Search results for: viability

Authors: Emma R. Arakelova, Stepan G. Grigoryan, Flora G. Arsenyan, Nelli S. Babayan, Ruzanna M. Grigoryan, Natalia K. Sarkisyan

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Novel nanosize zinc oxide composites of doxorubicin obtained by deposition of 180 nm thick zinc oxide film on the drug surface using DC-magnetron sputtering of a zinc target in the form of gels (PEO+Dox+ZnO and Starch+NaCMC+Dox+ZnO) were studied for drug delivery applications. The cancer specificity was revealed both in in vitro and in vivo models. The cytotoxicity of the test compounds was analyzed against human cancer (HeLa) and normal (MRC5) cell lines using MTT colorimetric cell viability assay. IC50 values were determined and compared to reveal the cancer specificity of the test samples. The mechanistic study of the most active compound was investigated using Flow cytometry analyzing of the DNA content after PI (propidium iodide) staining. Data were analyzed with Tree Star FlowJo software using cell cycle analysis Dean-Jett-Fox module. The in vivo anticancer activity estimation experiments were carried out on mice with inoculated ascitic Ehrlich’s carcinoma at intraperitoneal introduction of doxorubicin and its zinc oxide compositions. It was shown that the nanosize zinc oxide film deposition on the drug surface leads to the selective anticancer activity of composites at the cellular level with the range of selectivity index (SI) from 4 (Starch+NaCMC+Dox+ZnO) to 200 (PEO(gel)+Dox+ZnO) which is higher than that of free Dox (SI = 56). The significant increase in vivo antitumor activity (by a factor of 2-2.5) and decrease of general toxicity of zinc oxide compositions of doxorubicin in the form of the above mentioned gels compared to free doxorubicin were shown on the model of inoculated Ehrlich's ascitic carcinoma. Mechanistic studies of anticancer activity revealed the cytostatic effect based on the high level of DNA biosynthesis inhibition at considerable low concentrations of zinc oxide compositions of doxorubicin. The results of studies in vitro and in vivo behavior of PEO+Dox+ZnO and Starch+NaCMC+Dox+ZnO composites confirm the high potential of the nanosize zinc oxide composites as a vector delivery system for future application in cancer chemotherapy.

Keywords: anticancer activity, cancer specificity, doxorubicin, zinc oxide

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Authors: Rocio López, Miriam Tena, Montserrat Pérez, Rosario Solera

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Biotransformation of organic waste into biogas is considered an interesting alternative for the production of clean energy from renewable sources by reducing the volume and organic content of waste Anaerobic digestion is considered one of the most efficient technologies to transform waste into fertilizer and biogas in order to obtain electrical energy or biofuel within the concept of the circular economy. Currently, three types of anaerobic processes have been developed on a commercial scale: (1) single-stage process where sludge bioconversion is completed in a single chamber, (2) two-stage process where the acidogenic and methanogenic stages are separated into two chambers and, finally, (3) temperature-phase sequencing (TPAD) process that combines a thermophilic pretreatment unit prior to mesophilic anaerobic digestion. Two-stage processes can provide hydrogen and methane with easier control of the first and second stage conditions producing higher total energy recovery and substrate degradation than single-stage processes. On the other hand, co-digestion is the simultaneous anaerobic digestion of a mixture of two or more substrates. The technology is similar to anaerobic digestion but is a more attractive option as it produces increased methane yields due to the positive synergism of the mixtures in the digestion medium thus increasing the economic viability of biogas plants. The present study focuses on the energy recovery by anaerobic co-digestion of sewage sludge and waste from the aquaculture-fishing sector. The valorization is approached through the application of a temperature sequential phase process or TPAD technology (Temperature - Phased Anaerobic Digestion). Moreover, two-phase of microorganisms is considered. Thus, the selected process allows the development of a thermophilic acidogenic phase followed by a mesophilic methanogenic phase to obtain hydrogen (H₂) in the first stage and methane (CH₄) in the second stage. The combination of these technologies makes it possible to unify all the advantages of these anaerobic digestion processes individually. To achieve these objectives, a sequential study has been carried out in which the biochemical potential of hydrogen (BHP) is tested followed by a BMP test, which will allow checking the feasibility of the two-stage process. The best results obtained were high total and soluble COD yields (59.8% and 82.67%, respectively) as well as H₂ production rates of 12LH₂/kg SVadded and methane of 28.76 L CH₄/kg SVadded for TPAD.

Keywords: anaerobic co-digestion, TPAD, two-phase, BHP, BMP, sewage sludge, fish waste

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Authors: Marco Soto-Arriaza, Eduardo Cena-Ahumada, Jaime Melendez-Rojel

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Background: Liposomes have been widely used as a model of lipid bilayer to study the physicochemical properties of biological membrane, encapsulation, transport and release of different molecules. Furthermore, extensive research has focused on improving the efficiency in the transport of drugs, developing tools that improve the release of the encapsulated drug from liposomes. In this context, the enzymatic activity of PLA₂, despite having been shown to be an effective tool to promote the release of drugs from liposomes, is still an open field of research. Aim: The aim of the present study is to explore the effect of cholesterol (Cho) and amphipathic di- and tri-block copolymers, on calcein release mediated by enzymatic activity of PLA2 in Dipalmitoylphosphatidylcholine (DPPC) liposomes under physiological conditions. Methods: Different dispersions of DPPC, cholesterol, di-block POE₄₅-PCL₅₂ or tri-block PCL₁₂-POE₄₅-PCL₁₂ were prepared by the extrusion method after five freezing/thawing cycles; in Phosphate buffer 10mM pH 7.4 in presence of calcein. DPPC liposomes/Calcein were centrifuged at 15000rpm 10 min to separate free calcein. Enzymatic activity assays of PLA₂ were performed at 37°C using the TBS buffer pH 7.4. The size distribution, polydispersity, Z-potential and Calcein encapsulation of DPPC liposomes was monitored. Results: PLA₂ activity showed a slower kinetic of calcein release up to 20 mol% of cholesterol, evidencing a minimum at 10 mol% and then a maximum at 18 mol%. Regardless of the percentage of cholesterol, up to 18 mol% a one-hundred percentage release of calcein was observed. At higher cholesterol concentrations, PLA₂ showed to be inefficient or not to be involved in calcein release. In assays where copolymers were added in a concentration lower than their cmc, a similar behavior to those showed in the presence of Cho was observed, that is a slower kinetic in calcein release. In both experimental approaches, a one-hundred percentage of calcein release was observed. PLA₂ was shown to be sensitive to the 4-(4-Octadecylphenyl)-4-oxobutenoic acid inhibitor and calcium, reducing the release of calcein to 0%. Cell viability of HeLa cells decreased 7% in the presence of DPPC liposomes after 3 hours of incubation and 17% and 23% at 5 and 15 hours, respectively. Conclusion: Calcein release from DPPC liposomes, mediated by PLA₂ activity, depends on the percentage of cholesterol and the presence of copolymers. Both, cholesterol up to 20 mol% and copolymers below it cmc could be applied to the regulation of the kinetics of antitumoral drugs release without inducing cell toxicity per se.

Keywords: amphipathic copolymers, calcein release, cholesterol, DPPC liposome, phospholipase A₂

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Authors: Mohammad Noor Hisham Osman, Ahmed Razman Abdul Latiff, Zaidi Mat Daud, Zulkarnain Muhamad Sori

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The problem that Malaysian auditors do not issue going concern opinion (GC opinion) to seriously financially distressed companies is still a pressing issue. Policy makers, particularly the Financial Statement Review Committee (FSRC) of Malaysian Institute of Accountant, have raised this issue as early as in 2009. Similar problem happened in the US, UK, and many developing countries. It is important for auditors to issue GC opinion properly because such opinion is one signal about the viability of a company much needed by stakeholders. There are at least two unanswered questions or research gaps in the literature on determinants of GC opinion. Firstly, is client’s probability of failure associated with GC opinion issuance? Secondly, to what extent influential owners (management, family, and institution) moderate the association between client probability of failure and GC opinion issuance. The objective of this study is, therefore, twofold; (1) To examine the extent of the relationship between the probability of client failure and the issuance of GC opinion and (2) To examine the level of management, family, and institutional ownerships moderate the association between client probability of failure and the issuance of GC opinion. This study is quantitative in nature, and the sources of data are secondary (mainly company’s annual reports). A total of four hypotheses have been developed and tested on data accumulated from annual reports of seriously financially distressed Malaysian public listed companies. Data from 2006 to 2012 on a sample of 644 observations have been analyzed using panel logistic regression. It is found that certainty (rather than probability) of client failure affects the issuance of GC opinion. In addition, it is found that only the level of family ownership does positively moderate the relationship between client probability of failure and GC opinion issuance. This study is a contribution to auditing literature as its findings can enhance our understanding about audit quality; particularly on the variables that are associated with the issuance of GC opinion. The findings of this study shed light on the roles family owners in GC opinion issuance process, and this would open ways for the researcher to suggest measures that can be used to tackle the problem of auditors do not want to issue GC opinion to financially distressed clients. The measures to be suggested can be useful to policy makers in formulating future promulgations.

