Search results for: mitochondrial function

Authors: Aniekan Peter, ECS Naidu, Edidiong Akang, U. Offor, R. Kalhapure, A. A. Chuturgoon, T. Govender, O. O. Azu

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Introduction: Nano-drugs are novel innovations in the management of human immunodeficiency virus (HIV) pandemic, especially resistant strains of the virus in their sanctuary sites: testis and the brain. There are safety concerns to be addressed to achieve the full potential of this new drug delivery system. Aim of study: Our study was designed to investigate toxicity profile of Tenofovir Nanoparticle (TDF-N) synthesized by University of Kwazulu-Natal (UKZN) Nano-team for prevention and treatment of HIV infection. Methodology: Ten adult male Sprague-Dawley rats maintained at the Animal House of the Biomedical Resources Unit UKZN were used for the study. The animals were weighed and divided into two groups of 5 animal each. Control animals (A) were administered with normal saline. Therapeutic dose (4.3 mg/kg) of TDF-N was administered to group B. At the end of four weeks, animals were weighed and sacrificed. Liver and kidney were removed fixed in formal saline, processed and stained using H/E, PAS and MT stains for light microscopy. Serum was obtained for renal function test (RFT), liver function test (LFT) and full blood count (FBC) using appropriate analysers. Cellular measurements were done using ImageJ and Leica software 2.0. Data were analysed using graph pad 6, values < 0.05 were significant. Results: We reported no histological alterations in the liver, kidney, FBC, LFT and RFT between the TDF-N animals and saline control. There were no significant differences in weight, organo-somatic index and histological measurements in the treatment group when compared with saline control. Conclusion/recommendations: TDF-N is not toxic to the liver, kidney and blood cells in our study. More studies using human subjects is recommended.

Keywords: tenofovir nanoparticles, liver, kidney, blood cells

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Authors: E. Cenderelli, E. Bruno, G. Iacoviello, A. Lazzini

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The financial sector is exposed to the risk of cyber-attacks like any other industrial sector. Furthermore, the topic of CybeRisk (cyber risk) has become particularly relevant given that Information Technology (IT) attacks have increased drastically in recent years, and cannot be stopped by single organizations requiring a response at international and national level. IT risk is never a matter purely for the IT manager, although he clearly plays a key role. A bank's risk management function requires a thorough understanding of the evolving risks as well as the tools and practical techniques available to address them. Upon the request of European and national legislation regarding CybeRisk in the financial system, banks are therefore called upon to strengthen the operational model for CybeRisk management. This will require an important change with a more intense collaboration with the structures that deal with information security for the development of an ad hoc system for the evaluation and control of this type of risk. The aim of the work is to propose a framework for the management and control of CybeRisk that will bridge the gap in the literature regarding the understanding and consideration of CybeRisk as an integral part of business management. The IT function has a strong relevance in the management of CybeRisk, which is perceived mainly as operational risk, but with a positive tendency on the part of risk management to the identification of CybeRisk assessment methods that are increasingly complete, quantitative and able to better describe the possible impacts on the business. The paper provides answers to the research questions: Is it possible to define a CybeRisk governance structure able to support the comparison between risk and security? How can the relationships between IT assets be integrated into a cyberisk assessment framework to guarantee a system of protection and risks control? From a methodological point of view, this research uses a case study approach. The choice of “Monte dei Paschi di Siena” was determined by the specific features of one of Italy’s biggest lenders. It is chosen to use an intensive research strategy: an in-depth study of reality. The case study methodology is an empirical approach to explore a complex and current phenomenon that develops over time. The use of cases has also the advantage of allowing the deepening of aspects concerning the "how" and "why" of contemporary events, on which the scholar has little control. The research bases on quantitative data and qualitative information obtained through semi-structured interviews of an open-ended nature and questionnaires to directors, members of the audit committee, risk, IT and compliance managers, and those responsible for internal audit function and anti-money laundering. The added value of the paper can be seen in the development of a framework based on a mapping of IT assets from which it is possible to identify their relationships for purposes of a more effective management and control of cyber risk.

Keywords: bank, CybeRisk, information technology, risk management

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Authors: Dafna Knani, Sarit S. Sivan

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Aggrecan, one of the main components of the intervertebral disc (IVD), belongs to the family of proteoglycans (PGs) that are composed of glycosaminoglycan (GAG) chains covalently attached to a core protein. Its primary function is to maintain tissue hydration and hence disc height under the high loads imposed by muscle activity and body weight. Significant PG loss is one of the first indications of disc degeneration. A possible solution to recover disc functions is by injecting a synthetic hydrogel into the joint cavity, hence mimicking the role of PGs. One of the hydrogels proposed is GAG-analogues, based on sulfate-containing polymers, which are responsible for hydration in disc tissue. In the present work, we used molecular dynamics (MD) to study the effect of the hydrogel crosslinking (type and degree) on the swelling behavior of the suggested GAG-analogue biomimetics by calculation of cohesive energy density (CED), solubility parameter, enthalpy of mixing (ΔEmix) and the interactions between the molecules at the pure form and as a mixture with water. The simulation results showed that hydrophobicity plays an important role in the swelling of the hydrogel, as indicated by the linear correlation observed between solubility parameter values of the copolymers and crosslinker weight ratio (w/w); this correlation was found useful in predicting the amount of PEGDA needed for the desirable hydration behavior of (CS)₄-peptide. Enthalpy of mixing calculations showed that all the GAG analogs, (CS)₄ and (CS)₄-peptide are water-soluble; radial distribution function analysis revealed that they form interactions with water molecules, which is important for the hydration process. To conclude, our simulation results, beyond supporting the experimental data, can be used as a useful predictive tool in the future development of biomaterials, such as disc replacement.

Keywords: molecular dynamics, proteoglycans, enthalpy of mixing, swelling

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Authors: Marawan Abd Elbaset Mohamed, Hanan A. Ogaly, Rehab F. Abdel-Rahman, Ahmed-Farid O.A., Marwa S. Khattab, Reham M. Abd-Elsalam

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Liver fibrosis is a severe worldwide health concern due to various chronic liver disorders. Hepatic encephalopathy (HE) is one of its most common complications affecting liver and brain cognitive function. Beta-Carotene (B-Car) is an organic, strongly colored red-orange pigment abundant in fungi, plants, and fruits. The study attempted to know B-Car neuroprotective potential against thioacetamide (TAA)-induced neurotoxicity and cognitive decline in HE in rats. Hepatic encephalopathy was induced by TAA (100 mg/kg, i.p.) three times per week for two weeks. B-Car was given orally (10 or 20 mg/kg) daily for two weeks after TAA injections. Organ body weight ratio, Serum transaminase activities, liver’s antioxidant parameters, ammonia, and liver histopathology were assessed. Also, the brain’s mitogen-activated protein kinase (MAPK), nuclear factor kappa B (NF-κB), antioxidant parameters, adenosine triphosphate (ATP), adenosine monophosphate (AMP), norepinephrine (NE), dopamine (DA), serotonin (5-HT), 5-hydroxyindoleacetic acid (5-HIAA) cAMP response element-binding protein (CREB) expression and B-cell lymphoma 2 (Bcl-2) expression were measured. The brain’s cognitive functions (Spontaneous locomotor activity, Rotarod performance test, Object recognition test) were assessed. B-Car prevented alteration of the brain’s cognitive function in a dose-dependent manner. The histopathological outcomes supported these biochemical evidences. Based on these results, it could be established that B-Car could be assigned to treat the brain’s neurotoxicity consequences of HE via downregualtion of MAPK/NF-κB signaling pathways.

