Search results for: protein extraction

Authors: Sairah Bashir, Liu Yang, John Liggat, James Thomason

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Glass fibre is currently used as reinforcement in over 90% of all fibre-reinforced composites produced. The high rigidity and chemical resistance of these composites are required for optimum performance but unfortunately results in poor recyclability; when such materials are no longer fit for purpose, they are frequently deposited in landfill sites. Recycling technologies, for example, thermal treatment, can be employed to address this issue; temperatures typically between 450 and 600 °C are required to allow degradation of the rigid polymeric matrix and subsequent extraction of fibrous reinforcement. However, due to the severe thermal conditions utilised in the recycling procedure, glass fibres become too weak for reprocessing in second-life composite materials. In addition, more stringent legislation is being put in place regarding disposal of composite waste, and so it is becoming increasingly important to develop long-term recycling solutions for such materials. In particular, the development of a cost-effective method to regenerate strength of thermally recycled glass fibres will have a positive environmental effect as a reduced volume of composite material will be destined for landfill. This research study has demonstrated the positive impact of sodium hydroxide (NaOH) and potassium hydroxide (KOH) solution, prepared at relatively mild temperatures and at concentrations of 1.5 M and above, on the strength of heat-treated glass fibres. As a result, alkaline treatments can potentially be implemented to glass fibres that are recycled from composite waste to allow their reuse in second-life materials. The optimisation of the strength recovery process is being conducted by varying certain reaction parameters such as molarity of alkaline solution and treatment time. It is believed that deep V-shaped surface flaws exist commonly on severely damaged fibre surfaces and are effectively removed to form smooth, U-shaped structures following alkaline treatment. Although these surface flaws are believed to be present on glass fibres they have not in fact been observed, however, they have recently been discovered in this research investigation through analytical techniques such as AFM (atomic force microscopy) and SEM (scanning electron microscopy). Reaction conditions such as molarity of alkaline solution affect the degree of etching of the glass fibre surface, and therefore the extent to which fibre strength is recovered. A novel method in determining the etching rate of glass fibres after alkaline treatment has been developed, and the data acquired can be correlated with strength. By varying reaction conditions such as alkaline solution temperature and molarity, the activation energy of the glass etching process and the reaction order can be calculated respectively. The promising results obtained from NaOH and KOH treatments have opened an exciting route to strength regeneration of thermally recycled glass fibres, and the optimisation of the alkaline treatment process is being continued in order to produce recycled fibres with properties that match original glass fibre products. The reuse of such glass filaments indicates that closed-loop recycling of glass fibre reinforced composite (GFRC) waste can be achieved. In fact, the development of a closed-loop recycling process for GFRC waste is already underway in this research study.

Keywords: glass fibers, glass strengthening, glass structure and properties, surface reactions and corrosion

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Authors: Mandeep Singh Azad.Dibyendu Chakraborty, Vikas Vohra

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Introduction: Poonch is one of the remotest districts of the Jammu and Kashmir (UT) and situated on international borders. This native poultry population in these areas is quite hardy and thrives well in adverse climatic conditions. Till date, no local breed from this area (Jammu Province) has been characterized thus present study was undertaken with the main objectives of molecular characterization of ChB6 gene in local native chicken of Poonch region located at international borders between India and Pakistan. The chicken B-cell marker (ChB6) gene has been proposed as a candidate gene in regulating B-cell development. Material and Method: RNA was isolated by Blood RNA Purification Kit (HiPura) and Trizol method from whole blood samples. Positive PCR products with size 1110 bp were selected for further purification, sequencing and analysis. The amplified PCR product was sequenced by Sangers dideoxy chain termination method. The obtained sequence of ChB6 gene of Poonchi chicken were compared by MEGAX software. BioEdit software was used to construct phylogenic tree, and Neighbor Joining method was used to infer evolutionary history. In order to compute evolutionary distance Maximum Composite Likelihood method was used. Results: The positively amplified samples of ChB6 genes were then subjected to Sanger sequencing with “Primer Walking. The sequences were then analyzed using MEGA X and BioEdit software. The sequence results were compared with other reported sequence from different breed of chicken and with other species obtained from the NCBI (National Center for Biotechnology Information). ClustalW method using MEGA X software was used for multiple sequence alignment. The sequence results of ChB6 gene of Poonchi chicken was compared with Centrocercus urophasianus, G. gallus mRNA for B6.1 protein, G. gallus mRNA for B6.2, G. gallus mRNA for B6.3, Gallus gallus B6.1, Halichoeres bivittatus, Miniopterus fuliginosus Ferringtonia patagonica, Tympanuchus phasianellus. The genetic distances were 0.2720, 0.0000, 0.0245, 0.0212, 0.0147, 1.6461, 2.2394, 2.0070 and 0.2363 for ChB6 gene of Poonchi chicken sequence with other sequences in the present study respectively. Sequencing results showed variations between different species. It was observed that AT content were higher then GC content for ChB6 gene. The lower AT content suggests less thermostable. It was observed that there was no sequence difference within the Poonchi population for ChB6 gene. The high homology within chicken population indicates the conservation of ChB6 gene. The maximum difference was observed with Miniopterus fuliginosus (Eastern bent-wing bat) followed by Ferringtonia patagonica and Halichoeres bivittatus. Conclusion: Genetic variation is the essential component for genetic improvement. The results of immune related gene Chb6 shows between population genetic variability. Therefore, further association studies of this gene with some prevalent diseases in large population would be helpful to identify disease resistant/ susceptible genotypes in the indigenous chicken population.