Keywords: audit quality, auditing, auditor characteristics, going concern opinion, Malaysia

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Authors: Varsha Salian, Shama Rao, N. Narendra, B. Mohana Kumar

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Evolving evidence proposes the existence of a highly tumorigenic subpopulation of undifferentiated, self-renewing cancer stem cells, responsible for exhibiting resistance to conventional anti-cancer therapy, recurrence, metastasis and heterogeneous tumor formation. Importantly, the mechanisms exploited by cancer stem cells to resist chemotherapy are very less understood. Oral squamous cell carcinoma (OSCC) is one of the most regularly diagnosed cancer types in India and is associated commonly with alcohol and tobacco use. Therefore, the isolation and in vitro characterization of cancer stem-like cells from patients with OSCC is a critical step to advance the understanding of the chemoresistance processes and for designing therapeutic strategies. With this, the present study aimed to establish and characterize cancer stem-like cells in vitro from OSCC. The primary cultures of cancer stem-like cell lines were established from the tissue biopsies of patients with clinical evidence of an ulceroproliferative lesion and histopathological confirmation of OSCC. The viability of cells assessed by trypan blue exclusion assay showed more than 95% at passage 1 (P1), P2 and P3. Replication rate was performed by plating cells in 12-well plate and counting them at various time points of culture. Cells had a more marked proliferative activity and the average doubling time was less than 20 hrs. After being cultured for 10 to 14 days, cancer stem-like cells gradually aggregated and formed sphere-like bodies. More spheroid bodies were observed when cultured in DMEM/F-12 under low serum conditions. Interestingly, cells with higher proliferative activity had a tendency to form more sphere-like bodies. Expression of specific markers, including membrane proteins or cell enzymes, such as CD24, CD29, CD44, CD133, and aldehyde dehydrogenase 1 (ALDH1) is being explored for further characterization of cancer stem-like cells. To summarize the findings, the establishment of OSCC derived cancer stem-like cells may provide scope for better understanding the cause for recurrence and metastasis in oral epithelial malignancies. Particularly, identification and characterization studies on cancer stem-like cells in Indian population seem to be lacking thus provoking the need for such studies in a population where alcohol consumption and tobacco chewing are major risk habits.

Keywords: cancer stem-like cells, characterization, in vitro, oral squamous cell carcinoma

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Authors: Jean Yves Trepanier, Sami Ammar, Sagnik Banik

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Lattice Boltzmann Method has been advantageous in simulating complex boundary conditions and solving for fluid flow parameters by streaming and collision processes. This paper includes the study of three different test cases in a confined domain using the method of the Lattice Boltzmann model. 1. An SRT (Single Relaxation Time) approach in the Lattice Boltzmann model is used to simulate Lid Driven Cavity flow for different Reynolds Number (100, 400 and 1000) with a domain aspect ratio of 1, i.e., square cavity. A moment-based boundary condition is used for more accurate results. 2. A Thermal Lattice BGK (Bhatnagar-Gross-Krook) Model is developed for the Rayleigh Benard convection for both test cases - Horizontal and Vertical Temperature difference, considered separately for a Boussinesq incompressible fluid. The Rayleigh number is varied for both the test cases (10^3 ≤ Ra ≤ 10^6) keeping the Prandtl number at 0.71. A stability criteria with a precise forcing scheme is used for a greater level of accuracy. 3. The phase change problem governed by the heat-conduction equation is studied using the enthalpy based Lattice Boltzmann Model with a single iteration for each time step, thus reducing the computational time. A double distribution function approach with D2Q9 (density) model and D2Q5 (temperature) model are used for two different test cases-the conduction dominated melting and the convection dominated melting. The solidification process is also simulated using the enthalpy based method with a single distribution function using the D2Q5 model to provide a better understanding of the heat transport phenomenon. The domain for the test cases has an aspect ratio of 2 with some exceptions for a square cavity. An approximate velocity scale is chosen to ensure that the simulations are within the incompressible regime. Different parameters like velocities, temperature, Nusselt number, etc. are calculated for a comparative study with the existing works of literature. The simulated results demonstrate excellent agreement with the existing benchmark solution within an error limit of ± 0.05 implicates the viability of this method for complex fluid flow problems.

Keywords: BGK, Nusselt, Prandtl, Rayleigh, SRT

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Authors: Pawan Kumar, Pitamber Singh Negi

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Many of the regeneration failures in conifers are often being attributed to heavy insect attack and pathogens during the period of seed formation and under storage conditions. Conifer berries and seed insects occur throughout the known range of the hosts and also limit the production of seed for nursery stock. On occasion, even entire seed crops are lost due to insect attacks. The berry and seeds of both the species have been found to be infected with insects. Recently, heavy damage to the berry and seeds of Juniper and Chilgoza Pine was observed in the field as well as in stored conditions, leading to reduction in the viability of seeds to germinate. Both the species are under great threat and regeneration of the species is very low. Due to lack of adequate literature, the study on the damage potential of seed insects was urgently required to know the exact status of the insect-pests attacking seeds/berries of both the pine species so as to develop pest management practices against the insect pests attack. As both the species are also under threat and are fighting for survival, so the study is important to develop management practices for the insect-pests of seeds/berries of Juniper and Chilgoza pine so as to evaluate in the nursery, as these species form major vegetation of their distribution zones. A six-year study on the management of insect pests of seeds of Chilgoza revealed that seeds of this species are prone to insect pests mainly borers. During present investigations, it was recorded that cones of are heavily attacked only by Dioryctria abietella (Lepidoptera: Pyralidae) in natural conditions, but seeds which are economically important are heavily infected, (sometimes up to 100% damage was also recorded) by insect borer, Plodia interpunctella (Lepidoptera: Pyralidae) and is recorded for the first time ‘to author’s best knowledge’ infesting the stored Chilgoza seeds. Similarly, Juniper berries and seeds were heavily attacked only by a single borer, Homaloxestis cholopis (Lepidoptera: Lecithoceridae) recorded as a new report in natural habitat as well as in stored conditions. During the present investigation details of insect pest attack on Juniper and Chilgoza pine seeds and berries was observed and suitable management practices were also developed to contain the insect-pests attack.

Keywords: borer, chilgozapine, cones, conifer, Lepidoptera, juniper, management, seed

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Authors: Darshini Mahadevia, Kanika Gounder, Saumya Lathia

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Cities, an outcome of natural growth and migration, are ever-expanding due to urban sprawl. In the Global South, urban areas are experiencing a switch from public transport to private vehicles, coupled with intensified urban agglomeration, leading to frequent longer commutes by automobiles. This increase in travel distance and motorized vehicle kilometres lead to unsustainable cities. To achieve the nationally pledged GHG emission mitigation goal, the government is prioritizing a modal shift to low-carbon transport modes like mass transit and paratransit. Mixed land-use and urban densification are crucial for the economic viability of these projects. Informed by desktop assessment of mobility plans and in-person primary surveys, the paper explores the challenges around urban densification and travel patterns in two Indian cities of contrasting nature- Surat, a metropolitan industrial city with a 5.9 million population and a very compact urban form, and Udaipur, a heritage city attracting large international tourists’ footfall, with limited scope for further densification. Dense, mixed-use urban areas often improve access to basic services and economic opportunities by reducing distances and enabling people who don't own personal vehicles to reach them on foot/ cycle. But residents travelling on different modes end up contributing to similar trip lengths, highlighting the non-uniform distribution of land-uses and lack of planned transport infrastructure in the city and the urban-peri urban networks. Additionally, it is imperative to manage these densities to reduce negative externalities like congestion, air/noise pollution, lack of public spaces, loss of livelihood, etc. The study presents a comparison of the relationship between transport systems with the built form in both cities. The paper concludes with recommendations for managing densities in urban areas along with promoting low-carbon transport choices like improved non-motorized transport and public transport infrastructure and minimizing personal vehicle usage in the Global South.