Keywords: beta-carotene, liver injury, MAPK, NF-κB, rat, thioacetamide

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Authors: Jin Young Kim, Kwon Kyoo Kang

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The SGR1 (STAYGREEN1) protein is a critical regulator of plant leaves in chlorophyll degradation and senescence. The functions and mechanisms of tomato SGR1 action are poorly understood and worthy of further investigation. To investigate the function of the SGR1 gene, we generated a SGR1-knockout (KO) null line via clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-mediated gene editing and conducted RNA sequencing and gas chromatography tandem mass spectrometry (GC-MS/MS) analysis to identify the differentially expressed genes. The SlSGR1 (Solanum lycopersicum SGR1) knockout null line clearly showed a turbid brown color with significantly higher chlorophyll and carotenoid content compared to wild-type (WT) fruit. Differential gene expression analysis revealed 728 differentially expressed genes (DEGs) between WT and sgr1 #1-6 line, including 263 and 465 downregulated and upregulated genes, respectively, for which fold change was >2, and the adjusted p-value was <0.05. Most of the DEGs were related to photosynthesis and chloroplast function. In addition, the pigment, carotenoid changes in sgr1 #1-6 line was accumulated of key primary metabolites such as sucrose and its derivatives (fructose, galactinol, raffinose), glycolytic intermediates (glucose, G6P, Fru6P) and tricarboxylic acid cycle (TCA) intermediates (malate and fumarate). Taken together, the transcriptome and metabolite profiles of SGR1-KO lines presented here provide evidence for the mechanisms underlying the effects of SGR1 and molecular pathways involved in chlorophyll degradation and carotenoid biosynthesis.

Keywords: tomato, CRISPR/Cas9, null line, RNA-sequencing, metabolite profiling

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Authors: Zafar Iqbal, Nigel P. Grigg, K. Govindaraju, Nicola M. Campbell-Allen

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Quality Function Deployment (QFD) is structured approach. It has been used to improve the quality of products and process in a wide range of fields. Using this systematic tool, practitioners normally rank Voice of Customer ratings (VoCs) in order to produce Improvement Ratios (IRs) which become the basis for prioritising process / product design or improvement activities. In one matrix of the House of Quality (HOQ) competitors are rated. The method of obtaining improvement ratios (IRs) does not always integrate the competitors’ rating in a systematic way that fully utilises competitor rating information. This can have the effect of diverting QFD practitioners’ attention from a potentially important VOC to less important VOC. In order to enhance QFD analysis, we present a more systematic method for integrating competitor ratings, utilising the geometric mean of the customer rating matrix. In this paper we develop a new approach, based on the Analytic Hierarchy Process (AHP), in which we generating a matrix of multiple comparisons of all competitors, and derive a geometric mean for each competitor. For each VOC an improved IR is derived which-we argue herein - enhances the initial VOC importance ratings by integrating more information about competitor performance. In this way, our method can help overcome one of the possible shortcomings of QFD. We then use a published QFD example from literature as a case study to demonstrate the use of the new AHP-based IRs, and show how these can be used to re-rank existing VOCs to -arguably- better achieve the goal of customer satisfaction in relation VOC ratings and competitors’ rankings. We demonstrate how two dimensional AHP-based geometric mean derived from the multiple competitor comparisons matrix can be useful for analysing competitors’ rankings. Our method utilises an established methodology (AHP) applied within an established application (QFD), but in an original way (through the competitor analysis matrix), to achieve a novel improvement.

Keywords: quality function deployment, geometric mean, improvement ratio, AHP, competitors ratings

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Authors: Sajjad Shojaeidoust, Mohsen Ghanbarzadeh, Abdolhamid Habibi

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Exercise-induced bronchospasm (EIB) is a transitory condition of airflow obstruction that is associated with physical activities. It is noted that high ventilation can lead to an increase in the heat and reduce in the moisture in airways resistance of trachea. Also causes of pathophysiological mechanism are EIB. Accordingly, studying some parameters of pulmonary function (FVC, FEV1) among active people seems quintessential. The aim of this study was to analyze bronchospasm following the implementation of a program of maximum aerobic exercise in active men at Chamran University of Ahwaz. Method: In this quasi-experimental study, the population consisted of all students at Chamran University. Among from 55 participants, of which, 15 were randomly selected as the experimental group. In this study, the size of the maximum oxygen consumption was initially measured, and then, based on the maximum oxygen consumed, the active individuals were identified. After five minutes’ warm-up, Strand treadmill exercise test was taken (one session) and pulmonary parameters were measured at both pre- and post-tests (spirometer). After data normalization using KS and non-normality of the data, the Wilcoxon test was used to analyze the data. The significance level for all statistical surveys was considered p≤0/05. Results: The results showed that the ventilation factors and bronchospasm (FVC, FEV1) in the pre-test and post-test resulted in no significant difference among the active people (p≥0/05). Discussion and conclusion: Based on the results observed in this study, it appears that pulmonary indices in active individuals increased after aerobic test. The increase in this indicator in active people is due to increased volume and elasticity of the lungs as well. In other words, pulmonary index is affected by rib muscles. It is considered that progress over respiratory muscle strength and endurance has raised FEV1 in the active cases.