Keywords: ChB6, sequencing, ClustalW, genetic distance, poonchi chicken, SNP

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Authors: Helen S. Joyner (Melito), Mohammad Anvari

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Concentrated emulsions stabilized by proteins are systems of great importance in food, pharmaceutical and cosmetic products. Controlling emulsion rheology is critical for ensuring desired properties during formation, storage, and consumption of emulsion-based products. Studies on concentrated emulsions have focused on rheology of monodispersed systems. However, emulsions used for industrial applications are polydispersed in nature, and this polydispersity is regarded as an important parameter that also governs the rheology of the concentrated emulsions. Therefore, the objective of this study was to characterize rheological (small and large deformation behaviors) and microstructural properties of concentrated emulsions which were not truly monodispersed as usually encountered in food products such as margarines, mayonnaise, creams, spreads, and etc. The concentrated emulsions were prepared at different concentrations of fish gelatin (0.2, 0.4, 0.8% w/v in the whole emulsion system), oil-water ratio 80-20 (w/w), homogenization speed 10000 rpm, and 25oC. Confocal laser scanning microscopy (CLSM) was used to determine the microstructure of the emulsions. To prepare samples for CLSM analysis, FG solutions were stained by Fluorescein isothiocyanate dye. Emulsion viscosity profiles were determined using shear rate sweeps (0.01 to 100 1/s). The linear viscoelastic regions (LVRs) of the emulsions were determined using strain sweeps (0.01 to 100% strain) for each sample. Frequency sweeps were performed in the LVR (0.1% strain) from 0.6 to 100 rad/s. Large amplitude oscillatory shear (LAOS) testing was conducted by collecting raw waveform data at 0.05, 1, 10, and 100% strain at 4 different frequencies (0.5, 1, 10, and 100 rad/s). All measurements were performed in triplicate at 25oC. The CLSM results revealed that increased fish gelatin concentration resulted in more stable oil-in-water emulsions with homogeneous, finely dispersed oil droplets. Furthermore, the protein concentration had a significant effect on emulsion rheological properties. Apparent viscosity and dynamic moduli at small deformations increased with increasing fish gelatin concentration. These results were related to increased inter-droplet network connections caused by increased fish gelatin adsorption at the surface of oil droplets. Nevertheless, all samples showed shear-thinning and weak gel behaviors over shear rate and frequency sweeps, respectively. Lissajous plots, or plots of stress versus strain, and phase lag values were used to determine nonlinear behavior of the emulsions in LAOS testing. Greater distortion in the elliptical shape of the plots followed by higher phase lag values was observed at large strains and frequencies in all samples, indicating increased nonlinear behavior. Shifts from elastic-dominated to viscous dominated behavior were also observed. These shifts were attributed to damage to the sample microstructure (e.g. gel network disruption), which would lead to viscous-type behaviors such as permanent deformation and flow. Unlike the small deformation results, the LAOS behavior of the concentrated emulsions was not dependent on fish gelatin concentration. Systems with different microstructures showed similar nonlinear viscoelastic behaviors. The results of this study provided valuable information that can be used to incorporate concentrated emulsions in emulsion-based food formulations.