Keywords: India, low-carbon transport, travel behaviour, trip length, urban densification

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Authors: Bruno Baptista, Andreia S. R. Oliveira, Patricia V. Mendonca, Jorge F. J. Coelho, Fani Sousa

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Drug delivery systems are designed to allow adequate protection and controlled delivery of drugs to specific locations. These systems aim to reduce side effects and control the biodistribution profile of drugs, thus improving therapeutic efficacy. This study involved the synthesis of polymeric nanoparticles, based on amphiphilic diblock copolymers, comprising a biocompatible, poly (oligo (ethylene oxide) methyl ether methacrylate (POEOMA) as hydrophilic segment and a pH-sensitive block, the poly (2-diisopropylamino)ethyl methacrylate) (PDPA). The objective of this work was the development of polymeric pH-responsive nanoparticles to encapsulate and carry small RNAs as a model to further develop non-coding RNAs delivery systems with therapeutic value. The responsiveness of PDPA to pH allows the electrostatic interaction of these copolymers with nucleic acids at acidic pH, as a result of the protonation of the tertiary amine groups of this polymer at pH values below its pKa (around 6.2). Initially, the molecular weight parameters and chemical structure of the block copolymers were determined by size exclusion chromatography (SEC) and nuclear magnetic resonance (1H-NMR) spectroscopy, respectively. Then, the complexation with small RNAs was verified, generating polyplexes with sizes ranging from 300 to 600 nm and with encapsulation efficiencies around 80%, depending on the molecular weight of the polymers, their composition, and concentration used. The effect of pH on the morphology of nanoparticles was evaluated by scanning electron microscopy (SEM) being verified that at higher pH values, particles tend to lose their spherical shape. Since this work aims to develop systems for the delivery of non-coding RNAs, studies on RNA protection (contact with RNase, FBS, and Trypsin) and cell viability were also carried out. It was found that they induce some protection against constituents of the cellular environment and have no cellular toxicity. In summary, this research work contributes to the development of pH-sensitive polymers, capable of protecting and encapsulating RNA, in a relatively simple and efficient manner, to further be applied on drug delivery to specific sites where pH may have a critical role, as it can occur in several cancer environments.

Keywords: drug delivery systems, pH-responsive polymers, POEOMA-b-PDPA, small RNAs

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Authors: A. Díaz-Roa, P. I. Silva Junior, F. J. Bello

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Sarconesiopsis magellanica (Diptera: Calliphoridae) is a medically important necrophagous fly which is used for establishing the post-mortem interval. Dipterous maggots release diverse proteins and peptides contained in larval excretion and secretion (ES) products playing a key role in digestion. The most important mechanism for combating infection using larval therapy depends on larval ES. These larvae are protected against infection by a diverse spectrum of antimicrobial peptides (AMPs), one already known like lucifensin. Special interest in these peptides has also been aroused regarding understanding their role in wound healing since they degrade necrotic tissue and kill different bacteria during larval therapy. The action of larvae on wounds occurs through 3 mechanisms of action: removal of necrotic tissue, stimulation of granulation tissue, and antibacterial action of larval ES. Some components of the ES include calcium, urea, allantoin ammonium bicarbonate and reducing the viability of Gram positive and Gram negative bacteria. The Lucilia sericata fly larvae have been the most used, however, we need to evaluate new species that could potentially be similar or more effective than fly above. This study was thus aimed at identifying and characterizing S. magellanica AMPs contained in ES products for the first time and compared them with the common fly used L. sericata. These products were obtained from third-instar larvae taken from a previously established colony. For the first analysis, ES fractions were separate by Sep-Pak C18 disposable columns (first step). The material obtained was fractionated by RP-HPLC by using Júpiter C18 semi-preparative column. The products were then lyophilized and their antimicrobial activity was characterized by incubation with different bacterial strains. The first chromatographic analysis of ES from L. sericata gives 6 fractions with antimicrobial activity against Gram-positive bacteria Micrococus luteus, and 3 fractions with activity against Gram-negative bacteria Pseudomonae aeruginosa while the one from S. magellanica gaves 1 fraction against M. luteus and 4 against P. aeruginosa. Maybe one of these fractions could correspond to the peptide already known from L. sericata. These results show the first work for supporting further experiments aimed at validating S. magellanica use in larval therapy. We still need to search if we find some new molecules, by making mass spectrometry and ‘de novo sequencing’. Further studies are necessary to identify and characterize them to better understand their functioning.

Keywords: antimicrobial peptides, larval therapy, Lucilia sericata, Sarconesiopsis magellanica

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Authors: Elchin Hajiyev, Afet Memmedova Dadash, Sabina Hajiyeva, Aynur Karimova, Ramiz Aliyev

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Although potatoes have a wide distribution range, the yield potential of varieties varies greatly depending on the region. Our country is made up of agricultural regions with very different environmental characteristics.In this case, we cannot expect the introduced varieties to show the same adaptation to the different conditions of our country. For this reason, in our country, varieties with high general adaptability should be used, rather than varieties with special adaptability in certain areas. Soil salinization has become a global problem.Increased salinity has a serious impact on food security by reducing plant productivity. Plants have protective mechanisms of adaptation to salt stress, such as the synthesis of physiologically active substances, resistance to antioxidant stress and oxidation of membrane lipids. One of these substances is free proline. Our study revealed genetic variation in proline accumulation among samples exposed to stress factors.Changes in proline content under stress conditions were studied in 50 samples. There was wide variation across all treatments.The amount of proline varied between 7.2–37.7 μM/g under salinity conditions.The lowest rate was in the SF33 genotype (1.5 times more than the control (2.5 μM/g)).The highest level of proline under the influence of salt stress was in the SF45 genotype (7.25 times higher than the control (32.5 μM/g)). Our studies have found that the protective system reacts differently to the influence of stress factors. According to the results obtained on the amount of proline, adaptation mechanisms must be more actively activated to maintain metabolism and ensure viability in sensitive forms under the influence of stress factors. At high doses of the salt stressor, a tenfold increase in proline compared to the control indicates significant damage to the plant organism as a result of stress.To prevent damage to the body, the antioxidant system needs to quickly mobilize and work at full capacity in adverse conditions. An increase in the dose of the stress factor salt in our study caused a greater increase in the amount of free proline in plant tissues. Considering the functions of proline as an osmoprotector and antioxidant, it was found that increasing its amount is aimed at protecting the plant from the acute effects of stressors.

Keywords: genetic variability, potato, genotypes, proline, stress

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Authors: Ezzati Givi M., Hoveizi E., Nezhad Marani N.

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Platinum-based anticancer therapeutics are the most widely used drugs in clinical chemotherapy but have major limitations and various side effects in clinical applications. Gonadotoxicity and sterility is one of the most common complications for cancer survivors, which seem to be drug-specific and dose-related. Therefore, many efforts have been dedicated to discovering a new structure of platinum-based anticancer agents with improved therapeutic index, fewer side effects. In this regard, new Pt(II)-phosphane complexes containing heterocyclic thionate ligands (PCTL) have been synthesized, which show more potent antitumor activities in comparison to cisplatin. Cisplatin, the best leading metal-based antitumor drug in the field, induces testicular toxicity on Leydig and Sertoli cells leading to serious side effects such as azoospermia and infertility. Therefore in the present study, we aimed to investigate the cytotoxicity effect of PCTL on mice TM4 Sertoli cells with particular emphasis on the role of autophagy in comparison to cisplatin. In this study, an MTT assay was performed to evaluate the IC50 of PCTL and to analyze the TM3 Leydig cell's viability. Cells morphology was evaluated via invert microscope and Changing in morphology for nuclei swelling or autophagic vacuoles formation were assessed by DAPI and MDC staining. Testosterone production in the culture medium was measured using an ELISA kit. Finally, the expression of Autophagy-related genes, Atg5, Beclin1 and p62, were analyzed by qPCR. Based on the obtained results by MTT, the IC50 value of PCTL was 50 μM in TM3 cells and cytotoxic effects was in a dose- and time-dependent manner. Cells morphological changes investigated by inverted microscopy, DAPI, and MDC staining which showed the cytotoxic concentrations of PCTL was significantly higher than cisplatin in the treated TM3 Leydig cells. The results of PCR showed a lack of expression of the p62, Atg5 and Beclin1 gene in TM3 cells treated with PCTL in comparison to cisplatin and control groups. It should be noted that the effects of 25 μM PCTL concentration on TM3 cells have been associated with increased testosterone production and secretion, which requires further study to explain the possible causes and involved molecular mechanisms. The results of the study showed that the PCTL had less-lethal effects on TM3 cells in comparison to cisplatin and probably did not induce autophagy in TM3 cells.

Keywords: platinum-based anticancer agents, cisplatin, Leydig TM3 cells, autophagy

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Authors: Regiane A. Oliveira, Carlos E. Vaz Rossell, Rubens Maciel Filho

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Lactic acid, besides a valuable chemical may be considered a platform for other chemicals. In fact, the feasibility of hemicellulosic sugars as feedstock for lactic acid production process, may represent the drop of some of the barriers for the second generation bioproducts, especially bearing in mind the 5-carbon sugars from the pre-treatment of sugarcane bagasse. Bearing this in mind, the purpose of this study was to use the hemicellulosic liquor from sugarcane bagasse as a substrate to produce lactic acid by fermentation. To release of sugars from hemicellulose it was made a pre-treatment with a diluted sulfuric acid in order to obtain a xylose's rich liquor with low concentration of inhibiting compounds for fermentation (≈ 67% of xylose, ≈ 21% of glucose, ≈ 10% of cellobiose and arabinose, and around 1% of inhibiting compounds as furfural, hydroxymethilfurfural and acetic acid). The hemicellulosic sugars associated with 20 g/L of yeast extract were used in a fermentation process with Lactobacillus plantarum to produce lactic acid. The fermentation process pH was controlled with automatic injection of Ca(OH)2 to keep pH at 6.00. The lactic acid concentration remained stable from the time when the glucose was depleted (48 hours of fermentation), with no further production. While lactic acid is produced occurs the concomitant consumption of xylose and glucose. The yield of fermentation was 0.933 g lactic acid /g sugars. Besides, it was not detected the presence of by-products, what allows considering that the microorganism uses a homolactic fermentation to produce its own energy using pentose-phosphate pathway. Through facultative heterofermentative metabolism the bacteria consume pentose, as is the case of L. plantarum, but the energy efficiency for the cell is lower than during the hexose consumption. This implies both in a slower cell growth, as in a reduction in lactic acid productivity compared with the use of hexose. Also, L. plantarum had shown to have a capacity for lactic acid production from hemicellulosic hydrolysate without detoxification, which is very attractive in terms of robustness for an industrial process. Xylose from hydrolyzed bagasse and without detoxification is consumed, although the hydrolyzed bagasse inhibitors (especially aromatic inhibitors) affect productivity and yield of lactic acid. The use of sugars and the lack of need for detoxification of the C5 liquor from sugarcane bagasse hydrolyzed is a crucial factor for the economic viability of second generation processes. Taking this information into account, the production of second generation lactic acid using sugars from hemicellulose appears to be a good alternative to the complete utilization of sugarcane plant, directing molasses and cellulosic carbohydrates to produce 2G-ethanol, and hemicellulosic carbohydrates to produce 2G-lactic acid.

Keywords: fermentation, lactic acid, hemicellulosic sugars, sugarcane

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Authors: Kevin Kellner, Nga Lao, Orla Coleman, Paula Meleady, Niall Barron

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Chinese Hamster Ovary (CHO) cells are the prominent cell line used in biopharmaceutical production. To improve yields and find beneficial bioprocess phenotypes genetic engineering plays an essential role in recent research. The miR-23 cluster, specifically miR-24 and miR-27, was first identified as differentially expressed during hypothermic conditions suggesting a role in proliferation and productivity in CHO cells. In this study, we used sponge decoy technology to stably deplete the miRNA expression of the cluster. Furthermore, we implemented the CRISPR/Cas9 system to knockdown miRNA expression. Sponge constructs were designed for an imperfect binding of the miRNA target, protecting from RISC mediated cleavage. GuideRNAs for the CRISPR/Cas9 system were designed to target the seed region of the miRNA. The expression of mature miRNA and precursor were confirmed using RT-qPCR. For both approaches stable expressing mixed populations were generated and characterised in batch cultures. It was shown, that CRISPR/Cas9 can be implemented in CHO cells with achieving high knockdown efficacy of every single member of the cluster. Targeting of one miRNA member showed that its genomic paralog is successfully targeted as well. The stable depletion of miR-24 using CRISPR/Cas9 showed increased growth and specific productivity in a CHO-K1 mAb expressing cell line. This phenotype was further characterized using quantitative label-free LC-MS/MS showing 186 proteins differently expressed with 19 involved in proliferation and 26 involved in protein folding/translation. Targeting miR-27 in the same cell line showed increased viability in late stages of the culture compared to the control. To evaluate the phenotype in an industry relevant cell line; the miR-23 cluster, miR-24 and miR-27 were stably depleted in a Fc fusion CHO-S cell line which showed increased batch titers up to 1.5-fold. In this work, we highlighted that the stable depletion of the miR-23 cluster and its members can improve the bioprocess phenotype concerning growth and productivity in two different cell lines. Furthermore, we showed that using CRISPR/Cas9 is comparable to the traditional sponge decoy technology.

Keywords: Chinese Hamster ovary cells, CRISPR/Cas9, microRNAs, sponge decoy technology

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Authors: Saima Shaikh, Andre Brown, Wallace Enegbuma

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Earthquakes strike both developed and developing countries, causing tremendous damage and the loss of lives of millions of people, mainly due to the collapsing of buildings, particularly in poorer countries. Despite the socio-economic and technological restrictions, the poorer countries have adopted proven and established housing-strengthening techniques from affluent countries. Rural communities are aware of the earthquake-strengthening mechanisms for improving housing resilience, but owing to socio-economic and technological constraints, the seismic guidelines are rarely implemented, resulting in informal construction practice. Unregistered skilled laborers make substantial contributions to the informal construction sector, particularly in rural areas where knowledge is scarce. Laborers employ their local expertise in house construction; however, owing to a lack of seismic expertise in safe building procedures, the authorities' regulated seismic norms are not applied. From the perspective of seismic knowledge transformation in safe buildings practices, the study focuses on the feasibility of seismic guidelines implementation. The study firstly employs a literature review of massive-scale reconstruction after the 2005 earthquake in rural Pakistan. The 2005-earthquake damaged over 400,000 homes, killed 70,000 people and displaced 2.8 million people. The research subsequently corroborated the pragmatic approach using questionnaire field survey among the rural people in 2005-earthquake affected areas. Using the literature and the questionnaire survey, the research analyzing people's perspectives on technical acceptability, financial restrictions, and socioeconomic viability and examines the effectiveness of seismic knowledge transfer in safe buildings practices. The findings support the creation of a knowledge transfer framework in disaster mitigation and recovery planning, assisting rural communities and builders in minimising losses and improving response and recovery, as well as improving housing resilience and lowering vulnerabilities. Finally, certain conclusions are obtained in order to continue the resilience research. The research can be further applied in rural areas of developing countries having similar construction practices.

Keywords: earthquakes, knowledge transfer, resilience, informal construction practices

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Authors: Mercedes Del Rio Merino, Jaime Santacruz Astorqui, Paola Villoria Saez, Carmen Viñas Arrebola

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The lack of treatment of the waste from construction and demolition waste (CDW) is a problem that must be solved immediately. It is estimated that in the world not to use CDW generates an increase in the use of new materials close to 20% of the total value of the materials used. The problem is even greater in case these wastes are considered hazardous because the final deposition of them may also generate significant contamination. Therefore, the possibility of including CDW in the manufacturing of building materials, represents an interesting alternative to ensure their use and to reduce their possible risk. In this context and in the last years, many researches are being carried out in order to analyze the viability of using CDW as a substitute for the traditional raw material of high environmental impact. Even though it is true, much remains to be done, because these works generally characterize materials but not specific applications that allow the agents of the construction to have the guarantees required by the projects. Therefore, it is necessary the involvement of all the actors included in the life cycle of these new construction materials, and also to promote its use for, for example, definition of standards, tax advantages or market intervention is necessary. This paper presents the main findings reached in "Waste to resources (W2R)" project since it began in October 2014. The main goal of the project is to develop new materials, elements and construction systems, manufactured from CDW, to be used in improving the energy efficiency of buildings. Other objectives of the project are: to quantify the CDW generated in the energy rehabilitation works, specifically wastes from the building envelope; and to study the traceability of CDW generated and promote CDW reuse and recycle in order to get close to the life cycle of buildings, generating zero waste and reducing the ecological footprint of the construction sector. This paper determines the most important aspects to consider during the design of new constructive solutions, which improve the energy efficiency of buildings and what materials made with CDW would be the most suitable for that. Also, a survey to select best practices for reducing "close to zero waste" in refurbishment was done. Finally, several pilot rehabilitation works conform the parameters analyzed in the project were selected, in order to apply the results and thus compare the theoretical with reality. Acknowledgements: This research was supported by the Spanish State Secretariat for Research, Development and Innovation of the Ministry of Economy and Competitiveness under "Waste 2 Resources" Project (BIA2013-43061-R).

Keywords: building waste, construction and demolition waste, recycling, resources

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Authors: Manami Masuda, Mariko Era, Takayoshi Kawahara, Takahide Kanyama, Hiroshi Morita

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Objectives: Fatty acid salts are a type of anionic surfactant and are produced from fatty acids and alkali. Moreover, fatty acid salts are known to have potent antibacterial activities. Acanthamoeba is ubiquitously distributed in the environment including sea water, fresh water, soil and even from the air. Although generally free-living, Acanthamoeba can be an opportunistic pathogen, which could cause a potentially blinding corneal infection known as Acanthamoeba keratitis. So, in this study, we evaluated the anti-amoeba activity of fatty acid salts and fatty acids to Acanthamoeba castellanii ATCC 30010. Materials and Methods: The antibacterial activity of 9 fatty acid salts (potassium butyrate (C4K), caproate (C6K), caprylate (C8K), caprate (C10K), laurate (C12K), myristate (C14K), oleate (C18:1K), linoleate (C18:2K), linolenate (C18:3K)) tested on cells of Acanthamoeba castellanii ATCC 30010. Fatty acid salts (concentration of 175 mM and pH 10.5) were prepared by mixing the fatty acid with the appropriate amount of KOH. The amoeba suspension mixed with KOH with a pH adjusted solution was used as the control. Fatty acids (concentration of 175 mM) were prepared by mixing the fatty acid with Tween 80 (20 %). The amoeba suspension mixed with Tween 80 (20 %) was used as the control. The anti-amoeba method, the amoeba suspension (3.0 × 104 cells/ml trophozoites) was mixed with the sample of fatty acid potassium (final concentration of 175 mM). Samples were incubated at 30°C, for 10 min, 60 min, and 180 min and then the viability of A. castellanii was evaluated using plankton counting chamber and trypan blue stainings. The minimum inhibitory concentration (MIC) against Acanthamoeba was determined using the two-fold dilution method. The MIC was defined as the minimal anti-amoeba concentration that inhibited visible amoeba growth following incubation (180 min). Results: C8K, C10K, and C12K were the anti-amoeba effect of 4 log-unit (99.99 % growth suppression of A. castellanii) incubated time for 180 min against A. castellanii at 175mM. After the amoeba, the suspension was mixed with C10K or C12K, destroying the cell membrane had been observed. Whereas, the pH adjusted control solution did not exhibit any effect even after 180 min of incubation with A. castellanii. Moreover, C6, C8, and C18:3 were the anti-amoeba effect of 4 log-unit incubated time for 60 min. C4 and C18:2 exhibited a 4-log reduction after 180 min incubation. Furthermore, the minimum inhibitory concentration (MIC) was determined. The MIC of C10K, C12K and C4 were 2.7 mM. These results indicate that C10K, C12K and C4 have high anti-amoeba activity against A. castellanii and suggest C10K, C12K and C4 have great potential for antimi-amoeba agents.

Keywords: Fatty acid salts, anti-amoeba activities, Acanthamoeba, fatty acids

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Authors: Divya Kanchibhotla

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Of all the major health threats to emerge, none has challenged the very foundation of public health so profoundly as the rise of non-communicable diseases (NCD). Encouraging a holistic health approach encompasses the community’s well-being. Competent public health grounded with holistic health approach can lay a better foundation in the modern world. Yoga has been increasingly explored as an adjunct therapy to major disorders. This study explores the efficacy of Yoga as a tool for public health. A survey was administered to 5500 adults, and 300 teens were selected from 25 states of India. The study explored the differences in health, happiness, and sustainable living between Yoga-practitioners and Non-yoga practitioners. The study also explored the practice and habits of yoga practitioners (frequency, place, reasons to practice) and Health, Happiness, and Sustainable Living. The subjects were grouped based on age, education, experience in yoga (years of practice), and occupational background. The study population comprised of 54% males and 46% females. Majority of the respondents (59%) were from 18 to 30 years age group. The study indicated that 96.4% of the total respondents have heard of Yoga. However, only 46.8% of the total study population practice yoga (YP) and the rest 53.2% were non-practitioners (NP). From a perspective of how Yoga and health, 72.7% yoga practitioners asserted a peaceful and happy life, 71.9% yoga practitioners felt satisfaction in life, and 70.2 % yoga practitioners had satisfactory health. 61.9% of yoga practitioners report being vegetarian, not eating junk food, and not drinking alcohol than 38.1% Non-Practitioners population. 47% of yoga practitioners found themselves to be more sensitive to the environment compared to only 40% of non-practitioners. India has been witnessing an unprecedented rise in the NCDs, accounting for 61% deaths. The importance of yoga as an adjunct therapy for various disorders and diseases is gaining momentum across the globe. There are various studies on yoga that have indicated benefits of yoga as a unique holistic approach towards lifestyle and a consistent, complementary solution that could be adopted for long-term viability for a well being. The comprehensive study is the first of its kind that takes a holistic look at the prevalence of Yoga for public health in India. Our study is unique and stands out as it is detailed in its outlook with extensive coverage of almost the whole country (surveying 25 out of 29 states) and contemplates on the benefits to an individual at the grass-root level – physical, mental and social outlook. The insights from the study will enable the health care systems and grassroots organizations to make the holistic practice of Yoga accessible to spread sustainable living for a healthy community.

Keywords: non-communicable disease (NCD), sustainable development goal (SDG 3), public health, healthy living

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Authors: Albert Kwabena Osei Konadu

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Pterocarpus erinaceus, also known as Rosewood, is tropical wood, endemic in forest savannah transition zones within the middle and northern portion of Ghana. Its economic viability has made it increasingly popular and in high demand, leading to widespread conservation concerns. Ghana’s forest resource management regime for these ecozones is mainly on conservation and very little on resource utilization. Consequently, commercial logging management standards are at teething stage and not fully developed, leading to a deficiency in the monitoring of logging operations and quantification of harvested trees volumes. Tree information form (TIF); a volume estimation and tracking regime, has proven to be an effective, sustainable management tool for regulating timber resource extraction in the high forest zones of the country. This work aims to generate TIF that can track and capture requisite parameters to accurately estimate the volume of harvested rosewood within forest savannah transition zones. Tree information forms were created on three scenarios of individual billets, stacked billets and conveying vessel basis. These TIFs were field-tested to deduce the most viable option for the tracking and estimation of harvested volumes of rosewood using the smallian and cubic volume estimation formula. Overall, four districts were covered with individual billets, stacked billets and conveying vessel scenarios registering mean volumes of 25.83m3,45.08m3 and 32.6m3, respectively. These adduced volumes were validated by benchmarking to assigned volumes of the Forestry Commission of Ghana and known standard volumes of conveying vessels. The results did indicate an underestimation of extracted volumes under the quotas regime, a situation that could lead to unintended overexploitation of the species. The research revealed conveying vessels route is the most viable volume estimation and tracking regime for the sustainable management of the Pterocarpous erinaceus species as it provided a more practical volume estimate and data extraction protocol.

Keywords: convention on international trade in endangered species, cubic volume formula, forest transition savannah zones, pterocarpus erinaceus, smallian’s volume formula, tree information form

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Authors: Adetona Abbassa, Aliyu Shakirat B.

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The study area is part of Bosso local Government, enclosed within Longitude 6.25’ to 6.31’ and Latitude 9.35’ to 9.45’, an area of 16x8 km², within the basement region of central Nigeria. The region is a host to Nigerian Airforce base 12 (NAF 12quick response) and its staff quarters, the headquarters of Bosso local government, the Independent National Electoral Commission’s two offices, four government secondary schools, six primary schools and Minna international airport. The area suffers an acute shortage of water from November when rains stop to June when rains commence within North Central Nigeria. A way of addressing this problem is a reconnaissance method to delineate possible fractures and fault lines that exists within the region by sampling the Aeromagnetic data and using an appropriate analytical algorithm to delineate these fractures. This is followed by an appropriate ground truthing method that will confirm if the fracture is connected to underground water movement. The first vertical derivative for structural analysis, reveals a set of lineaments labeled AA’, BB’, CC’, DD’, EE’ and FF’ all trending in the Northeast – Southwest directions. AA’ is just below latitude 9.45’ above Maikunkele village, cutting off the upper part of the field, it runs through Kangwo, Nini, Lawo and other communities. BB’ is at Latitude 9.43’ it truncated at about 2Km before Maikunkele and Kuyi. CC’ is around 9.40’ sitting below Maikunkele runs down through Nanaum. DD’ is from Latitude 9.38’; interestingly no community within this region where the fault passes through. A result from the three sites where Vertical Electrical Sounding was carried out reveals three layers comprised of topsoil, intermediate Clay formation and weathered/fractured or fresh basement. The depth to basement map was also produced, depth to the basement from the ground surface with VES A₂, B5, D₂ and E₁ to be relatively deeper with depth values range between 25 to 35 m while the shallower region of the area has a depth range value between 10 to 20 m. Hence, VES A₂, A₅, B₄, B₅, C₂, C₄, D₄, D₅, E₁, E₃, and F₄ are high conductivity zone that are prolific for groundwater potential. The depth range of the aquifer potential zones is between 22.7 m to 50.4 m. The result from site C is quite unique though the 3 layers were detected in the majority of the VES points, the maximum depth to the basement in 90% of the VES points is below 8 km, only three VES points shows considerably viability, which are C₆, E₂ and F₂ with depths of 35.2 m and 38 m respectively but lack of connectivity will be a big challenge of chargeability.

Keywords: lithology, aeromagnetic, aquifer, geoelectric, iso-resistivity, basement, vertical electrical sounding(VES)

Procedia PDF Downloads 139

Authors: Iva Blazkova, Amitava Moulick, Vedran Milosavljevic, Pavel Kopel, Marketa Vaculovicova, Vojtech Adam, Rene Kizek

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Doxorubicin is one of the most commonly used and the most effective chemotherapeutic drugs. This antracycline drug isolated from the bacteria Streptomyces peuceticus var. caesius is sold under the trade name Adriamycin (hydroxydaunomycin, hydroxydaunorubicin). Doxorubicin is used in single therapy to treat hematological malignancies (blood cancers, leukaemia, lymphoma), many types of carcinoma (solid tumors) and soft tissue sarcomas. It has many serious side effects like nausea and vomiting, hair lost, myelosupression, oral mucositis, skin reactions and redness, but the most serious one is the cardiotoxicity. Because of the risk of heart attack and congestive heart failure, the total dose administered to patients has to be accurately monitored. With the aim to lower the side effects and to targeted delivery of doxorubicin into the tumor tissue, the different nanoparticles are studied. The drug can be bound on a surface of nanoparticle, encapsulated in the inner cavity, or incorporated into the structure of nanoparticle. Among others, carbon nanoparticles (graphene, carbon nanotubes, fullerenes) are highly studied. Besides the number of inorganic nanoparticles, a great potential exhibit also organic ones mainly lipid-based and polymeric nanoparticle. The aim of this work was to perform a toxicity study of free doxorubicin compared to doxorubicin conjugated with various nanotransporters. The effect of liposomes, fullerenes, graphene, and carbon nanotubes on the toxicity was analyzed. As a first step, the binding efficacy of between doxorubicin and the nanotransporter was determined. The highest efficacy was detected in case of liposomes (85% of applied drug was encapsulated) followed by graphene, carbon nanotubes and fullerenes. For the toxicological studies, the chicken embryos incubated under controlled conditions (37.5 °C, 45% rH, rotation every 2 hours) were used. In 7th developmental day of chicken embryos doxorubicin or doxorubicin-nanotransporter complex was applied on the chorioallantoic membrane of the eggs and the viability was analyzed every day till the 17th developmental day. Then the embryos were extracted from the shell and the distribution of doxorubicin in the body was analyzed by measurement of organs extracts using laser induce fluorescence detection. The chicken embryo mortality caused by free doxorubicin (30%) was significantly lowered by using the conjugation with nanomaterials. The highest accumulation of doxorubicin and doxorubicin nanotransporter complexes was observed in the liver tissue

Keywords: doxorubicin, chicken embryos, nanotransporters, toxicity

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Authors: S. Murr, S. Phillips

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One of the biggest challenges entering a market with a carsharing or any other shared mobility (SM) service is sound investment decision-making. To support this process, the authors think that a city index evaluating different criteria is necessary. The goal of such an index is to benchmark cities along a set of external measures to answer the main two challenges: financially viability and the understanding of its specific requirements. The authors have consulted several shared mobility projects and industry experts to create such a Shared Mobility City Index (SMCI). The current proposal of the SMCI consists of 11 individual index measures: general data (demographics, geography, climate and city culture), shared mobility landscape (current SM providers, public transit options, commuting patterns and driving culture) and political vision and goals (vision of the Mayor, sustainability plan, bylaws/tenders supporting SM). To evaluate the suitability of the index, 16 cities on the East Coast of North America were selected and secondary research was conducted. The main sources of this study were census data, organisational records, independent press releases and informational websites. Only non-academic sources where used because the relevant data for the chosen cities is not published in academia. Applying the index measures to the selected cities resulted in three major findings. Firstly, density (city area divided by number of inhabitants) is not an indicator for the number of SM services offered: the city with the lowest density has five bike and carsharing options. Secondly, there is a direct correlation between commuting patterns and how many shared mobility services are offered. New York, Toronto and Washington DC have the highest public transit ridership and the most shared mobility providers. Lastly, except one, all surveyed cities support shared mobility with their sustainability plan. The current version of the shared mobility index is proving a practical tool to evaluate cities, and to understand functional, political, social and environmental considerations. More cities will have to be evaluated to refine the criteria further. However, the current version of the index can be used to assess cities on their suitability for shared mobility services and will assist investors deciding which city is a financially viable market.

Keywords: carsharing, transportation, urban planning, shared mobility city index

Procedia PDF Downloads 303

Authors: Calister Wingang Makebe, Wilson Ambindei Agwanande, Emmanuel Jong Nso, P. Nisha

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Traditional liquid-state fermentation processes of Annona muricata L. juice can result in fluctuating product quality and quantity due to difficulties in control and scale up. This work describes a laboratory-scale batch fermentation process to produce a probiotic Annona muricata L. enzymatically extracted juice, which was modeled using the Doehlert design with independent extraction factors being incubation time, temperature, and enzyme concentration. It aimed at a better understanding of the traditional process as an initial step for future optimization. Annona muricata L. juice was fermented with L. acidophilus (NCDC 291) (LA), L. casei (NCDC 17) (LC), and a blend of LA and LC (LCA) for 72 h at 37 °C. Experimental data were fitted into mathematical models (Monod, Logistic and Luedeking and Piret models) using MATLAB software, to describe biomass growth, sugar utilization, and organic acid production. The optimal fermentation time was obtained based on cell viability, which was 24 h for LC and 36 h for LA and LCA. The model was particularly effective in estimating biomass growth, reducing sugar consumption, and lactic acid production. The values of the determination coefficient, R2, were 0.9946, 0.9913 and 0.9946, while the residual sum of square error, SSE, was 0.2876, 0.1738 and 0.1589 for LC, LA and LCA, respectively. The growth kinetic parameters included the maximum specific growth rate, µm, which was 0.2876 h-1, 0.1738 h-1 and 0.1589 h-1 as well as the substrate saturation, Ks, with 9.0680 g/L, 9.9337 g/L and 9.0709 g/L respectively for LC, LA and LCA. For the stoichiometric parameters, the yield of biomass based on utilized substrate (YXS) was 50.7932, 3.3940 and 61.0202, and the yield of product based on utilized substrate (YPS) was 2.4524, 0.2307 and 0.7415 for LC, LA, and LCA, respectively. In addition, the maintenance energy parameter (ms) was 0.0128, 0.0001 and 0.0004 with respect to LC, LA and LCA. With the kinetic model proposed by Luedeking and Piret for lactic acid production rate, the growth associated, and non-growth associated coefficients were determined as 1.0028 and 0.0109, respectively. The model was demonstrated for batch growth of LA, LC, and LCA in Annona muricata L. juice. The present investigation validates the potential of Annona muricata L. based medium for heightened economical production of a probiotic medium.

Keywords: L. acidophilus, L. casei, fermentation, modelling, kinetics

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Authors: Vimal Kumar Dewangan, T. S. Sampath Kumar, Mukesh Doble, Viju Daniel Varghese

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The apatite-based, i.e., calcium-deficient hydroxyapatite (CDHAp) bone cement is well-known potential bone graft/substitute in orthopaedics due to its similar chemical composition with natural bone minerals. Therefore, an easy approach was attempted to fabricate the apatite-based (CDHAp) bone cement with improved injectability, bioresorbability, and macroporosity. In this study, the desired bone cement was developed by mixing the solid phase (consisting of wet chemically synthesized nanocrystalline hydroxyapatite and commercially available (synthetic) tricalcium phosphate) and the liquid phase (consisting of cement binding accelerator with few biopolymers in a dilute acidic solution) along with a liquid porogen as polysorbate or a solid porogen as mannitol (for comparison) in an optimized liquid-to-powder ratio. The fabricated cement sets within clinically preferred setting time (≤20 minutes) are better injectable (>70%) and also stable at ~7.3-7.4 (physiological pH). The CDHAp phased bone cement was resulted by immersing the fabricated after-set cement in phosphate buffer solution and other similar artificial body fluids and incubated at physiological conditions for seven days, confirmed through the X-ray diffraction and Fourier transform-infrared spectroscopy analyses. The so-formed synthetic apatite-based bone cement holds the acceptable compressive strength (within the range of trabecular bone) with average interconnected pores size falls in a macropores range (~50-200μm) inside the cement, verified by scanning electron microscopy (SEM), mercury intrusion porosimetry and micro-CT analysis techniques. Also, it is biodegradable (degrades ~19-22% within 10-12 weeks) when incubated in artificial body fluids under physiological conditions. The biocompatibility study of the bone cement, when incubated with MG63 cells, shows a significant increase in the cell viability after 3rd day of incubation compared with the control, and the cells were well-attached and spread completely on the surface of the bone cement, confirmed through SEM and fluorescence microscopy analyses. With this all, we can conclude that the developed synthetic macroporous apatite-based bone cement may have the potential to become promising material used as a trabecular bone substitute.

Keywords: calcium deficient hydroxyapatite, synthetic apatite-based bone cement, injectability, macroporosity, trabecular bone substitute

Procedia PDF Downloads 87

Authors: Christoph Hacker, Zeynep Karahaliloglu, Gunnar Seide, Emir Baki Denkbas, Thomas Gries

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A wound dressing material is designed to facilitate wound healing and minimize scarring. An ideal wound dressing material should protect the wound from any contaminations of exogeneous microorganism. In addition, the dressing material should provide a moist environment through extraction of body fluid from the wound area. Recently, wound dressing electrospun nanofibrous membranes are produced by electrospinning from a polymer solution or a polymer melt. These materials have a great potential as dressing materials for wound healing because of superior properties such as high surface-to-volume ratio, high porosity with excellent pore interconnectivity. Melt electrospinning is an attractive tissue engineering scaffold manufacturing process which eliminated the health risk posed by organic solvents used in electrospinning process and reduced the production costs. In this study, antibacterial wound dressing materials were prepared from TPU (Elastollan 1185A) by a melt-electrospinning technique. The electrospinning parameters for an efficient melt-electrospinning process of TPU were optimized. The surface of the fibers was modified with poly(ethylene glycol) (PEG) by radio-frequency glow discharge plasma deposition method and with silver nanoparticles (nAg) to improve their wettability and antimicrobial properties. TPU melt-electrospun mats were characterized using SEM, DSC, TGA and XPS. The cell viability and proliferation on modified melt-electrospun TPU mats were evaluated using a mouse fibroblast cell line (L929). Antibacterial effects of theirs against both Staphylococcus aureus strain and Escherichia coli were investigated by disk-diffusion method. TPU was successfully processed into a porous, fibrous network of beadless fibers in the micrometer range (4.896±0.94 µm) with a voltage of 50 kV, a working distance of 6 cm, a temperature of the thermocouple and hot coil of 225–230ºC, and a flow rate of 0.1 mL/h. The antibacterial test indicated that PEG-modified nAg-loaded TPU melt-electrospun structure had excellent antibacterial effects and cell study results demonstrated that nAg-loaded TPU mats had no cytotoxic effect on the fibroblast cells. In this work, the surface of a melt-electrospun TPU mats was modified via PEG monomer and then nAg. Results showed melt-electrospun TPU mats modified with PEG and nAg have a great potential for use as an antibacterial wound dressing material and thus, requires further investigation.

Keywords: melt electrospinning, nanofiber, silver nanoparticles, wound dressing

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Authors: Peter C. St. John, Michael F. Crowley, Yannick J. Bomble

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Production of chemicals from engineered organisms in a batch culture typically involves a trade-off between productivity, yield, and titer. However, strategies for strain design typically involve designing mutations to achieve the highest yield possible while maintaining growth viability. Such approaches tend to follow the principle of designing static networks with minimum metabolic functionality to achieve desired yields. While these methods are computationally tractable, optimum productivity is likely achieved by a dynamic strategy, in which intracellular fluxes change their distribution over time. One can use multi-stage fermentations to increase either productivity or yield. Such strategies would range from simple manipulations (aerobic growth phase, anaerobic production phase), to more complex genetic toggle switches. Additionally, some computational methods can also be developed to aid in optimizing two-stage fermentation systems. One can assume an initial control strategy (i.e., a single reaction target) in maximizing productivity - but it is unclear how close this productivity would come to a global optimum. The calculation of maximum theoretical yield in metabolic engineering can help guide strain and pathway selection for static strain design efforts. Here, we present a method for the calculation of a maximum theoretical productivity of a batch culture system. This method follows the traditional assumptions of dynamic flux balance analysis: that internal metabolite fluxes are governed by a pseudo-steady state and external metabolite fluxes are represented by dynamic system including Michealis-Menten or hill-type regulation. The productivity optimization is achieved via dynamic programming, and accounts explicitly for an arbitrary number of fermentation stages and flux variable changes. We have applied our method to succinate production in two common microbial hosts: E. coli and A. succinogenes. The method can be further extended to calculate the complete productivity versus yield Pareto surface. Our results demonstrate that nearly optimal yields and productivities can indeed be achieved with only two discrete flux stages.

Keywords: A. succinogenes, E. coli, metabolic engineering, metabolite fluxes, multi-stage fermentations, succinate

Procedia PDF Downloads 215

Authors: Shih-Huang Pan, Tsuyoshi Miyazaki, Minoru Otani, Santhanamoorthi Nachimuthu, Jyh-Chiang Jiang

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Ammonia (NH₃) is crucial in renewable energy and agriculture, yet its traditional production via the Haber-Bosch process faces challenges due to the inherent inertness of nitrogen (N₂) and the need for high temperatures and pressures. The electrocatalytic nitrogen reduction (ENRR) presents a more sustainable option, functioning at ambient conditions. However, its advancement is limited by selectivity and efficiency challenges due to the competing hydrogen evolution reaction (HER). The critical roles of protonation of N-species and HER highlight the necessity of selecting optimal catalysts and solvents to enhance ENRR performance. Notably, transition metal oxides, with their adjustable electronic states and excellent chemical and thermal stability, have shown promising ENRR characteristics. In this study, we use density functional theory (DFT) methods to investigate the ENRR mechanisms on IrO₂ (110), a material known for its tunable electronic properties and exceptional chemical and thermal stability. Employing the constant electrode potential (CEP) model, where the electrode - electrolyte interface is treated as a polarizable continuum with implicit solvation, and adjusting electron counts to equalize work functions in the grand canonical ensemble, we further incorporate the advanced 3D Reference Interaction Site Model (3D-RISM) to accurately determine the ENRR limiting potential across various solvents and pH conditions. Our findings reveal that the limiting potential for ENRR on IrO₂ (110) is significantly more favorable than for HER, highlighting the efficiency of the IrO₂ catalyst for converting N₂ to NH₃. This is supported by the optimal *NH₃ desorption energy on IrO₂, which enhances the overall reaction efficiency. Microkinetic simulations further predict a promising NH₃ production rate, even at the solution's boiling point¸ reinforcing the catalytic viability of IrO₂ (110). This comprehensive approach provides an atomic-level understanding of the electrode-electrolyte interface in ENRR, demonstrating the practical application of IrO₂ in electrochemical catalysis. The findings provide a foundation for developing more efficient and selective catalytic strategies, potentially revolutionizing industrial NH₃ production.

Keywords: density functional theory, electrocatalyst, nitrogen reduction reaction, electrochemistry

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Authors: Debasmita Misra, Arthur Nash

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Energy security and sufficiency enables the economic development and welfare of a nation or a society. Currently, the global energy system is dominated by fossil fuels, which is a non-renewable energy resource, which renders vulnerability to energy security. Hence, many nations have begun augmenting their energy system with renewable energy resources, such as solar, wind, biomass and hydro. However, with climate change, how sustainable are some of the renewable energy resources in the future is a matter of concern. Geothermal energy resources have been underexplored or underexploited in global renewable energy production and security, although it is gaining attractiveness as a renewable energy resource. The question is, whether geothermal energy resources are more sustainable than other renewable energy resources. High-temperature reservoirs (> 220 °F) can produce electricity from flash/dry steam plants as well as binary cycle production facilities. Most of the world’s high enthalpy geothermal resources are within the seismo-tectonic belt. However, exploration for geothermal energy is of great importance in conventional geothermal systems in order to improve its economic viability. In recent years, there has been an increase in the use and development of several exploration methods for geo-thermal resources, such as seismic or electromagnetic methods. The thermal infrared band of the Landsat can reflect land surface temperature difference, so the ETM+ data with specific grey stretch enhancement has been used to explore underground heat water. Another way of exploring for potential power is utilizing fairway play analysis for sites without surface expression and in rift zones. Utilizing this type of analysis can improve the success rate of project development by reducing exploration costs. Identifying the basin distribution of geologic factors that control the geothermal environment would help in identifying the control of resource concentration aside from the heat flow, thus improving the probability of success. The first step is compiling existing geophysical data. This leads to constructing conceptual models of potential geothermal concentrations which can then be utilized in creating a geodatabase to analyze risk maps. Geospatial analysis and other GIS tools can be used in such efforts to produce spatial distribution maps. The goal of this paper is to discuss how climate change may impact renewable energy resources and how could a synthesized analysis be developed for geothermal resources to ensure sustainable and cost effective exploitation of the resource.

Keywords: exploration, geothermal, renewable energy, sustainable

Procedia PDF Downloads 154

Authors: Azieva A. M., Yastremsky E. V., Kirillova D. A., Patsaev T. D., Sharikov R. V., Kamyshinsky R. A., Lukanina K. I., Sharikova N. A., Grigoriev T. E., Vasiliev A. L.

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Adhesive properties of scaffolds, which predominantly depend on the chemical and structural features of their surface, play the most important role in tissue engineering. The basic requirements for such scaffolds are biocompatibility, biodegradation, high cell adhesion, which promotes cell proliferation and differentiation. In many cases, synthetic polymers scaffolds have proven advantageous because they are easy to shape, they are tough, and they have high tensile properties. The regeneration of nerve tissue still remains a big challenge for medicine, and neural stem cells provide promising therapeutic potential for cell replacement therapy. However, experiments with stem cells have their limitations, such as low level of cell viability and poor control of cell differentiation. Whereas the study of already differentiated neuronal cell culture obtained from newborn mouse brain is limited only to cell adhesion. The growth and implantation of neuronal culture requires proper scaffolds. Moreover, the polymer scaffolds implants with neuronal cells could demand specific morphology. To date, it has been proposed to use numerous synthetic polymers for these purposes, including polystyrene, polylactic acid (PLA), polyglycolic acid, and polylactide-glycolic acid. Tissue regeneration experiments demonstrated good biocompatibility of PLA scaffolds, despite the hydrophobic nature of the compound. Problem with poor wettability of the PLA scaffold surface could be overcome in several ways: the surface can be pre-treated by poly-D-lysine or polyethyleneimine peptides; roughness and hydrophilicity of PLA surface could be increased by plasma treatment, or PLA could be combined with natural fibers, such as collagen or chitosan. This work presents a study of adhesion of both induced pluripotent stem cells (iPSCs) and mouse primary neuronal cell culture on the polylactide scaffolds of various types: oriented and non-oriented fibrous nonwoven materials and sponges – with and without the effect of plasma treatment and composites with collagen and chitosan. To evaluate the effect of different types of PLA scaffolds on the neuronal differentiation of iPSCs, we assess the expression of NeuN in differentiated cells through immunostaining. iPSCs more effectively differentiate into neurons on PLA scaffolds with high adhesive properties for primary neuronal cells.

Keywords: PLA scaffold, neurons, neuronal differentiation, stem cells, polylactid

Procedia PDF Downloads 84

Authors: Palak Patel

Abstract:

Mumbai is one of the largest cities of the country with a population of 12.44 million over 437 Sq.km, and it is known as financial hub of India. In early 20th century, with the expansion of industrialization and growth of port, a huge demand for housing was created. In response to this, government enacted rent controls. Over a period of time, due to rent controls, the existing rental housing stock has deteriorated. Therefore, in last 25 years, government has been focusing on redevelopment of these rental buildings, also called ‘Cessed buildings’, in order to provide better standard of living to the tenants and also, to supply new housing units in the market. In India, developers are the main players in the housing market as they are the supplier of maximum dwelling units in the market. Hence, government attempts are inclined toward facilitating developers for the cessed building redevelopment projects by incentivizing them through making special provisions in the development control regulations. This research focuses on the entire process of redevelopment by the developers and issues faced by the related stakeholders in the same to reduce the stress on housing. It also highlights the loopholes in the current system and inefficient functioning of the process. The research was carried out by interviewing various developers, tenants and landlords in the island city who have already gone through redevelopment. From the case studies, it is very evident that redevelopment is undoubtedly a huge profit making business. In some cases, developers make profit of almost double the amount of the investment. But yet, satisfactory results are not seen on ground. It clearly indicates that there are some issues faced by developers which have not been addressed. Some of these issues include cumbersome legal procedures, negotiations with landlords and tenants, congestion and narrow roads, small size of the plots, informal practicing of ‘Pagdi system’ and financial viability of the project. This research recommends the up gradation of the existing cessed buildings by sharing the repairing and maintenance cost between landlords and tenants and also, income levels of tenants can be traced and housing vouchers or incentives can be provided to those who actual need it so that landlord does not have to subsidize the tenants. For redevelopment, the current interventions are generalized in nature as it does not take on ground issues into the consideration. There is need to identify local issues and give area specific solutions. And also, government should play a role of mediator to ensure all the stakeholders are satisfied and project gets completed on time.

Keywords: cessed buildings, developers, government’s interventions, redevelopment, rent controls, tenants

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