Keywords: aerobic active maximum, bronchospasm, pulmonary function, spirometer

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Authors: Walid A. Attia, Taher M. Bazan, Fawzy Eltohamy, Mahmoud Fathy

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System performance evaluation is an essential stage in the design of high-resolution satellite telescopes prior to the development process. In this paper, a system performance evaluation of a very high-resolution satellite telescope is investigated. The evaluated system has a Korsch optical scheme design. This design has been discussed in another paper with respect to three-mirror anastigmat (TMA) scheme design and the former configuration showed better results. The investigated system is based on the Korsch optical design integrated with a time-delay and integration charge coupled device (TDI-CCD) sensor to achieve a ground sampling distance (GSD) of 25 cm. The key performance metrics considered are the spatial resolution, the signal to noise ratio (SNR) and the total modulation transfer function (MTF) of the system. In addition, the national image interpretability rating scale (NIIRS) metric is assessed to predict the image quality according to the modified general image quality equation (GIQE). Based on the orbital, optical and detector parameters, the estimated GSD is found to be 25 cm. The SNR has been analyzed at different illumination conditions of target albedos, sun and sensor angles. The system MTF has been computed including diffraction, aberration, optical manufacturing, smear and detector sampling as the main contributors for evaluation the MTF. Finally, the system performance evaluation results show that the computed MTF value is found to be around 0.08 at the Nyquist frequency, the SNR value was found to be 130 at albedo 0.2 with a nadir viewing angles and the predicted NIIRS is in the order of 6.5 which implies a very good system image quality.

Keywords: modulation transfer function, national image interpretability rating scale, signal to noise ratio, satellite telescope performance evaluation

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Authors: Gustavo Axel Elizalde-Velázquez

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Metformin is the first line of oral therapy to treat type II diabetes and is also employed as a treatment for other indications, such as polycystic ovary syndrome, cancer, and COVID-19. Recent data suggest it is the aspirin of the 21st century due to its antioxidant and anti-aging effects. However, increasingly current articles indicate its long-term consumption generates mitochondrial impairment. Up to date, it is known metformin increases the biogenesis of Alzheimer's amyloid peptides via up-regulating BACE1 transcription, but further information related to brain damage after its consumption is missing. Bearing in mind the above, this work aimed to establish whether or not chronic exposure to metformin may alter swimming behavior and induce neurotoxicity in Danio rerio adults. For this purpose, 250 Danio rerio grown-ups were assigned to six tanks of 50 L of capacity. Four of the six systems contained 50 fish, while the remaining two had 25 fish (≈1 male:1 female ratio). Every system with 50 fish was allocated one of the three metformin treatment concentrations (1, 20, and 40 μg/L), with one system as the control treatment. Systems with 25 fish, on the other hand, were used as positive controls for acetylcholinesterase (10 μg/L of Atrazine) and oxidative stress (3 μg/L of Atrazine). After four months of exposure, a mean of 32 fish (S.D. ± 2) per group of MET treatment survived, which were used for the evaluation of behavior with the Novel Tank test. Moreover, after the behavioral assessment, we aimed to collect the blood and brains of all fish from all treatment groups. For blood collection, fish were anesthetized with an MS-222 solution (150 mg/L), while for brain gathering, fish were euthanized using the hypothermic shock method (2–4 °C). Blood was employed to determine CASP3 activity and the percentage of apoptotic cells with the TUNEL assay, and brains were used to evaluate acetylcholinesterase activity, oxidative damage, and gene expression. After chronic exposure, MET-exposed fish exhibited less swimming activity when compared to control fish. Moreover, compared with the control group, MET significantly inhibited the activity of AChE and induced oxidative damage in the brain of fish. Concerning gene expression, MET significantly upregulated the expression of Nrf1, Nrf2, BAX, p53, BACE1, APP, PSEN1, and downregulated CASP3 and CASP9. Although MET did not overexpress the CASP3 gene, we saw a meaningful rise in the activity of this enzyme in the blood of fish exposed to MET compared to the control group, which we then confirmed by a high number of apoptotic cells in the TUNEL assay. To the best of our understanding, this is the first study that delivers evidence of oxidative impairment, apoptosis, AChE alteration, and overexpression of B- amyloid-related genes in the brain of fish exposed to metformin.

Keywords: AChE inhibition, CASP3 activity, NovelTank test, oxidative damage, TUNEL assay

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Authors: Yu Menshikov

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In this paper the influence of errors of function derivatives in initial time which have been obtained by experiment (uncontrollable inaccuracy) to the results of inverse problem solution was investigated. It was shown that these errors distort the inverse problem solution as a rule near the beginning of interval where the solution are analyzed. Several methods for remove the influence of uncontrollable inaccuracy have been suggested.

Keywords: inverse problems, filtration, uncontrollable inaccuracy

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Authors: Samson Mil'shtein, Dhawal Asthana, Benjamin Sullivan

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The wide energy gap of GaN is the major parameter justifying the design and fabrication of high-power electronic components made of this material. However, the existence of a piezo-electrics in nature sheet charge at the AlGaN/GaN interface complicates the control of carrier injection into the intrinsic channel of GaN HEMTs (High Electron Mobility Transistors). As a result, most of the transistors created as R&D prototypes and all of the designs used for mass production are D-mode devices which introduce challenges in the design of integrated circuits. This research presents the design and modeling of an E-mode GaN HEMT with a very low turn-on voltage. The proposed device includes two critical elements allowing the transistor to achieve zero conductance across the channel when Vg = 0V. This is accomplished through the inclusion of an extremely thin, 2.5nm intrinsic Ga₀.₇₄Al₀.₂₆N spacer layer. The added spacer layer does not create piezoelectric strain but rather elastically follows the variations of the crystal structure of the adjacent GaN channel. The second important factor is the design of a gate metal with a high work function. The use of a metal gate with a work function (Ni in this research) greater than 5.3eV positioned on top of n-type doped (Nd=10¹⁷cm⁻³) Ga₀.₇₄Al₀.₂₆N creates the necessary built-in potential, which controls the injection of electrons into the intrinsic channel as the gate voltage is increased. The 5µm long transistor with a 0.18µm long gate and a channel width of 30µm operate at Vd=10V. At Vg =1V, the device reaches the maximum drain current of 0.6mA, which indicates a high current density. The presented device is operational at frequencies greater than 10GHz and exhibits a stable transconductance over the full range of operational gate voltages.

Keywords: compound semiconductors, device modeling, enhancement mode HEMT, gallium nitride

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Authors: Nino Pailodze, Malkhaz Sulashvili, Vladimer Kekenadze, Tea Khutsishvili, Irma Makharashvili, Aleksandre Kekenadze

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In the nearest future among the important problems which Georgia has solve the most important is economic stability, that bases on fiscal policy and the proper definition of the its directions. The main source of the Budget revenue is the national income. The State uses taxes, loans and emission in order to create national income, were the principal weapon are taxes. As well as fiscal function of the fulfillment of the budget, tax systems successfully implement economic and social development and the regulatory functions of foreign economic relations. A tax is a mandatory, unconditional monetary payment to the budget made by a taxpayer in accordance with this Code, based on the necessary, nonequivalent and gratuitous character of the payment. Taxes shall be national and local. National taxes shall be the taxes provided for under this Code, the payment of which is mandatory across the whole territory of Georgia. Local taxes shall be the taxes provided for under this Code, introduced by normative acts of local self-government representative authorities (within marginal rates), the payment of which is mandatory within the territory of the relevant self-governing unit. National taxes have the leading role in tax systems, but also the local taxes have an importance role in tax systems. Exactly in the means of local taxes, the most part of the budget is formatted. National taxes shall be: income tax, profit tax, value added tax (VAT), excise tax, import duty, property tax shall be a local tax The property tax is one of the significant taxes in Georgia. The paper deals with the taxation mechanism that has been operated in Georgia. The above mention has the great influence in financial accounting. While comparing foreign legislation towards Georgian legislation we discuss the opportunity of using their experience. Also, we suggested recommendations in order to improve the tax system in financial accounting. In addition to accounting, which is regulated according the International Accounting Standards we have tax accounting, which is regulated by the Tax Code, various legal orders / regulations of the Minister of Finance. The rules are controlled by the tax authority, Revenue Service. The tax burden from the tax values are directly related to expenditures of the state from the emergence of the first day. Fiscal policy of the state is as well as expenditure of the state and decisions of taxation. In order to get the best and the most effective mobilization of funds, Government’s primary task is to decide the kind of taxation rules. Tax function is to reveal the substance of the act. Taxes have the following functions: distribution or the fiscal function; Control and regulatory functions. Foreign tax systems evolved in the different economic, political and social conditions influence. The tax systems differ greatly from each other: taxes, their structure, typing means, rates, the different levels of fiscal authority, the tax base, the tax sphere of action, the tax breaks.

Keywords: international accounting standards, financial accounting, tax systems, financial obligations

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Authors: Su-Lien Liao

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This article explores the language in the four plays of Samuel Beckett–Waiting for Godot, Endgame, Krapp’s Last Tape, and Footfalls. It considers the way in which Beckett uses language, especially through fragmentation utterances, repetitions, monologues, contradictions, and silence. It discusses the function of language in modern society, in the theater of the absurd, and in the plays. Paradoxically enough, his plays attempts to communicate the incommunicability of language.

Keywords: language, Samuel Beckett, theater of the absurd, foreign language teaching

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Authors: S. Serhal, Y. Melinge, D. Rangeard, F. Hage Chehadeh

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Filling of estuaries is an international problem that can cause economic and environmental damage. This work aims the study of the rheological structuring mechanisms of natural sedimentary liquid-solid mixture in estuaries in order to better understand their filling. The estuary of the Rance river, located in Brittany, France is particularly targeted by the study. The aim is to provide answers on the rheological behavior of natural sediments by detecting structural factors influencing the rheological parameters. So we can better understand the fillings estuarine areas and especially consider sustainable solutions of ‘cleansing’ of these areas. The sediments were collected from the trap of Lyvet in Rance estuary. This trap was created by the association COEUR (Comité Opérationnel des Elus et Usagers de la Rance) in 1996 in order to facilitate the cleansing of the estuary. It creates a privileged area for the deposition of sediments and consequently makes the cleansing of the estuary easier. We began our work with a preliminary study to establish the trend of the rheological behavior of the suspensions and to specify the dormant phase which precedes the beginning of the biochemical reactivity of the suspensions. Then we highlight the visco-plastic character at younger age using the Kinexus rheometer, plate-plate geometry. This rheological behavior of suspensions is represented by the Bingham model using dynamic yield stress and viscosity which can be a function of volume fraction, granular extent, and chemical reactivity. The evolution of the viscosity as a function of the solid volume fraction is modeled by the Krieger-Dougherty model. On the other hand, the analysis of the dynamic yield stress showed a fairly functional link with the solid volume fraction.

Keywords: estuaries, rheological behavior, sediments, Kinexus rheometer, Bingham model, viscosity, yield stress

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Authors: Vivek Hokam, Vishrut Landge

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An ideal maintenance program for an industrial road network is one that would maintain all sections at a sufficiently high level of functional and structural conditions. However, due to various constraints such as budget, manpower and equipment, it is not possible to carry out maintenance on all the needy industrial road sections within a given planning period. A rational and systematic priority scheme needs to be employed to select and schedule industrial road sections for maintenance. Priority analysis is a multi-criteria process that determines the best ranking list of sections for maintenance based on several factors. In priority setting, difficult decisions are required to be made for selection of sections for maintenance. It is more important to repair a section with poor functional conditions which includes uncomfortable ride etc. or poor structural conditions i.e. sections those are in danger of becoming structurally unsound. It would seem therefore that any rational priority setting approach must consider the relative importance of functional and structural condition of the section. The maintenance priority index and pavement performance models tend to focus mainly on the pavement condition, traffic criteria etc. There is a need to develop the model which is suitably used with respect to limited budget provisions for maintenance of pavement. Linear programming is one of the most popular and widely used quantitative techniques. A linear programming model provides an efficient method for determining an optimal decision chosen from a large number of possible decisions. The optimum decision is one that meets a specified objective of management, subject to various constraints and restrictions. The objective is mainly minimization of maintenance cost of roads in industrial area. In order to determine the objective function for analysis of distress model it is necessary to fix the realistic data into a formulation. Each type of repair is to be quantified in a number of stretches by considering 1000 m as one stretch. A stretch considered under study is having 3750 m length. The quantity has to be put into an objective function for maximizing the number of repairs in a stretch related to quantity. The distress observed in this stretch are potholes, surface cracks, rutting and ravelling. The distress data is measured manually by observing each distress level on a stretch of 1000 m. The maintenance and rehabilitation measured that are followed currently are based on subjective judgments. Hence, there is a need to adopt a scientific approach in order to effectively use the limited resources. It is also necessary to determine the pavement performance and deterioration prediction relationship with more accurate and economic benefits of road networks with respect to vehicle operating cost. The infrastructure of road network should have best results expected from available funds. In this paper objective function for distress model is determined by linear programming and deterioration model considering overloading is discussed.

Keywords: budget, maintenance, deterioration, priority

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Authors: Spira A., Marabelle A., Kientop D., Moser E., Mumm J.

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Background: Both interleukin-2 (IL-2) and interleukin-10 (IL-10) have been extensively studied for their stimulatory function on T cells and their potential to obtain sustainable tumor control in RCC, melanoma, lung, and pancreatic cancer as monotherapy, as well as combination with PD-1 blockers, radiation, and chemotherapy. While approved, IL-2 retains significant toxicity, preventing its widespread use. The significant efforts undertaken to uncouple IL-2 toxicity from its anti-tumor function have been unsuccessful, and early phase clinical safety observed with PEGylated IL-10 was not met in a blinded Phase 3 trial. Deka Biosciences has engineered a novel molecule coupling wild-type IL-2 to a high affinity variant of Epstein Barr Viral (EBV) IL-10 via a scaffold (scFv) that binds to epidermal growth factor receptors (EGFR). This patented molecule, termed DK210 (EGFR), is retained at high levels within the tumor microenvironment for days after dosing. In addition to overlapping and non-redundant anti-tumor function, IL-10 reduces IL-2 mediated cytokine release syndrome risks and inhibits IL-2 mediated T regulatory cell proliferation. Methods: DK210 (EGFR) is being evaluated in an open-label, dose-escalation (Phase 1) study with 5 (0.025-0.3 mg/kg) monotherapy dose levels and (expansion cohorts) in combination with PD-1 blockers, or radiation or chemotherapy in patients with advanced solid tumors overexpressing EGFR. Key eligibility criteria include 1) confirmed progressive disease on at least one line of systemic treatment, 2) EGFR overexpression or amplification documented in histology reports, 3) at least a 4 week or 5 half-lives window since last treatment, and 4) excluding subjects with long QT syndrome, multiple myeloma, multiple sclerosis, myasthenia gravis or uncontrolled infectious, psychiatric, neurologic, or cancer disease. Plasma and tissue samples will be investigated for pharmacodynamic and predictive biomarkers and genetic signatures associated with IFN-gamma secretion, aiming to select subjects for treatment in Phase 2. Conclusion: Through successful coupling of wild-type IL-2 with a high affinity IL-10 and targeting directly to the tumor microenvironment, DK210 (EGFR) has the potential to harness IL-2 and IL-10’s known anti-cancer promise while reducing immunogenicity and toxicity risks enabling safe concomitant cytokine treatment with other anti-cancer modalities.

Keywords: cytokine, EGFR over expression, interleukine-2, interleukine-10, clinical trial

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Authors: Neha Bhardwaj

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In this paper, we consider positive linear operators and study the Voronovskaya type result of the operator then obtain an error estimate in terms of the higher order modulus of continuity of the function being approximated and its A-statistical convergence. Also, we compute the corresponding rate of A-statistical convergence for the linear positive operators.

Keywords: Poisson distribution, Voronovskaya, modulus of continuity, a-statistical convergence

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Authors: Adamu S. Salawu, Ibrahim O. Isah

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Direct solution to several forms of fourth-order ordinary differential equations is not easily obtained without first reducing them to a system of first-order equations. Thus, numerical methods are being developed with the underlying techniques in the literature, which seeks to approximate some classes of fourth-order initial value problems with admissible error bounds. Multistep methods present a great advantage of the ease of implementation but with a setback of several functions evaluation for every stage of implementation. However, hybrid methods conventionally show a slightly higher order of truncation for any k-step linear multistep method, with the possibility of obtaining solutions at off mesh points within the interval of solution. In the light of the foregoing, we propose the continuous form of a hybrid multistep method with Chebyshev polynomial as a basis function for the numerical integration of fourth-order initial value problems of ordinary differential equations. The basis function is interpolated and collocated at some points on the interval [0, 2] to yield a system of equations, which is solved to obtain the unknowns of the approximating polynomial. The continuous form obtained, its first and second derivatives are evaluated at carefully chosen points to obtain the proposed block method needed to directly approximate fourth-order initial value problems. The method is analyzed for convergence. Implementation of the method is done by conducting numerical experiments on some test problems. The outcome of the implementation of the method suggests that the method performs well on problems with oscillatory or trigonometric terms since the approximations at several points on the solution domain did not deviate too far from the theoretical solutions. The method also shows better performance compared with an existing hybrid method when implemented on a larger interval of solution.

Keywords: Chebyshev polynomial, collocation, hybrid multistep method, initial value problems, interpolation

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Authors: Ashok K. Gupta

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Over the past few decades, since the bio-engineering revolution, autologous cell therapy (ACT) has become a rapidly evolving field. Currently, this form of therapy has broad applications in modern medicine and plastic surgery, ranging from the treatment/improvement of wound healing to life-saving operations. A study was conducted on 50 patients having to disfigure, and deform post burn scars and was treated by injection of extracted, refined adipose tissue grafts with their unique stem cell properties. To compare the outcome, a control of 20 such patients was treated with conventional skin or soft-tissue flaps or skin grafting, and a control of 10 was treated with more advanced microsurgical techniques such as Pre-fabricated flaps/pre laminated flaps / free flaps. Assessment of fat volume and survival post- follow up period was done by radiological aid, using MRI and clinically (Survival of the autograft and objective parameters for scar elasticity were evaluated skin elasticity parameters 3 to 9 months postoperatively). Recently, an enzyme that is involved in collagen crosslinking in fibrotic tissue, lysyl hydroxylase (LH2), was identified. This enzyme is normally active in bone and cartilage but hardly in the skin. It has been found that this enzyme is highly expressed in scar tissue and subcutaneous fat; this is in contrast to the dermis, where the enzyme is hardly expressed. Adipose tissue-derived stem cell injections are an effective method in the treatment of various extensive post-burn scar deformities that makes it possible to re-create the lost sub-dermal tissue for improvement in the function of involved joint movements.

Keywords: adipose tissue-derived stem cell injections, treatment of various extensive post-burn scar deformities, re-create the lost sub-dermal tissue, improvement in function of involved joint movements

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Authors: Londeka Ntuli, Frasia Oosthuizen

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Background: Tenefovir disiproxil fumarate (TDF) is a nephrotoxic drug and has been proven to contribute to renal insufficiency necessitating intensive monitoring and management of adverse effects arising from prolonged exposure to the drug. TDF is one of the preferred first-line drugs used in combination therapy in most regions. There are estimated 300 000 patients being initiated on the Efavirenz/TDF/Emtricitabine first-line regimen annually in South Africa. It is against this background that this study aims to investigate the effects of TDF on renal sufficiency of HIV positive patients. Methodology: A retrospective quantitative study was conducted, analysing clinical charts of HIV positive patient’s older than 18 years of age and on a TDF-containing regimen for more than 1 year. Data were obtained from the analysis of patient files and was transcribed into Microsoft® Excel® spreadsheet. Extracted data were coded, categorised and analysed using STATA®. Results: A total of 275 patient files were included in this study. Renal function started decreasing after 3 months of treatment (with 93.5% patients having a normal EGFR), and kept on decreasing as time progressed with only 39.6% normal renal function at year 4. Additional risk factors for renal insufficiency included age below 25, female gender, and additional medication. Conclusion: It is clear from this study that the use of TDF necessitates intensive monitoring and management of adverse effects arising from prolonged exposure to the drug. The findings from this study generated pertinent information on the safety profile of the drug TDF in a resource-limited setting of a public health institution. The appropriate management is of tremendous importance in the South African context where the majority of HIV positive individuals are on the TDF containing regimen; thus it is beneficial to ascertain the possible level of toxicities these patients may be experiencing.

Keywords: renal insufficiency, tenefovir, HIV, risk factors

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Authors: Sharmeen Rahman, Daniel Schmidt, Jane Hughes

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Paratya australiensis Kemp (Decapoda: Atyidae) is a widely distributed indigenous freshwater shrimp, highly abundant in eastern Australia. This species has been considered as a model stream organism to study genetics, dispersal, biology, behaviour and evolution in Atyids. Paratya has a filter feeding and scavenging habit which plays a significant role in the formation of lotic community structure. It has been shown to reduce periphyton and sediment from hard substrates of coastal streams and hence acts as a strongly-interacting ecosystem macroconsumer. Besides, Paratya is one of the major food sources for stream dwelling fishes. Paratya australiensis is a cryptic species complex consisting of 9 highly divergent mitochondrial DNA lineages. Among them, one lineage has been observed to favour upstream sites at higher altitudes, with cooler water temperatures. This study aims to identify local adaptation in upstream and downstream populations of this lineage in three streams in the Conondale Range, North-eastern Brisbane, Queensland, Australia. Two populations (up and down stream) from each stream have been chosen to test for local adaptation, and a parallel pattern of adaptation is expected across all streams. Six populations each consisting of 24 individuals were sequenced using the Restriction Site Associated DNA-seq (RAD-seq) technique. Genetic markers (SNPs) were developed using double digest RAD sequencing (ddRAD-seq). These were used for de novo assembly of Paratya genome. De novo assembly was done using the STACKs program and produced 56, 344 loci for 47 individuals from one stream. Among these individuals, 39 individuals shared 5819 loci, and these markers are being used to test for local adaptation using Fst outlier tests (Arlequin) and Bayesian analysis (BayeScan) between up and downstream populations. Fst outlier test detected 27 loci likely to be under selection and the Bayesian analysis also detected 27 loci as under selection. Among these 27 loci, 3 loci showed evidence of selection at a significance level using BayeScan program. On the other hand, up and downstream populations are strongly diverged at neutral loci with a Fst =0.37. Similar analysis will be done with all six populations to determine if there is a parallel pattern of adaptation across all streams. Furthermore, multi-locus among population covariance analysis will be done to identify potential markers under selection as well as to compare single locus versus multi-locus approaches for detecting local adaptation. Adaptive genes identified in this study can be used for future studies to design primers and test for adaptation in related crustacean species.

Keywords: Paratya australiensis, rainforest streams, selection, single nucleotide polymorphism (SNPs)

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Authors: Amir Mirzaei

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The powertrain system is one of the most basic and essential components in a machine. The occurrence of motion is practically impossible without the presence of this system. When power is generated by the engine, it is transmitted by the powertrain system to the wheels, which are the last parts of the system. Powertrain system has different components according to the type of use and design. When the force generated by the engine reaches to the wheels, the amount of frictional force between the tire and the ground determines the amount of traction and non-slip or the amount of slip. At various levels, such as icy, muddy, and snow-covered ground, the amount of friction coefficient between the tire and the ground decreases dramatically and considerably, which in turn increases the amount of force loss and the vehicle traction decreases drastically. This condition is caused by the phenomenon of slipping, which, in addition to the waste of energy produced, causes the premature wear of driving tires. It also causes the temperature of the transmission oil to rise too much, as a result, causes a reduction in the quality and become dirty to oil and also reduces the useful life of the clutches disk and plates inside the transmission. this issue is much more important in road construction and mining machinery than passenger vehicles and is always one of the most important and significant issues in the design discussion, in order to overcome. One of these methods is the automatic spin regulator system which is abbreviated as ASR. The importance of this method and its structure and function have solved one of the biggest challenges of the powertrain system in the field of construction and mining machinery. That this research is examined.

Keywords: automatic spin regulator, ASR, methods of reducing slipping, methods of preventing the reduction of the useful life of clutches disk and plate, methods of preventing the premature dirtiness of transmission oil, method of preventing the reduction of the useful life of tires

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Authors: Fatemeh Abbasi, Sahand Daneshvar

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Productivity is one of the essential goals of companies to improve performance, which as a strategy-oriented method, determines the basis of the company's economic growth. The history of productivity goes back centuries, but most researchers defined productivity as the relationship between a product and the factors used in production in the early twentieth century. Productivity as the optimal use of available resources means that "more output using less input" can increase companies' economic growth and prosperity capacity. Also, having a quality life based on economic progress depends on productivity growth in that society. Therefore, productivity is a national priority for any developed country. There are several methods for calculating productivity growth measurements that can be divided into parametric and non-parametric methods. Parametric methods rely on the existence of a function in their hypotheses, while non-parametric methods do not require a function based on empirical evidence. One of the most popular non-parametric methods is Data Envelopment Analysis (DEA), which measures changes in productivity over time. The DEA evaluates the productivity of decision-making units (DMUs) based on mathematical models. This method uses multiple inputs and outputs to compare the productivity of similar DMUs such as banks, government agencies, companies, airports, Etc. Non-parametric methods are themselves divided into the frontier and non frontier approaches. The Malmquist productivity index (MPI) proposed by Caves, Christensen, and Diewert (1982), the Hicks–Moorsteen productivity index (HMPI) proposed by Bjurek (1996), or the Luenberger productivity indicator (LPI) proposed by Chambers (2002) are powerful tools for measuring productivity changes over time. This study will compare the Malmquist, Hicks–Moorsteen, and Luenberger indices theoretically and empirically based on DEA models and review their strengths and weaknesses.

Keywords: data envelopment analysis, Hicks–Moorsteen productivity index, Leuenberger productivity indicator, malmquist productivity index

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Authors: Xingyu Peng, Qingyuan Hu, Xuebin Zhu, Xi Yuan

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Neutron spectrometry has contributed much to the development of nuclear physics since 1932 and has also become an importance tool in several other fields, notably nuclear technology, fusion plasma diagnostics and radiation protection. Compared with neutron fluxes, neutron spectra can provide more detailed information on the internal physical process of neutron sources, such as fast neutron reactors, fusion plasma, fission-fusion hybrid reactors, and so on. However, high performance neutron spectrometer is not so commonly available as it requires the use of large and complex instrumentation. This work describes the development and characterization of a compact magnetic proton recoil (MPR) spectrometer for high-resolution measurements of fast neutron spectra. The compact MPR spectrometer is featured by its large recoil angle, small size permanent analysis magnet, short beam transport line and dual-purpose detector array for both steady state and pulsed neutron spectra measurement. A 3-dimensional electromagnetic particle transport code is developed to simulate the response function of the spectrometer. Simulation results illustrate that the performance of the spectrometer is mainly determined by n-p recoil foil and proton apertures, and an overall energy resolution of 3% is achieved for 14 MeV neutrons. Dedicated experiments using alpha source and mono-energetic neutron beam are employed to verify the simulated response function of the compact MPR spectrometer. These experimental results show a good agreement with the simulated ones, which indicates that the simulation code possesses good accuracy and reliability. The compact MPR spectrometer described in this work is a valuable tool for fast neutron spectra measurements for the fission or fusion devices.

Keywords: neutron spectrometry, magnetic proton recoil spectrometer, neutron spectra, fast neutron

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Authors: Saeedeh Eshkil, Seyedeh Farnaz Mousavi, Hamid Reza Kianifar, Seyed Java Sayyedi, Mehdi Sohrabi, Elham Bakhtiari, Morteza Mashoughi, Ezzat Khodashenas

Abstract:

Background: The effect of the SPARK physical education program on lung function in cystic fibrosis patients is not yet determined.).SPARK is Sports, play and active recreation for kids, including moving skills, aerobic games, jogging or walking, aerobic dance and jump rope. Regarding the high prevalence of cystic fibrosis and its destructive effects on the lungs, the aim of this study is to evaluate lung function and quality of life before and after undergoing the SPARK physical education program in children with cystic fibrosis. Method: In this quasi-experimental study, all patients with cystic fibrosis aged 6-18 years referred to the cystic fibrosis clinic of Dr. Sheikh Hospital were enrolled. The patients went under 12 weeks of SPARK training program (3 sessions per week, each session 45 minutes). The quality of life questionnaire ( Cystic Fibrosis Questionnaire includes self-examination, parental ) for patients with cystic fibrosis and spirometry indices (FEV1, FVC, FEV1/FVC, FEF25-75) was filled out before and after intervention for all patients. Results The mean and standard deviation of patients' age were 9.85±2.67 years, and 65% of patients were female. The FEV1 was significantly different before and after the SPARK physical education program (P=0.03), and the respiratory component of quality of life significantly increased after intervention (P=0.002). The overall score of quality of life from parents’ point of view was 2.87 ± 0.38, which increased to 2.99 ± 0.38 after the intervention. Conclusion: The SPARK training program may improve the spirometric parameters in children with cystic fibrosis. It also had a significant effect on improving the quality of life of patients, especially in the respiratory component.

Keywords: cystic fibrosis, pediatrics, SPARK motor program, spirometry

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Authors: Pirta Palola, Richard Bailey, Lisa Wedding

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Societies and economies worldwide fundamentally depend on natural capital. Alarmingly, natural capital assets are quickly depreciating, posing an existential challenge for humanity. The development of robust natural capital accounting systems is essential for transitioning towards sustainable economic systems and ensuring sound management of capital assets. However, the accurate, equitable and comprehensive estimation of natural capital asset stocks and their accounting values still faces multiple challenges. In particular, the representation of socio-cultural values held by groups or communities has arguably been limited, as to date, the valuation of natural capital assets has primarily been based on monetary valuation methods and assumptions of individual rationality. People relate to and value the natural environment in multiple ways, and no single valuation method can provide a sufficiently comprehensive image of the range of values associated with the environment. Indeed, calls have been made to improve the representation of multiple types of value (instrumental, intrinsic, and relational) and diverse ontological and epistemological perspectives in environmental valuation. This study addresses this need by establishing a novel valuation framework, Environmental Value Functions (EVF), that allows for the integration of multiple types of value in natural capital accounting systems. The EVF framework is based on the estimation and application of value functions, each of which describes the relationship between the value and quantity (or quality) of an ecosystem component of interest. In this framework, values are estimated in terms of change relative to the current level instead of calculating absolute values. Furthermore, EVF was developed to also support non-marginalist conceptualizations of value: it is likely that some environmental values cannot be conceptualized in terms of marginal changes. For example, ecological resilience value may, in some cases, be best understood as a binary: it either exists (1) or is lost (0). In such cases, a logistic value function may be used as the discriminator. Uncertainty in the value function parameterization can be considered through, for example, Monte Carlo sampling analysis. The use of EVF is illustrated with two conceptual examples. For the first time, EVF offers a clear framework and concrete methodology for the representation of multiple types of value in natural capital accounting systems, simultaneously enabling 1) the complementary use and integration of multiple valuation methods (monetary and non-monetary); 2) the synthesis of information from diverse knowledge systems; 3) the recognition of value incommensurability; 4) marginalist and non-marginalist value analysis. Furthermore, with this advancement, the coupling of EVF and ecosystem modeling can offer novel insights to the study of spatial-temporal dynamics in natural capital asset values. For example, value time series can be produced, allowing for the prediction and analysis of volatility, long-term trends, and temporal trade-offs. This approach can provide essential information to help guide the transition to a sustainable economy.

Keywords: economics of biodiversity, environmental valuation, natural capital, value function

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Authors: Sri Sai Ramya Bojedla, Falguni Pati

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Nature provides the best of solutions to humans. One such incredible gift to regenerative medicine is silk. The literature has publicized a long appreciation for silk owing to its incredible physical and biological assets. Its bioactive nature, unique mechanical strength, and processing flexibility make us curious to explore further to apply it in the clinics for the welfare of mankind. In this study, Antheraea mylitta and Bombyx mori silk fibroin microfibers are developed by two economical and straightforward steps via degumming and hydrolysis for the first time, and a bioactive composite is manufactured by mixing silk fibroin microfibers at various concentrations with polycaprolactone (PCL), a biocompatible, aliphatic semi-crystalline synthetic polymer. Reconstructive surgery in any part of the body except for the maxillofacial region deals with replacing its function. But answering both the aesthetics and function is of utmost importance when it comes to facial reconstruction as it plays a critical role in the psychological and social well-being of the patient. The main concern in developing adequate bone graft substitutes or a scaffold is the noteworthy variation in each patient's bone anatomy. Additionally, the anatomical shape and size will vary based on the type of defect. The advent of additive manufacturing (AM) or 3D printing techniques to bone tissue engineering has facilitated overcoming many of the restraints of conventional fabrication techniques. The acquired patient's CT data is converted into a stereolithographic (STL)-file which is further utilized by the 3D printer to create a 3D scaffold structure in an interconnected layer-by-layer fashion. This study aims to address the limitations of currently available materials and fabrication technologies and develop a customized biomaterial implant via 3D printing technology to reconstruct complex form, function, and aesthetics of the facial anatomy. These composite scaffolds underwent structural and mechanical characterization. Atomic force microscopic (AFM) and field emission scanning electron microscopic (FESEM) images showed the uniform dispersion of the silk fibroin microfibers in the PCL matrix. With the addition of silk, there is improvement in the compressive strength of the hybrid scaffolds. The scaffolds with Antheraea mylitta silk revealed higher compressive modulus than that of Bombyx mori silk. The above results of PCL-silk scaffolds strongly recommend their utilization in bone regenerative applications. Successful completion of this research will provide a great weapon in the maxillofacial reconstructive armamentarium.

Keywords: compressive modulus, 3d printing, maxillofacial reconstruction, natural fiber reinforced composites, silk fibroin microfibers

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Authors: S. Daneshi, G. Shahcheraghi, F. Ghaffarinejad

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Back ground: Osteoarthritis (OA) is the most common form of arthritis, affecting millions of people around the world during the aging process. Knee joint proprioception sense decrease with OA and Total Knee Arthroplasty (TKA) surgery may affect them. We investigated two parameters of proprioception sense (the joint position sense and kinesthesia) and standing balance in affected limbs before and after TKA, in patient with Knee OA. Methods and Materials: In this Analytic study, 10 patients who were candidate for TKA during two months in Dena Hospital of Shiraz, selected for further analysis. All of cases were female in range of 55-70 years old. Participants assessed before and two weeks after TKA using three instruments: electrogoniometer and continuous passive motion (CPM) to assess Knee joint position sense and kinesthesia in 20 and 45 degrees; and chronometer to assess duration of standing balance on affected leg with open and closed eyes. Results: To examine differences between before and after of TKA scorings Willcoxon Signed Rank and Mann-Whitney was performed which indicated no significant differences between knee joint position sense and kinesthesia in 20 and 45 degrees (P>0.05) and no significant differences between Standing Balance in a patient with knee OA before and after TKA (P>0.05). Conclusion: The study indicates that, OA can affect proprioception sense and standing balance but TKA doesn’t have any effect on these parameters. Intra articular structures such as cruciate ligaments and mines are responsible for proprioception sense in normal knee joint. Since in severe knee OA the number of mechanoreceptors in these intra articular structures decrease and their function reduce more than normal knee joint, so the anterior cruciate ligaments (ACL) become defected, thus after TKA surgery which this ligament is removed no significant change was found in proprioception sense. As a result of involving proprioception sense, muscles strength and the function of vestibular system in balance, standing balance did not show significant difference before and after TKA.

Keywords: knee joint, proprioception sense, standing balance, rehabilitation sciences

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Authors: Aneri Mehta, Krunal Mehta

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Financial administration is part of government which deals with collection, preservation and distribution of public funds, with the coordination of public revenue and expenditure, with the management of credit operation on behalf of the state and with the general control of the financial affairs of public households. The researcher has taken the prime body of the local self government viz. Municipal Corporation. However, the number of municipal corporations in India has rapidly increased in recent years. Countries 27% of the total population are living in urban area & in recent it increasing very fast. People are moving very fast from rural area to urban area. Their demand, awareness is increasing day by day. The Municipal Corporations render many services for the development of the urban area. Thus, researcher has taken a step to know the accounting practices of the municipal corporations of Gujarat state (AMC & BMC ). The research will try to show you the status of finance of municipal corporations. Article 243(w) of the constitution of India envisaged that the state government maybe, by law , endow the municipalities with such powers and authorities as may be necessary to enable them to function as institution of self government and such law may contain provision for devolution of powers and responsibilities upon municipalities subjects to such condition as may be specified there in with respect to (i) the peroration of plans for economic development and social justice and (ii) the performance of the function and the implementation of schemes as may be entrusted to them including those in relation to the matters listed in the twelfth schedule. The three tier structure of the Indian Government i.e. Union, State & Local Self Government is the scenario of the Indian constitution. Local Self Government performs or renders many services under the direct control of state government. They (local bodies) possess autonomy within its limited sphere, raise revenue through local taxation and spend its income on local services.

Keywords: financial administration, urban local bodies, local self government, constitution

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Authors: Chang-Hui Shen, Paulina Konarzewska

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Cell homeostasis is regulated by vacuolar activity and it has been shown that lipid composition of the vacuole plays an important role in vacuolar function. The major phosphoinositide species present in the vacuolar membrane include phosphatidylinositol 3,5-biphosphate (PI(3,5)P₂) which is generated from PI(3)P controlled by Fab1p. Deletion of FAB1 gene reduce the synthesis of PI(3,5)P₂ and thus result in enlarged or fragmented vacuoles, with neutral vacuolar pH due to reduced vacuolar H⁺-ATPase activity. These mutants also exhibited poor growth at high extracellular pH and in the presence of CaCl₂. Conversely, VPS34 regulates the synthesis of PI(3)P from phosphatidylinositol (PI), and the lack of Vps34p results in the reduction of vacuolar activity. Although the cellular observations are clear, it is still unknown about the molecular mechanism between the phospholipid biosynthesis pathway and vacuolar activity. Since both VPS34 and FAB1 are important in vacuolar activity, we hypothesize that the molecular mechanism of vacuolar function might be regulated by the transcriptional regulators of phospholipid biosynthesis. In this study, we study the role of the major phospholipid biosynthesis transcription factor, INO2, in the regulation of vacuolar activity. We first performed qRT-PCR to examine the effect of Ino2p on the expression of VPS34 and FAB1. Our results showed that VPS34 was upregulated in the presence of inositol for both WT and ino2Δ cells. However, FAB1 was only upregulated significantly in ino2Δ cells. This indicated that Ino2p might be the negative regulator for FAB1 expression. Next, growth sensitivity experiment showed that WT, vma3Δ, and ino2Δ grew well in growth medium buffered to pH 5.5 containing 10 mM CaCl₂. As cells were switched to growth medium buffered to pH 7 containing CaCl₂ WT, ino2Δ and opi1Δ showed growth reduction, whereas vma3Δ was completely nonviable. As the concentration of CaCl₂ was increased to 60 mM, ino2Δ cells showed moderate growth reduction compared to WT. This result suggests that ino2Δ cells have better vacuolar activity. Microscopic analysis and vacuolar acidification were employed to further elucidate the importance of INO2 in vacuolar homeostasis. Analysis of vacuolar morphology indicated that WT and vma3Δ cells displayed vacuoles that occupied a small area of the cell when grown in media buffered to pH 5.5. Whereas, ino2Δ displayed fragmented vacuoles. On the other hand, all strains grown in media buffered to pH 7, exhibited enlarged vacuoles that occupied most of the cell’s surface. This indicated that the presence of INO2 may play negative effect in vacuolar morphology when cells are grown in media buffered to pH 5.5. Furthermore, vacuolar acidification assay showed that only vma3Δ cells displayed notably less acidic vacuoles as cells were grown in media buffered to pH 5.5 and pH 7. Whereas, ino2Δ cells displayed more acidic pH compared to WT at pH7. Taken together, our results demonstrated the molecular mechanism of the vacuolar activity regulated by the phospholipid biosynthesis transcription factors Ino2p. Ino2p negatively regulates vacuolar activity through the expression of FAB1.

Keywords: vacuole, phospholipid, homeostasis, Ino2p, FAB1

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