Keywords: concentrated emulsion, fish gelatin, microstructure, rheology

Procedia PDF Downloads 250

Authors: A. Casale, J. Alessi, C. N. Bianchi, G. Bozzini, M. Brunoldi, V. Cappanera, P. Corvisiero, G. Fanciulli, D. Grosso, N. Magnoli, A. Mandich, C. Melchiorre, C. Morri, P. Povero, N. Stasi, M. Taiuti, G. Viano, M. Wurtz

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This work describes a real-time algorithm for detecting, tracking and classifying single motorboats, developed using the acoustic data recorded by a hydrophone array within the framework of EU LIFE + project ARION (LIFE09NAT/IT/000190). The project aims to improve the conservation status of bottlenose dolphins through a real-time simultaneous monitoring of their population and surface ship traffic. A Passive Acoustic Monitoring (PAM) system is installed on two autonomous permanent marine buoys, located close to the boundaries of the Marine Protected Area (MPA) of Portofino (Ligurian Sea- Italy). Detecting surface ships is also a necessity in many other sensible areas, such as wind farms, oil platforms, and harbours. A PAM system could be an effective alternative to the usual monitoring systems, as radar or active sonar, for localizing unauthorized ship presence or illegal activities, with the advantage of not revealing its presence. Each ARION buoy consists of a particular type of structure, named meda elastica (elastic beacon) composed of a main pole, about 30-meter length, emerging for 7 meters, anchored to a mooring of 30 tons at 90 m depth by an anti-twist steel wire. Each buoy is equipped with a floating element and a hydrophone tetrahedron array, whose raw data are send via a Wi-Fi bridge to a ground station where real-time analysis is performed. Bottlenose dolphin detection algorithm and ship monitoring algorithm are operating in parallel and in real time. Three modules were developed and commissioned for ship monitoring. The first is the detection algorithm, based on Time Difference Of Arrival (TDOA) measurements, i.e., the evaluation of angular direction of the target respect to each buoy and the triangulation for obtaining the target position. The second is the tracking algorithm, based on a Kalman filter, i.e., the estimate of the real course and speed of the target through a predictor filter. At last, the classification algorithm is based on the DEMON method, i.e., the extraction of the acoustic signature of single vessels. The following results were obtained; the detection algorithm succeeded in evaluating the bearing angle with respect to each buoy and the position of the target, with an uncertainty of 2 degrees and a maximum range of 2.5 km. The tracking algorithm succeeded in reconstructing the real vessel courses and estimating the speed with an accuracy of 20% respect to the Automatic Identification System (AIS) signals. The classification algorithm succeeded in isolating the acoustic signature of single vessels, demonstrating its temporal stability and the consistency of both buoys results. As reference, the results were compared with the Hilbert transform of single channel signals. The algorithm for tracking multiple targets is ready to be developed, thanks to the modularity of the single ship algorithm: the classification module will enumerate and identify all targets present in the study area; for each of them, the detection module and the tracking module will be applied to monitor their course.

Keywords: acoustic-noise, bottlenose-dolphin, hydrophone, motorboat

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Authors: Swati Gautam, Salman Akhtar, S. P. Jaiswar, Amita Jain

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Background: FGTB is now a major global health problem mostly in developing countries including India. In humans, Mycobacterium Tuberculosis (M.tb) is a causating agent of infection. High index of suspicion is required for early diagnosis due to asymptomatic presentation of FGTB disease. In macrophages Toll Like Receptor-2 (TLR-2) is one which mediated host’s immune response to M.tb. The expression of TLR-2 on macrophages is important to determine the fate of innate immune responses to M.tb. TLR-2 have two work. First its high expression on macrophages worsen the outer of infection and another side, it maintains M.tb to its dormant stage avoids activation of M.tb from latent phase. Single Nucleotide Polymorphism (SNP) of TLR-2 gene plays an important role in susceptibility to TB among different populations and subsequently, in the development of infertility. Methodology: This Case-Control study was done in the Department of Obs and Gynae and Department of Microbiology at King George’s Medical University, U.P, Lucknow, India. Total 300 subjects (150 Cases and 150 Controls) were enrolled in the study. All subjects were enrolled only after fulfilling the given inclusion and exclusion criteria. Inclusion criteria: Age 20-35 years, menstrual-irregularities, positive on Acid-Fast Bacilli (AFB), TB-PCR, (LJ/MGIT) culture in Endometrial Aspiration (EA). Exclusion criteria: Koch’s active, on ATT, PCOS, and Endometriosis fibroid women, positive on Gonococal and Chlamydia. Blood samples were collected in EDTA tubes from cases and healthy control women (HCW) and genomic DNA extraction was carried out by salting-out method. Genotyping of TLR2 genetic variants (Arg753Gln and Arg677Trp) were performed by using single amplification refractory mutation system (ARMS) PCR technique. PCR products were analyzed by electrophoresis on 1.2% agarose gel and visualized by gel-doc. Statistical analysis of the data was performed using the SPSS 16.3 software and computing odds ratio (OR) with 95% CI. Linkage Disequiliribium (LD) analysis was done by SNP stats online software. Results: In TLR-2 (Arg753Gln) polymorphism significant risk of FGTB observed with GG homozygous mutant genotype (OR=13, CI=0.71-237.7, p=0.05), AG heterozygous mutant genotype (OR=13.7, CI=0.76-248.06, p=0.03) however, G allele (OR=1.09, CI=0.78-1.52, p=0.67) individually was not associated with FGTB. In TLR-2 (Arg677Trp) polymorphism a significant risk of FGTB observed with TT homozygous mutant genotype (OR= 0.020, CI=0.001-0.341, p < 0.001), CT heterozygous mutant genotype (OR=0.53, CI=0.33-0.86, p=0.014) and T allele (OR=0.463, CI=0.32-0.66, p < 0.001). TT mutant genotype was only found in FGTB cases and frequency of CT heterozygous more in control group as compared to FGTB group. So, CT genotype worked as protective mutation for FGTB susceptibility group. In haplotype analysis of TLR-2 genetic variants, four possible combinations, i.e. (G-T, A-C, G-C, and A-T) were obtained. The frequency of haplotype A-C was significantly higher in FGTB cases (0.32). Control group did not show A-C haplotype and only found in FGTB cases. Conclusion: In conclusion, study showed a significant association with both genetic variants of TLR-2 of FGTB disease. Moreover, the presence of specific associated genotype/alleles suggest the possibility of disease severity and clinical approach aimed to prevent extensive damage by disease and also helpful for early detection of disease.

