Search results for: biological treatment

Authors: M. I. Tyletc, O. I. Podgornya, T. G. Shaposhnikova, S. V. Shabelnikov, A. G. Mittenberg, M. A. Daugavet

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Ascidiacea is the most numerous class of the Tunicata subtype. These chordates' distinctive feature of the anatomical structure is a tunic consisting of cellulose fibrils, protein molecules, and single cells. The mechanisms of the tunic formation are not known in detail; tunic formation could be used as the model system for studying the interaction of cells with the extracellular matrix. Our model species is the ascidian Styela rustica, which is prevalent in benthic communities of the White Sea. As previously shown, the tunic formation involves morula blood cells, which contain the major 48 kDa protein p48. P48 participation in the tunic formation was proved using antibodies against the protein. The nature of the protein and its function remains unknown. The current research aims to determine the amino acid sequence of p48, as well as to clarify its role in the tunic formation. The peptides that make up the p48 amino acid sequence were determined by mass spectrometry. A search for peptides in protein sequence databases identified sequences homologous to p48 in Styela clava, Styela plicata, and Styela canopus. Based on sequence alignment, their level of similarity was determined as 81-87%. The correspondent sequence of ascidian Styela canopus was used for further analysis. The Styela rustica p48 sequence begins with a signal peptide, which could indicate that the protein is secretory. This is consistent with experimentally obtained data: the contents of morula cells secreted in the tunic matrix. The isoelectric point of p48 is 9.77, which is consistent with the experimental results of acid electrophoresis of morula cell proteins. However, the molecular weight of the amino acid sequence of ascidian Styela canopus is 103 kDa, so p48 of Styela rustica is a shorter homolog. The search for conservative functional domains revealed the presence of two Ca-binding EGF-like domains, thrombospondin (TSP1) and tyrosinase domains. The p48 peptides determined by mass spectrometry fall into the region of the sequence corresponding to the last two domains and have amino acid substitutions as compared to Styela canopus homolog. The tyrosinase domain (pfam00264) is known to be part of the phenoloxidase enzyme, which participates in melanization processes and the immune response. The thrombospondin domain (smart00209) interacts with a wide range of proteins, and is involved in several biological processes, including coagulation, cell adhesion, modulation of intercellular and cell-matrix interactions, angiogenesis, wound healing and tissue remodeling. It can be assumed that the tyrosinase domain in p48 plays the role of the phenoloxidase enzyme, and TSP1 provides a link between the extracellular matrix and cell surface receptors, and may also be responsible for the repair of the tunic. The results obtained are consistent with experimental data on p48. The domain organization of protein suggests that p48 is an enzyme involved in the tunic tunning and is an important regulator of the organization of the extracellular matrix.

Keywords: ascidian, p48, thrombospondin, tyrosinase, tunic, tunning

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Authors: Shrestha Sharma, Jasjeet K. Sahni, Javed Ali, Sanjula Baboota

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Introduction: Rutin is a naturally occurring strong antioxidant molecule belonging to bioflavonoid category. Due to its free radical scavenging properties, it has been found to be beneficial in the treatment of various diseases including inflammation, cancer, diabetes, allergy, cardiovascular disorders and various types of microbial infections. Despite its beneficial effects, it suffers from the problem of low aqueous solubility which is responsible for low oral bioavailability. The aim of our study was to optimize and characterize self-nanoemulsifying drug delivery system (SNEDDS) of rutin using Box-Behnken design (BBD) combined with a desirability function. Further various antioxidant, pharmacokinetic and pharmacodynamic studies were performed for the optimized rutin SNEDDS formulation. Methodologies: Selection of oil, surfactant and co-surfactant was done on the basis of solubility/miscibility studies. Sefsol+ Vitamin E, Solutol HS 15 and Transcutol P were selected as oil phase, surfactant and co-surfactant respectively. Optimization of SNEDDS formulations was done by a three-factor, three-level (33)BBD. The independent factors were Sefsol+ Vitamin E, Solutol HS15, and Transcutol P. The dependent variables were globule size, self emulsification time (SEF), % transmittance and cumulative percentage drug released. Various response surface graphs and contour plots were constructed to understand the effect of different factor, their levels and combinations on the responses. The optimized Rutin SNEDDS formulation was characterized for various parameters such as globule size, zeta potential, viscosity, refractive index , % Transmittance and in vitro drug release. Ex vivo permeation studies and pharmacokinetic studies were performed for optimized formulation. Antioxidant activity was determined by DPPH and reducing power assays. Anti-inflammatory activity was determined by using carrageenan induced rat paw oedema method. Permeation of rutin across small intestine was assessed using confocal laser scanning microscopy (CLSM). Major findings:The optimized SNEDDS formulation consisting of Sefsol+ Vitamin E - Solutol HS15 -Transcutol HP at proportions of 25:35:17.5 (w/w) was prepared and a comparison of the predicted values and experimental values were found to be in close agreement. The globule size and PDI of optimized SNEDDS formulation was found to be 16.08 ± 0.02 nm and 0.124±0.01 respectively. Significant (p˂0.05) increase in percentage drug release was achieved in the case of optimized SNEDDS formulation (98.8 %) as compared to rutin suspension. Furthermore, pharmacokinetic study showed a 2.3-fold increase in relative oral bioavailability compared with that of the suspension. Antioxidant assay results indicated better efficacy of the developed formulation than the pure drug and it was found to be comparable with ascorbic acid. The results of anti-inflammatory studies showed 72.93 % inhibition for the SNEDDS formulation which was significantly higher than the drug suspension 46.56%. The results of CLSM indicated that the absorption of SNEDDS formulation was considerably higher than that from rutin suspension. Conclusion: Rutin SNEDDS have been successfully prepared and they can serve as an effective tool in enhancing oral bioavailability and efficacy of Rutin.

Keywords: rutin, oral bioavilability, pharamacokinetics, pharmacodynamics

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Authors: Tatsiana Ihnatovich, Darya Nizheharodava, Mikalai Halabarodzka, Tatsiana Savitskaya, Marina Zafranskaya

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Liver fibrosis is a complex of histological changes resulting from chronic liver disease accompanied by an excessive production and deposition of extracellular matrix components in the hepatic parenchyma. Liver fibrosis is a serious medical and social problem. Hepatic stellate cells (HSCs) make a significant contribution to the extracellular matrix deposition due to liver injury. Mesenchymal stem cells (MSCs) have a pronounced anti-inflammatory, regenerative and immunomodulatory effect; they are able to differentiate into hepatocytes and induce apoptosis of activated HSCs that opens the prospect of their use for preventing the excessive fibro-formation and the development of liver cirrhosis. The aim of the study is to evaluate the effect of MSCs therapy on the expression of fibrogenesis markers genes in liver tissue and HSCs cultures of rats with experimental liver cirrhosis (ELC). Materials and methods: ELC was induced by the common bile duct ligation (CBDL) in female Wistar rats (n = 19) with an average body weight of 250 (220 ÷ 270) g. Animals from the control group (n = 10) were sham-operated. On the 56th day after the CBDL, the rats of the experimental (n = 12) and the control (n = 5) groups received intraportal MSCs in concentration of 1×106 cells/animal (previously obtained from rat’s bone marrow) or saline, respectively. The animals were taken out of the experiment on the 21st day. HSCs were isolated by sequential liver perfusion in situ with following disaggregation, enzymatic treatment and centrifugation of cell suspension on a two-stage density gradient. The expression of collagen type I (Col1a1) and type III (Col3a1), matrix metalloproteinase type 2 (MMP2) and type 9 (MMP9), tissue inhibitor of matrix metalloproteinases type 1 (TIMP1), transforming growth factor β type 1 (TGFβ1) and type 3 (TGFβ3) was determined by real-time polymerase chain reaction. Statistical analysis was performed using Statistica 10.0. Results: In ELC rats compared to sham-operated animals, a significant increase of all studied markers expression was observed. The administration of MSCs led to a significant decrease of all detectable markers in the experimental group compared to rats without cell therapy. In ELC rats, an increased MMP9/TIMP1 ratio after cell therapy was also detected. The infusion of MSCs in the sham-operated animals did not lead to any changes. In the HSCs from ELC animals, the expression of Col1a1 and Col3a1 exceeded the similar parameters of the control group (p <0.05) and statistically decreased after the MSCs administration. The correlation between Col3a1 (Rs = 0.51, p <0.05), TGFβ1 (Rs = 0.6, p <0.01), and TGFβ3 (Rs = 0.75, p <0.001) expression in HSCs cultures and liver tissue has been found. Conclusion: Intraportal administration of MSCs to rats with ELC leads to a decreased Col1a1 and Col3a1, MMP2 and MMP9, TIMP1, TGFβ1 and TGFβ3 expression. The correlation between the expression of Col3a1, TGFβ1 and TGFβ3 in liver tissue and in HSCs cultures indicates the involvement of activated HSCs in the fibrogenesis that allows considering HSCs to be the main cell therapy target in ELC.

Keywords: cell therapy, experimental liver cirrhosis, hepatic stellate cells, mesenchymal stem cells

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Authors: Marie Guittonny-LarchevêQue

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In boreal Canada, industrial activities such as forestry, peat extraction and metal mines often occur nearby. At closure, mine waste storage facilities have to be reclaimed. On tailings storage facilities, tree plantations can achieve rapid restoration of forested landscapes. However, trees poorly grow in mine tailings and organic amendments like peat are required to improve tailings’ structure and nutrients. Canada is a well-known producer of horticultural quality peat, but some lower quality peats coming from areas adjacent to the reclaimed mines could allow successful revegetation. In particular, hemic peat coming from the bottom of peat-bogs is more decomposed than fibric peat and is less valued for horticulture. Moreover, forest peat is sometimes excavated and piled by the forest industry after cuttings to stimulate tree regeneration on the exposed mineral soil. The objective of this project was to compare the ability of peats of differing quality and origin to improve tailings structure, nutrients and tree development. A greenhouse experiment was conducted along one growing season in 2016 with a complete randomized block design combining 8 repetitions (blocks) x 2 tree species (Populus tremuloides and Pinus banksiana) x 6 substrates (tailings, commercial horticultural peat, and mixtures of tailings with commercial peat, forest peat, local fibric peat, or local hemic peat) x 2 fertilization levels (with or without mineral fertilization). The used tailings came from a gold mine and were low in sulfur and trace metals. The commercial peat had a slightly acidic pH (around 6) while other peats had a clearly acidic pH (around 3). However, mixing peat with slightly alkaline tailings resulted in a pH close to 7 whatever the tested peats. The macroporosity of mixtures was intermediate between the low values of tailings (4%) and the high values of commercial peat alone (34%). Seedling survival was lower on tailings for poplar compared to all other treatments, with or without fertilization. Survival and growth were similar among all treatments for pine. Fertilization had no impact on the maximal height and diameter of poplar seedlings but changed the relative performance of the substrates. When not fertilized, poplar seedlings grown in commercial peat were the highest and largest, and the smallest and slenderest in tailings, with intermediate values in mixtures. When fertilized, poplar seedlings grown in commercial peat were smaller and slender compared to all other substrates. However for this species, foliar, shoot, and root biomass production was the greatest in commercial peat and the lowest in tailings compared to all mixtures, whether fertilized or not. The mixture with local fibric peat provided the seedlings with the lowest foliar N concentrations compared to all other substrates whatever the species or the fertilization treatment. At the short-term, the performance of all the tested peats were close when mixed to tailings, showing that peats of lower quality could be valorized instead of using horticultural peat. These results demonstrate that intersectorial synergies in accordance with the principles of circular economy may be developed in boreal Canada between local industries around the reclamation of mine waste dumps.

Keywords: boreal trees, mine spoil, mine revegetation, intersectorial synergies

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Authors: Esther Gumboh

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The police play a critical role in society and are an integral aspect of the rule of law. Equally, respect for human rights is an integral part of professional policing. In view of the vast powers that the police enjoy and the attendant risk of abuse and resulting human rights violations, the need for police accountability and civilian police oversight is internationally and regionally recognised. Policing oversight springs from the duty to investigate human rights violations. Those implicated in perpetrating or covering up violations must be disciplined or prosecuted to ensure effective accountability. Police accountability is particularly important in Malawi given the dark history of policing in the country during the 30-year dictatorial era under President Kamuzu Banda. Described as one of the most repressive regimes in Africa, the Banda administration was characterised by gross state-sponsored violence, repressive policing and human rights violations. Indeed, the police were involved in various forms of human rights abuse including arbitrary arrests and unlawful detentions, torture, and excessive use of force in conducting arrests and public order policing. This situation flourished within a culture of police impunity bolstered in part by the absence of clear oversight mechanisms for police accountability. In turn, there was immense public mistrust of the police. Unsurprisingly, the criminal justice system was one of the priority areas for reform when Malawi adopted its first democratic Constitution in 1994. Section 153 of the Constitution envisions a police service that is, for all intents and purposes, there to provide for the protection of public safety and the rights of persons in Malawi according to the prescriptions of the Constitution and any other law. This position reflects the view that the duty to protect and promote human rights is not incompatible with effective policing. Despite this, the police continue to engage in questionable behaviour in public order policing, excessive use of force, deaths in police custody, ill-treatment, torture and other forms of abuse including sexual abuse. Perpetrators of abuses are occasionally punished, but investigations are often delayed, abandoned, or remain inconclusive. Police accountability remains largely elusive. Commendably, the law does subject the police to significant oversight both internally and externally. However, until 2010, Malawi lacked a wholly independent civilian oversight mechanism specifically mandated to monitor the activities of the Malawi Police Service and held it accountable. This void has since been filled by the Independent Complaints Commission established under the Police Act. This is a positive development that reiterates Malawi’s commitment to the investigation of human rights violations by the police and to ending police impunity. This contribution examines the legal framework for this Commission to project the effectiveness of the Commission. While the framework looks promising on various fronts, there are potential challenges that lie ahead. Malawi must pre-emptively deal with these challenges carefully if the Commission is to have any practical significance in transforming police accountability in the country. Drawing on lessons from other jurisdictions like South Africa, the paper makes recommendations for legislative reform to strengthen the Commission’s framework.

Keywords: civilian policing oversight, Malawi, police, police accountability, policing, policing oversight

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Authors: Simran Baweja, Bhavika Kalal, Surajit Maity

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Photoinduced tautomerization reactions have been the centre of attention among scientific community over past several decades because of their significance in various biological systems. 7-azaindole (7AI) is considered as a model system for DNA base pairing and to understand the role of such tautomerization reactions in mutations. To the best of our knowledge, extensive studies have been carried on 7-azaindole and its solvent clusters exhibiting proton/ hydrogen transfer in both solution as well as gas phase. Derivatives of above molecule, like 2,7- and 2,6-diazaindoles are proposed to have even better photophysical properties due to the presence of -aza group on the 2nd position. However, there are a few studies in the solution phase which suggest the relevance of these molecules, but there are no experimental studies reported in the gas phase yet. In our current investigation, we present the first gas phase spectroscopic data of 2,7-diazaindole (2,7-DAI) and its solvent cluster (2,7-DAI-H2O). In this, we have employed state-of-the-art laser spectroscopic methods such as fluorescence excitation (LIF), dispersed fluorescence (DF), resonant two-photon ionization time of flight mass spectrometry (2C-R2PI), photoionization efficiency spectroscopy (PIE), IR-UV double resonance spectroscopy i.e. fluorescence-dip infrared spectroscopy (FDIR) and resonant ion-dip infrared spectroscopy (IDIR) to understand the electronic structure of the molecule. The origin band corresponding to S1 ← S0 transition of the bare 2,7-DAI is found to be positioned at 33910 cm-1 whereas the origin band corresponding to S1 ← S0 transition of the 2,7-DAI-H2O is positioned at 33074 cm-1. The red shifted transition in case of solvent cluster suggests the enhanced feasibility of excited state hydrogen/ proton transfer. The ionization potential for the 2,7-DAI molecule is found to be 8.92 eV, which is significantly higher that the previously reported 7AI (8.11 eV) molecule, making it a comparatively complex molecule to study. The ionization potential is reduced by 0.14 eV in case of 2,7-DAI-H2O (8.78 eV) cluster compared to that of 2,7-DAI. Moreover, on comparison with the available literature values of 7AI, we found the origin band of 2,7-DAI and 2,7-DAI-H2O to be red shifted by -729 and -280 cm-1 respectively. The ground and excited state N-H stretching frequencies of the 27DAI molecule were determined using fluorescence-dip infrared spectra (FDIR) and resonant ion dip infrared spectroscopy (IDIR), obtained at 3523 and 3467 cm-1, respectively. The lower value of vNH in the electronic excited state of 27DAI implies the higher acidity of the group compared to the ground state. Moreover, we have done extensive computational analysis, which suggests that the energy barrier in excited state reduces significantly as we increase the number of catalytic solvent molecules (S= H2O, NH3) as well as the polarity of solvent molecules. We found that the ammonia molecule is a better candidate for hydrogen transfer compared to water because of its higher gas-phase basicity. Further studies are underway to understand the excited state dynamics and photochemistry of such N-rich chromophores.

Keywords: photoinduced tautomerization reactions, gas phse spectroscopy, ), IR-UV double resonance spectroscopy, resonant two-photon ionization time of flight mass spectrometry (2C-R2PI)

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Authors: Olugbenga S. Alebiosu, Olusola H. Adekanmbi, Oluwatoyin T. Ogundipe

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Pollen and spores have been identified as major airborne bio-particles inducing respiratory disorders such as asthma, allergic rhinitis and atopic dermatitis among hypersensitive individuals. An aeropalynological study was conducted within a one year sampling period with a view to investigating the monthly depositional rate of atmospheric pollen and spores; influence of the immediate vegetation on airborne pollen distribution; allergenic potentials of dominant atmospheric pollen types at selected study locations in Bauchi and Taraba states, Northwestern Nigeria. A tauber-like pollen trap was employed in aerosampling with the sampler positioned at a height of 5 feet above the ground, followed by a monthly collection of the recipient solution for the sampling period. The collected samples were subjected to acetolysis treatment, examined microscopically with the identification of pollen grains and spores using reference materials and published photomicrographs. Plants within the surrounding vegetation were enumerated. Crude protein contents extracted from pollen types found to be commonly dominant at both study locations; Senna siamea, Terminalia cattapa, Panicum maximum and Zea mays were used to sensitize Musmusculus. Histopathological studies of bronchi and lung sections from certain dead M.musculus in the test groups was conducted. Blood samples were collected from the pre-orbital vein of M.musculus and processed for serological and haematological (differential and total white blood cell counts) studies. ELISA was used in determining the levels of serological parameters: IgE and cytokines (TNF-, IL-5, and IL-13). Statistical significance was observed in the correlation between the levels of serological and haematological parameters elicited by each test group, differences between the levels of serological and haematological parameters elicited by each test group and those of the control, as well as at varying sensitization periods. The results from this study revealed dominant airborne pollen types across the study locations; Syzygiumguineense, Tridaxprocumbens, Elaeisguineensis, Mimosa sp., Borreria sp., Terminalia sp., Senna sp. and Poaceae. Nephrolepis sp., Pteris sp. and a trilete fern also produced spores. This study also revealed that some of the airborne pollen types were produced by local plants at the study locations. Bronchi sections of M.musculus after first and second sensitizations, as well as lung section after first sensitization with Senna siamea, showed areas of necrosis. Statistical significance was recorded in the correlation between the levels of some serological and haematological parameters produced by each test group and those of the control, as well as at certain sensitization periods. The study revealed some candidate pollen allergens at the study locations allergy sufferers and also established a complexity of interaction between immune cells, IgE and cytokines at varied periods of mice sensitization and forming a paradigm of human immune response to different pollen allergens. However, it is expedient that further studies should be conducted on these candidate pollen allergens for their allergenicity potential in humans within their immediate environment.

Keywords: airborne, hypersensitive, mus musculus, pollen allergens, respiratory, tauber-like

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Authors: Leigha C. Crout

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This paper will investigate the categorical disregard of the People’s Republic of China (PRC) in establishing and maintaining a baseline standard of civil guarantees for economic migrant women and their dependents. In light of the relative forward strides in terms of policy facilitating the ascension of female workers in China, this oft-invisible subgroup of women remains neglected from the modern-day “iron rice bowl” of the self-identified communist state. This study is being undertaken to rectify the absence of data on this subject and provide a baseline for future studies on the matter, as the human rights of migrants has become an established facet of transnational dialogue and debate. The basic methodology of this research will consist of the evaluation of China’s compliance with its own national guidelines, and the eight international human rights law treaties it has ratified. Data will be extracted and cross-checked from a number of relevant sources to monitor the extent of compliance, including but by no means limited to the United Nations Human Rights Council (UNHRC) Universal Periodic Review (UPR) reports and responses, submissions and responses of international human rights treaty bodies, local and international nongovernmental organizations (NGOs) and their annual reports, and articles and commentaries authored by specialists on the modern state and implementation of Chinese law. Together, these data will illuminate the vast network of compliance that has forced many migrant women to work within situations of extreme economic precarity. The structure will proceed as follows: first, an outline of the current status of migrant workers and the enforcement of stipulated protections will be provided; next, the analysis of the oft-debated regulations directing and the outline of mandatory services guaranteed to external and internal migrants; and finally, a conclusion incorporating various recommendations to improve transparency and gradually decrease the amount of migrant work turned forced labor that typifies the economic migrant experience, especially in the case of women. The internal and international migrant workers in China are bound by different and uncomplimentary systems. The first, which governs Chinese citizens moving to different regions or provinces to find more sustainable employment (internal migrants), is called the hukou (or huji) residency system. This law enforces strict regulation of the movement of peoples, while ensuring that residents of urban areas receive preferential benefits to those received by their so-called “agricultural” resident counterparts. Given the overwhelming presence of the Communist Party of China throughout the vast state, the management of internal migrants and the disregard for foreign domestic workers is, at minimum, a surprising oversight. This paper endeavors to provide a much-needed foundation for future commentary and discussion on the treatment of female migrant workers and their families in the People’s Republic of China.

Keywords: female migrant worker’s rights, the People’s Republic of China, forced labor, Hukou residency system

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Authors: Yakup Ulusu, Faruk Ozel, Numan Eczacioglu, Abdurrahman Ozen, Sabriye Acikgoz

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GFP discovered in the mid-1970s, has been used as a marker after replicated genetic study by scientists. In biotechnology, cell, molecular biology, the GFP gene is frequently used as a reporter of expression. In modified forms, it has been used to make biosensors. Many animals have been created that express GFP as an evidence that a gene can be expressed throughout a given organism. Proteins labeled with GFP identified locations are determined. And so, cell connections can be monitored, gene expression can be reported, protein-protein interactions can be observed and signals that create events can be detected. Additionally, monitoring GFP is noninvasive; it can be detected by under UV-light because of simply generating fluorescence. Moreover, GFP is a relatively small and inert molecule, that does not seem to treat any biological processes of interest. The synthesis of GFP has some steps like, to construct the plasmid system, transformation in E. coli, production and purification of protein. GFP carrying plasmid vector pBAD–GFPuv was digested using two different restriction endonuclease enzymes (NheI and Eco RI) and DNA fragment of GFP was gel purified before cloning. The GFP-encoding DNA fragment was ligated into pET28a plasmid using NheI and Eco RI restriction sites. The final plasmid was named pETGFP and DNA sequencing of this plasmid indicated that the hexa histidine-tagged GFP was correctly inserted. Histidine-tagged GFP was expressed in an Escherichia coli BL21 DE3 (pLysE) strain. The strain was transformed with pETGFP plasmid and grown on LuiraBertoni (LB) plates with kanamycin and chloramphenicol selection. E. coli cells were grown up to an optical density (OD 600) of 0.8 and induced by the addition of a final concentration of 1mM isopropyl-thiogalactopyranoside (IPTG) and then grown for additional 4 h. The amino-terminal hexa-histidine-tag facilitated purification of the GFP by using a His Bind affinity chromatography resin (Novagen). Purity of GFP protein was analyzed by a 12 % sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The concentration of protein was determined by UV absorption at 280 nm (Varian Cary 50 Scan UV/VIS spectrophotometer). Synthesis of GFP-Polymer composite nanofibers was produced by using GFP solution (10mg/mL) and polymer precursor Polyvinylpyrrolidone, (PVP, Mw=1300000) as starting materials and template, respectively. For the fabrication of nanofibers with the different fiber diameter; a sol–gel solution comprising of 0.40, 0.60 and 0.80 g PVP (depending upon the desired fiber diameter) and 100 mg GFP in 10 mL water: ethanol (3:2) mixtures were prepared and then the solution was covered on collecting plate via electro spinning at 10 kV with a feed-rate of 0.25 mL h-1 using Spellman electro spinning system. Results show that GFP-based nano-fiber can be used plenty of biomedical applications such as bio-imaging, bio-mechanic, bio-material and tissue engineering.

Keywords: biomaterial, GFP, nano-fibers, protein expression

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Authors: Irfan Anjum Manarvi, Fawzi Aljassir

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Human bones have been a keen area of research over a long time in the field of biomechanical engineering. Medical professionals, as well as engineering academics and researchers, have investigated various bones by using medical, mechanical, and materials approaches to discover the available body of knowledge. Their major focus has been to establish properties of these and ultimately develop processes and tools either to prevent fracture or recover its damage. Literature shows that mechanical professionals conducted a variety of tests for hardness, deformation, and strain field measurement to arrive at their findings. However, they considered these results accuracy to be insufficient due to various limitations of tools, test equipment, difficulties in the availability of human bones. They proposed the need for further studies to first overcome inaccuracies in measurement methods, testing machines, and experimental errors and then carry out experimental or theoretical studies. Finite Element analysis is a technique which was developed for the aerospace industry due to the complexity of design and materials. But over a period of time, it has found its applications in many other industries due to accuracy and flexibility in selection of materials and types of loading that could be theoretically applied to an object under study. In the past few decades, the field of biomechanical engineering has also started to see its applicability. However, the work done in the area of Tarsals, metatarsals and phalanges using this technique is very limited. Therefore, present research has been focused on using this technique for analysis of these critical bones of the human body. This technique requires a 3-dimensional geometric computer model of the object to be analyzed. In the present research, a 3d laser scanner was used for accurate geometric scans of individual tarsals, metatarsals, and phalanges from a typical human foot to make these computer geometric models. These were then imported into a Finite Element Analysis software and a length refining process was carried out prior to analysis to ensure the computer models were true representatives of actual bone. This was followed by analysis of each bone individually. A number of constraints and load conditions were applied to observe the stress and strain distributions in these bones under the conditions of compression and tensile loads or their combination. Results were collected for deformations in various axis, and stress and strain distributions were observed to identify critical locations where fracture could occur. A comparative analysis of failure properties of all the three types of bones was carried out to establish which of these could fail earlier which is presented in this research. Results of this investigation could be used for further experimental studies by the academics and researchers, as well as industrial engineers, for development of various foot protection devices or tools for surgical operations and recovery treatment of these bones. Researchers could build up on these models to carryout analysis of a complete human foot through Finite Element analysis under various loading conditions such as walking, marching, running, and landing after a jump etc.

Keywords: tarsals, metatarsals, phalanges, 3D scanning, finite element analysis

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Authors: Simone Raimondi De Souza, Glaucia Maria Moraes De Oliveira, Ronir Raggio Luiz, Glorimar Rosa

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Introduction: Obesity is among the main risk factors for cardiovascular disease (CVD). Genesis is multifactorial, including genetic, hormonal and environmental factors disorders, among which inadequate feeding pattern, for which nutritional counseling strategies have proven effective. The consumption of beta-glucans (soluble fibers that reportedly promote satiety) present in oat bran can be an effective strategy for preventing and treating obesity. Other benefits have been observed with oat bran consumption, such as reduction of hypercholesterolemia and hyperglycemia, two other risk factors for CVD. Objectives: To analyze the effect of oat bran consumption associated with nutritional counseling in reducing body mass index (BMI), blood cholesterol, glucose profile, waist and neck circumference in obese individuals, and to evaluate the change in eating pattern. Methods: clinical trial, randomized, double-blind, placebo-controlled, lasting 90 days with adults of both genders, with BMI ≥30kg/m2. The study was approved by the Ethics in Research involving human beings in a public institute of cardiology, in Rio de Janeiro, Brazil. Individuals were invited to participate and accepted formally by signing the Terms of Consent. Participants were randomized into oat bran group (gOB) or placebo group (gPCB) and received, respectively: morning prepared consisting of 40g oat bran, 30g of skimmed milk powder and 1g sweetener sucralose; refined flour 40g rice, 30g of milk powder and 1g sweetener sucralose. The Ten Steps to Healthy Eating, of Brazilian Ministry of Health were used to support the nutritional counseling. Variables analyzed: gender; age; BMI, waist circumference (WC) neck circumference (NC); systolic blood pressure (SBP); diastolic blood pressure (DBP); food consumption, total cholesterol (TC), LDL-cholesterol (LDL-c), HDL-cholesterol (HDL-c), non-HDL cholesterol (nHDLc), triglycerides (TG), fasting glucose (FG), fasting insulin (FI) and HOMA-IR. Dietary intake was assessed by 24-hour dietary recall. The Diet Quality Index revised for the Brazilian population (IQD-R) assessed quality of feeding pattern. Statistical analyzes were performed using SPSS version 21, considering statistically significant p-value less than 0.05. Results: A total of 38 participants were included, age = 50 ± 7,6years, 63% women. 19 subjects were placed in gOB and 19 in gPCB. After intervention, statistically significant reductions were observed in the following parameters: in gOB: IQD-R, TC, LDL-c, nHDL-c, FI, SBP, DBP, BMI, WC, NC; in gPCB: IQD-R, LDL-c, SBP, DBP, BMI, WC, NC. No statistically significant differences were observed in the results between groups. Conclusion: Our results reinforce nutritional counseling as important strategy for prevention and treatment of obesity and suggest that inclusion of oat bran in daily diet can bring additional benefits controlling risk factors for CVD. More studies are needed to establish all benefits of oat bran to human health as well as the ideal daily dose for consumption.

Keywords: oat bran, cardiovascular disease, nutritional counseling, obesity

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Authors: V. Vsevolozhskaia

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Since 1990, energy-efficient development concepts have constituted both a turning point in civil engineering and a challenge for an environmentally friendly future. Energy and water currently play an essential role in the sustainable economic growth of the world in general and Russia in particular: the efficiency of the water supply system is the second most important parameter for energy consumption according to the British assessment method, while the water-energy nexus has been identified as a focus for accelerating sustainable growth and developing effective, innovative solutions. The activities considered in this study were aimed at organizing and executing the renovation of the property in residential buildings located in St. Petersburg, specifically buildings with local or federal historical heritage status under the control of the St. Petersburg Committee for the State Inspection and Protection of Historic and Cultural Monuments (KGIOP) and UNESCO. Even after reconstruction, these buildings still fall into energy efficiency class D. Russian Government Resolution No. 87 on the structure and required content of project documentation contains a section entitled ‘Measures to ensure compliance with energy efficiency and equipment requirements for buildings, structures, and constructions with energy metering devices’. Mention is made of the need to install collectors and meters, which only calculate energy, neglecting the main purpose: to make buildings more energy-efficient, potentially even energy efficiency class A. The least-explored aspects of energy-efficient technology in the Russian Federation remain the water balance and the possibility of implementing rain and meltwater collection systems. These modern technologies are used exclusively for new buildings due to a lack of government directive to create project documentation during the planning of major renovations and reconstruction that would include the collection and reuse of rainwater. Energy-efficient technology for rain and meltwater collection is currently applied only to new buildings, even though research has proved that using rainwater is safe and offers a huge step forward in terms of eco-efficiency analysis and water innovation. Where conservation is mandatory, making changes to protected sites is prohibited. In most cases, the protected site is the cultural heritage building itself, including the main walls and roof. However, the installation of a second water supply system and collection of rainwater would not affect the protected building itself. Water efficiency in St. Petersburg is currently considered only from the point of view of the installation that regulates the flow of the pipeline shutoff valves. The development of technical guidelines for the use of grey- and/or rainwater to meet the needs of residential buildings during reconstruction or renovation is not yet complete. The ideas for water treatment, collection and distribution systems presented in this study should be taken into consideration during the reconstruction or renovation of residential cultural heritage buildings under the protection of KGIOP and UNESCO. The methodology applied also has the potential to be extended to other cultural heritage sites in northern countries and lands with an average annual rainfall of over 600 mm to cover average toilet-flush needs.

Keywords: cultural heritage, energy efficiency, renovation, rainwater collection, reconstruction, water management, water supply

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Authors: Grzegorz Raniszewski, Zbigniew Kolacinski, Lukasz Szymanski, Slawomir Wiak, Lukasz Pietrzak, Dariusz Koza

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One of the most promising area for carbon nanotubes (CNTs) application is medicine. One of the most devastating diseases is cancer. Carbon nanotubes may be used as carriers of a slowly released drug. It is possible to use of electromagnetic waves to destroy cancer cells by the carbon nanotubes (CNTs). In our research we focused on thermal ablation by ferromagnetic carbon nanotubes (Fe-CNTs). In the cancer cell hyperthermia functionalized carbon nanotubes are exposed to radio frequency electromagnetic field. Properly functionalized Fe-CNTs join the cancer cells. Heat generated in nanoparticles connected to nanotubes warm up nanotubes and then the target tissue. When the temperature in tumor tissue exceeds 316 K the necrosis of cancer cells may be observed. Several techniques can be used for Fe-CNTs synthesis. In our work, we use high-temperature methods where arc-discharge is applied. Low-temperature systems are microwave plasma with assisted chemical vapor deposition (MPCVD) and hybrid physical-chemical vapor deposition (HPCVD). In the arc discharge system, the plasma reactor works with a pressure of He up to 0,5 atm. The electric arc burns between two graphite rods. Vapors of carbon move from the anode, through a short arc column and forms CNTs which can be collected either from the reactor walls or cathode deposit. This method is suitable for the production of multi-wall and single-wall CNTs. A disadvantage of high-temperature methods is a low purification, short length, random size and multi-directional distribution. In MPCVD system plasma is generated in waveguide connected to the microwave generator. Then containing carbon and ferromagnetic elements plasma flux go to the quartz tube. The additional resistance heating can be applied to increase the reaction effectiveness and efficiency. CNTs nucleation occurs on the quartz tube walls. It is also possible to use substrates to improve carbon nanotubes growth. HPCVD system involves both chemical decomposition of carbon containing gases and vaporization of a solid or liquid source of catalyst. In this system, a tube furnace is applied. A mixture of working and carbon-containing gases go through the quartz tube placed inside the furnace. As a catalyst ferrocene vapors can be used. Fe-CNTs may be collected then either from the quartz tube walls or on the substrates. Low-temperature methods are characterized by higher purity product. Moreover, carbon nanotubes from tested CVD systems were partially filled with the iron. Regardless of the method of Fe-CNTs synthesis the final product always needs to be purified for applications in medicine. The simplest method of purification is an oxidation of the amorphous carbon. Carbon nanotubes dedicated for cancer cell thermal ablation need to be additionally treated by acids for defects amplification on the CNTs surface what facilitates biofunctionalization. Application of ferromagnetic nanotubes for cancer treatment is a promising method of fighting with cancer for the next decade. Acknowledgment: The research work has been financed from the budget of science as a research project No. PBS2/A5/31/2013

Keywords: arc discharge, cancer, carbon nanotubes, CVD, thermal ablation

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Authors: Deinzel R. Uezono, Vivien Fe F. Fadrilan-Camacho, Bianca Margarita L. Medina, Antonio Domingo R. Reario, Trisha M. Salcedo, Luke Wesley P. Borromeo

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The environment of health care workers is considered one of the most hazardous settings due to the nature of their work. In developing countries especially, the Philippines, this continues to be overlooked in terms of programs and services on occupational health and safety (OHS). One possible reason for this is the existing information gap on OHS which limits data comparability and impairs effective monitoring and assessment of interventions. To address this gap, there is a need to determine the current conditions of Filipino health care workers in their workplace. This descriptive cross-sectional study assessed the occupational health and safety conditions of health care workers in barangay health centers in a selected city in Metro Manila, Philippines by: (1) determining the hazards present in the workplace; (2) determining the most common self-reported medical problems; and (3) describing the elements of an OHS system based on the six building blocks of health system. Assessment was done through walkthrough survey, self-administered questionnaire, and key informant interview. Data analysis was done using Epi Info 7 and NVivo 11. Results revealed different health hazards present in the workplace particularly biological hazards (exposure to sick patients and infectious specimens), physical hazards (inadequate space and/or lighting), chemical hazards (toxic reagents and flammable chemicals), and ergonomic hazards (activities requiring repetitive motion and awkward posture). Additionally, safety hazards (improper capping of syringe and lack of fire safety provisions) were also observed. Meanwhile, the most commonly self-reported chronic diseases among health care workers (N=336) were hypertension (20.24%, n=68) and diabetes (12.50%, n=42). Top commonly self-reported symptoms were colds (66.07%, n=222), coughs (63.10%, n=212), headache (55.65%, n=187), and muscle pain (50.60%, n=170) while other diseases were influenza (16.96%, n=57) and UTI (15.48%, n=52). In terms of the elements of the OHS system, a general policy on occupational health and safety was found to be lacking and in effect, an absence of health and safety committee overseeing the implementing and monitoring of the policy. No separate budget specific for OHS programs and services was also found to be a limitation. As a result, no OHS personnel and trainings/seminar were identified. No established information system for OHS was in place. In conclusion, health and safety hazards were observed to be present across the barangay health centers visited in a selected city in Metro Manila. Medical conditions identified as most commonly self-reported were hypertension and diabetes for chronic diseases; colds, coughs, headache, and muscle pain for medical symptoms; and influenza and UTI for other diseases. As for the elements of the occupational health and safety system, there was a lack in the general components of the six building blocks of the health system.

Keywords: health hazards, occupational health and safety, occupational health and safety system, safety hazards

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Authors: Vrushali Guhe, Shailza Singh

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Cutaneous leishmaniasis is neglected tropical disease present worldwide caused by the protozoan parasite Leishmania major, the therapeutic armamentarium for leishmaniasis are showing several limitations as drugs are showing toxic effects with increasing resistance by a parasite. Thus identification of novel therapeutic targets is of paramount importance. Previous studies have shown that autophagy, a cellular process, can either facilitate infection or aid in the elimination of the parasite, depending on the specific parasite species and host background in leishmaniasis. In the present study, our objective was to target the essential autophagy protein ATG8, which plays a crucial role in the survival, infection dynamics, and differentiation of the Leishmania parasite. ATG8 in Leishmania major and its homologue, LC3, in Homo sapiens, act as autophagic markers. Present study manifested the crucial role of ATG8 protein as a potential target for combating Leishmania major infection. Through bioinformatics analysis, we identified non-conserved motifs within the ATG8 protein of Leishmania major, which are not present in LC3 of Homo sapiens. Against these two non-conserved motifs, we generated a peptide library of 60 peptides on the basis of physicochemical properties. These peptides underwent a filtering process based on various parameters, including feasibility of synthesis and purification, compatibility with Selective Reaction Monitoring (SRM)/Multiple reaction monitoring (MRM), hydrophobicity, hydropathy index, average molecular weight (Mw average), monoisotopic molecular weight (Mw monoisotopic), theoretical isoelectric point (pI), and half-life. Further filtering criterion shortlisted three peptides by using molecular docking and molecular dynamics simulations. The direct interaction between ATG8 and the shortlisted peptides was confirmed through Surface Plasmon Resonance (SPR) experiments. Notably, these peptides exhibited the remarkable ability to penetrate the parasite membrane and exert profound effects on Leishmania major. The treatment with these peptides significantly impacted parasite survival, leading to alterations in the cell cycle and morphology. Furthermore, the peptides were found to modulate autophagosome formation, particularly under starved conditions, suggesting their involvement in disrupting the regulation of autophagy within Leishmania major. In vitro, studies demonstrated that the selected peptides effectively reduced the parasite load within infected host cells. Encouragingly, these findings were corroborated by in vivo experiments, which showed a reduction in parasite burden upon peptide administration. Additionally, the peptides were observed to affect the levels of LC3II within host cells. In conclusion, our findings highlight the efficacy of these novel peptides in targeting Leishmania major’s ATG8 and disrupting parasite survival. These results provide valuable insights into the development of innovative therapeutic strategies against leishmaniasis via targeting autophagy protein ATG8 of Leishmania major.

Keywords: ATG8, leishmaniasis, surface plasmon resonance, MD simulation, molecular docking, peptide designing, therapeutics

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Authors: Mitali Saha, Soma Das

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The fabrication of nanoscale materials for use in chemical sensing, biosensing and biological analyses has proven a promising avenue in the last few years. Cholesterol has aroused considerable interest in recent years on account of its being an important parameter in clinical diagnosis. There is a strong positive correlation between high serum cholesterol level and arteriosclerosis, hypertension, and myocardial infarction. Enzyme-based electrochemical biosensors have shown high selectivity and excellent sensitivity, but the enzyme is easily denatured during its immobilization procedure and its activity is also affected by temperature, pH, and toxic chemicals. Besides, the reproducibility of enzyme-based sensors is not very good which further restrict the application of cholesterol biosensor. It has been demonstrated that carbon nanotubes could promote electron transfer with various redox active proteins, ranging from cytochrome c to glucose oxidase with a deeply embedded redox center. In continuation of our earlier work on the synthesis and applications of carbon and metal based nanoparticles, we have reported here the synthesis of carbon nanotubes (CCNT) by burning coconut oil under insufficient flow of air using an oil lamp. The soot was collected from the top portion of the flame, where the temperature was around 6500C which was purified, functionalized and then characterized by SEM, p-XRD and Raman spectroscopy. The SEM micrographs showed the formation of tubular structure of CCNT having diameter below 100 nm. The XRD pattern indicated the presence of two predominant peaks at 25.20 and 43.80, which corresponded to (002) and (100) planes of CCNT respectively. The Raman spectrum (514 nm excitation) showed the presence of 1600 cm-1 (G-band) related to the vibration of sp2-bonded carbon and at 1350 cm-1 (D-band) responsible for the vibrations of sp3-bonded carbon. A nonenzymatic cholesterol biosensor was then fabricated on an insulating Teflon material containing three silver wires at the surface, covered by CCNT, obtained from coconut oil. Here, CCNTs worked as working as well as counter electrodes whereas reference electrode and electric contacts were made of silver. The dimensions of the electrode was 3.5 cm×1.0 cm×0.5 cm (length× width × height) and it is ideal for working with 50 µL volume like the standard screen printed electrodes. The voltammetric behavior of cholesterol at CCNT electrode was investigated by cyclic voltammeter and differential pulse voltammeter using 0.001 M H2SO4 as electrolyte. The influence of the experimental parameters on the peak currents of cholesterol like pH, accumulation time, and scan rates were optimized. Under optimum conditions, the peak current was found to be linear in the cholesterol concentration range from 1 µM to 50 µM with a sensitivity of ~15.31 μAμM−1cm−2 with lower detection limit of 0.017 µM and response time of about 6s. The long-term storage stability of the sensor was tested for 30 days and the current response was found to be ~85% of its initial response after 30 days.

Keywords: coconut oil, CCNT, cholesterol, biosensor

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Authors: Noor S. Nasri, Eric C. A. Tatt, Usman D. Hamza, Jibril Mohammed, Husna M. Zain

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Climate change has becoming a global environmental issue that may trigger irreversible changes in the environment with catastrophic consequences for human, animals and plants on our planet. Methane, carbon dioxide and nitrous oxide are the greenhouse gases (GHG) and as the main factor that significantly contributes to the global warming. Mainly carbon dioxide be produced and released to atmosphere by thermal industrial and power generation sectors. Methane is dominant component of natural gas releases significant of thermal heat, and the gaseous pollutants when homogeneous thermal combustion takes place at high temperature. Heterogeneous catalytic Combustion (HCC) principle is promising technologies towards environmental friendly energy production should be developed to ensure higher yields with lower pollutants gaseous emissions and perform complete combustion oxidation at moderate temperature condition as comparing to homogeneous high thermal combustion. Hence the principle has become a very interesting alternative total oxidation for the treatment of pollutants gaseous emission especially NOX product formation. Noble metals are dispersed on a support-porous HCC such as γ- Al2O3, TiO2 and ThO2 to increase thermal stability of catalyst and to increase to effectiveness of catalytic combustion. Support-porous HCC material to be selected based on factors of the surface area, porosity, thermal stability, thermal conductivity, reactivity with reactants or products, chemical stability, catalytic activity, and catalyst life. γ- Al2O3 with high catalytic activity and can last longer life of catalyst, is commonly used as the support for Pd catalyst at low temperatures. Sustainable and renewable support-material of bio-mass char was derived from agro-industrial waste material and used to compare with those the conventional support-porous material. The abundant of biomass wastes generated in palm oil industries is one potential source to convert the wastes into sustainable material as replacement of support material for catalysts. Objective of this study was to compare the kinetic rate of reaction the combustion of methane on Palladium (Pd) based catalyst with Al2O3 support and bio-char (Bc) support derived from shell kernel. The 2wt% Pd was prepared using incipient wetness impregnation method and the HCC performance was accomplished using tubular quartz reactor with gas mixture ratio of 3% methane and 97% air. Material characterization was determined using TGA, SEM, and BET surface area. The methane porous-HCC conversion was carried out by online gas analyzer connected to the reactor that performed porous-HCC. BET surface area for prepared 2 wt% Pd/Bc is smaller than prepared 2wt% Pd/ Al2O3 due to its low porosity between particles. The order of catalyst activity based on kinetic rate on reaction of catalysts in low temperature is prepared 2wt% Pd/Bc > calcined 2wt% Pd/ Al2O3 > prepared 2wt% Pd/ Al2O3 > calcined 2wt% Pd/Bc. Hence the usage of agro-industrial bio-mass waste material can enhance the sustainability principle.

Keywords: catalytic-combustion, environmental, support-bio-char material, sustainable and renewable material

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Authors: Juliana Chiarini Balbino Fernandes

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This article aims to investigate the Spanish didactic book, entitled ‘El Calculo y La Medida en El Primer Grado de La Escuela Decroly’, written by Dr. O. Decroly and A. Hamaide, published in Madrid, in the year 1934. In addition to analyzing how mathematical knowledge is present in the proposed Centers of Interest. The textbooks, in addition to pedagogical tools, reflect a certain moment in society and allow the analysis of the theoretical-methodological proposal that can be implemented by the teacher. The study proposed here will be carried out by the lens of Cultural History, supported by Roger Chartier (1991) and by the concepts on textbooks, based on Alain Choppin (2004). The textbook selected for this study exposes a program of ideas associated with the method of Centers of Interest and arithmetic is linked to these interests. In the first courses (six to eight years), most centers can be considered to correspond to occasional calls, as they take advantage of events that arise spontaneously to work with observation, measurement, association and expression exercises. The program of ideas associated with Centers of Interest addresses the biological and social aspects of children, as long as they can express their needs for activities and games, satisfying the natural curiosity. Still, the program of associated ideas offers occasions for problems whose data are taken in observation exercises and concrete expressions (manuals, drawings). In the method applied at the school of L'Ermitage, school created by Decroly in Belgium in 1907, observation, is the basis of each center of interest. It offers the chance to compare and measure. To observe is more than to perceive; it is also to establish relations between the graded aspects of the same object, to seek relations between different intensities; is to verify successions, special and temporary relationships; is to make comparisons, to notice differences and similarities in block or datable (analysis), is to establish a bridge between the world and the thought. To make the observation more precise, it is important to compare, measure, and resort to considered objects as natural units of measure. Measurement and calculation are, therefore, quite naturally subject to observation. Thus, it is possible to make the child enter into the interest in the calculation, linking it to the observation. It was observed that the Centers of Interest, according to Decroly, should respond to the concerns and attend to the motivations of the students and the teaching of arithmetical must obey a logical seriation, considering the interest and the experience of the children. The teaching of arithmetical should not be limited to the schedule, it should cover every quantitative aspect that arises in the other disciplines. The feeling of unity is established in observation, association and expression, which coordinate a whole program of cultural activities, concentrating it around a central idea.

Keywords: didactic book, centers of interest, mathematical knowledge, primary education

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Authors: Geraldine G. Granados Vazquez

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Every human being is exposed to the risk of dying; wherein some of them are more susceptible than others depending on the cause. Therefore, the cause could be the hazard to die that a group or individual has, making this irreversible damage the condition of vulnerability. Risk is a dynamic concept; which means that it depends on the environmental, social, economic and political conditions. Thus vulnerability may only be evaluated in terms of relative parameters. This research is focusing specifically on building a model that evaluate the risk or propensity of death in past urban societies in connection with the everyday life of individuals, considering that death can be a consequence of two coexisting issues: hazard and the deterioration of the resistance to destruction. One of the most important discussions in bioarchaeology refers to health and life conditions in ancient groups; the researchers are looking for more flexible models that evaluate these topics. In that way, this research proposes a theoretical-methodological model that assess the vulnerability of death in past urban groups. This model pretends to be useful to evaluate the risk of death, considering their sociohistorical context, and their intrinsic biological features. This theoretical and methodological model, propose four areas to assess vulnerability. The first three areas use statistical methods or quantitative analysis. While the last and fourth area, which corresponds to the embodiment, is based on qualitative analysis. The four areas and their techniques proposed are a) Demographic dynamics. From the distribution of age at the time of death, the analysis of mortality will be performed using life tables. From here, four aspects may be inferred: population structure, fertility, mortality-survival, and productivity-migration, b) Frailty. Selective mortality and heterogeneity in frailty can be assessed through the relationship between characteristics and the age at death. There are two indicators used in contemporary populations to evaluate stress: height and linear enamel hypoplasias. Height estimates may account for the individual’s nutrition and health history in specific groups; while enamel hypoplasias are an account of the individual’s first years of life, c) Inequality. Space reflects various sectors of society, also in ancient cities. In general terms, the spatial analysis uses measures of association to show the relationship between frail variables and space, d) Embodiment. The story of everyone leaves some evidence on the body, even in the bones. That led us to think about the dynamic individual's relations in terms of time and space; consequently, the micro analysis of persons will assess vulnerability from the everyday life, where the symbolic meaning also plays a major role. In sum, using some Mesoamerica examples, as study cases, this research demonstrates that not only the intrinsic characteristics related to the age and sex of individuals are conducive to vulnerability, but also the social and historical context that determines their state of frailty before death. An attenuating factor for past groups is that some basic aspects –such as the role they played in everyday life– escape our comprehension, and are still under discussion.

Keywords: bioarchaeology, frailty, Mesoamerica, vulnerability

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Authors: Alaa Fezai, Anuj Sharma, Wolfgang Mueller-Hirsch, André Zimmermann

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Viscoelastic materials have been widely used in the automotive industry over the last few decades with different functionalities. Besides their main application as a simple and efficient surface damping treatment, they may ensure optimal operating conditions for on-board electronics as thermal interface or sealing layers. The dynamic behavior of viscoelastic materials is generally dependent on many environmental factors, the most important being temperature and strain rate or frequency. Prior to the reliability analysis of systems including viscoelastic layers, it is, therefore, crucial to accurately predict the dynamic and lifetime behavior of these materials. This includes the identification of the dynamic material parameters under critical temperature and frequency conditions along with a precise damage localization and identification methodology. The goal of this work is twofold. The first part aims at applying an inverse viscoelastic material-characterization approach for a wide frequency range and under different temperature conditions. For this sake, dynamic measurements are carried on a single lap joint specimen using an electrodynamic shaker and an environmental chamber. The specimen consists of aluminum beams assembled to adapter plates through a viscoelastic adhesive layer. The experimental setup is reproduced in finite element (FE) simulations, and frequency response functions (FRF) are calculated. The parameters of both the generalized Maxwell model and the fractional derivatives model are identified through an optimization algorithm minimizing the difference between the simulated and the measured FRFs. The second goal of the current work is to guarantee an on-line detection of the damage, i.e., delamination in the viscoelastic bonding of the described specimen during frequency monitored end-of-life testing. For this purpose, an inverse technique, which determines the damage location and size based on the modal frequency shift and on the change of the mode shapes, is presented. This includes a preliminary FE model-based study correlating the delamination location and size to the change in the modal parameters and a subsequent experimental validation achieved through dynamic measurements of specimen with different, pre-generated crack scenarios and comparing it to the virgin specimen. The main advantage of the inverse characterization approach presented in the first part resides in the ability of adequately identifying the material damping and stiffness behavior of soft viscoelastic materials over a wide frequency range and under critical temperature conditions. Classic forward characterization techniques such as dynamic mechanical analysis are usually linked to limitations under critical temperature and frequency conditions due to the material behavior of soft viscoelastic materials. Furthermore, the inverse damage detection described in the second part guarantees an accurate prediction of not only the damage size but also its location using a simple test setup and outlines; therefore, the significance of inverse numerical-experimental approaches in predicting the dynamic behavior of soft bonding layers applied in automotive electronics.

Keywords: damage detection, dynamic characterization, inverse approaches, vibration testing, viscoelastic layers

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Authors: Saloni Khandelwal

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The Aravalli hills are one of the oldest and most distinctive mountain chains of peninsular India spanning in around 692 Km. More than 60% of it falls in the state of Rajasthan and influences ecological equilibrium in about 30% of the state. Because of natural and human-induced activities, physical gaps in the Aravallis are increasing, new gaps are coming up, and its physical structure is changing. There are no strict regulations to protect and monitor the Aravallis and no comprehensive research and study has been done for the enhancement of ecosystem functions of these ranges. Through this study, various factors leading to Aravalli’s degradation are identified and its impacts on selected areas are analyzed. A literature study is done to identify factors responsible for the degradation. To understand the severity of the problem at the lowest level, two tehsils from different districts in Rajasthan, which are the most affected due to illegal mining and increasing physical gaps are selected for the study. Case-1 of three-gram panchayats in Kishangarh Tehsil of Ajmer district focuses on the expanding physical gaps in the Aravalli range, and case-2 of three-gram panchayats in Tijara Tehsil of Alwar district focuses on increasing illegal mining in the Aravalli range. For measuring the degradation, physical, biological and social indicators are identified through literature review and for both the cases analysis is done on the basis of these indicators. Primary survey and focus group discussions are done with villagers, mining owners, illegal miners, and various government officials to understand dependency of people on the Aravalli and its importance to them along with the impact of degradation on their livelihood and environment. From the analysis, it has been found that green cover is continuously decreasing in both cases, dense forest areas do not exist now, the groundwater table is depleting at a very fast rate, soil is losing its moisture resulting in low yield and shift in agriculture. Wild animals which were easily seen earlier are now extinct. Cattles of villagers are dependent on the forest area in the Aravalli range for food, but with a decrease in fodder, their cattle numbers are decreasing. There is a decrease in agricultural land and an increase in scrub and salt-affected land. Analysis of various national and state programmes, acts which were passed to conserve biodiversity has been done showing that none of them is helping much to protect the Aravalli. For conserving the Aravalli and its forest areas, regional level and local level initiatives are required and are proposed in this study. This study is an attempt to formulate conservation and management strategies for the Aravalli range. These strategies will help in improving biodiversity which can lead to the revival of its ecosystem functions. It will also help in curbing the pollution at the regional and local level. All this will lead to the sustainable development of the region.

Keywords: Aravalli, ecosystem, LULC, Rajasthan

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Authors: Shewit Kidane, Abebe Getahun, Wassie Anteneh, Admassu Demeke, Peter Goethals

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We have reviewed major findings of scientific studies on Lake Tana fish resources and their threats. The aim was to provide summarized information for all concerned bodies and international readers to get full and comprehensive picture about the lake’s fish resource and conservation problems. The Lake Tana watershed comprise 28 fish species, of which 21 are endemic. Moreover, Lake Tana is the one among the top 250 lake regions of global importance for biodiversity and it is world recognized migratory birds wintering site. Lake Tana together with its adjacent wetlands provide directly and indirectly a livelihood for more than 500,000 people. However, owing to anthropogenic activities, the lake ecosystem as well as fish and attributes of the fisheries sector are severely degraded. Fish species in Lake Tana are suffering due to illegal fishing, damming, habitat/breeding ground degradation, wastewater disposal, introduction of exotic species, and lack of implementing fisheries regulations. Currently, more than 98% of fishers in Lake Tana are using the most destructive monofilament. Indeed, dams, irrigation schemes and hydropower are constructed in response to the emerging development need only. Mitigation techniques such as construction of fish ladders for the migratory fishes are the most forgotten. In addition, water resource developers are likely unaware of both the importance of the fisheries and the impact of dam construction on fish. As a result, the biodiversity issue is often missed. Besides, Lake Tana wetlands, which play vital role to sustain biodiversity, are not wisely utilised in the sense of the Ramsar Convention’s definition. Wetlands are considered as unhealthy and hence wetland conversion for the purpose of recession agriculture is still seen as advanced mode of development. As a result, many wetlands in the lake watershed are shrinking drastically over time and Cyprus papyrus, one of the characteristic features of Lake Tana, has dramatically declined in its distribution with some local extinction. Furthermore, the recently introduced water hyacinth (Eichhornia crassipes) is creating immense problems on the lake ecosystem. Moreover, currently, 1.56 million tons of sediment have deposited into the lake each year and wastes from the industries and residents are directly discharged into the lake without treatment. Recently, sign of eutrophication is revealed in Lake Tana and most coarsely, the incidence of cyanobacteria genus Microcystis was reported from the Bahir Dar Gulf of Lake Tana. Thus, the direct dependency of the communities on the lake water for drinking as well as to wash their body and clothes and its fisheries make the problem worst. Indeed, since it is home to many endemic migratory fish, such kind of unregulated developmental activities could be detrimental to their stocks. This can be best illustrated by the drastic stock reduction (>75% in biomass) of the world unique Labeobarbus species. So, unless proper management is put in place, the anthropogenic impacts can jeopardize the aquatic ecosystems. Therefore, in order to sustainably use the aquatic resources and fulfil the needs of the local people, every developmental activity and resource utilization should be carried out adhering to the available policies.

Keywords: anthropogenic impacts, dams, endemic fish, wetland degradation

Procedia PDF Downloads 218

Authors: Farrah Putri Salmanida, Mei-Li Wu, Rika Wahyuningtyas, Wen-Bin Chung, Hso-Chi Chaung, Ko-Tung Chang

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Within the field of immunotherapy, oncolytic virotherapy (OVT) employs dual approaches that directly eliminate tumor cells while preserving healthy ones and indirectly reprogram the tumor microenvironment (TME) to elicit antitumor responses. Within the TME, tumor associated macrophages (TAMs) manifest characteristics akin to those of anti-inflammatory M2 macrophages, thus earning the designation of M2-like TAMs. In prior research, two antigens denoted as A1 (g6Ld10T) and A3 (ORF6L5), derived from a complete sequence of ORF5 with partial sequence of ORF6 in Porcine Reproductive and Respiratory Syndrome Virus Genotype 2 (PRRSV-2), demonstrated the capacity to repolarize M2-type porcine alveolar macrophages (PAMs) into M1 phenotypes. In this study, we sought for utilizing OVT strategies by introducing A1 or A3 on TAMs to endow them with the anti-tumor traits of M1 macrophages while retaining their capacity to target cancer cells. Upon exposing human THP-1-derived M2 macrophages to a cross-species test with 2 µg/ml of either A1 or A3 for 24 hours, real time PCR revealed that A3, but not A1, treated cells exhibited upregulated gene expressions of M1 markers (CCR7, IL-1ß, CCL2, Cox2, CD80). These cells reacted to virus-derived antigen, as evidenced by increased expression of pattern-recognition receptors TLR3, TLR7, and TLR9, subsequently providing feedback in the form of type I interferon responses like IFNAR1, IFN-ß, IRF3, IRF7, OAS1, Mx1, and ISG15. Through an MTT assay, only after 15 µg/ml of A3 treatment could the cell viability decrease, with a predicted IC50 of 16.96 µg/ml. Interestingly, A3 caused dose-dependent toxicity to a rat C6 glial cancer cell line even at doses as low as 2.5 µg/ml and reached its IC50 at 9.419 µg/ml. Using Annexin V/7AAD staining and PCR test, we deduced that a significant proportion of C6 cells were undergoing the early apoptosis phase predominantly through the intrinsic apoptosis cascade involving Bcl-2 family proteins. Following this stage, we conducted a test on A3’s repolarization ability, which revealed a significant rise in M1 gene expression markers, such as TNF, CD80, and IL-1ß, in M2-like TAMs generated in vitro from murine RAW264.7 macrophages grown with conditioned medium of 4T1 breast cancer cells. This was corroborated by the results of transcriptome analysis, which revealed that the primary subset among the top 10 to top 30 significantly upregulated differentially expressed genes (DEGs) dominantly consisted of M1 macrophages profiles, including Ccl3, Ccl4, Csf3, TNF, Bcl6b, Stc1, and Dusp2. Our findings unveiled the remarkable potential of the PRRSV-derived antigen A3 to repolarize macrophages while also being capable of selectively inducing apoptosis in cancerous cells. While further in vivo study is needed for A3, it holds promise as an adjuvant by its dual effects in cancer therapy modalities.

Keywords: cancer cell apoptosis, interferon responses, macrophage repolarization, recombinant protein

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Authors: Pampita Chakraborty, Sukumar Mukherjee

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This study was done to determine the bacteriological profile and antibiotic resistance of the isolates and to find out the potential risk factors for infection with multidrug-resistant organisms. Diabetic foot ulcer is a major medical, social, economic problem and a leading cause of morbidity and mortality, especially in the developing countries like India. 25 percent of all diabetic patients develop a foot ulcer at some point in their lives which is highly susceptible to infections and that spreads rapidly, leading to overwhelming tissue destruction and subsequent amputation. Infection with multidrug resistant organisms (MDRO) may increase the cost of management and may cause additional morbidity and mortality. Proper management of these infections requires appropriate antibiotic selection based on culture and antimicrobial susceptibility testing. Early diagnosis of microbial infections is aimed to institute the appropriate antibacterial therapy initiative to avoid further complications. A total of 200 Type 2 Diabetic Mellitus patients with infection were admitted at GD Hospital and Diabetes Institute, Kolkata. 60 of them who developed ulcer during the year 2013 were included in this study. A detailed clinical history and physical examination were carried out for every subject. Specimens for microbiological studies were obtained from ulcer region. Gram-negative bacilli were tested for extended spectrum Beta-lactamase (ESBL) production by double disc diffusion method. Staphylococcal isolates were tested for susceptibility to oxacillin by screen agar method and disc diffusion. Potential risk factors for MDRO-positive samples were explored. Gram-negative aerobes were most frequently isolated, followed by gram-positive aerobes. Males were predominant in the study and majority of the patients were in the age group of 41-60 years. The presence of neuropathy was observed in 80% cases followed by peripheral vascular disease (73%). Proteus spp. (22) was the most common pathogen isolated, followed by E.coli (17). Staphylococcus aureus was predominant amongst the gram-positive isolates. S.aureus showed a high rate of resistance to antibiotic tested (63.6%). Other gram-positive isolates were found to be highly resistant to erythromycin, tetracycline and ciprofloxacin, 40% each. All isolates were found to be sensitive to Vancomycin and Linezolid. ESBL production was noted in Proteus spp and E.coli. Approximately 70 % of the patients were positive for MDRO. MDRO-infected patients had poor glycemic control (HbA1c 11± 2). Infection with MDROs is common in diabetic foot ulcers and is associated with risk factors like inadequate glycemic control, the presence of neuropathy, osteomyelitis, ulcer size and increased the requirement for surgical treatment. There is a need for continuous surveillance of resistant bacteria to provide the basis for empirical therapy and reduce the risk of complications.

Keywords: diabetic foot ulcer, bacterial infection, multidrug-resistant organism, extended spectrum beta-lactamase

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Authors: Luis Castiella, Mariano Rueda, Catalina Palacio

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The world’s rapid urbanisation represents an excellent opportunity to implement initiatives that are oriented towards a country’s general development. However, this phenomenon has created considerable pressure on current urban models, pushing them nearer to a crisis. As a result, several factors usually associated with underdevelopment have been steadily rising. Moreover, actions taken by public authorities have not been able to keep up with the speed of urbanisation, which has impeded them from meeting the demands of society, responding with reactionary policies instead of with coordinated, organised efforts. In contrast, the concept of a Smart City which emerged around two decades ago, in principle, represents a city that utilises innovative technologies to remedy the everyday issues of the citizen, empowering them with the newest available technology and information. This concept has come to adopt a wider meaning, including human and social capital, as well as productivity, economic growth, quality of life, environment and participative governance. These developments have also disrupted the management of institutions such as academia, which have become key in generating scientific advancements that can solve pressing problems, and in forming a specialised class that is able to follow up on these breakthroughs. In this light, the Ministry of Modernisation of the Argentinian Nation has created a model that is rooted in the concept of a ‘Smart City’. This effort considered all the dimensions that are at play in an urban environment, with careful monitoring of each sub-dimensions in order to establish the government’s priorities and improving the effectiveness of its operations. In an attempt to ameliorate the overall efficiency of the country’s economic and social development, these focused initiatives have also encouraged citizen participation and the cooperation of the private sector: replacing short-sighted policies with some that are coherent and organised. This process was developed gradually. The first stage consisted in building the model’s structure; the second, at applying the method created on specific case studies and verifying that the mechanisms used respected the desired technical and social aspects. Finally, the third stage consists in the repetition and subsequent comparison of this experiment in order to measure the effects on the ‘treatment group’ over time. The first trial was conducted on 717 municipalities and evaluated the dimension of Governance. Results showed that levels of governmental maturity varied sharply with relation to size: cities with less than 150.000 people had a strikingly lower level of governmental maturity than cities with more than 150.000 people. With the help of this analysis, some important trends and target population were made apparent, which enabled the public administration to focus its efforts and increase its probability of being successful. It also permitted to cut costs, time, and create a dynamic framework in tune with the population’s demands, improving quality of life with sustained efforts to develop social and economic conditions within the territorial structure.

Keywords: composite index, comprehensive model, smart cities, strategic framework

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Authors: Hilal T. Sasmazel, Seda Surucu

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Skin tissue engineering is a promising field for the treatment of skin defects using scaffolds. This approach involves the use of living cells and biomaterials to restore, maintain, or regenerate tissues and organs in the body by providing; (i) larger surface area for cell attachment, (ii) proper porosity for cell colonization and cell to cell interaction, and (iii) 3-dimensionality at macroscopic scale. Recent studies on this area mainly focus on fabrication of scaffolds that can closely mimic the natural extracellular matrix (ECM) for creation of tissue specific niche-like environment at the subcellular scale. Scaffolds designed as ECM-like architectures incorporating into the host with minimal scarring/pain and facilitate angiogenesis. This study is related to combining of synthetic PCL and natural chitosan polymers to form 3D PCL/Chitosan core-shell structures for skin tissue engineering applications. Amongst the polymers used in tissue engineering, natural polymer chitosan and synthetic polymer poly(ε-caprolactone) (PCL) are widely preferred in the literature. Chitosan has been among researchers for a very long time because of its superior biocompatibility and structural resemblance to the glycosaminoglycan of bone tissue. However, the low mechanical flexibility and limited biodegradability properties reveals the necessity of using this polymer in a composite structure. On the other hand, PCL is a versatile polymer due to its low melting point (60°C), ease of processability, degradability with non-enzymatic processes (hydrolysis) and good mechanical properties. Nevertheless, there are also several disadvantages of PCL such as its hydrophobic structure, limited bio-interaction and susceptibility to bacterial biodegradation. Therefore, it became crucial to use both of these polymers together as a hybrid material in order to overcome the disadvantages of both polymers and combine advantages of those. The scaffolds here were fabricated by using electrospinning technique and the characterizations of the samples were done by contact angle (CA) measurements, scanning electron microscopy (SEM), transmission electron microscopy (TEM) and X-Ray Photoelectron spectroscopy (XPS). Additionally, gas permeability test, mechanical test, thickness measurement and PBS absorption and shrinkage tests were performed for all type of scaffolds (PCL, chitosan and PCL/chitosan core-shell). By using ImageJ launcher software program (USA) from SEM photographs the average inter-fiber diameter values were calculated as 0.717±0.198 µm for PCL, 0.660±0.070 µm for chitosan and 0.412±0.339 µm for PCL/chitosan core-shell structures. Additionally, the average inter-fiber pore size values exhibited decrease of 66.91% and 61.90% for the PCL and chitosan structures respectively, compare to PCL/chitosan core-shell structures. TEM images proved that homogenous and continuous bead free core-shell fibers were obtained. XPS analysis of the PCL/chitosan core-shell structures exhibited the characteristic peaks of PCL and chitosan polymers. Measured average gas permeability value of produced PCL/chitosan core-shell structure was determined 2315±3.4 g.m-2.day-1. In the future, cell-material interactions of those developed PCL/chitosan core-shell structures will be carried out with L929 ATCC CCL-1 mouse fibroblast cell line. Standard MTT assay and microscopic imaging methods will be used for the investigation of the cell attachment, proliferation and growth capacities of the developed materials.

Keywords: chitosan, coaxial electrospinning, core-shell, PCL, tissue scaffold

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Authors: V. K. Tewari, M. Hussain, H. K. D. Gupta

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Background: Nitric Oxide Donars (NODs) (intrathecal sodium nitroprusside (ITSNP) and oral tadalafil 20mg post ITSNP) has been studied in this context in cerebral palsy patients for fast recovery. This work proposes two mechanisms for acute cases and one mechanism for chronic cases, which are interrelated, for physiological recovery. a) Retrograde Neurotransmission (acute cases): 1) Normal excitatory impulse: at the synaptic level, glutamate activates NMDA receptors, with nitric oxide synthetase (NOS) on the postsynaptic membrane, for further propagation by the calcium-calmodulin complex. Nitric oxide (NO, produced by NOS) travels backward across the chemical synapse and binds the axon-terminal NO receptor/sGC of a presynaptic neuron, regulating anterograde neurotransmission (ANT) via retrograde neurotransmission (RNT). Heme is the ligand-binding site of the NO receptor/sGC. Heme exhibits > 10,000-fold higher affinity for NO than for oxygen (the 10,000-fold effect) and is completed in 20 msec. 2) Pathological conditions: normal synaptic activity, including both ANT and RNT, is absent. A NO donor (SNP) releases NO from NOS in the postsynaptic region. NO travels backward across a chemical synapse to bind to the heme of a NO receptor in the axon terminal of a presynaptic neuron, generating an impulse, as under normal conditions. b) Vasopasm: (acute cases) Perforators show vasospastic activity. NO vasodilates the perforators via the NO-cAMP pathway. c) Long-Term Potentiation (LTP): (chronic cases) The NO–cGMP-pathway plays a role in LTP at many synapses throughout the CNS and at the neuromuscular junction. LTP has been reviewed both generally and with respect to brain regions specific for memory/learning. Aims/Study Design: The principles of “generation of impulses from the presynaptic region to the postsynaptic region by very potent RNT (10,000-fold effect)” and “vasodilation of arteriolar perforators” are the basis of the authors’ hypothesis to treat cerebral palsy cases. Case-control prospective study. Materials and Methods: The experimental population included 82 cerebral palsy patients (10 patients were given control treatments without NOD or with 5% dextrose superfusion, and 72 patients comprised the NOD group). The mean time for superfusion was 5 months post-cerebral palsy. Pre- and post-NOD status was monitored by Gross Motor Function Classification System for Cerebral Palsy (GMFCS), MRI, and TCD studies. Results: After 7 days in the NOD group, the mean change in the GMFCS score was an increase of 1.2 points mean; after 3 months, there was an increase of 3.4 points mean, compared to the control-group increase of 0.1 points at 3 months. MRI and TCD documented the improvements. Conclusions: NOD (ITSNP boosts up the recovery and oral tadalafil maintains the recovery to a well-desired level) acts swiftly in the treatment of CP, acting within 7 days on 5 months post-cerebral palsy either of the three mechanisms.

Keywords: cerebral palsy, intrathecal sodium nitroprusside, oral tadalafil, perforators, vasodilations, retrograde transmission, the 10, 000-fold effect, long-term potantiation

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Authors: Charlotte Jeavons, Charitini Stavropoulous, Nicolas Drey

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Background: Over one-third of five-year-olds and almost half of all eight-year-olds in the UK have obvious caries experience that can be detected by visual screening techniques. School-based caries preventions programs to apply fluoride varnish to young children’s teeth operate in many areas in the UK. Their aim is to reduce dental caries in children. The Department of Health guidance (2009) on consent states information must be provided to parents to enable informed autonomous decision-making prior to any treatment involving their young children. Fluoride varnish schemes delivered in primary schools use letters for this purpose. Parents are expected to return these indicating their consent or refusal. A large proportion of parents do not respond. In the absence of positive consent, these children are excluded from the program. Non-response is more common in deprived areas creating inequality. The reason for this is unknown. The consent process used is underpinned by the ethical theory of deontology that is prevalent in clinical dentistry and widely accepted in bio-ethics. Objective: To investigate parents’ views, understanding and experience of the fluoride varnish program taking place in their child’s school, including their views about the practical consent arrangements. Method: Schools participating in the fluoride varnish scheme operating in Enfield, North London, were asked to take part. Parents with children in nursery, reception, or year one were invited to participate via semi-structured interviews and focus groups. Thematic analysis was conducted. Findings: 40 parents were recruited from eight schools. The global theme of ‘trust’ was identified as the strongest influence on parental responses. Six themes were identified; protecting children from harm is viewed by parents as their role, parents have the capability to decide but lack confidence, sharing responsibility for their child’s oral health with the State is welcomed by a parent, existing relationships within parents’ social networks strongly influences consent decisions, official dental information is not communicated effectively, sending a letter to parents’ and excluding them from meeting dental practitioners is ineffective. The information delivered via a letter was not strongly identified by parents as influencing their response. Conclusions: Personal contact with the person(s) providing information and requesting consent has a greater impact on parental consent responses than written information provided alone. This demonstrates that traditional bio-ethical ideas about rational decision-making where emotions are transcended and interference is not justified unless preventing harm to an unaware person are outdated. Parental decision-making is relational and the consent process should be adapted to reflect this. The current system that has a deontology view of decision making at its core impoverishes parental autonomy and may, ultimately, increase dental inequalities as a result.

Keywords: consent, decision, ethics, fluoride, parents

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Authors: Lina Paola Orozco Marin, Yuliet Montoya Osorio, John Bustamante Osorno

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Ischemic events can culminate in acute myocardial infarction by irreversible cardiac lesions that cannot be restored due to the limited regenerative capacity of the heart. Cell therapy seeks to replace these injured or necrotic cells by transplanting healthy and functional cells. The therapeutic alternatives proposed by tissue engineering and cardiovascular regenerative medicine are the use of biomaterials to mimic the native extracellular medium, which is full of proteins, proteoglycans, and glycoproteins. The selected biomaterials must provide structural support to the encapsulated cells to avoid their migration and death in the host tissue. In this context, the present research work focused on developing a natural thermosensitive hydrogel, its physical and chemical characterization, and the determination of its biocompatibility in vitro. The hydrogel was developed by mixing hydrolyzed bovine and porcine collagen at 2% w/v, chitosan at 2.5% w/v, and beta-glycerolphosphate at 8.5% w/w and 10.5% w/w in magnetic stirring at 4°C. Once obtained, the thermosensitivity and gelation time were determined, incubating the samples at 37°C and evaluating them through the inverted tube method. The morphological characterization of the hydrogels was carried out through scanning electron microscopy. Chemical characterization was carried out employing infrared spectroscopy. The biocompatibility was determined using the MTT cytotoxicity test according to the ISO 10993-5 standard for the hydrogel’s precursors using the fetal human ventricular cardiomyocytes cell line RL-14. The RL-14 cells were also seeded on the top of the hydrogels, and the supernatants were subculture at different periods to their observation under a bright field microscope. Four types of thermosensitive hydrogels were obtained, which differ in their composition and concentration, called A1 (chitosan/bovine collagen/beta-glycerolphosphate 8.5%w/w), A2 (chitosan/porcine collagen/beta-glycerolphosphate 8.5%), B1 (chitosan/bovine collagen/beta-glycerolphosphate 10.5%) and B2 (chitosan/porcine collagen/beta-glycerolphosphate 10.5%). A1 and A2 had a gelation time of 40 minutes, and B1 and B2 had a gelation time of 30 minutes at 37°C. Electron micrographs revealed a three-dimensional internal structure with interconnected pores for the four types of hydrogels. This facilitates the exchange of nutrients, oxygen, and the exit of metabolites, allowing to preserve a microenvironment suitable for cell proliferation. In the infrared spectra, it was possible to observe the interaction that occurs between the amides of polymeric compounds with the phosphate groups of beta-glycerolphosphate. Finally, the biocompatibility tests indicated that cells in contact with the hydrogel or with each of its precursors are not affected in their proliferation capacity for a period of 16 days. These results show the potential of the hydrogel to increase the cell survival rate in the cardiac cell therapies under investigation. Moreover, the results lay the foundations for its characterization and biological evaluation in both in vitro and in vivo models.

Keywords: cardiac cell therapy, cardiac ischemia, natural polymers, thermosensitive hydrogel

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Authors: Karim Hrratha, Khaled Essalahb, Christophe Morellc, Henry Chermettec, Salima Boughdiria

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Among the carbon-carbon bond formation types, allylation of active methylene compounds with cyclic Baylis-Hillman (BH) alcohols is a reliable and widely used method. This reaction is a very attractive tool in organic synthesis of biological and biodiesel compounds. Thus, in view of an insistent and peremptory request for an efficient and straightly method for synthesizing the desired product, a thorough analysis of various aspects of the reaction processes is an important task. The product afforded by the reaction of active methylene with BH alcohols depends largely on the experimental conditions, notably on the catalyst properties. All experiments reported that catalysis is needed for this reaction type because of the poor ability of alcohol hydroxyl group to be as a suitable leaving group. Within the catalysts, several transition- metal based have been used such as palladium in the presence of acid or base and have been considered as reliable methods. Furthemore, acid catalysts such as BF3.OEt2, BiX3 (X= Cl, Br, I, (OTf)3), InCl3, Yb(OTf)3, FeCl3, p-TsOH and H-montmorillonite have been employed to activate the C-C bond formation through the alkylation of active methylene compounds. Interestingly a report of a smoothly process for the ability of 4-imethyaminopyridine(DMAP) to catalyze the allylation reaction of active methylene compounds with cyclic Baylis-Hillman (BH) alcohol appeared recently. However, the reaction mechanism remains ambiguous, since the C- allylation process leads to an unexpected product (noted P1), corresponding to a direct allylation instead of conjugate allylation, which involves the most electrophilic center according to the electron withdrawing group CO effect. The main objective of the present theoretical study is to better understand the role of the DMAP catalytic activity as well as the process leading to the end- product (P1) for the catalytic reaction of a cyclic BH alcohol with active methylene compounds. For that purpose, we have carried out computations of a set of active methylene compounds varying by R1 and R2 toward the same alcohol, and we have attempted to rationalize the mechanisms thanks to the acid–base approach, and conceptual DFT tools such as chemical potential, hardness, Fukui functions, electrophilicity index and dual descriptor, as these approaches have shown a good prediction of reactions products.The present work is then organized as follows: In a first part some computational details will be given, introducing the reactivity indexes used in the present work, then Section 3 is dedicated to the discussion of the prediction of the selectivity and regioselectivity. The paper ends with some concluding remarks. In this work, we have shown, through DFT method at the B3LYP/6-311++G(d,p) level of theory that: The allylation of active methylene compounds with cyclic BH alcohol is governed by orbital control character. Hence the end- product denoted P1 is generated by direct allylation.

Keywords: DFT calculation, gas phase pKa, theoretical mechanism, orbital control, charge control, Fukui function, transition state

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