were computed. For univariate analysis, Chi-square/Fisher Exact test was used to find an association between clients’ satisfaction in public and private sectors. Ten different multivariate models were made. Variables were checked for multi-collinearity, confounding, and interaction, and then advanced logistic regression was used to explore the relationship between client satisfaction and dependent outcome after adjusting for all known confounding factors and results are presented as OR and AOR (95% CI). Results: Multivariate analyses showed that clients were less satisfied in contraceptive provision from private sector as compared to public sector (AOR 0.92,95% CI 0.63-1.68) even though the result was not statistically significant. Clients were more satisfied from private sector as compared to the public sector with respect to other determinants of quality-of-care (follow-up care (AOR 3.29, 95% CI 1.95-5.55), infection prevention (AOR 2.41, 95% CI 1.60-3.62), counseling services (AOR 2.01, 95% CI 1.27-3.18, timely treatment (AOR 3.37, 95% CI 2.20-5.15), attitude of staff (AOR 2.23, 95% CI 1.50-3.33), punctuality of staff (AOR 2.28, 95% CI 1.92-4.13), timely referring (AOR 2.34, 95% CI 1.63-3.35), staff cooperation (AOR 1.75, 95% CI 1.22-2.51) and complications handling (AOR 2.27, 95% CI 1.56-3.29).

Keywords: client satisfaction, family planning, public private partnership, quality of care

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Authors: Mayada Mazher, Ahmed Moustafa, Ahmed Abdellatif

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Background: Autophagy is a eukaryotic, highly conserved catabolic process implicated in many pathophysiologies such as wound healing. Autophagy-associated genes serve as a scaffolding platform for signal transduction of macrophage polarization during the inflammatory phase of wound healing and tissue repair process. In the current study, we report a model for the interplay between autophagy-associated genes and macrophages polarization associated genes. Methods: In silico analysis was performed on 249 autophagy-related genes retrieved from the public autophagy database and gene expression data retrieved from Gene Expression Omnibus (GEO); GSE81922 and GSE69607 microarray data macrophages polarization 199 DEGS. An integrated protein-protein interaction network was constructed for autophagy and macrophage gene sets. The gene sets were then used for GO terms pathway enrichment analysis. Common transcription factors for autophagy and macrophages' polarization were identified. Finally, microRNAs enriched in both autophagy and macrophages were predicated. Results: In silico prediction of common transcription factors in DEGs macrophages and autophagy gene sets revealed a new role for the transcription factors, HOMEZ, GABPA, ELK1 and REL, that commonly regulate macrophages associated genes: IL6,IL1M, IL1B, NOS1, SOC3 and autophagy-related genes: Atg12, Rictor, Rb1cc1, Gaparab1, Atg16l1. Conclusions: Autophagy and macrophages' polarization are interdependent cellular processes, and both autophagy-related proteins and macrophages' polarization related proteins coordinate in tissue remodelling via transcription factors and microRNAs regulatory network. The current work highlights a potential new role for transcription factors HOMEZ, GABPA, ELK1 and REL in wound healing.

Keywords: autophagy related proteins, integrated network analysis, macrophages polarization M1 and M2, tissue remodelling

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Authors: Mohd Farooq Naqshbandi

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The four fractions of aqueous extract of Sesame Seeds (Sesamum indicum L.) were studied for invitro DNA fragmentation, cell migration, and cellular apoptosis on SW480 and HTC116 human colorectal cancer cell lines. The seeds of Sesamum indicum were extracted with six solvents, including Methanol, Ethanol, Aqueous, Chloroform, Acetonitrile, and Hexane. The aqueous extract (IC₅₀ value 154 µg/ml) was found to be the most active in terms of cytotoxicity with SW480 human colorectal cancer cell lines. Further fractionation of this aqueous extract on flash chromatography gave four fractions. These four fractions were studied for anticancer and DNA binding studies. Cell viability was assessed by colorimetric assay (MTT). IC₅₀ values for all these four fractions ranged from 137 to 548 µg/mL for the HTC116 cancer cell line and 141 to 402 µg/mL for the SW480 cancer cell line. The four fractions showed good anticancer and DNA binding properties. The DNA binding constants ranged from 10.4 ×10⁴ 5 to 28.7 ×10⁴, showing good interactions with DNA. The DNA binding interactions were due to intercalative and π-π electron forces. The results indicate that aqueous extract fractions of sesame showed inhibition of cell migration of SW480 and HTC116 human colorectal cancer cell lines and induced DNA fragmentation and apoptosis. This was demonstrated by calculating the low wound closure percentage in cells treated with these fractions as compared to the control (80%). Morphological features of nuclei of cells treated with fractions revealed chromatin compression, nuclear shrinkage, and apoptotic body formation, which indicate cell death by apoptosis. The flow cytometer of fraction-treated cells of SW480 and HTC116 human colorectal cancer cell lines revealed death due to apoptosis. The results of the study indicate that aqueous extract of sesame seeds may be used to treat colorectal cancer.