Keywords: ARMS, EDTA, FGTB, TLR

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Authors: Anastasia Constantinou, Stephen Morris

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Background: The adoption of synchronous and asynchronous telemedicine modalities for primary care consultations has exponentially increased since the COVID-19 pandemic. However, there is limited understanding of how virtual consultations influence healthcare resource utilization and other quality measures including safety, timeliness, efficiency, patient and provider satisfaction, cost-effectiveness and environmental impact. Aim: Quantify the rate of follow-up visits, emergency department visits, hospitalizations, request for investigations and prescriptions and comment on the effect on different quality measures associated with different telemedicine modalities used for primary care services and primary care referrals to secondary care Design and setting: Systematic review in primary care Methods: A systematic search was carried out across three databases (Medline, PubMed and Scopus) between August and November 2023, using terms related to telemedicine, general practice, electronic referrals, follow-up, use and efficiency and supported by citation searching. This was followed by screening according to pre-defined criteria, data extraction and critical appraisal. Narrative synthesis and metanalysis of quantitative data was used to summarize findings. Results: The search identified 2230 studies; 50 studies are included in this review. There was a prevalence of asynchronous modalities in both primary care services (68%) and referrals from primary care to secondary care (83%), and most of the study participants were females (63.3%), with mean age of 48.2. The average follow-up for virtual consultations in primary care was 28.4% (eVisits: 36.8%, secure messages 18.7%, videoconference 23.5%) with no significant difference between them or F2F consultations. There was an average annual reduction of primary care visits by 0.09/patient, an increase in telephone visits by 0.20/patient, an increase in ED encounters by 0.011/patient, an increase in hospitalizations by 0.02/patient and an increase in out of hours visits by 0.019/patient. Laboratory testing was requested on average for 10.9% of telemedicine patients, imaging or procedures for 5.6% and prescriptions for 58.7% of patients. When looking at referrals to secondary care, on average 36.7% of virtual referrals required follow-up visit, with the average rate of follow-up for electronic referrals being higher than for videoconferencing (39.2% vs 23%, p=0.167). Technical failures were reported on average for 1.4% of virtual consultations to primary care. When using carbon footprint estimates, we calculate that the use of telemedicine in primary care services can potentially provide a net decrease in carbon footprint by 0.592kgCO2/patient/year. When follow-up rates are taken into account, we estimate that virtual consultations reduce carbon footprint for primary care services by 2.3 times, and for secondary care referrals by 2.2 times. No major concerns regarding quality of care, or patient satisfaction were identified. 5/7 studies that addressed cost-effectiveness, reported increased savings. Conclusions: Telemedicine provides quality, cost-effective, and environmentally sustainable care for patients in primary care with inconclusive evidence regarding the rates of subsequent healthcare utilization. The evidence is limited by heterogeneous, small-scale studies and lack of prospective comparative studies. Further research to identify the most appropriate telemedicine modality for different patient populations, clinical presentations, service provision (e.g. used to follow-up patients instead of initial diagnosis) as well as further education for patients and providers alike on how to make best use of this service is expected to improve outcomes and influence practice.

Keywords: telemedicine, healthcare utilisation, digital interventions, environmental impact, sustainable healthcare

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Authors: Berna Çalışkan

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The integrated water management is a colloborative approach to planning that brings together institutions that influence all elements of the water cycle, waterways, watershed characteristics, wetlands, ponds, lakes, floodplain areas, stream channel structure. It encourages collaboration where it will be beneficial and links between water planning and other planning processes that contribute to improving sustainable urban development and liveability. Hydraulic considerations can influence the selection of a highway corridor and the alternate routes within the corridor. widening a roadway, replacing a culvert, or repairing a bridge. Because of this, the type and amount of data needed for planning studies can vary widely depending on such elements as environmental considerations, class of the proposed highway, state of land use development, and individual site conditions. The extraction of drainage networks provide helpful preliminary drainage data from the digital elevation model (DEM). A case study was carried out using the Arc Hydro extension within ArcGIS in the study area. It provides the means for processing and presenting spatially-referenced Stream Model. Study area’s flow routing, stream levels, segmentation, drainage point processing can be obtained using DEM as the 'Input surface raster'. These processes integrate the fields of hydrologic, engineering research, and environmental modeling in a multi-disciplinary program designed to provide decision makers with a science-based understanding, and innovative tools for, the development of interdisciplinary and multi-level approach. This research helps to manage transport project planning and construction phases to analyze the surficial water flow, high-level streams, wetland sites for development of transportation infrastructure planning, implementing, maintenance, monitoring and long-term evaluations to better face the challenges and solutions associated with effective management and enhancement to deal with Low, Medium, High levels of impact. Transport projects are frequently perceived as critical to the ‘success’ of major urban, metropolitan, regional and/or national development because of their potential to affect significant socio-economic and territorial change. In this context, sustaining and development of economic and social activities depend on having sufficient Water Resources Management. The results of our research provides a workflow to build a stream network how can classify suitability map according to stream levels. Transportation projects establish, develop, incorporate and deliver effectively by selecting best location for reducing construction maintenance costs, cost-effective solutions for drainage, landslide, flood control. According to model findings, field study should be done for filling gaps and checking for errors. In future researches, this study can be extended for determining and preventing possible damage of Sensitive Areas and Vulnerable Zones supported with field investigations.

Keywords: water resources management, hydro tool, water protection, transportation

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Authors: Abha Gupta, Deepak K. Mishra

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The focus of food security studies in India has been on the adequacy of calories and its linkage with poverty level. India currently being undergoing a massive demographic and epidemiological transition has demonstrated a decline in average physical activity with improved mechanization and urbanization. Food consumption pattern is also changing with decreasing intake of coarse cereals and a marginal increase in the consumption of fruits, vegetables and meat products resulting into a nutrition transition in the country. However, deficiency of essential micronutrients such as vitamins and minerals is rampant despite their growing importance in fighting back with lifestyle and other modern diseases. The calorie driven studies can hardly tackle the complex problem of malnutrition. This paper fills these research lacuna and analyses mean intake of different major and micro-nutrients among different socio-economic groups and adequacy of these nutrients from recommended dietary allowance. For the purpose, a cross-sectional survey covering 304 households selected through proportional stratified random sampling was conducted in six villages of Aligarh district of the state of Uttar Pradesh, India. Data on quantity consumed of 74 food items grouped into 10 food categories with a recall period of seven days was collected from the households and converted into energy, protein, fat, carbohydrate, calcium, iron, thiamine, riboflavin, niacin and vitamin C using standard guidelines of National Institute of Nutrition. These converted nutrients were compared with recommended norms given by National Nutrition Monitoring Bureau. Per capita nutrient adequacy was calculated by dividing mean nutrient intake by the household size and then by comparing it with recommended norm. Findings demonstrate that source of both macro and micro-nutrients are mainly cereals followed by milk, edible oil and sugar items. Share of meat in providing essential nutrients is very low due to vegetarian diet. Vegetables, pulses, nuts, fruits and dry fruits are a poor source for most of the nutrients. Further analysis evinces that intake of most of the nutrients is higher than the recommended norm. Riboflavin is the only vitamin whose intake is less than the standard norm. Poor group, labour, small farmers, Muslims, scheduled caste demonstrate comparatively lower intake of all nutrients than their counterpart groups, though, they get enough macro and micro-nutrients significantly higher than the norm. One of the major reasons for higher intake of most of the nutrients across all socio-economic groups is higher consumption of monotonous diet based on cereals and milk. Most of the nutrients get their major share from cereals particularly wheat and milk intake. It can be concluded from the analysis that although there is adequate intake of most of the nutrients in the diet of rural population yet their source is mainly cereals and milk products depicting a monotonous diet. Hence, more efforts are needed to diversify the diet by giving more focus to the production of other food items particularly fruits, vegetables and pulse products. Awareness among the population, more accessibility and incorporating food items other than cereals in government social safety programmes are other measures to improve food security in India.

Keywords: hidden hunger, India, nutrients, recommended norm

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Authors: M. Skibinska, A. Rajewska-Rager, M. Dmitrzak-Weglarz, N. Lepczynska, P. Sibilski, P. Kapelski, J. Pawlak, J. Twarowska-Hauser

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Introduction: Neurotrophic factors have been implicated in neuropsychiatric disorders. Brain-Derived Neurotrophic Factor (BDNF) influences neuron differentiation in development as well as synaptic plasticity and neuron survival in adulthood. BDNF is widely studied in mood disorders and has been proposed as a biomarker for depression. BDNF is synthesized as precursor protein – proBDNF. Both forms are biologically active and exert opposite effects on neurons. Aim: The aim of the study was to examine the serum levels of BDNF and proBDNF in unipolar and bipolar young patients below 24 years old during hypo/manic, depressive episodes and in remission compared to healthy control group. Methods: In a prospective 2 years follow-up study, we investigated alterations in levels of BDNF and proBDNF in 79 patients (23 males, mean age 19.08, SD 3.3 and 56 females, mean age 18.39, SD 3.28) diagnosed with mood disorders: unipolar and bipolar disorder compared with 35 healthy control subjects (7 males, mean age 20.43, SD 4.23 and 28 females, mean age 21.25, SD 2.11). Clinical characteristics including mood, comorbidity, family history, and treatment, were evaluated during control visits and clinical symptoms were rated using the Hamilton Depression Rating Scale and Young Mania Rating Scale. Serum BDNF and proBDNF concentrations were determined by Enzyme-Linked Immunosorbent Assays (ELISA) method. Serum BDNF and proBDNF levels were analysed with covariates: sex, age, age > 18 and < 18 years old, family history of affective disorders, drug-free vs. medicated status. Normality of the data was tested using Shapiro-Wilk test. Levene’s test was used to calculate homogeneity of variance. Non-parametric Tests: Mann-Whitney U test, Kruskal-Wallis ANOVA, Friedman’s ANOVA, Wilcoxon signed rank test, Spearman correlation coefficient were applied in analyses The statistical significance level was set at p < 0.05. Results: BDNF and proBDNF serum levels did not differ between patients at baseline and controls as well as comparing patients in acute episode of depression/hypo/mania at baseline and euthymia (at month 3 or 6). Comparing BDNF and proBDNF levels between patients in euthymia and control group no differences have been found. Increased BDNF level in women compared to men at baseline (p=0.01) have been observed. BDNF level at baseline was negatively correlated with depression and mania occurence at 24 month (p=0.04). BDNF level at 12 month was negatively correlated with depression and mania occurence at 12 month (p=0.01). Correlation of BDNF level with sex have been detected (p=0.01). proBDNF levels at month 3, 6 and 12 negatively correlated with disease status (p=0.02, p=0.008, p=0.009, respectively). No other correlations of BDNF and proBDNF levels with clinical and demographical variables have been detected. Discussion: Our results did not show any differences in BDNF and proBDNF levels between depression, mania, euthymia, and controls. Imbalance in BDNF/proBDNF signalling may be involved in pathogenesis of mood disorders. Further studies on larger groups are recommended. Grant was founded by National Science Center in Poland no 2011/03/D/NZ5/06146.

Keywords: bipolar disorder, Brain-Derived Neurotrophic Factor (BDNF), proBDNF, unipolar depression

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Authors: Deon Bezuidenhout

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Pre-seeding with autologous endothelial cells improves the long-term patency of synthetic vascular grafts levels obtained with autografts, but is limited to a single centre due to resource, time and other constraints. Spontaneous in vivo endothelialization would obviate the need for pre-seeding, but has been shown to be absent in man due to limited transanastomotic and fallout healing, and the lack of transmural ingrowth due to insufficient porosity. Two types of graft scaffolds with increased interconnected porosity for improved tissue ingrowth and healing are thus proposed and described. Foam-type polyurethane (PU) scaffolds with small, medium and large, interconnected pores were made by phase inversion and spherical porogen extraction, with and without additional surface modification with covalently attached heparin and subsequent loading with and delivery of growth factors. Fibrillar scaffolds were made either by standard electrospinning using degradable PU (Degrapol®), or by dual electrospinning using non-degradable PU. The latter process involves sacrificial fibres that are co-spun with structural fibres and subsequently removed to increased porosity and pore size. Degrapol samples were subjected to in vitro degradation, and all scaffold types were evaluated in vivo for tissue ingrowth and vascularization using rat subcutaneous model. The foam scaffolds were additionally evaluated in a circulatory (rat infrarenal aortic interposition) model that allows for the grafts to be anastomotically and/or ablumenally isolated to discern and determine endothelialization mode. Foam-type grafts with large (150 µm) pores showed improved subcutaneous healing in terms of vascularization and inflammatory response over smaller pore sizes (60 and 90µm), and vascularization of the large porosity scaffolds was significantly increased by more than 70% by heparin modification alone, and by 150% to 400% when combined with growth factors. In the circulatory model, extensive transmural endothelialization (95±10% at 12 w) was achieved. Fallout healing was shown to be sporadic and limited in groups that were ablumenally isolated to prevent transmural ingrowth (16±30% wrapped vs. 80±20% control; p<0.002). Heparinization and GF delivery improved both mural vascularization and lumenal endothelialization. Degrapol electrospun scaffolds showed decrease in molecular mass and corresponding tensile strength over the first 2 weeks, but very little decrease in mass over the 4w test period. Studies on the effect of tissue ingrowth with and without concomitant degradation of the scaffolds, are being used to develop material models for the finite element modelling. In the case of the dual-spun scaffolds, the PU fibre fraction could be controlled shown to vary linearly with porosity (P = −0.18FF +93.5, r2=0.91), which in turn showed inverse linear correlation with tensile strength and elastic modulus (r2 > 0.96). Calculated compliance and burst pressures of the scaffolds increased with fibre fraction, and compliances matching the human popliteal artery (5-10 %/100 mmHg), and high burst pressures (> 2000 mmHg) could be achieved. Increasing porosity (76 to 82 and 90%) resulted in increased tissue ingrowth from 33±7 to 77±20 and 98±1% after 28d. Transmural endothelialization of highly porous foamed grafts is achievable in a circulatory model, and the enhancement of porosity and tissue ingrowth may hold the key the development of spontaneously endothelializing electrospun grafts.

Keywords: electrospinning, endothelialization, porosity, scaffold, vascular graft

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Authors: Yaser Ghasemi Aryan, Firoozeh Moghiminejad, Mohammadreza Shahraki

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Background: The cooperative institution is considered the first and most essential pillar of strengthening social capital, whose sustainability is the main guarantee of survival and continued participation of local communities in natural resource management projects. The Village Development Group and the Microcredit Fund are two important social and economic institutions in the implementation of the International Project for the Restoration of Degraded Forest Lands (RFLDL) in Sarayan City, South Khorasan Province, which has learned positive lessons from the participation of the beneficiaries in the implementation. They have brought more effective projects to deal with desertification. However, the low activity or liquidation of some of these institutions has become one of the important challenges and concerns of project executive experts. The current research was carried out with the aim of explaining the main components of the instability of these institutions. Materials and Methods: This research is descriptive-analytical in terms of method, practical in terms of purpose, and the method of collecting information is two documentary and survey methods. The statistical population of the research included all the members of the village development groups and microcredit funds in the target villages of the RFLDL project of Sarayan city, based on the Kochran formula and matching with the Karjesi and Morgan table. Net people were selected as a statistical sample. After confirming the validity of the expert's opinions, the reliability of the questionnaire was 0.83, which shows the appropriate reliability of the researcher-made questionnaire. Data analysis was done using SPSS software. Results: The results related to the extraction of obstacles to the stability of social and economic networks were classified and prioritized in the form of 5 groups of social-cultural, economic, administrative, educational-promotional and policy-management factors. Based on this, in the socio-cultural factors, the items ‘not paying attention to the structural characteristics and composition of groups’, ‘lack of commitment and moral responsibility in some members of the group,’ and ‘lack of a clear pattern for the preservation and survival of groups’, in the disciplinary factors, The items ‘Irregularity in holding group meetings’ and ‘Irregularity of members to participate in meetings’, in the economic factors of the items "small financial capital of the fund’, ‘the low amount of loans of the fund’ and ‘the fund's inability to conclude contracts and attract capital from other sources’, in the educational-promotional factors of the items ‘non-simultaneity of job training with the granting of loans to create jobs’ and ‘insufficient training for the effective use of loans and job creation’ and in the policy-management factors of the item ‘failure to provide government facilities for support From the funds, they had the highest priority. Conclusion: In general, the results of this research show that policy-management factors and social factors, especially the structure and composition of social and economic institutions, are the most important obstacles to their sustainability. Therefore, it is suggested to form cooperative institutions based on network analysis studies in order to achieve the appropriate composition of members.

Keywords: cooperative institution, social capital, network analysis, participation, Sarayan.

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Authors: Rusudan Chagelishvili, Nino Sulava, Tamar Beridze, Nana Rezesidze, Nikoloz Tatuashvili

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The South Caucasus is one of the most important centers of prehistoric metallurgy, known for its Colchian bronze culture. Modern Lechkhumi – historical Mountainous Colchis where the existence of prehistoric metallurgy is confirmed by the discovery of many artifacts is a part of this area. Studies focused on prehistoric smelting sites, related artefacts, and ore deposits have been conducted during last ten years in Lechkhumi. More than 20 prehistoric smelting sites and artefacts associated with metallurgical activities (ore roasting furnaces, slags, crucible, and tuyères fragments) have been identified so far. Within the framework of integrated studies was established that these sites were operating in 13-9 centuries B.C. and used for copper smelting. Palynological studies of slags revealed that chestnut (Castanea sativa) and hornbeam (Carpinus sp.) wood were used as smelting fuel. Geological exploration-analytical studies revealed that copper ore mining, processing, and smelting sites were distributed close to each other. Despite recent complex data, the signs of prehistoric mines (trenches) haven’t been found in this part of the study area so far. Since 2018 the archaeological-geological exploration has been focused on the southern part of Lechkhumi and covered the areas of villages Okureshi and Opitara. Several copper smelting sites (Okureshi 1 and 2, Opitara 1), as well as a Colchian Bronze culture settlement, have been identified here. Three mine workings have been found in the narrow gorge of the river Rtkhmelebisgele in the vicinities of the village Opitara. In order to establish a link between the Opitara-Okureshi archaeometallurgical sites, Late Bronze Age settlements, and mines, various scientific analytical methods -mineralized rock and slags petrography and atomic absorption spectrophotometry (AAS) analysis have been applied. The careful examination of Opitara mine workings revealed that there is a striking difference between the mine #1 on the right bank of the river and mines #2 and #3 on the left bank. The first one has all characteristic features of the Soviet period mine working (e. g. high portal with angular ribs and roof showing signs of blasting). In contrast, mines #2 and #3, which are located very close to each other, have round-shaped portals/entrances, low roofs, and fairly smooth ribs and are filled with thick layers of river sediments and collapsed weathered rock mass. A thorough review of the publications related to prehistoric mine workings revealed some striking similarities between mines #2 and #3 with their worldwide analogues. Apparently, the ore extraction from these mines was conducted by fire-setting applying primitive tools. It was also established that mines are cut in Jurassic mineralized volcanic rocks. Ore minerals (chalcopyrite, pyrite, galena) are related to calcite and quartz veins. The results obtained through the petrochemical and petrography studies of mineralized rock samples from Opitara mines and prehistoric slags are in complete correlation with each other, establishing the direct link between copper mining and smelting within the study area. Acknowledgment: This work was supported by the Shota Rustaveli National Science Foundation of Georgia (grant # FR-19-13022).

Keywords: archaeometallurgy, Mountainous Colchis, mining, ore minerals

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Authors: Prem Kumar, Anuj Tyagi, Harsh Panwar, Vaneet Inder Kaur

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Though aquaculture started as an occupation for poor and weak farmers for livelihood, it has now acquired the shape of one of the biggest industry to grow live protein in the form of aquatic organisms. Industrialization of the aquaculture sector has led to intensification resulting in stress on aquatic organisms and frequent disease outbreaks leading to huge economic impacts. Indiscriminate use of antibiotics as growth promoter and prophylactic agent in aquaculture has resulted in rapid emergence and spread of antibiotic resistance in bacterial pathogens. Over the past few years, use of probiotics (as an alternative of antibiotics) in aquaculture has gained attention due to their immunostimulant and growth promoting properties. It has now well known that after administration, a probiotic bacterium has to compete and establish itself against native microbiota to show its eventual beneficial properties. Due to their non-fish origin, commercial probiotics sometimes may display poor probiotic functionalities and antagonistic effects. