Search results for: reverse transcription

Authors: Michal Kielbik, Izabela Szulc-Kielbik, Anna Brzostek, Jaroslaw Dziadek, Magdalena Klink

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The goal of many research groups in the world is to find new components that are important for survival of mycobacteria in the host cells. Mycobacterium tuberculosis (Mtb) possesses a number of enzymes degrading cholesterol that are considered to be an important factor for its survival and persistence in host macrophages. One of them - cholesterol oxidase (ChoD), although not being essential for cholesterol degradation, is discussed as a virulence compound, however its involvement in macrophages’ response to Mtb is still not sufficiently determined. The recognition of tubercle bacilli antigens by pathogen recognition receptors is crucial for the initiation of the host innate immune response. An important receptor that has been implicated in the recognition and/or uptake of Mtb is Toll-like receptor type 2 (TLR2). Engagement of TLR2 results in the activation and phosphorylation of intracellular signaling proteins including IRAK-1 and -4, TRAF-6, which in turn leads to the activation of target kinases and transcription factors responsible for bactericidal and pro-inflammatory response of macrophages. The aim of these studies was a detailed clarification of the role of Mtb cholesterol oxidase as a virulence factor affecting the TLR2 signaling pathway in human macrophages. As human macrophages the THP-1 differentiated cells were applied. The virulent wild-type Mtb strain (H37Rv), its mutant lacking a functional copy of gene encoding cholesterol oxidase (∆choD), as well as complimented strain (∆choD–choD) were used. We tested the impact of Mtb strains on the expression of TLR2-depended signaling proteins (mRNA level, cytosolic level and phosphorylation status). The cytokine and bactericidal response of THP-1 derived macrophages infected with Mtb strains in relation to TLR2 signaling pathway dependence was also determined. We found that during the 24-hours of infection process the wild-type and complemented Mtb significantly reduced the cytosolic level and phosphorylation status of IRAK-4 and TRAF-6 proteins in macrophages, that was not observed in the case of ΔchoD mutant. Decreasement of TLR2-dependent signaling proteins, induced by wild-type Mtb, was not dependent on the activity of proteasome. Blocking of TLR2 expression, before infection, effectively prevented the induced by wild-type strain reduction of cytosolic level and phosphorylation of IRAK-4. None of the strains affected the surface expression of TLR2. The mRNA level of IRAK-4 and TRAF-6 genes were significantly increased in macrophages 24 hours post-infection with either of tested strains. However, the impact of wild-type Mtb strain on both examined genes was significantly stronger than its ΔchoD mutant. We also found that wild-type strain stimulated macrophages to release high amount of immunosuppressive IL-10, accompanied by low amount of pro-inflammatory IL-8 and bactericidal nitric oxide in comparison to mutant lacking cholesterol oxidase. The influence of wild-type Mtb on this type of macrophages' response strongly dependent on fully active IRAK-1 and IRAK-4 signaling proteins. In conclusion, Mtb using cholesterol oxidase causes the over-activation of TLR2 signaling proteins leading to the reduction of their cytosolic level and activity resulting in the modulation of macrophages response to allow its intracellular survival. Supported by grant: 2014/15/B/NZ6/01565, National Science Center, Poland

Keywords: Mycobacterium tuberculosis, cholesterol oxidase, macrophages, TLR2-dependent signaling pathway

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Authors: Bekim Berisha, Sebastian Hirsiger, Pavel Hora

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Advanced material models involving several sets of model parameters require a big experimental effort. As models are getting more and more complex like e.g. the so called “Homogeneous Anisotropic Hardening - HAH” model for description of the yielding behavior in the 2D/3D stress space, the number and complexity of the required experiments are also increasing continuously. In the context of sheet metal forming, these requirements are even more pronounced, because of the anisotropic behavior or sheet materials. In addition, some of the experiments are very difficult to perform e.g. the plane stress biaxial compression test. Accordingly, tensile tests in at least three directions, biaxial tests and tension-compression or shear-reverse shear experiments are performed to determine the parameters of the macroscopic models. Therefore, determination of the macroscopic model parameters based on virtual experiments is a very promising strategy to overcome these difficulties. For this purpose, in the framework of multiscale material modeling, a dislocation density based crystal plasticity model in combination with a FFT-based spectral solver is applied to perform virtual experiments. Modeling of the plastic behavior of metals based on crystal plasticity theory is a well-established methodology. However, in general, the computation time is very high and therefore, the computations are restricted to simplified microstructures as well as simple polycrystal models. In this study, a dislocation density based crystal plasticity model – including an implementation of the backstress – is used in a spectral solver framework to generate virtual experiments for three deep drawing materials, DC05-steel, AA6111-T4 and AA4045 aluminum alloys. For this purpose, uniaxial as well as multiaxial loading cases, including various pre-strain histories, has been computed and validated with real experiments. These investigations showed that crystal plasticity modeling in the framework of Representative Volume Elements (RVEs) can be used to replace most of the expensive real experiments. Further, model parameters of advanced macroscopic models like the HAH model can be determined from virtual experiments, even for multiaxial deformation histories. It was also found that crystal plasticity modeling can be used to model anisotropic hardening more accurately by considering the backstress, similar to well-established macroscopic kinematic hardening models. It can be concluded that an efficient coupling of crystal plasticity models and the spectral solver leads to a significant reduction of the amount of real experiments needed to calibrate macroscopic models. This advantage leads also to a significant reduction of computational effort needed for the optimization of metal forming process. Further, due to the time efficient spectral solver used in the computation of the RVE models, detailed modeling of the microstructure are possible.

Keywords: anisotropic hardening, crystal plasticity, micro structure, spectral solver

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Authors: Khiem M. Nguyen, Ming C. Yang

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Chloroplast pigments are extremely important during photosynthesis since they play essential roles in light absorption and energy transfer. Therefore, understanding the efficiency of chlorophyll (Chl) biosynthesis could facilitate enhancement in photo-assimilates accumulation, and ultimately, in crop yield. The Chl-deficient mutants have been used extensively to study the Chl biosynthetic pathways and the biogenesis of the photosynthetic apparatus. Rice (Oryza sativa L.) is one of the most leading food crops, serving as staple food for many parts of the world. To author’s best knowledge, Chl b–lacking rice has been found; however the molecular mechanism of Chl biosynthesis still remains unclear compared to wild-type rice. In this study, the ultrastructure analysis, photosynthetic properties, and transcriptome profile of wild-type rice (Norin No.8, N8) and its Chl b-lacking mutant (Chlorina 1, C1) were examined. The finding concluded that total Chl content and Chl b content in the C1 leaves were strongly reduced compared to N8 leaves, suggesting that reduction in the total Chl content contributes to leaf color variation at the physiological level. Plastid ultrastructure of C1 possessed abnormal thylakoid membranes with loss of starch granule, large number of vesicles, and numerous plastoglobuli. The C1 rice also exhibited thinner stacked grana, which was caused by a reduction in the number of thylakoid membranes per granum. Thus, the different Chl a/b ratio of C1 may reflect the abnormal plastid development and function. Transcriptional analysis identified 23 differentially expressed genes (DEGs) and 671 transcription factors (TFs) that were involved in Chl metabolism, chloroplast development, cell division, and photosynthesis. The transcriptome profile and DEGs revealed that the gene encoding PsbR (PSII core protein) was down-regulated, therefore suggesting that the lower in light-harvesting complex proteins are responsible for the lower photosynthetic capacity in C1. In addition, expression level of cell division protein (FtsZ) genes were significantly reduced in C1, causing chloroplast division defect. A total of 19 DEGs were identified based on KEGG pathway assignment involving Chl biosynthesis pathway. Among these DEGs, the GluTR gene was down-regulated, whereas the UROD, CPOX, and MgCH genes were up-regulated. Observation through qPCR suggested that later stages of Chl biosynthesis were enhanced in C1, whereas the early stages were inhibited. Plastid structure analysis together with transcriptomic analysis suggested that the Chl a/b ratio was amplified both by the reduction in Chl contents accumulation, owning to abnormal chloroplast development, and by the enhanced conversion of Chl b to Chl a. Moreover, the results indicated the same Chl-cycle pattern in the wild-type and C1 rice, indicating another Chl b degradation pathway. Furthermore, the results demonstrated that normal grana stacking, along with the absence of Chl b and greatly reduced levels of Chl a in C1, provide evidence to support the conclusion that other factors along with LHCII proteins are involved in grana stacking. The findings of this study provide insight into the molecular mechanisms that underlie different Chl a/b ratios in rice.

Keywords: Chl-deficient mutant, grana stacked, photosynthesis, RNA-Seq, transcriptomic analysis

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Authors: Wisam H. Benamer, Ehab A. Elfallah, Mohamed A. Elshaari, Farag A. Elshaari

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A challenge for laboratories is to provide bacterial identification and antibiotic sensitivity results within a short time. Hence, advancement in the required technology is desirable to improve timing, accuracy and quality. Even with the current advances in methods used for both phenotypic and genotypic identification of bacteria the need is there to develop method(s) that enhance the outcome of bacteriology laboratories in accuracy and time. The hypothesis introduced here is based on the assumption that the chromosome of any bacteria contains unique sequences that can be used for its identification and typing. The outcome of a pilot study designed to test this hypothesis is reported in this manuscript. Methods: The complete chromosome sequences of several bacterial species were downloaded to use as search targets for unique sequences. Visual basic and SQL server (2014) were used to generate a complete set of 18-base long primers, a process started with reverse translation of randomly chosen 6 amino acids to limit the number of the generated primers. In addition, the software used to scan the downloaded chromosomes using the generated primers for similarities was designed, and the resulting hits were classified according to the number of similar chromosomal sequences, i.e., unique or otherwise. Results: All primers that had identical/similar sequences in the selected genome sequence(s) were classified according to the number of hits in the chromosomes search. Those that were identical to a single site on a single bacterial chromosome were referred to as unique. On the other hand, most generated primers sequences were identical to multiple sites on a single or multiple chromosomes. Following scanning, the generated primers were classified based on ability to differentiate between medically important bacterial and the initial results looks promising. Conclusion: A simple strategy that started by generating primers was introduced; the primers were used to screen bacterial genomes for match. Primer(s) that were uniquely identical to specific DNA sequence on a specific bacterial chromosome were selected. The identified unique sequence can be used in different molecular diagnostic techniques, possibly to identify bacteria. In addition, a single primer that can identify multiple sites in a single chromosome can be exploited for region or genome identification. Although genomes sequences draft of isolates of organism DNA enable high throughput primer design using alignment strategy, and this enhances diagnostic performance in comparison to traditional molecular assays. In this method the generated primers can be used to identify an organism before the draft sequence is completed. In addition, the generated primers can be used to build a bank for easy access of the primers that can be used to identify bacteria.

Keywords: bacteria chromosome, bacterial identification, sequence, primer generation

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Authors: E. M. Condori-Peñaloza, S. S. Costa

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Eleusine indica L, known as goose-grass, is considered a troublesome weed that can cause important economic losses in the agriculture worldwide. However, this grass is used as a medicinal plant in some regions of Brazil to treat influenza and pneumonia. In Africa and Asia, it is used to treat malaria and as diuretic, anti-helminthic, among other uses. Despite its therapeutic potential, little is known about the chemical composition and bioactive compounds of E. indica. Hitherto, two major flavonoids, schaftoside and vitexin, were isolated from aerial part of the species and showed inhibitory activity on lung neutrophil influxes in mice, suggesting a beneficial effect on airway inflammation. Therefore, the aim of this study was to analyze the chemical profile of aqueous extracts from aerial parts of Eleusine indica specimens using high performance liquid chromatography (HPLC-DAD) and 1H nuclear magnetic resonance spectroscopy (NMR), with emphasis on phenolic compounds. Specimens of E. indica were collected in Minas Gerais state, Brazil. Aerial parts of fresh plants were extracted by decoction (10% p/v). After spontaneous precipitation of the aqueous extract at 10-12°C for 24 hours, the supernatant obtained was frozen and lyophilized. After that, 1 g of the extract was dissolved into 25 mL of water and fractionated on a reverse phase chromatography column (RP-2), eluted with a gradient of H2O/EtOH. Five fractions were obtained. The extract and fractions had their chemical profile analyzed by using HPLC-DAD (C-18 column: 20 μL, 256 -365 nm; gradient water 0.01% phosphoric acid/ acetonitrile. The extract was also analyzed by NMR (1H, 500 MHz, D2O) in order to access its global chemical composition. HPLC-DAD analyses of crude extract allowed the identification of ten phenolic compounds. Fraction 1, eluted with 100% water, was poor in phenolic compounds and no major peak was detected. In fraction 2, eluted with 100% water, it was possible to observe one major peak at retention time (RT) of 23.75 minutes compatible with flavonoid; fraction 3, also eluted with 100% water, showed four peaks at RT= 21.47, 23.52, 24.33 and 25.84 minutes, all of them compatible with flavonoid. In fraction 4, eluted with 50%/ethanol/50% water, it was possible to observe 3 peaks compatible with flavonoids at RT=24.65, 26.81, 27.49 minutes, and one peak (28.83 min) compatible with a phenolic acid derivative. Finally, in fraction 5, eluted with 100% ethanol, no phenolic substance was detected. The UV spectra of all flavonoids detected were compatible with the flavone subclass (λ= 320-345 nm). The 1H NMR spectra of aerial parts extract showed signals in three regions: δ 0.8-3.0 ppm (aliphatic compounds), δ 3.0-5.5 ppm corresponding to carbohydrates (signals most abundant and overlapped), and δ 6.0-8.5 ppm (aromatic compounds). Signals compatible with flavonoids (rings A and B) could also be detected in the crude extract spectra. These results suggest the presence of several flavonoids in E. indica, which reinforces their therapeutic potential. The pharmacological activities of Eleusine indica extracts and fractions will be further evaluated.

Keywords: flavonoids, HPLC, NMR, phenolic compounds

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Authors: Mohammad Hassan Ziraksaz

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Fuel atomizers are used in a wide range of IC engines, turbojets and a variety of liquid propellant rocket engines. As the fuel spray fully develops its characters approach their ultimate amounts. Fuel spray characters such as SMD, injection pressure, mass flow rate, droplet velocity and spray cone angle play important roles to atomize the liquid fuel to finely atomized fuel droplets and finally form the fine fuel spray. Well performed, fully developed, fine spray without any defections, brings the idea of finding an experimental relation between the main effective spray characters. Extracting an experimental relation between SMD and other fuel spray physical characters in swirl fuel atomizers is the main scope of this experimental work. Droplet velocity, fuel mass flow rate, SMD and spray cone angle are the parameters which are measured. A set of twelve reverse engineering atomizers without any spray defections and a set of eight original atomizers as referenced well-performed spray are contributed in this work. More than 350 tests, mostly repeated, were performed. This work shows that although spray cone angle plays a very effective role in spray formation, after formation, it smoothly approaches to an almost constant amount while the other characters are changed to create fine droplets. Therefore, the work to find the relation between the characters is focused on SMD, droplet velocity, fuel mass flow rate, and injection pressure. The process of fuel spray formation begins in 5 Psig injection pressures, where a tiny fuel onion attaches to the injector tip and ended in 250 Psig injection pressure, were fully developed fine fuel spray forms. Injection pressure is gradually increased to observe how the spray forms. In each step, all parameters are measured and recorded carefully to provide a data bank. Various diagrams have been drawn to study the behavior of the parameters in more detail. Experiments and graphs show that the power equation can best show changes in parameters. The SMD experimental relation with pressure P, fuel mass flow rate Q ̇ and droplet velocity V extracted individually in pairs. Therefore, the proportional relation of SMD with other parameters is founded. Now it is time to find an experimental relation including all the parameters. Using obtained proportional relation, replacing the parameters with experimentally measured ones and drawing the graphs of experimental SMD versus proportion SMD (〖SMD〗_P), a correctional equation and consequently the final experimental equation is obtained. This experimental equation is specified to use for swirl fuel atomizers and the use of this experimental equation in different conditions shows about 3% error, which is expected to achieve lower error and consequently higher accuracy by increasing the number of experiments and increasing the accuracy of data collection.

Keywords: droplet velocity, experimental relation, mass flow rate, SMD, swirl fuel atomizer

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Authors: Chao Yang, Jinchuan Zhang, Yongqiang Xiong

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The Lower Cambrian Niutitang Formation shale (NFS) deposited in the marine deep-shelf environment in Southeast Upper Yangtze (SUY), possess excellent source rock basis for shale gas generation, however, it is currently challenged by being over-mature with strong tectonic deformations, leading to much uncertainty of gas-bearing potential. With emphasis on the shale gas enrichment of the NFS, analyses were made based on the regional gas-bearing differences obtained from field gas-desorption testing of 18 geological survey wells across the study area. Results show that the NFS bears low gas content of 0.2-2.5 m³/t, and the eastern region of SUY is higher than the western region in gas content. Moreover, the methane fraction also presents the similar regional differentiation with the western region less than 10 vol.% while the eastern region generally more than 70 vol.%. Through the analysis of geological theory, the following conclusions are drawn: Depositional environment determines the gas-enriching zones. In the western region, the Dengying Formation underlying the NFS in unconformity contact was mainly plateau facies dolomite with caves and thereby bears poor gas-sealing ability. Whereas the Laobao Formation underling the NFS in eastern region was a set of siliceous rocks of shelf-slope facies, which can effectively prevent the shale gas from escaping away from the NFS. The tectonic conditions control the gas-enriching bands in the SUY, which is located in the fold zones formed by the thrust of the Southern China plate towards to the Sichuan Basin. Compared with the western region located in the trough-like folds, the eastern region at the fold-thrust belts was uplifted early and deformed weakly, resulting in the relatively less mature level and relatively slight tectonic deformation of the NFS. Faults determine whether shale gas can be accumulated in large scale. Four deep and large normal faults in the study area cut through the Niutitang Formation to the Sinian strata, directly causing a large spillover of natural gas in the adjacent areas. For the secondary faults developed within the shale formation, the reverse faults generally have a positive influence on the shale accumulation while the normal faults perform the opposite influence. Overall, shale gas enrichment targets of the NFS, are the areas with certain thickness of siliceous rocks at the basement of the Niutitang Formation, and near the margin of the paleouplift with less developed faults. These findings provide direction for shale gas exploration in South China, and also provide references for the areas with similar geological conditions all over the world.

Keywords: over-mature marine shale, shale gas accumulation, structure-complicated area, Southeast Upper Yangtze

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Authors: Michael Josef Schwerer

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Background: Analyzing any aircraft accident is mandatory based on the regulations of the International Civil Aviation Organization and the respective country’s criminal prosecution authorities. Legal medicine investigations are unavoidable when fatalities involve the flight crew or when doubts arise concerning the pilot’s aeromedical health status before the event. As a result of frequently tremendous blunt and sharp force trauma along with the impact of the aircraft to the ground, consecutive blast or fire exposition of the occupants or putrefaction of the dead bodies in cases of delayed recovery, relevant findings can be masked or destroyed and therefor being inaccessible in standard pathology practice comprising just forensic autopsy and histopathology. Such cases are of considerable risk of remaining unsolved without legal consequences for those responsible. Further, no lessons can be drawn from these scenarios to improve flight safety and prevent future mishaps. Aims and Methods: To learn from previously unsolved aircraft accidents, re-evaluations of the investigation files and modern molecular pathology studies were performed. Genetic testing involved predominantly PCR-based analysis of gene regulation, studying DNA promotor methylations, RNA transcription and posttranscriptional regulation. In addition, the presence or absence of infective agents, particularly DNA- and RNA-viruses, was studied. Technical adjustments of molecular genetic procedures when working with archived sample material were necessary. Standards for the proper interpretation of the respective findings had to be settled. Results and Discussion: Additional molecular genetic testing significantly contributes to the quality of forensic pathology assessment in aviation mishaps. Previously undetected cardiotropic viruses potentially explain e.g., a pilot’s sudden incapacitation resulting from cardiac failure or myocardial arrhythmia. In contrast, negative results for infective agents participate in ruling out concerns about an accident pilot’s fitness to fly and the aeromedical examiner’s precedent decision to issue him or her an aeromedical certificate. Care must be taken in the interpretation of genetic testing for pre-existing diseases such as hypertrophic cardiomyopathy or ischemic heart disease. Molecular markers such as mRNAs or miRNAs, which can establish these diagnoses in clinical patients, might be misleading in-flight crew members because of adaptive changes in their tissues resulting from repeated mild hypoxia during flight, for instance. Military pilots especially demonstrate significant physiological adjustments to their somatic burdens in flight, such as cardiocirculatory stress and air combat maneuvers. Their non-pathogenic alterations in gene regulation and expression will likely be misinterpreted for genuine disease by inexperienced investigators. Conclusions: The growing influence of molecular pathology on legal medicine practice has found its way into aircraft accident investigation. As appropriate quality standards for laboratory work and data interpretation are provided, forensic genetic testing supports the medico-legal analysis of aviation mishaps and potentially reduces the number of unsolved events in the future.

Keywords: aviation medicine, aircraft accident investigation, forensic pathology, molecular pathology

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Authors: Thanthrige Thiunuwan Priyathilaka, Don Anushka Sandaruwan Elvitigala, Bong-Soo Lim, Hyung-Bok Jeong, Jehee Lee

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Toll like receptors (TLRs) are a phylogeneticaly conserved family of pattern recognition receptors, which participates in the host immune responses against various pathogens and pathogen derived mitogen. TLR21, a non-mammalian type, is almost restricted to the fish species even though those can be identified rarely in avians and amphibians. Herein, this study was carried out to identify and characterize TLR21 from rock bream (Oplegnathus fasciatus) designated as RbTLR21, at transcriptional and genomic level. In this study, the full length cDNA and genomic sequence of RbTLR21 was identified using previously constructed cDNA sequence database and BAC library, respectively. Identified RbTLR21 sequence was characterized using several bioinformatics tools. The quantitative real time PCR (qPCR) experiment was conducted to determine tissue specific expressional distribution of RbTLR21. Further, transcriptional modulation of RbTLR21 upon the stimulation with Streptococcus iniae (S. iniae), rock bream iridovirus (RBIV) and Edwardsiella tarda (E. tarda) was analyzed in spleen tissues. The complete coding sequence of RbTLR21 was 2919 bp in length which can encode a protein consisting of 973 amino acid residues with molecular mass of 112 kDa and theoretical isoelectric point of 8.6. The anticipated protein sequence resembled a typical TLR domain architecture including C-terminal ectodomain with 16 leucine rich repeats, a transmembrane domain, cytoplasmic TIR domain and signal peptide with 23 amino acid residues. Moreover, protein folding pattern prediction of RbTLR21 exhibited well-structured and folded ectodomain, transmembrane domain and cytoplasmc TIR domain. According to the pair wise sequence analysis data, RbTLR21 showed closest homology with orange-spotted grouper (Epinephelus coioides) TLR21with 76.9% amino acid identity. Furthermore, our phylogenetic analysis revealed that RbTLR21 shows a close evolutionary relationship with its ortholog from Danio rerio. Genomic structure of RbTLR21 consisted of single exon similar to its ortholog of zebra fish. Sevaral putative transcription factor binding sites were also identified in 5ʹ flanking region of RbTLR21. The RBTLR 21 was ubiquitously expressed in all the tissues we tested. Relatively, high expression levels were found in spleen, liver and blood tissues. Upon induction with rock bream iridovirus, RbTLR21 expression was upregulated at the early phase of post induction period even though RbTLR21 expression level was fluctuated at the latter phase of post induction period. Post Edwardsiella tarda injection, RbTLR transcripts were upregulated throughout the experiment. Similarly, Streptococcus iniae induction exhibited significant upregulations of RbTLR21 mRNA expression in the spleen tissues. Collectively, our findings suggest that RbTLR21 is indeed a homolog of TLR21 family members and RbTLR21 may be involved in host immune responses against bacterial and DNA viral infections.

Keywords: rock bream, toll like receptor 21 (TLR21), pattern recognition receptor, genomic characterization

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Authors: Lam Hong Lan, Gabor Sarlos

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It is hard to search for best practices in promotion for local fashion brands in Vietnam as the industry is still very young. Local fashion start-ups have grown quickly in the last five years, thanks in part to the internet and social media. However, local designer/owners can face a huge challenge when competing with international brands in the Vietnamese market – and few local case studies are available for guidance. In response, this paper studied how local small- to medium-sized enterprises (SMEs) promote to their target customers in order to compete with international brands. Knowledge of both successful and unsuccessful approaches generated by this study is intended to both contribute to the academic literature on local fashion in Vietnam as well as to help local designers to learn from and improve their brand-building strategy. The primary study featured qualitative data collection via semi-structured depth interviews. Transcription and data analysis were conducted manually in order to identify success factors that local brands should consider as part of their promotion strategy. Purposive sampling of SMEs identified five designers in Ho Chi Minh City (the biggest city in Vietnam) and three designers in Hanoi (the second biggest) as interviewees. Participant attributes included: born in the 1980s or 1990s; familiar with internet and social media; designer/owner of a successful local fashion brand in the key middle market and/or mass market segments (which are crucial to the growth of local brands). A secondary study was conducted using social listening software to gather further qualitative data on what were considered to be successful or unsuccessful approaches to local fashion brand promotion on social media. Both the primary and secondary studies indicated that local designers had maximized their promotion budget by using owned media and earned media instead of paid media. Findings from the qualitative interviews indicate that internet and social media have been used as effective promotion platforms by local fashion start-ups. Facebook and Instagram were the most popular social networks used by the SMEs interviewed, and these social platforms were believed to offer a more affordable promotional strategy than traditional media such as TV and/or print advertising. Online stores were considered an important factor in helping the SMEs to reach customers beyond the physical store. Furthermore, a successful online store allowed some SMEs to reduce their business rental costs by maintaining their physical store in a cheaper, less central city area as opposed to a more traditional city center store location. In addition, the small comparative size of the SMEs allowed them to be more attentive to their customers, leading to higher customer satisfaction and rate of return. In conclusion, this study found that these kinds of cost savings helped the SMEs interviewed to focus their scarce resources on producing unique, high-quality collections in order to differentiate themselves from international brands. Facebook and Instagram were the main platforms used for promotion and brand-building. The main challenge to this promotion strategy identified by the SMEs interviewed was to continue to find innovative ways to maximize the impact of a limited marketing budget.

Keywords: Vietnam, SMEs, fashion brands, promotion, marketing, social listening

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Authors: Iman Janghorban Esfahani

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Due to the increased use of irrigation in agriculture, the importance and need for highly reliable water pumping systems have significantly increased. The pumping of the groundwater is essential to provide water for both drip and furrow irrigation to increase the agricultural yield, especially in arid regions that suffer from scarcities of surface water. The most common irrigation pumping systems (IPS) consume conventional energies through the use of electric motors and generators or connecting to the electricity grid. Due to the shortage and transportation difficulties of fossil fuels, and unreliable access to the electricity grid, especially in the rural areas, and the adverse environmental impacts of fossil fuel usage, such as greenhouse gas (GHG) emissions, the need for renewable energy sources such as photovoltaic systems (PVS) as an alternative way of powering irrigation pumping systems is urgent. Integration of the photovoltaic systems with irrigation pumping systems as the Photovoltaic Powered-Irrigation Pumping System (PVP-IPS) can avoid fossil fuel dependency and the subsequent greenhouse gas emissions, as well as ultimately lower energy costs and improve efficiency, which made PVP-IPS systems as an environmentally and economically efficient solution for agriculture irrigation in every region. The greatest problem faced by integration of PVP with IPS systems is matching the intermittence of the energy supply with the dynamic water demand. The best solution to overcome the intermittence is to incorporate a storage system into the PVP-IPS to provide water-on-demand as a highly reliable stand-alone irrigation pumping system. The water storage tank (WST) is the most common storage device for PVP-IPS systems. In the integrated PVP-IPS with a water storage tank (PVP-IPS-WST), a water storage tank stores the water pumped by the IPS in excess of the water demand and then delivers it when demands are high. The Freshwater pinch analysis (FWaPA) as an alternative to mathematical modeling was used by other researchers for retrofitting the off-grid battery less photovoltaic-powered reverse osmosis system. However, the Freshwater pinch analysis has not been used to integrate the photovoltaic systems with irrigation pumping system with water storage tanks. In this study, FWaPA graphical and numerical tools were used for retrofitting an existing PVP-IPS system located in Salahadin, Republic of Iraq. The plant includes a 5 kW submersible water pump and 7.5 kW solar PV system. The Freshwater Composite Curve as the graphical tool and Freashwater Storage Cascade Table as the numerical tool were constructed to determine the minimum required outsourced water during operation, optimal amount of delivered electricity to the water pump, and optimal size of the water storage tank for one-year operation data. The results of implementing the FWaPA on the case study show that the PVP-IPS system with a WST as the reliable system can reduce outsourced water by 95.41% compare to the PVP-IPS system without storage tank.

Keywords: irrigation, photovoltaic, pinch analysis, pumping, solar energy

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Authors: Malika Imoula, Rachid Saci, Renee Gatignol

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A numerical investigation is carried out to analyze vortex flows in a free surface cylinder, driven by the independent rotation and differentially heated boundaries. As a basic uncontrolled isothermal flow, we consider configurations which exhibit steady axisymmetric toroidal type vortices which occur at the free surface; under given rates of the bottom disk uniform rotation and for selected aspect ratios of the enclosure. In the isothermal case, we show that sidewall differential rotation constitutes an effective kinematic means of flow control: the reverse flow regions may be suppressed under very weak co-rotation rates, while an enhancement of the vortex patterns is remarked under weak counter-rotation. However, in this latter case, high rates of counter-rotation reduce considerably the strength of the meridian flow and cause its confinement to a narrow layer on the bottom disk, while the remaining bulk flow is diffusion dominated and controlled by the sidewall rotation. The main control parameters in this case are the rotational Reynolds number, the cavity aspect ratio and the rotation rate ratio defined. Then, the study proceeded to consider the sensitivity of the vortex pattern, within the Boussinesq approximation, to a small temperature gradient set between the ambient fluid and an axial thin rod mounted on the cavity axis. Two additional parameters are introduced; namely, the Richardson number Ri and the Marangoni number Ma (or the thermocapillary Reynolds number). Results revealed that reducing the rod length induces the formation of on-axis bubbles instead of toroidal structures. Besides, the stagnation characteristics are significantly altered under the combined effects of buoyant-thermocapillary convection. Buoyancy, induced under sufficiently high Ri, was shown to predominate over the thermocapillay motion; causing the enhancement (suppression) of breakdown when the rod is warmer (cooler) than the ambient fluid. However, over small ranges of Ri, the sensitivity of the flow to surface tension gradients was clearly evidenced and results showed its full control over the occurrence and location of breakdown. In particular, detailed timewise evolution of the flow indicated that weak thermocapillary motion was sufficient to prevent the formation of toroidal patterns. These latter detach from the surface and undergo considerable size reduction while moving towards the bulk flow before vanishing. Further calculations revealed that the pattern reappears with increasing time as steady bubble type on the rod. However, in the absence of the central rod and also in the case of small rod length l, the flow evolved into steady state without any breakdown.

Keywords: buoyancy, cylinder, surface tension, toroidal vortex

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Authors: Yasinta Ratna Esti Wulandari, Vivitri Dewi Prasasty, Adrianus Rio, Cindy Geniola

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Mulberry is a plant that widely cultivated around the world, mostly for silk industry. In recent years, the study showed that the mulberry leaves have an anti-diabetic effect which mostly comes from the compound known as 1-deoxynojirimycin (DNJ). DNJ is a very potent α-glucosidase inhibitor. It will decrease the degradation rate of carbohydrates in digestive tract, leading to slower glucose absorption and reducing the post-prandial glucose level significantly. The mulberry leaves also known as the best source of DNJ. Since then, the DNJ in mulberry leaves had received a considerable attention, because of the increased number of diabetic patients and the raise of people awareness to find a more natural cure for diabetic. The DNJ content in mulberry leaves varied depend on the mulberry species, leaf’s age, and the plant’s growth environment. Few of the mulberry varieties that were cultivated in Indonesiaare Morus alba var. kanva-2, M. alba var. multicaulis, M. bombycis var. lembang, and M. cathayana. The lack of data concerning phytochemicals contained in the Indonesian mulberry leaves are restraining their use in the medicinal field. The aim of this study is to fully utilize the use of mulberry leaves cultivated in Indonesia as a medicinal herb in local, national, or global community, by determining the DNJ and other phytochemical contents in them. This study used eight leaf samples which are the young leaves and mature leaves of both Morus alba var. kanva-2, M. alba var. multicaulis, M. bombycis var. lembang, and M. cathayana. The DNJ content was analyzed using reverse phase high performance liquid chromatography (HPLC). The stationary phase was silica C18 column and the mobile phase was acetonitrile:acetic acid 0.1% 1:1 with elution rate 1 mL/min. Prior to HPLC analysis the samples were derivatized with FMOC to ensure the DNJ detectable by VWD detector at 254 nm. Results showed that the DNJ content in samples are ranging from 2.90-0.07 mg DNJ/ g leaves, with the highest content found in M. cathayana mature leaves (2.90 ± 0.57 mg DNJ/g leaves). All of the mature leaf samples also found to contain higher amount of DNJ from their respective young leaf samples. The phytochemicals in leaf samples was tested using qualitative test. Result showed that all of the eight leaf samples contain alkaloids, phenolics, flavonoids, tannins, and terpenes. The presence of this phytochemicals contribute to the therapeutic effect of mulberry leaves. The pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS) analysis was also performed to the eight samples to quantitatively determine their phytochemicals content. The pyrolysis temperature was set at 400 °C, with capillary column Phase Rtx-5MS 60 × 0.25 mm ID stationary phase and helium gas mobile phase. Few of the terpenes found are known to have anticancer and antimicrobial properties. From all the results, all of four samples of mulberry leaves which are cultivated in Indonesia contain DNJ and various phytochemicals like alkaloids, phenolics, flavonoids, tannins, and terpenes which are beneficial to our health.

Keywords: Morus, 1-deoxynojirimycin, HPLC, Py-GC-MS

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Authors: Irina P. Starodubtseva, Aleksandr N. Pavlenko

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Quenching is a term generally accepted for the process of rapid cooling of a solid that is overheated above the thermodynamic limit of the liquid superheat. The main objective of many previous studies on quenching is to find a way to reduce the total time of the transient process. Computational experiments were performed to simulate quenching by a falling liquid nitrogen film of an extremely overheated vertical copper plate with a structured capillary-porous coating. The coating was produced by directed plasma spraying. Due to the complexities in physical pattern of quenching from chaotic processes to phase transition, the mechanism of heat transfer during quenching is still not sufficiently understood. To our best knowledge, no information exists on when and how the first stable liquid-solid contact occurs and how the local contact area begins to expand. Here we have more models and hypotheses than authentically established facts. The peculiarities of the quench front dynamics and heat transfer in the transient process are studied. The created numerical model determines the quench front velocity and the temperature fields in the heater, varying in space and time. The dynamic pattern of the running quench front obtained numerically satisfactorily correlates with the pattern observed in experiments. Capillary-porous coatings with straight and reverse orientation of crests are investigated. The results show that the cooling rate is influenced by thermal properties of the coating as well as the structure and geometry of the protrusions. The presence of capillary-porous coating significantly affects the dynamics of quenching and reduces the total quenching time more than threefold. This effect is due to the fact that the initialization of a quench front on a plate with a capillary-porous coating occurs at a temperature significantly higher than the thermodynamic limit of the liquid superheat, when a stable solid-liquid contact is thermodynamically impossible. Waves present on the liquid-vapor interface and protrusions on the complex micro-structured surface cause destabilization of the vapor film and the appearance of local liquid-solid micro-contacts even though the average integral surface temperature is much higher than the liquid superheat limit. The reliability of the results is confirmed by direct comparison with experimental data on the quench front velocity, the quench front geometry, and the surface temperature change over time. Knowledge of the quench front velocity and total time of transition process is required for solving practically important problems of nuclear reactors safety.

Keywords: capillary-porous coating, heat transfer, Leidenfrost phenomenon, numerical simulation, quenching

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Authors: Hui-Chieh Li

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Circular economy promotes greater resource productivity and avoids pollution through greater recycling and re-use which bring benefits for both the environment and the economy. The concept is contrast to a linear economy which is ‘take, make, dispose’ model of production. A well-design reverse logistics service strategy could enhance the willingness of recycling of the users and reduce the related logistics cost as well as carbon emissions. Moreover, the recycle brings the manufacturers most advantages as it targets components for closed-loop reuse, essentially converting materials and components from worn-out product into inputs for new ones at right time and right place. This study considers demand-supply interaction, time-dependent recycle demand, time-dependent surplus value of recycled product and constructs models on recycle service strategy for the recyclable waste collector. A crucial factor in optimizing a recycle service strategy is consumer demand. The study considers the relationships between consumer demand towards recycle and product characteristics, surplus value and user behavior. The study proposes a recycle service strategy which differs significantly from the conventional and typical uniform service strategy. Periods with considerable demand and large surplus product value suggest frequent and short service cycle. The study explores how to determine a recycle service strategy for recyclable waste collector in terms of service cycle frequency and duration and vehicle type for all service cycles by considering surplus value of recycled product, time-dependent demand, transportation economies and demand-supply interaction. The recyclable waste collector is responsible for the collection of waste product for the manufacturer. The study also examines the impacts of utilization rate on the cost and profit in the context of different sizes of vehicles. The model applies mathematical programming methods and attempts to maximize the total profit of the distributor during the study period. This study applies the binary logit model, analytical model and mathematical programming methods to the problem. The model specifically explores how to determine a recycle service strategy for the recycler by considering product surplus value, time-dependent recycle demand, transportation economies and demand-supply interaction. The model applies mathematical programming methods and attempts to minimize the total logistics cost of the recycler and maximize the recycle benefits of the manufacturer during the study period. The study relaxes the constant demand assumption and examines how service strategy affects consumer demand towards waste recycling. Results of the study not only help understanding how the user demand for recycle service and product surplus value affects the logistics cost and manufacturer’s benefits, but also provide guidance such as award bonus and carbon emission regulations for the government.

Keywords: circular economy, consumer demand, product surplus value, recycle service strategy

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Authors: Kassahun Tilahun

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The purpose of the study was two-fold: to examine the socio-demographic and psychological predictors of well-being and formulate a socio-culturally sound approach explaining the meaning and experience of psychological well-being among Ethiopian adults. Ryan and Deci’s Self-Determination Theory was duly considered as a theoretical framework of the study. The study followed a sequential explanatory mixed method design. Both quantitative and qualitative data were obtained, via scales and open-ended questionnaires, from 438 civil servants working in Addis Ababa. 30 interviews were also conducted to gain further information. An in-depth analysis of the reliability and validity of instruments was made before employing them to the main study. The results showed that adults were better off in both their scores of psychological and subjective well-being. Besides, adults’ well-being was found to be quite a function of their gender, age, marital status, educational level and household income. Males had a healthier psychological well-being status than females, where as females were better in their subjective well-being. A significant difference in psychological well-being was also observed between emerging and young adults, in favor of the young; and between cohabitated and married adults, married being advantageous. A significant difference in subjective well-being measures was also noticed among single, cohabitated and married adults, in favor of the married adults in all measures. The finding revealed that happiness level of adults decrease as their educational status increases while the reverse is true to psychological well-being. Besides, as adults’ household income boosts, so do their psychological well-being and satisfaction in life. The regression analysis also produced significant independent contributions of household income to overall well-being of adults. As such, subjective well-being was significantly predicted by dummy variable of sex and marital status. Likewise, the agreeableness, conscientiousness, neuroticism and openness dimensions of personality were notable significant predictors of adults’ psychological well-being where as extraversion and agreeableness were significant predictors of their subjective well-being. Religiosity was also a significant predictor of adults’ psychological well-being. Besides, adults’ well-being was significantly predicted by the interaction between conscientiousness and religiosity. From goal pursuit dimensions, attainment of extrinsic life goals was a significant predictor of both psychological and subjective well-being. Importance and attainment of intrinsic life goals also significantly predicts adults’ psychological well-being. Finally, the subjective well-being of adults was significantly predicted by environmental mastery, positive relations with others, self-acceptance and overall psychological well-being scores of adults. The thematic analysis identified five major categories of themes, which are essential in explaining the psychological well-being of Ethiopian adults. These were; socio-cultural harmony, social cohesion, security, competence and accomplishment, and the self. Detailed discussion on the rational for including these themes was made and appropriate implications were proposed. Researchers are encouraged to expand the findings of this research and in turn develop a suitable approach taping the psychological well-being of adults living in countries like Ethiopia.

Keywords: psychological well-being, subjective well-being, adulthood, Ethiopia

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Authors: Veena Easvaradoss, Ebenezer Joseph, Sumathi Chandrasekaran, Sweta Jain, Aparna Anna Mathai, Senta Christy

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Working memory refers to a cognitive processing space where information is received, managed, transformed, and briefly stored. It is an operational process of transforming information for the execution of cognitive tasks in different and new ways. Many class room activities require children to remember information and mentally manipulate it. While the impact of chess training on intelligence and academic performance has been unequivocally established, its impact on working memory needs to be studied. This study, funded by the Cognitive Science Research Initiative, Department of Science & Technology, Government of India, analyzed the effect of one-year chess training on the working memory of children. A pretest–posttest with control group design was used, with 52 children in the experimental group and 50 children in the control group. The sample was selected from children studying in school (grades 3 to 9), which included both the genders. The experimental group underwent weekly chess training for one year, while the control group was involved in extracurricular activities. Working memory was measured by two subtests of WISC-IV INDIA. The Digit Span Subtest involves recalling a list of numbers of increasing length presented orally in forward and in reverse order, and the Letter–Number Sequencing Subtest involves rearranging jumbled alphabets and numbers presented orally following a given rule. Both tasks require the child to receive and briefly store information, manipulate it, and present it in a changed format. The Children were trained using Winning Moves curriculum, audio- visual learning method, hands-on- chess training and recording the games using score sheets, analyze their mistakes, thereby increasing their Meta-Analytical abilities. They were also trained in Opening theory, Checkmating techniques, End-game theory and Tactical principles. Pre equivalence of means was established. Analysis revealed that the experimental group had significant gains in working memory compared to the control group. The present study clearly establishes a link between chess training and working memory. The transfer of chess training to the improvement of working memory could be attributed to the fact that while playing chess, children evaluate positions, visualize new positions in their mind, analyze the pros and cons of each move, and choose moves based on the information stored in their mind. If working-memory’s capacity could be expanded or made to function more efficiently, it could result in the improvement of executive functions as well as the scholastic performance of the child.

Keywords: chess training, cognitive development, executive functions, school children, working memory

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Authors: Blanca A. Pichardo, Victor H. Lopez, Melchor Salazar, Rafael Garcia, Alberto Ruiz

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Clad pipes in comparison to plain carbon steel pipes offer the oil and gas industry high corrosion resistance, reduction in economic losses due to pipeline failures and maintenance, lower labor risk, prevent pollution and environmental damage due to hydrocarbons spills caused by deteriorated pipelines. In this context, it is paramount to establish reliable welding procedures to join bimetallic plates or pipes. Thus, the aim of this work is to study the microstructure and mechanical behavior of clad plates welded by the gas metal arc welding (GMAW) process. A clad of 316L stainless steel was deposited onto API 5L X-60 plates by overlay welding with the GMAW process. Welding parameters were, 22.5 V, 271 A, heat input 1,25 kJ/mm, shielding gas 98% Ar + 2% O₂, reverse polarity, torch displacement speed 3.6 mm/s, feed rate 120 mm/s, electrode diameter 1.2 mm and application of an electromagnetic field of 3.5 mT. The overlay welds were subjected to macro-structural and microstructural characterization. After manufacturing the clad plates, a single V groove joint was machined with a 60° bevel and 1 mm root face. GMA welding of the bimetallic plates was performed in four passes with ER316L-Si filler for the root pass and an ER70s-6 electrode for the subsequent welding passes. For joining the clad plates, an electromagnetic field was applied with 2 purposes; to improve the microstructural characteristics and to assist the stability of the electric arc during welding in order to avoid magnetic arc blow. The welds were macro and microstructurally characterized and the mechanical properties were also evaluated. Vickers microhardness (100 g load for 10 s) measurements were made across the welded joints at three levels. The first profile, at the 316L stainless steel cladding, was quite even with a value of approximately 230 HV. The second microhardness profile showed high values in the weld metal, ~400 HV, this was due to the formation of a martensitic microstructure by dilution of the first welding pass with the second. The third profile crossed the third and fourth welding passes and an average value of 240 HV was measured. In the tensile tests, yield strength was between 400 to 450 MPa with a tensile strength of ~512 MPa. In the Charpy impact tests, the results were 86 and 96 J for specimens with the notch in the face and in the root of the weld bead, respectively. The results of the mechanical properties were in the range of the API 5L X-60 base material. The overlap welding process used for cladding is not suitable for large components, however, it guarantees a metallurgical bond, unlike the most commonly used processes such as thermal expansion. For welding bimetallic plates, control of the temperature gradients is key to avoid distortions. Besides, the dissimilar nature of the bimetallic plates gives rise to the formation of a martensitic microstructure during welding.

Keywords: clad pipe, dissimilar welding, gas metal arc welding, magnetic fields

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Authors: Karthyayani Rajamani, Yi-Chun Lin, Tung-Chou Wen, Jeanne Hsieh, Yi-Maun Subeq, Jen-Wei Liu, Po-Cheng Lin, Horng-Jyh Harn, Shinn-Zong Lin, Tzyy-Wen Chiou

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Assuring cell quality is an essential parameter for the success of stem cell therapy, utilization of various components to improve this potential has been the primary goal of stem cell research. The aim of this study was not only to demonstrate the capacity of trans-cinnamaldehyde (TC) to reverse stress-induced senescence but also improve the therapeutic abilities of stem cells. Because of the availability and the promising application potential in regenerative medicine, adipose-derived stem cells (ADSCs) were chosen for the study. We found that H2O2 treatment resulted in the expression of senescence characteristics in the ADSCs, including decreased proliferation rate, increased senescence-associated- β-galactosidase (SA-β-gal) activity, decreased SIRT1 (silent mating type information regulation 2 homologs) expression and decreased telomerase activity. However, TC treatment was sufficient to rescue or reduce the effects of H2O2 induction, ultimately leading to an increased proliferation rate, a decrease in the percentage of SA-β-gal positive cells, upregulation of SIRT1 expression, and increased telomerase activity of the senescent ADSCs at the cellular level. Further recently it was observed that the ADSCs were treated with TC without induction of senescence, all the before said positives were observed. Moreover, a chemically induced liver fibrosis animal model was used to evaluate the functionality of these rescued cells in vivo. Liver dysfunction was established by injecting 200 mg/kg thioacetamide (TAA) intraperitoneally into Wistar rats every third day for 60 days. The experimental rats were separated into groups; normal group (rats without TAA induction), sham group (without ADSC transplantation), positive control group (transplanted with normal ADSCs); H2O2 group (transplanted with H2O2 -induced senescent ADSCs), H2O2+TC group (transplanted with ADSCs pretreated with H2O2 and then further treated with TC) and TC group (ADSC treated with TC without H2O2 treatment). In the transplantation group, 1 × 106 human ADSCs were introduced into each rat via direct liver injection. Based on the biochemical analysis and immunohistochemical staining results, it was determined that the therapeutic effects on liver fibrosis by the induced senescent ADSCs (H2O2 group) were not as significant as those exerted by the normal ADSCs (the positive control group). However, the H2O2+TC group showed significant reversal of liver damage when compared to the H2O2 group 1 week post-transplantation. Further ADSCs without H2O2 treatment but with just TC treatment performed much better than all the groups. These data confirmed that the TC treatment had the potential to improve the therapeutic effect of ADSCs. It is therefore suggested that TC has potential applications in maintaining stem cell quality and could possibly aid in the treatment of senescence-related disorders.

Keywords: senescence, SIRT1, adipose derived stem cells, liver fibrosis

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Authors: Gifty Kumadey, Albert Tchey Agbenyegah

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(i) Overview and research goal(s): This study aims to address research gaps in the Ghanaian pharmaceutical industry by examining the impact of environmental supply chain management (ESCM) practices on environmental and operational performance. Previous studies have provided inconclusive evidence on the relationship between ESCM practices and environmental and operational performance. The research aims to provide a clearer understanding of the impact of ESCM practices on environmental and operational performance in the context of the Ghanaian pharmaceutical industry. Limited research has been conducted on ESCM practices in developing countries, particularly in Africa. The study aims to bridge this gap by examining the drivers and barriers specific to the pharmaceutical industry in Ghana. The research aims to analyze the impact of ESCM practices on the achievement of Sustainable Development Goals (SDGs) in the Ghanaian pharmaceutical industry, focusing on SDGs 3, 12, 13, and 17. It also explores the potential for partnerships and collaborations to advance ESCM practices in the pharmaceutical industry. The research hypotheses suggest that pressure from stakeholder positively influences the adoption of ESCM practices in the Ghanaian pharmaceutical industry. By addressing these goals, the study aims to contribute to sustainable development initiatives and offer practical recommendations to enhance ESCM A practices in the industry. (ii) Research methods and data: This study uses a quantitative research design to examine the drivers and barriers to environmental supply chain management in the pharmaceutical industry in Accra.The sample size is approximately 150 employees, with senior and middle-level managers from pharmaceutical industry of Ghana. A purposive sampling technique is used to select participants with relevant knowledge and experience in environmental supply chain management. Data will be collected using a structured questionnaire using Likert scale responses. Descriptive statistics will be used to analyze the data and provide insights into current practices and their impact on environmental and operational performance. (iii) Preliminary results and conclusions: Main contributions: Identifying drivers/barriers to ESCM in Ghana's pharmaceutical industry, evaluating current ESCM practices, examining impact on performance, providing practical insights, contributing to knowledge on ESCM in Ghanaian context. The research contributes to SDGs 3, 9, and 12 by promoting sustainable practices and responsible consumption in the industry. The study found that government rules and regulations are the most critical drivers for ESCM adoption, with senior managers playing a significant role. However, employee and competitor pressures have a lesser impact. The industry has made progress in implementing certain ESCM practices, but there is room for improvement in areas like green distribution and reverse logistics. The study emphasizes the importance of government support, management engagement, and comprehensive implementation of ESCM practices in the industry. Future research should focus on overcoming barriers and challenges to effective ESCM implementation.

Keywords: environmental supply chain, sustainable development goal, ghana pharmaceutical industry, government regulations

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Authors: Igor Guatelli

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The universe of aesthetic perception entails impasses about sensitive divergences that each text or visual object may be subjected to. If approached through intertextuality that is not based on the misleading notion of kinships or similarities a priori admissible, the possibility of anachronistic, heterogeneous - and non-diachronic - assemblies can enhance the emergence of interval movements, intermediate, and conflicting, conducive to a method of reading, interpreting, and assigning meaning that escapes the rigid antinomies of the mere being and non-being of things. In negative, they operate in a relationship built by the lack of an adjusted meaning set by their positive existences, with no remainders; the generated interval becomes the remnant of each of them; it is the opening that obscures the stable positions of each one. Without the negative of absence, of that which is always missing or must be missing in a text, concept, or image made positive by history, nothing is perceived beyond what has been already given. Pairings or binary oppositions cannot lead only to functional syntheses; on the contrary, methodological disturbances accumulated by the approximation of signs and entities can initiate a process of becoming as an opening to an unforeseen other, transformation until a moment when the difficulties of [re]conciliation become the mainstay of a future of that sign/entity, not envisioned a priori. A counter-history can emerge from these unprecedented, misadjusted approaches, beginnings of unassigned injunctions and disjunctions, in short, difficult alliances that open cracks in a supposedly cohesive history, chained in its apparent linearity with no remains, understood as a categorical historical imperative. Interstices are minority fields that, because of their opening, are capable of causing opacity in that which, apparently, presents itself with irreducible clarity. Resulting from an incomplete and maladjusted [at the least dual] marriage between the signs/entities that originate them, this interval may destabilize and cause disorder in these entities and their own meanings. The interstitials offer a hyphenated relationship: a simultaneous union and separation, a spacing between the entity’s identity and its otherness or, alterity. One and the other may no longer be seen without the crack or fissure that now separates them, uniting, by a space-time lapse. Ontological, semantic shifts are caused by this fissure, an absence between one and the other, one with and against the other. Based on an improbable approximation between some conceptual and semantic shifts within the design production of architect Rem Koolhaas and the textual production of the philosopher Jacques Derrida, this article questions the notion of unity, coherence, affinity, and complementarity in the process of construction of thought from these ontological, epistemological, and semiological fissures that rattle the signs/entities and their stable meanings. Fissures in a thought that is considered coherent, cohesive, formatted are the negativity that constitutes the interstices that allow us to move towards what still remains as non-identity, which allows us to begin another story.

Keywords: clearing, interstice, negative, remnant, spectrum

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Authors: Lesibana Rafapa

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Accounts exist of circumcision-anchored cultural initiation in central Africa, East Africa, Southern Africa, North Africa, and West Africa -straddling states like Botswana, Kenya, Lesotho, Malawi, Senegal, South Africa, Zambia, and Zimbabwe. This attests to the continent-wide spread of this cultural practice. In this paper, the writer relates the cultural aspect of circumcision-subsuming initiation among black African cultural groups across the continent to the notion that African cultures are varied yet subscribe to a common central concept. The premise of the paper is that the common practice of initiation for both male and female children that have to be initiated by adults to the tradition and customs of a people coincides with such a central concept. The practice of traditional initiation is as broad as to encompass aspects of spirituality, morality, and social organisation, in the nature of the central concept of which it is a trans-sectional part. Cultural initiation, sometimes referred to as traditional circumcision, constitutes culture-determined rites of passage for the initiates. The study’s aim, the findings of which are presented in this paper, was to probe gender equality in the development and promotion of the cultural practice of initiation. The researcher intended to demonstrate how in South Africa, female circumcision is treated equally or marginalised in efforts of the democratic government to regulate and strengthen the practice of circumcision as part of its broader liberation programme meant to reverse politico-cultural bondage experienced during apartheid rule that the present black regime helped bring to an end. It is argued that the failure to regard female circumcision as equal to its male counterpart is a travesty of the black government’s legislation and policies espousing equality and the protection and empowerment of vulnerable and previously marginalised population groups that include black women. The writer did a desk-top study of the history and characteristics of female circumcision among the black Northern Sotho, VaTsonga, and VhaVenda cultural groups of the Limpopo Province, stretching north to the border of South Africa with Zimbabwe, as well as literature on how political and other authorities exert efforts to preserve and empower the practice. The findings were that male initiation is foregrounded and totalised to represent the practice of initiation as a whole, at the expense of its female counterpart facing marginalisation and unequal regard. It is outlined in this paper how such impoverishment of an otherwise woman-empowering cultural practice deprives hitherto black cultures that suffered brutal repression during apartheid of a fuller recovery much needed in the democratic era. The writer applies some aspects of postcolonial theory and some tropes of feminism in the discussion of an uneven status of cultural circumcision at the hands of present day powers that be.

Keywords: African cultures, female circumcision, gender equality, women empowerment

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Authors: Sundru Manjulata Devi, Prakash M. Halami

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The immemorial use of fermented foods from vegetables, dairy and other biological sources are of great demand in India because of their health benefits. However, the diversity of Lactobacillus plantarum group (LPG) of vegetable origin has not been revealed yet, particularly with reference to their probiotic functionalities. In the present study, the different species of probiotic Lactobacillus plantarum group (LPG) i.e., L. plantarum subsp. plantarum MTCC 5422 (from fermented cereals), L. plantarum subsp. argentoratensis FG16 (from fermented bamboo shoot) and L. paraplantarum MTCC 9483 (from fermented gundruk) (as characterized by multiplex recA PCR assay) were considered to investigate their relative expression of MUB domains of mub gene (mucin binding protein) by Real time PCR. Initially, the allelic variation in the mub gene was assessed and found to encode three different variants (Type I, II and III). All the three types had 8, 9 and 10 MUB domains respectively (as analysed by Pfam database) and were found to be responsible for adhesion of bacteria to the host intestinal epithelial cells. These domains either get inserted or deleted during speciation or evolutionary events and lead to divergence. The reverse transcriptase qPCR analysis with mubLPF1+R1 primer pair supported variation in amplicon sizes with 300, 500 and 700 bp among different LPG strains. The relative expression of these MUB domains significantly unregulated in the presence of 1% mucin in overnight grown cultures. Simultaneously, the mub gene expressed efficiently by 7 fold in the culture L. paraplantarum MTCC 9483 with 10 MUB domains. An increase in the expression levels for L. plantarum subsp. plantarum MTCC 5422 and L. plantarum subsp. argentoratensis FG16 (MCC 2974) with 9 and 8 repetitive domains was around 4 and 2 fold, respectively. The detection and expression of an integrase (int) gene in the upstream region of mub gene reveals the excision and integration of these repetitive domains. Concurrently, an in vitro adhesion assay to mucin and exclusion of pathogens (such as Listeria monocytogenes and Micrococcus leuteus) was investigated and observed that the L. paraplantarum MTCC 9483 with more adhesion domains has more ability to adhere to mucin and inhibited the growth of pathogens. The production and expression of plantaricin-like bacteriocin (plnNC8 type) in MTCC 9483 suggests the pathogen inhibition. Hence, the expression of MUB domains can act as potential biomarkers in the screening of a novel probiotic LPG strain with adherence property. The present study provides a platform for an easy, rapid, less time consuming, low-cost methodology for the detection of potential probiotic bacteria. It was known that the traditional practices followed in the preparation of fermented bamboo shoots/gundruk/cereals of Indian foods contain different kinds of neutraceuticals for functional food and novel compounds with health promoting factors. In future, a detailed study of these food products can add more nutritive value, consumption and suitable for commercialization.

Keywords: adhesion gene, fermented foods, MUB domains, probiotics

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Authors: Fahimeh Palizban, Farshad Noravesh, Amir Hossein Saeidian, Mahya Mehrmohamadi

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In the recent decade, the emergence of liquid biopsy has significantly improved cancer monitoring and detection. Dying cells, including those originating from tumors, shed their DNA into the blood and contribute to a pool of circulating fragments called cell-free DNA. Accordingly, identifying the tissue origin of these DNA fragments from the plasma can result in more accurate and fast disease diagnosis and precise treatment protocols. Open chromatin regions are important epigenetic features of DNA that reflect cell types of origin. Profiling these features by DNase-seq, ATAC-seq, and histone ChIP-seq provides insights into tissue-specific and disease-specific regulatory mechanisms. There have been several studies in the area of cancer liquid biopsy that integrate distinct genomic and epigenomic features for early cancer detection along with tissue of origin detection. However, multimodal analysis requires several types of experiments to cover the genomic and epigenomic aspects of a single sample, which will lead to a huge amount of cost and time. To overcome these limitations, the idea of predicting OCRs from WGS is of particular importance. In this regard, we proposed a computational approach to target the prediction of open chromatin regions as an important epigenetic feature from cell-free DNA whole genome sequence data. To fulfill this objective, local sequencing depth will be fed to our proposed algorithm and the prediction of the most probable open chromatin regions from whole genome sequencing data can be carried out. Our method integrates the signal processing method with sequencing depth data and includes count normalization, Discrete Fourie Transform conversion, graph construction, graph cut optimization by linear programming, and clustering. To validate the proposed method, we compared the output of the clustering (open chromatin region+, open chromatin region-) with previously validated open chromatin regions related to human blood samples of the ATAC-DB database. The percentage of overlap between predicted open chromatin regions and the experimentally validated regions obtained by ATAC-seq in ATAC-DB is greater than 67%, which indicates meaningful prediction. As it is evident, OCRs are mostly located in the transcription start sites (TSS) of the genes. In this regard, we compared the concordance between the predicted OCRs and the human genes TSS regions obtained from refTSS and it showed proper accordance around 52.04% and ~78% with all and the housekeeping genes, respectively. Accurately detecting open chromatin regions from plasma cell-free DNA-seq data is a very challenging computational problem due to the existence of several confounding factors, such as technical and biological variations. Although this approach is in its infancy, there has already been an attempt to apply it, which leads to a tool named OCRDetector with some restrictions like the need for highly depth cfDNA WGS data, prior information about OCRs distribution, and considering multiple features. However, we implemented a graph signal clustering based on a single depth feature in an unsupervised learning manner that resulted in faster performance and decent accuracy. Overall, we tried to investigate the epigenomic pattern of a cell-free DNA sample from a new computational perspective that can be used along with other tools to investigate genetic and epigenetic aspects of a single whole genome sequencing data for efficient liquid biopsy-related analysis.

Keywords: open chromatin regions, cancer, cell-free DNA, epigenomics, graph signal processing, correlation clustering

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Authors: Marek Vondra, Petr Bobák

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Vacuum evaporation is a reliable and well-proven technology with a wide application range which is frequently used in food, chemical or pharmaceutical industries. Recently, numerous remarkable studies have been carried out to investigate utilization of this technology in the area of wastewater treatment. One of the most successful applications of vacuum evaporation principal is connected with seawater desalination. Since 1950’s, multi-stage flash distillation (MSF) has been the leading technology in this field and it is still irreplaceable in many respects, despite a rapid increase in cheaper reverse-osmosis-based installations in recent decades. MSF plants are conveniently operated in countries with a fluctuating seawater quality and at locations where a sufficient amount of waste heat is available. Nowadays, most of the MSF research is connected with alternative heat sources utilization and with hybridization, i.e. merging of different types of desalination technologies. Some of the studies are concerned with basic principles of the static flash phenomenon, but only few scientists have lately focused on the fundamentals of continuous multi-stage evaporation. Limited measurement possibilities at operating plants and insufficiently equipped experimental facilities may be the reasons. The aim of the presented study was to design, construct and test an up-to-date test rig with an advanced measurement system which will provide real time monitoring options of all the important operational parameters under various conditions. The whole system consists of a conventionally designed MSF unit with 8 evaporation chambers, versatile heating circuit for different kinds of feed water (e.g. seawater, waste water), sophisticated system for acquisition and real-time visualization of all the related quantities (temperature, pressure, flow rate, weight, conductivity, pH, water level, power input), access to a wide spectrum of operational media (salt, fresh and softened water, steam, natural gas, compressed air, electrical energy) and integrated transparent features which enable a direct visual control of selected physical mechanisms (water evaporation in chambers, water level right before brine and distillate pumps). Thanks to the adjustable process parameters, it is possible to operate the test unit at desired operational conditions. This allows researchers to carry out statistical design and analysis of experiments. Valuable results obtained in this manner could be further employed in simulations and process modeling. First experimental tests confirm correctness of the presented approach and promise interesting outputs in the future. The presented experimental apparatus enables flexible and efficient research of the whole MSF process.

Keywords: design of experiment, multi-stage flash distillation, test rig, vacuum evaporation

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Authors: Partha Sarathi Nayek, Abhiparna Dasgupta, Maheshreddy Gade

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Earth retaining structure plays a vital role in stabilizing unstable road cuts and slopes in the mountainous region. The retaining structures located in seismically active regions like the Himalayas may experience moderate to severe earthquakes. An earthquake produces two kinds of ground motion: permanent quasi-static displacement (fault rapture) on the fault rupture plane and transient vibration, traveling a long distance. There has been extensive research work to understand the dynamic behavior of retaining structures subjected to transient ground motions. However, understanding the effect caused by fault rapture phenomena on retaining structures is limited. The presence of shallow crustal active faults and natural slopes in the Himalayan region further highlights the need to study the response of retaining structures subjected to fault rupture phenomena. In this paper, an attempt has been made to understand the dynamic response of the cantilever retaining wall subjected to surface fault rupture. For this purpose, a 2D finite element model consists of a retaining wall, backfill and foundation have been developed using Abaqus 6.14 software. The backfill and foundation material are modeled as per the Mohr-Coulomb failure criterion, and the wall is modeled as linear elastic. In this present study, the interaction between backfill and wall is modeled as ‘surface-surface contact.’ The entire simulation process is divided into three steps, i.e., the initial step, gravity load step, fault rupture step. The interaction property between wall and soil and fixed boundary condition to all the boundary elements are applied in the initial step. In the next step, gravity load is applied, and the boundary elements are allowed to move in the vertical direction to incorporate the settlement of soil due to the gravity load. In the final step, surface fault rupture has been applied to the wall-backfill system. For this purpose, the foundation is divided into two blocks, namely, the hanging wall block and the footwall block. A finite fault rupture displacement is applied to the hanging wall part while the footwall bottom boundary is kept as fixed. Initially, a numerical analysis is performed considering the reverse fault mechanism with a dip angle of 45°. The simulated result is presented in terms of contour maps of permanent displacements of the wall-backfill system. These maps highlighted that surface fault rupture can induce permanent displacement in both horizontal and vertical directions, which can significantly influence the dynamic behavior of the wall-backfill system. Further, the influence of fault mechanism, dip angle, and surface fault rupture position is also investigated in this work.

Keywords: surface fault rupture, retaining wall, dynamic response, finite element analysis

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Authors: Anita Bajpai, Sarah Duan, Ashlee Erskine, Shehzein Khan, Raymond Kramer

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Introduction: This research focuses on exploring the beneficial effects if any, of Vitamin-D in reducing the need for supplemental oxygen among hospitalized COVID-19 patients. Two questions are investigated – Q1)Doeshaving a healthy level of baselineVitamin-D 25-OH (≥ 30ng/ml) help,andQ2) does administering Vitamin-D therapy after-the-factduring inpatient hospitalization help? Methods/Study Design: This is a comprehensive, retrospective, observational study of all inpatients at RUHS from March through December 2020 who tested positive for COVID-19 based on real-time reverse transcriptase–polymerase chain reaction assay of nasal and pharyngeal swabs and rapid assay antigen test. To address Q1, we looked atall N1=182 patients whose baseline plasma Vitamin-D 25-OH was known and who needed supplemental oxygen. Of this, a total of 121 patients had a healthy Vitamin-D level of ≥30 ng/mlwhile the remaining 61 patients had low or borderline (≤ 29.9ng/ml)level. Similarly, for Q2, we looked at a total of N2=893 patients who were given supplemental oxygen, of which713 were not given Vitamin-D and 180 were given Vitamin-D therapy. The numerical value of the maximum amount of oxygen flow rate(dependent variable) administered was recorded for each patient. The mean values and associated standard deviations for each group were calculated. Thesetwo sets of independent data served as the basis for independent, two-sample t-Test statistical analysis. To be accommodative of any reasonable benefitof Vitamin-D, ap-value of 0.10(α< 10%) was set as the cutoff point for statistical significance. Results: Given the large sample sizes, the calculated statistical power for both our studies exceeded the customary norm of 80% or better (β< 0.2). For Q1, the mean value for maximumoxygen flow rate for the group with healthybaseline level of Vitamin-D was 8.6 L/min vs.12.6L/min for those with low or borderline levels, yielding a p-value of 0.07 (p < 0.10) with the conclusion that those with a healthy level of baseline Vitamin-D needed statistically significant lower levels of supplemental oxygen. ForQ2, the mean value for a maximum oxygen flow rate for those not administered Vitamin-Dwas 12.5 L/min vs.12.8L/min for those given Vitamin-D, yielding a p-valueof 0.87 (p > 0.10). We thereforeconcludedthat there was no statistically significant difference in the use of oxygen therapy between those who were or were not administered Vitamin-D after-the-fact in the hospital. Discussion/Conclusion: We found that patients who had healthy levels of Vitamin-D at baseline needed statistically significant lower levels of supplemental oxygen. Vitamin-D is well documented, including in a recent article in the Lancet, for its anti-inflammatory role as an adjuvant in the regulation of cytokines and immune cells. Interestingly, we found no statistically significant advantage for giving Vitamin-D to hospitalized patients. It may be a case of “too little too late”. A randomized clinical trial reported in JAMA also did not find any reduction in hospital stay of patients given Vitamin-D. Such conclusions come with a caveat that any delayed marginal benefits may not have materialized promptly in the presence of a significant inflammatory condition. Since Vitamin-D is a low-cost, low-risk option, it may still be useful on an inpatient basis until more definitive findings are established.

Keywords: COVID-19, vitamin-D, supplemental oxygen, vitamin-D in primary care

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Authors: Valentina V. Goftman, Vladislav A. Pankratov, Alexey V. Markin, Tangi Aubert, Zeger Hens, Sarah De Saeger, Irina Yu. Goryacheva

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The most important requirements for biochemical applicability of quantum dots (QDs) are: 1) the surface cap should render intact or improved optical properties; 2) mono-dispersion and good stability in aqueous phase in a wide range of pH and ionic strength values; 3) presence of functional groups, available for bioconjugation; 4) minimal impact from the environment on the QDs’ properties and, vice versa, minimal influence of the QDs’ components on the environment; and 5) stability against chemical/biochemical/physical influence. The latter is especially important for in vitro and in vivo applications. For example, some physical intracellular delivery strategies (e.g., electroporation) imply a rapid high-voltage electric field impulse in order to temporarily generate hydrophilic pores in the cell plasma membrane, necessary for the passive transportation of QDs into the cell. In this regard, it is interesting to investigate how different capping layers, which can provide high stability and sufficient fluorescent properties of QDs in a water solution, behave under these abnormal conditions. In this contribution, hydrophobic core-shell CdSe/CdS/CdZnS/ZnS QDs (λem=600 nm), produced by means of the Successive Ion Layer Adsorption and Reaction (SILAR) technique, were transferred to a water solution using two of the most commonly used methods: (i) encapsulation in an amphiphilic brush polymer based on poly(maleic anhydride-alt-1-octadecene) (PMAO) modified with polyethylene glycol (PEG) chains and (ii) silica covering. Polymer encapsulation preserves the initial ligands on the QDs’ surface owing to the hydrophobic attraction between the hydrophobic groups of the amphiphilic molecules and the surface hydrophobic groups of the QDs. This covering process allows maintaining the initial fluorescent properties, but it leads to a considerable increase of the QDs’ size. However, covering with a silica shell, by means of the reverse microemulsion method, allows maintaining both size and fluorescent properties of the initial QDs. The obtained water solutions of polymer covered and silica-coated QDs in three different concentrations were exposed to a low-voltage electric field for a short time and the fluorescent properties were investigated. It is shown that the PMAO-PEG polymer acquires some additional charges in the presence of the electric field, which causes repulsion between the polymer and the QDs’ surface. This process destroys the homogeneity of the whole amphiphilic shell and it dramatically decreases the fluorescent properties (dropping to 10% from its initial value) because of the direct contact of the QDs with the strongly oxidative environment (water). In contrast, a silica shell possesses dielectric properties which allow retaining 90% of its initial fluorescence intensity, even after a longer electric impact. Thus, silica shells are clearly a preferable covering for bio-application of QDs, because – besides the high uniform morphology, controlled size and biocompatibility – it allows protecting QDs from oxidation, even under the influence of an electric field.

Keywords: electric field, polymer coating, quantum dots, silica covering, stability

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Authors: Katarzyna Drela, Miroslaw Wielgos, Mikolaj Wrobel, Barbara Lukomska

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Mesenchymal stem cells (MSCs) have a great potential in regenerative medicine. Since the initial number of isolated MSCs is limited, in vitro propagation is often required to reach sufficient numbers of cells for therapeutic applications. During long-term culture MSCs may undergo genetic or epigenetic alterations that subsequently increase the probability of spontaneous malignant transformation. Thus, factors that influence genomic stability of MSCs following long-term expansions need to be clarified before cultured MSCs are employed for clinical application. The aim of our study was to investigate the potential for spontaneous transformation of human neonatal cord blood (HUCB-MSCs) and adult bone marrow (BM-MSCs) derived MSCs. Materials and Methods: HUCB-MSCs and BM-MSCs were isolated by standard Ficoll gradient centrifugations method. Isolated cells were initially plated in high density 106 cells per cm2. After 48 h medium were changed and non-adherent cells were removed. The malignant transformation of MSCs in vitro was evaluated by morphological changes, proliferation rate, ability to enter cell senescence, the telomerase expression and chromosomal abnormality. Proliferation of MSCs was analyzed with WST-1 reduction method and population doubling time (PDT) was calculated at different culture stages. Then the expression pattern of genes characteristic for mesenchymal or epithelial cells, as well as transcriptions factors were examined by RT-PCR. Concomitantly, immunocytochemical analysis of gene-related proteins was employed. Results: Our studies showed that MSCs from all bone marrow isolations ultimately entered senescence and did not undergo spontaneous malignant transformation. However, HUCB-MSCs from one of the 15 donors displayed an increased proliferation rate, failed to enter senescence, and exhibited an altered cell morphology. In this sample we observed two different cell phenotypes: one mesenchymal-like exhibited spindle shaped morphology and express specific mesenchymal surface markers (CD73, CD90, CD105, CD166) with low proliferation rate, and the second one with round, densely package epithelial-like cells with significantly increased proliferation rate. The PDT of epithelial-like populations was around 1day and 100% of cells were positive for proliferation marker Ki-67. Moreover, HUCB-MSCs showed a positive expression of human telomerase reverse transcriptase (hTERT), cMYC and exhibit increased number of CFU during the long-term culture in vitro. Furthermore, karyotype analysis revealed chromosomal abnormalities including duplications. Conclusions: Our studies demonstrate that HUCB-MSCs are susceptible to spontaneous malignant transformation during long-term culture. Spontaneous malignant transformation process following in vitro culture has enormous effect on the biosafety issues of future cell-based therapies and regenerative medicine regimens.

Keywords: mesenchymal stem cells, spontaneous, transformation, long-term culture

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Authors: Rahaba Marima, Clement Penny

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The Health-related quality of life (HRQoL) for HIV positive patients has improved since the introduction of the highly active antiretroviral treatment (HAART). However, in the present HAART era, HIV co-morbidities such as lung cancer, a non-AIDS (NAIDS) defining cancer have been documented to be on the rise. Under normal physiological conditions, cells grow, repair and proliferate through the cell-cycle as cellular homeostasis is important in the maintenance and proper regulation of tissues and organs. Contrarily, the deregulation of the cell-cycle is a hallmark of cancer, including lung cancer. The association between lung cancer and the use of HAART components such as Efavirenz (EFV) is poorly understood. This study aimed at elucidating the effects of EFV on the cell-cycle genes’ expression in lung cancer. For this purpose, the human cell-cycle gene array composed of 84 genes was evaluated on both normal lung fibroblasts (MRC-5) cells and adenocarcinoma (A549) lung cells, in response to 13µM EFV or 0.01% vehicle. The ±2 up or down fold change was used as a basis of target selection, with p < 0.05. Additionally, RT-qPCR was done to validate the gene array results. Next, In-silico bio-informatics tools, Search Tool for the Retrieval of Interacting Genes/Proteins (STRING), Reactome, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and Ingenuity Pathway Analysis (IPA) were used for gene/gene interaction studies as well as to map the molecular and biological pathways influenced by the identified targets. Interestingly, the DNA damage response (DDR) pathway genes such as p53, Ataxia telangiectasia mutated and Rad3 related (ATR), Growth arrest and DNA damage inducible alpha (GADD45A), HUS1 checkpoint homolog (HUS1) and Role of radiation (RAD) genes were shown to be upregulated following EFV treatment, as revealed by STRING analysis. Additionally, functional enrichment analysis by the KEGG pathway revealed that most of the differentially expressed gene targets function at the cell-cycle checkpoint such as p21, Aurora kinase B (AURKB) and Mitotic Arrest Deficient-Like 2 (MAD2L2). Core analysis by IPA revealed that p53 downstream targets such as survivin, Bcl2, and cyclin/cyclin dependent kinases (CDKs) complexes are down-regulated, following exposure to EFV. Furthermore, Reactome analysis showed a significant increase in cellular response to stress genes, DNA repair genes, and apoptosis genes, as observed in both normal and cancerous cells. These findings implicate the genotoxic effects of EFV on lung cells, provoking the DDR pathway. Notably, the constitutive expression of this pathway (DDR) often leads to uncontrolled cell proliferation and eventually tumourigenesis, which could be the attribute of HAART components’ (such as EFV) effect on human cancers. Targeting the cell-cycle and its regulation holds a promising therapeutic intervention to the potential HAART associated carcinogenesis, particularly lung cancer.

Keywords: cell-cycle, DNA damage response, Efavirenz, lung cancer

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