Keywords: Sesamum indicum, cell migration inhibition, apoptosis induction, anticancer activity, colorectal cancer

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Authors: Sudha Summarwar, Sudesh Agarwal, Deepali Lall, Nalini Kataria, Jyotsana Pandey

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Assessment of antioxidant status is an effective tool to appraise the presence of oxidative stress. A combination of assays can be used to evaluate the antioxidant status like serum catalase (CAT), superoxide dismutase (SOD) and monoamine oxidase (MAO). In human medicine pregnancy is known to be associated with oxidative stress. Oxidative stress produces harmful effects to the developing foetus. Several metabolic changes occur in the maternal body to meet the demand of energy of developing foetus. Due to these changes susceptibility of maternal body increases to oxidative stress. There is paucity of research work on this aspect in nondescript goats. Therefore, the present study was intended to appraise the oxidative stress in pregnant and non-pregnant non-descript goat. Blood samples were collected for serum separation in otherwise healthy pregnant and non-pregnant nondescript goats. Mean values of serum CAT, SOD and MAO were found on a higher side (p≤0.05) with serum SOD values showing a rise of 2.5 times higher than the control healthy value. Correlations among all the three parameters were found to be highly significant (p≤0.01) especially greatest in youngest group of pregnant animals. Illustration of result enlightened the veracity of bumped up production of free radicals in pregnant animals. Technical savoir-faire of oxidative stress supervision is essential for upholding of health status of foetus. The upshot of present study undoubtedly implied the development of oxidative stress in pregnant goats on the basis of altered antioxidant status. These findings conclude that initially the oxidative stress due to pregnancy is critically combated by the intricate defensive mechanism of natural antioxidant system of the body. It appears that this imbalance between oxidant and antioxidant must be checked in time to prevent cellular damage by regularly appraising the antioxidant status through laboratory methods.

Keywords: antioxidant, oxidative stress, pregnancy, serum catalase

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Authors: Živilė Jurgelėnė, Vitalijus Karabanovas, Augustas Morkvėnas, Reda Dzingelevičienė, Nerijus Dzingelevičius, Saulius Raugelė, Boguslaw Buszewski

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The short- and long-term effects of COVID-19 antiviral drug molnupiravir and micro/nanoplastics on the early development of Salmo trutta were investigated using accumulation and exposure studies. Salmo trutta were used as standardized test organisms in toxicity studies of COVID-19 waste contaminants. The 2D/3D imaging was performed using confocal fluorescence spectral imaging microscopy to assess the uptake, bioaccumulation, and distribution of molnupiravir and micro/nanoplastics complex in live fish. Our study results demonstrated that molnupiravir may interact with a micro/nanoplastics and modify their spectroscopic parameters and toxicity to S. trutta embryos and larvae. The 0.2 µm size microplastics at a concentration of 10 mg/L were found to be stable in aqueous media than 0.02 µm, and 2 µm sizes polymeric particles. This study demonstrated that polymeric particles can adsorb molnupiravir that are present in mixtures and modify the accumulation of molnupiravir in Salmo trutta embryos and larvae. In addition, 2D/3D confocal fluorescence imaging showed that the single polymeric particle hardly accumulates and couldn't penetrate outer tissues of the tested organism. However, co-exposure micro/nanoplastics and molnupiravir could significantly enhance the polymeric particles capability of accumulating on surface tissues and penetrating surface tissue of fish in early development. Exposure to molnupiravir at 2 g/L concentration and co-exposure to micro/nanoplastics and molnupiravir did not bring about survival changes in in the early stages of Salmo trutta development, but we observed the reduction in heart rate and decrease in gill ventilation. The statistical analysis confirmed that micro/nanoplastics used in combination with molnupiravir enhance the toxicity of the latter micro/nanoplastics to embryos and larvae. This research has received funding from the European Regional Development Fund (project No 13.1.1-LMT-K-718-05-0014) under a grant agreement with the Research Council of Lithuania (LMTLT), and it was funded as part of the European Union’s measure in response to the COVID-19 pandemic.

Keywords: fish, micro/nanoplastics, molnupiravir, toxicity

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Authors: Chien Y. Lin, Jung Y. Huang, Leu-Wei Lo

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A growing body of data reveals that the membrane cholesterol molecules can alter the signaling pathways of living cells. However, the understanding about how membrane cholesterol modulates receptor proteins is still lacking. Single-molecule tracking can effectively probe into the microscopic environments and thermal fluctuations of receptor proteins in a living cell. In this study we applies single-molecule optical tracking on ligand-induced dimerization process of EGFRs in the plasma membranes of two cancer cell lines (HeLa and A431) and one normal endothelial cell line (MCF12A). We tracked individual EGFR and dual receptors, diffusing in a correlated manner in the plasma membranes of live cells. We developed an energetic model by integrating the generalized Langevin equation with the Cahn-Hilliard equation to help extracting important information from single-molecule trajectories. From the study, we discovered that ligand-bound EGFRs move from non-raft areas into lipid raft domains. This ligand-induced motion is a common behavior in both cancer and normal cells. By manipulating the total amount of membrane cholesterol with methyl-β-cyclodextrin and the local concentration of membrane cholesterol with nystatin, we further found that the amount of cholesterol can affect the stability of EGFR dimers. The EGFR dimers in the plasma membrane of normal cells are more sensitive to the local concentration changes of cholesterol than EGFR dimers in the cancer cells. Our method successfully captures dynamic interactions of receptors at the single-molecule level and provides insight into the functional architecture of both the diffusing EGFR molecules and their local cellular environment.

Keywords: membrane proteins, single-molecule tracking, Cahn-Hilliard equation, EGFR dimers

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Authors: Birgul Hizlar, Sibel Karakaya

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Nowadays, consumers are more and more aware of the benefits beyond basic nutrition provided by food and food compounds. Between these, carotenoids have been demonstrated to exhibit multiple health benefits (for example, some types of cancer, cardiovascular diseases, eye disorders, among others). However, carotenoid bioaccessibility and bioavailability is generally rather low due to their specific localization in plant tissue and lipophilic nature. This situation is worldwide issue, since both developed and developing countries have their interest and benefits in increasing the uptake of carotenoids from the human diet. Recently, a new class of foods designed to improve the bioaccessibility/bioavailability of orally administered bioactive compounds is introduced: excipient foods. Excipient foods are specially designed foods which are prepared depending on the physicochemical properties of target bioactive compounds and increasing the bioavailability or bioaccessibility of bioactive compound. In this study, effects of food matrix (greating, boiling and mashing) and different excipient foods (olive oil, lemon juice, whey curd and dried artichoke leaf powder) on bioaccessibility of β-carotene in carrot were investigated by means of simulating in vitro gastrointestinal (GI) digestion. β-carotene contents of grated, boiled and mashed (after boiling process) carrots were 79.28, 147.63 and 151.19 μg/g respectively. No significant differences among boiled and mashed samples indicated that mashing process had no effect on the release of β-carotene from the food matrix (p > 0.05). On the contrary, mashing causes significant increase in the β-carotene bioaccessibility (p < 0.05). The highest β-carotene content was found in the mashed carrots incorporated with olive oil and lemon juice (C2). However, no significant differences between that sample and C1 (mashed carrot with lemon juice, olive oil, dried artichoke leaf powder), C3 (mashed carrot with addition of olive oil, lemon juice, whey curd) and). Similarly, the highest β-carotene bioaccessibility (50.26%) was found mashed C3 sample (p < 0.05). The increase in the bioaccessibility was approximately 5 fold and 50 fold when compared to grated and mashed samples containing olive oil, lemon juice and whey curd. The results demonstrate that both, food matrix and excipient foods, are able to increase the bioaccessibility of β-carotene.

Keywords: bioaccessibility, carotenoids, carrot, β-carotene

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Authors: Sani Ismaila, Shafiu Yau, Abubakar Salisu, Buhari Salisu, Sharifat Balogun, Mustapha Abubakar, Biobaku Khalid, Agaie Bello

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Over the years, Japanese quail egg has been in use in the management of diseases. The objective of this study was to evaluate the analgesic, anti-inflammatory and gastroprotective effects of raw Quail egg (yolk + albumin) in rats. Pain was assessed in rats by recording the latent period and writing reflex, anti-inflammatory effect was determined using both motility and compression test, while the gastro-protective effects were assessed by observing the histology of the stomach after diclofenac-induced gastric ulcers and subsequent treatment with the quail egg, Rats were randomly assigned into 4 groups; Groups I: were the control non-treated (NT), Group II were treated with Tramadol 50 mg/kg/Os (TMD) or Indomethacin (IND) 5mg/kg/Os (positive control for the writhing reflex determination), while group III and IV were treated with 3 and 6g/kg of raw quail egg respectively). Groups treated with quail egg in both doses showed a significant increase in the latent period (p <0 .05) when compared to the control NT, but lower than the group treated with tramadol at 20mins interval (p<0.05). Writing reflexes decrease in groups II, III, and IV compared to the NT group (p < 0.05). While motility increases significantly (p < 0.05) in groups II, compared to I (p<0.05). Control non-treated rats showed a quicker and extensive response to compression using the Vanier calliper on the inflamed paw compared to groups II-IV (p < 0.05). Histological studies of the stomach revealed sloughing of the epithelia, cellular infiltration with micro abscesses in the non-treated, while groups treated concurrently with quail egg showed proliferation of the glandular epithelia and goblet cells, and those treated 30 minutes before diclofenac administration showed proliferation of glands and thickening of the squamous epithelia. This study showed that quail egg has analgesic, anti-inflammatory and gastro-protective potentials and can be used as adjuvant treatment whenever COX-2 enzymes inhibitors are indicated.

Keywords: analgesia, anti-inflammatory, gastroprotective effect, japanese quail egg

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Authors: Sellidj Abdelaziz, Lebaili Soltane

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A cobalt-based alloy type Co-Cr-Ni-WC was deposited by plasma transferred arc projection (PTA) on a stainless steel valve. The alloy is characterized at the equilibrium by a solid solution Co (γ) mainly dendritic, and eutectic carbides M₇C₃ and ηM₆C. At the deposit/substrate interface, this microstructure is modified by the fast cooling mode of the alloy when applied in the liquid state on the relatively cold steel substrate. The structure formed in this case is heterogeneous and metastable phases can occur and evolve over temperature service. Coating properties and reliability are directly related to microstructures formed during deposition. We were interested more particularly in this microstructure formed during the solidification of the deposit in the region of the interface joining the soldered couple and its evolution during cyclic heat treatments at temperatures similar to those of the thermal environment of the valve. The characterization was carried out by SEM-EDS microprobe CAMECA, XRD, and micro hardness profiles. The deposit obtained has a linear and regular appearance that is free of cracks and with little porosity. The morphology of the microstructure represents solidification stages that are relatively fast with a temperature gradient high at the beginning of the interface by forming a plane front solid solution Co (γ). It gradually changes with the decreasing temperature gradient by getting farther from the junction towards the outer limit of the deposit. The matrix takes the forms: cellular, mixed (cells and dendrites) and dendritic. Dendritic growth is done according to primary ramifications in the direction of the heat removal which takes place in the direction perpendicular to the interface, towards the external surface of the deposit, following secondary and tertiary undeveloped arms. The eutectic carbides M₇C₃ and ηM₆C formed are very thin and are located in the intercellular and interdendritic spaces of the solid solution Co (γ).

Keywords: Co-Ni-Cr-W-C alloy, solid deposit, microstructure, carbides, cyclic heat treatment

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Authors: Ikram Elsiddig, Yacouba Djamila, Amna Hamad

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Abstract—The worldwide increasing of using herbs in form of medicine with or without prescription medications potentiates the interactions between herbal products and conventional medicines; due to more research for herb-drug interactions are needed. for a long time hyperglycemia had been treated with several medicinal plants. A. sativum, belonging to the Liliaceae family is well known for its medicinal uses in African traditional medicine, it used for treating of many human diseases mainly diabetes, high cholesterol and high blood pressure. The purpose of this study is to determine the interaction effect between A. sativum bulb extracts and metformin drug used in diabetes treatment. The in vitro and in vivo evaluation were conducted by glucose reuptake using isolated rats hemidiaphgrams tissue and by estimate glucose tolerance in glucose-loaded wistar albino rats. The results showed that, petroleum ether, chloroform and ethyl acetate extracts were found to have activity of glucose uptake in isolated rats hemidiaphgrams of 24.11 mg/g, 19.07 mg/g and 15.66 mg/g compared to metformin drug of 17 mg/g. These activity were reducded to 17.8 mg/g, 13.59 mg/g and 14.46 mg/g after combination with metformin, metformin itself reduced to 13.59 mg/g, 14.46 mg/g and 12.71 mg/g in comination with chloroform and ethyl acetate. These decrease in activity could be due to herbal–drug interaction between the extracts of A. sativum bulb and metformin drug. The interaction between A. sativum extract and metformin was also shown by in vivo study on the induced hyperglycemic rats. The glucose level after administered of 200 mg/kg was found to be increase with 47.2 % and 17.7% at first and second hour compared to the increase of blood glucose in the control group of 82.6% and76.7%.. At fourth hour the glucose level was became less than normal with 3.4% compared to control which continue to increase with 68.2%. Dose of 400 mg/kg at first hour showed increase in blood glucose of 31.5 %, at second and fourth hours the glucose level was became less than normal with decrease of 3.2 % and 30.4%. After combination the activity was found to be less than that of extract at both high and low dose, whereas, at first and second hour, the glucose level was found to be increase with 50.4% and 21.2%, at fourth hour the glucose level was became less than normal with 14%. Therefore A. sativum could be a potential source for anti-diabetic when it used alone, and it is significant important to use the garlic extract alone instead of combined with Metformin drug in diabetes- treatment.

Keywords: Antagonistic, Garlic, Metformin, Synergistic

Procedia PDF Downloads 182

Authors: Simona Capossela, Ramona Schaniel, Singer Franziska, Aquino Fournier Catharine, Daniel Stekhoven, Jivko Stoyanov

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A spinal cord injury (SCI) is a traumatic life event that often results in ageing associated health conditions such as muscle mass decline, adipose tissue increase, decline in immune function, frailty, systemic chronic inflammation, and psychological distress and depression. Psychological, oxidative, and metabolic stressors may facilitate accelerated ageing in the SCI population with reduced life expectancy. Research designs using biomarkers of aging and stress are needed to elucidate the role of psychological distress in accelerated aging. The aim of this project is a feasibility pilot study to observe changes in stress biomarkers and correlate them with aging markers in SCI patients during their first rehabilitation (longitudinal cohort study). Biological samples were collected in the SwiSCI (Swiss Spinal Cord Injury Cohort Study) Biobank in Nottwil at 4 weeks±12 days after the injury (T1) and at the end of the first rehabilitation (discharge, T4). The "distress thermometer" is used as a selfassessment tool for psychological distress. Stress biomarkers, as cortisol and protein carbonyl content (PCC), and markers of cellular aging, such as telomere lengths, will be measured. 2 Preliminary results showed that SCI patients (N= 129) are still generally distressed at end of rehabilitation, however we found a statistically significant (p< 0.001) median decrease in distress from 6 (T1) to 5 (T4) during the rehabilitation. In addition, an explorative transcriptomics will be conducted on N=50 SCI patients to compare groups of persons with SCI who have different trajectories of selfreported distress at the beginning and end of the first rehabilitation after the trauma. We identified 4 groups: very high chronic stress (stress thermometer values above 7 at T1 and T4; n=14); transient stress (high to low; n=14), low stress (values below 5 at T1 and T4; n=14), increasing stress (low to high; n=8). The study will attempt to identify and address issues that may occur in relation to the design and conceptualization of future study on stress and aging in the SCI population.

Keywords: stress, aging, spinal cord injury, biomarkers

Procedia PDF Downloads 106

Authors: Umar Hayatu Sidik

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Application of liquid base petroleum fuel (petrol and diesel) for transportation fuel causes emissions of greenhouse gases (GHGs), while natural gas (NG) reduces the emissions of greenhouse gases (GHGs). At present, compression and liquefaction are the most matured technology used for transportation system. For transportation use, compression requires high pressure (200–300 bar) while liquefaction is impractical. A relatively low pressure of 30-40 bar is achievable by adsorbed natural gas (ANG) to store nearly compressed natural gas (CNG). In this study, adsorbents for high-pressure adsorption of methane (CH4) was prepared from coconut shells and polyetheretherketone (PEEK) using potassium hydroxide (KOH) and microwave-assisted activation. Design expert software version 7.1.6 was used for optimization and prediction of preparation conditions of the adsorbents for CH₄ adsorption. Effects of microwave power, activation time and quantity of PEEK on the adsorbents performance toward CH₄ adsorption was investigated. The adsorbents were characterized by Fourier transform infrared spectroscopy (FTIR), thermogravimetric (TG) and derivative thermogravimetric (DTG) and scanning electron microscopy (SEM). The ideal CH4 adsorption capacities of adsorbents were determined using volumetric method at pressures of 5, 17, and 35 bar at an ambient temperature and 5 oC respectively. Isotherm and kinetics models were used to validate the experimental results. The optimum preparation conditions were found to be 15 wt% amount of PEEK, 3 minutes activation time and 300 W microwave power. The highest CH4 uptake of 9.7045 mmol CH4 adsorbed/g adsorbent was recorded by M33P15 (300 W of microwave power, 3 min activation time and 15 wt% amount of PEEK) among the sorbents at an ambient temperature and 35 bar. The CH4 equilibrium data is well correlated with Sips, Toth, Freundlich and Langmuir. Isotherms revealed that the Sips isotherm has the best fit, while the kinetics studies revealed that the pseudo-second-order kinetic model best describes the adsorption process. In all scenarios studied, a decrease in temperature led to an increase in adsorption of both gases. The adsorbent (M33P15) maintained its stability even after seven adsorption/desorption cycles. The findings revealed the potential of coconut shell-PEEK as CH₄ adsorbents.

Keywords: adsorption, desorption, activated carbon, coconut shells, polyetheretherketone

Procedia PDF Downloads 67

Authors: Muñiz-Lino Marcos Agustín, Vazquez Borbolla Jessica, Licéaga-Escalera Carlos

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The ectomesenchymal tissues involved in tooth development and their remnants are the origin of different odontogenic lesions, including tumors and cysts of the jaws, with a wide range of clinical behaviors. Dentigerous cyst (DC) represents approximately 20% of all cases of odontogenic cysts, and it has been demonstrated that it can develop benign and malignant odontogenic tumors. DC is characterized by bone destruction of the area surrounding the crown of a tooth which has not erupted and it contain is liquid. The treatment of odontogenic tumors and cysts usually are partial or total removal of the jaw, causing important secondary co-morbidities. However, molecules implicated in DC pathogenesis as well in its development to odontogenic tumors remains unknown. A cellular model may be useful to study these molecules, but that model has not been established yet. Here, we reported the establishment of a cell culture derived from a dentigerous cyst. This cell line was named DeCy-1. In spite of its ectomesenchymal morphology, DeCy-1 cells express epithelial markers such as cytokeratins 5, 6, and 8. Furthermore, these cells express the ODAM protein, which is present in odontogenesis and in dental follicle, indicating that DeCy-1 cells derived from odontogenic epithelium. Analysis by electron microscopy of this cell line showed that it has a high vesicular activity, suggesting that DeCy-1 could secrete molecules that may be involved in DC pathogenesis. Thus, secreted proteins were analyzed by PAGE-SDS, where we observed approximately 11 bands. In addition, the capacity of these secretions to degrade proteins was analyzed by gelatin substrate zymography. A degradation band of about 62 kDa was found in these assays. Western blot assays suggested that the matrix metalloproteinase 2 (MMP-2) is responsible of this protease activity. Thus, our results indicate that the establishment of a cell line derived from DC is a useful in vitro model to study the biology of this odontogenic lesion and its participation in the development of odontogenic tumors.

Keywords: dentigerous cyst, MMP20, cancer, cell culture

Procedia PDF Downloads 136

Authors: A. Ghribi, M. Bagane

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The discharge of dye in industrial effluents is of great concern because their presence and accumulation have a toxic or carcinogenic effect on living species. The removals of such compounds at such low levels are a difficult problem. Physicochemical technique such as coagulation, flocculation, ozonation, reverse osmosis and adsorption on activated carbon, manganese oxide, silica gel and clay are among the methods employed. The adsorption process is an effective and attractive proposition for the treatment of dye contaminated wastewater. Activated carbon adsorption in fixed beds is a very common technology in the treatment of water and especially in processes of decolouration. However, it is expensive and the powdered one is difficult to be separated from aquatic system when it becomes exhausted or the effluent reaches the maximum allowable discharge level. The regeneration of exhausted activated carbon by chemical and thermal procedure is also expensive and results in loss of the sorbent. Dye molecules also have very high affinity for clay surfaces and are readily adsorbed when added to clay suspension. The elimination of the organic dye by clay was studied by serval researchers. The focus of this research was to evaluate the adsorption potential of the raw clay in removing congo red from aqueous solutions using a laboratory fixed-bed column. The continuous sorption process was conducted in this study in order to simulate industrial conditions. The effect of process parameters, such as inlet flow rate, adsorbent bed height and initial adsorbate concentration on the shape of breakthrough curves was investigated. A glass column with an internal diameter of 1.5 cm and height of 30 cm was used as a fixed-bed column. The pH of feed solution was set at 7.Experiments were carried out at different bed heights (5-20 cm), influent flow rates (1.6- 8 mL/min) and influent congo red concentrations (10-50 mg/L). The obtained results showed that the adsorption capacity increases with the bed depth and the initial concentration and it decreases at higher flow rate. The column regeneration was possible for four adsorption–desorption cycles. The clay column study states the value of the excellent adsorption capacity for the removal of congo red from aqueous solution. Uptake of congo red through a fixed-bed column was dependent on the bed depth, influent congo red concentration and flow rate.

Keywords: adsorption, breakthrough curve, clay, congo red, fixed bed column, regeneration

Procedia PDF Downloads 334

Authors: M. Varmaziar

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Magnesium (Mg) is an essential mineral that plays a crucial role in the human body. Certain types of foods, including nuts, grains, fruits, vegetables, and whole grains, are rich sources of magnesium. Mg serves as an essential cofactor for numerous enzymatic reactions, including energy metabolism, cellular growth, glycolysis, and protein synthesis. The Mg-ATP complex serves as an energy source and is vital for many physiological functions, including nerve conduction, muscle contraction, and blood pressure regulation. Despite the vital role of magnesium in energy metabolism, maintaining adequate magnesium intake is often overlooked among the general population and athletes. The aim of this study was to investigate the effect of magnesium supplementation on the physical activities of field athletes. Field athletes were divided into two groups: those who consumed magnesium supplements and those who received a placebo. These two groups received either 500 mg of magnesium oxide or a placebo daily for 8 weeks. At the beginning and end of the study, athletes completed ISI questionnaires and physical activity assessments. Nutritional analyses were performed using N4 software, and statistical analyses were conducted using SPSS19 software. The results of this study revealed a significant difference between the two study groups. Athletes who received magnesium supplements experienced less fatigue related to field athletic activities and muscle soreness. In contrast, athletes who received the placebo reported more significant fatigue and muscle soreness. A concerning finding in these results is that the performance of athletic activities may be at risk with low magnesium levels. Therefore, magnesium is essential for maintaining health and plays a crucial role in athletic performance. Consuming a variety of magnesium-rich foods ensures that individuals receive an adequate amount of this essential nutrient in their diet. The consumption of these foods improves performance parameters in athletic exercises.

Keywords: athletic performance, effect, field athletes, magnesium supplement

Procedia PDF Downloads 81

Authors: P. R. Chauhan, S. K. Tyagi

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Adsorption refrigeration technology offers a sustainable and energy-efficient cooling alternative over traditional refrigeration technologies for meeting the fast-growing cooling demands. With its ability to utilize natural refrigerants, low-grade heat sources, and modular configurations, it has the potential to revolutionize the cooling industry. Despite these benefits, the commercial viability of this technology is hampered by several fundamental limiting constraints, including its large size, low uptake capacity, and poor performance as a result of deficient heat and mass transfer characteristics. The primary cause of adequate heat and mass transfer characteristics and magnitude of exergy loss in various real processes of adsorption cooling system can be assessed by the entropy generation rate analysis, i. e. Second law of Thermodynamics. Therefore, this article presents the second law of thermodynamic-based investigation in terms of entropy generation rate (EGR) to identify the energy losses in various processes of the HPCC-based adsorption system using MATLAB R2021b software. The adsorption technology-based cooling system consists of two beds made up of silica gel and arranged in a single stage, while the water is employed as a refrigerant, coolant, and hot fluid. The variation in process-wise EGR is examined corresponding to cycle time, and a comparative analysis is also presented. Moreover, the EGR is also evaluated in the external units, such as the heat source and heat sink unit used for regeneration and heat dump, respectively. The research findings revealed that the combination of adsorber and desorber, which operates across heat reservoirs with a higher temperature gradient, shares more than half of the total amount of EGR. Moreover, the EGR caused by the heat transfer process is determined to be the highest, followed by a heat sink, heat source, and mass transfer, respectively. in case of heat transfer process, the operation of the valve is determined to be responsible for more than half (54.9%) of the overall EGR during the heat transfer. However, the combined contribution of the external units, such as the source (18.03%) and sink (21.55%), to the total EGR, is 35.59%. The analysis and findings of the present research are expected to pinpoint the source of the energy waste in HPCC based adsorption cooling systems.

Keywords: adsorption cooling cycle, heat transfer, mass transfer, entropy generation, silica gel-water

Procedia PDF Downloads 107

Authors: L. Meksem Amara, M. Ferfar, N. Meksem, M. R. Djebar

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The wheat is the cereal the most consumed in the world. In Algeria, the production of this cereal covers only 20 in 25 % of the needs for the country, the rest being imported. To improve the efficiency and the productivity of the durum wheat, the farmers turn to the use of pesticides: weed-killers, fungicides and insecticides. However this use often entrains losses of products more at least important contaminating the environment and all the food chain. Weed-killers are substances developed to control or destroy plants considered unwanted. That they are natural or produced by the human being (molecule of synthesis), the absorption and the metabolization of weed-killers by plants cause the death of these plants. In this work, we set as goal the evaluation of the effect of a weed-killer sulfonylurea, the CossackOD with various concentrations (0, 2, 4 and 9 µg) on variety of Triticum durum: Cirta. We evaluated the plant growth by measuring the leaves and root length, compared with the witness as well as the content of proline and analyze the level of one of the antioxydative enzymes: catalase, after 14 days of treatment. Sulfonylurea is foliar and root weed-killers inhibiting the acetolactate synthase: a vegetable enzyme essential to the development of the plant. This inhibition causes the ruling of the growth then the death. The obtained results show a diminution of the average length of leaves and roots this can be explained by the fact that the ALS inhibitors are more active in the young and increasing regions of the plant, what inhibits the cellular division and talks a limitation of the foliar and root’s growth. We also recorded a highly significant increase in the proline levels and a stimulation of the catalase activity. As a response to increasing the herbicide concentrations a particular increases in antioxidative mechanisms in wheat cultivar Cirta suggest that the high sensitivity of Cirta to this sulfonylurea herbicide is related to the enhanced production and oxidative damage of reactive oxygen species.

Keywords: sulfonylurea, triticum durum, oxydative stress, toxicity

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Authors: Sweta Pandey, Samridhi Dhyani, Susmita Chaudhuri

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Klebsiella pneumoniae causes a wide range of infections, including urinary tract infections, sepsis, bacteremia, pneumonia, and liver abscesses. The emergence of multi-drug resistance in this bacterium led to a major setback for clinical management. WHO also endorsed a need for finding alternative therapy to antibiotics for the treatment of these infections. Development of vaccines and passive antibody therapy has been proven as a potent alternative to antibiotics in the case of MDR, XDR, and PDR Klebsiella infections. Siderophore receptors have been demonstrated to be overexpressed for the internalization of iron siderophore complexes during infections in most Gram-negative bacteria. For the present study, immune response to siderophore receptors to establish this protein as a potential immunogen for the development of therapeutic leads was explored. Clinical strains of Klebsiella pneumoniae were grown in iron-deficient conditions, and the iron-regulated outer membrane proteins were extracted and characterized through mass spectrometry for specific identification. The gene for identified protein was cloned in pET- 28a vector and expressed in E. coli. The native protein and the recombinant protein were isolated and purified and used as antigens for the generation of immune response in BALB/c mice. The native protein of Klebsiella pneumoniae grown in iron-deficient conditions was identified as FepA (Ferrienterobactin receptor) and other siderophore receptors. This 80 kDa protein generated an immune response in BALB/c mice. The antiserum from mice after subsequent booster doses was collected and showed binding with FepA protein in western blot and phagocytic uptake of the K. pneumoniae in the presence antiserum from immunized mice also observed from the animal studies after bacterial challenge post immunisation in mice have shown bacterial clearance. The antiserum from mice showed binding and clearance of the Klebsiella pneumoniae bacteria in vitro and in vivo. These antigens used for generating an active immune response in mice can further be used for therapeutic monoclonal antibody development against Klebsiella pneumoniae infections.

Keywords: antiserum, FepA, Klebsiella pneumoniae, multi drug resistance, siderophore receptor

Procedia PDF Downloads 102

Authors: Maryam Parsian, Pelin Mutlu, Ufuk Gunduz

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Two-dimensional cell culture affords simplicity and low cost, but it has serious limitations; lacking cell-cell and cell-matrix interactions that are present in tissues. Cancer cells grown in 3D culture systems have distinct phenotypes of adhesion, growth, migration, invasion as well as profiles of gene and protein expression. These interactions cause the 3D-cultured cells to acquire morphological and cellular characteristics relevant to in vivo tumors. Palbociclib is a chemotherapeutic agent for the treatment of ER-positive and HER-negative metastatic breast cancer. Poly-amidoamine (PAMAM) dendrimer is a well-defined, special three-dimensional structure and has a multivalent surface and internal cavities that can play an essential role in drug delivery systems. In this study, palbociclib is loaded onto the magnetic PAMAM dendrimer. Hanging droplet method was used in order to form 3D spheroids. The possible toxic effects of both free drug and drug loaded nanoparticles were evaluated in 2D and 3D MCF-7, MD-MB-231 and SKBR-3 breast cancer cell culture models by performing MTT cell viability and Alamar Blue assays. MTT analysis was performed with six different doses from 1000 µg/ml to 25 µg/ml. Drug unloaded PAMAM dendrimer did not demonstrate significant toxicity on all breast cancer cell lines. The results showed that 3D spheroids are clearly less sensitive than 2D cell cultures to free palbociclib. Also, palbociclib loaded PAMAM dendrimers showed more toxic effect than free palbociclib in all cell lines at 2D and 3D cultures. The results suggest that the traditional cell culture method (2D) is insufficient for mimicking the actual tumor tissue. The response of the cancer cells to anticancer drugs is different in the 2D and 3D culture conditions. This study showed that breast cancer cells are more resistant to free palbociclib in 3D cultures than in 2D cultures. However, nanoparticle loaded drugs can be more cytotoxic when compared to free drug.

Keywords: 2D and 3D cell culture, breast cancer, palbociclibe, PAMAM magnetic nanoparticles

Procedia PDF Downloads 149

Authors: R. A. C. H. Seneviratne, M. Gunawardana, R. P. N. P. Rajapakse

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To address the micronutrient deficiencies in the Asian region, the World Food Program in its current mandate highlights the requirement of employing efficient fortification of micronutrients in rice, under the program 'Scaling-up Rice Fortification in Asia'. The current industrial methods of rice fortification with micronutrients are not promising due to poor permeation or retention of fortificants. This study was carried out to develop a method to improve fortification of micronutrients in rice by removing the air barriers for diffusing micronutrients through the husk. For the purpose, soaking stage of paddy was coupled with vacuum (- 0.6 bar) for different time periods. Both long and short grain varieties of paddy (BG 352 and BG 358, respectively) initially tested for water uptake during hot soaking (70 °C) under vacuum (28.5 and 26.15%, respectively) were significantly (P < 0.05) higher than that of non-vacuum conditions (25.24 and 25.45% respectively), exhibiting the effectiveness of water diffusion into the rice grains through the cleared pores under negative pressure. To fortify the selected micronutrients (iron and zinc), paddy was vacuum-soaked in Fe2+ or Zn2+ solutions (500 ppm) separately for one hour, and continued soaking for another 3.5 h without vacuum. Significantly (P<0.05) higher amounts of Fe2+ and Zn2+ were observed throughout the soaking period, in both short and long grain varieties of rice compared to rice treated without vacuum. To achieve the recommended limits of World Food Program standards for fortified iron (40-48 mg/kg) and zinc (60-72 mg/kg) in rice, soaking was done with different concentrations of Fe2+ or Zn2+ for varying time periods. For both iron and zinc fortifications, hot soaking (70 °C) in 400 ppm solutions under vacuum (- 0.6 bar) during the first hour followed by 2.5 h under atmospheric pressure exhibited the optimum fortification (Fe2+: 46.59±0.37 ppm and Zn2+: 67.24±1.36 ppm) with a greater significance (P < 0.05) compared to the controls (Fe2+: 38.84±0.62 ppm and Zn2+: 52.55±0.55 ppm). This finding was further confirmed by the XRF images, clearly showing a greater fixation of Fe2+ and Zn2+ in the rice grains under vacuum treatment. Moreover, there were no significant (P>0.05) differences among both Fe2+ and Zn2+ contents in fortified rice even after polishing and washing, confirming their greater retention. A seven point hedonic scale showed that the overall acceptability for both iron and zinc fortified rice were significantly (P < 0.05) higher than the parboiled rice without fortificants. With all the drawbacks eliminated, per kilogram cost will be less than US$ 1 for both iron and zinc fortified rice. The new method of rice fortification studied and developed in this research, can be claimed as the best method in comparison to other rice fortification methods currently deployed.

Keywords: fortification, vacuum assisted diffusion, micronutrients, parboiling

Procedia PDF Downloads 253

Authors: Soultana Konstantinidou, Tiziana Schmidt, Elena Landi, Alessandro De Carli, Giovanni Maltinti, Darius Witt, Alicja Dziadosz, Agnieszka Lindstaedt, Michele Lai, Mauro Pistello, Valentina Cappello, Luciana Dente, Chiara Gabellini, Piotr Barski, Vittoria Raffa

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CRISPR/Cas9 technology has gained the interest of researchers in the field of biotechnology for genome editing. Since its discovery as a microbial adaptive immune defense, this system has been widely adopted and is acknowledged for having a variety of applications. However, critical barriers related to safety and delivery are persisting. Here, we propose a new concept of genome engineering, which is based on a nano-formulation of Cas9. The Cas9 enzyme was conjugated to a gold nanoparticle (AuNP-Cas9). The AuNP-Cas9 maintained its cleavage efficiency in vitro, to the same extent as the ribonucleoprotein, including non-conjugated Cas9 enzyme, and showed high gene editing efficiency in vivo in zebrafish embryos. Since CRISPR/Cas9 technology is extensively used in cancer research, melanoma was selected as a validation target. Cell studies were performed in A375 human melanoma cells. Particles per se had no impact on cell metabolism and proliferation. Intriguingly, the AuNP-Cas9 internalized spontaneously in cells and localized as a single particle in the cytoplasm and organelles. More importantly, the AuNP-Cas9 showed a high nuclear localization signal. The AuNP-Cas9, overcoming the delivery difficulties of Cas9, could be used in cellular biology and localization studies. Taking advantage of the plasmonic properties of gold nanoparticles, this technology could potentially be a bio-tool for combining gene editing and photothermal therapy in cancer cells. Further work will be focused on intracellular interactions of the nano-formulation and characterization of the optical properties.

Keywords: CRISPR/Cas9, gene editing, gold nanoparticles, nanotechnology

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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

Procedia PDF Downloads 292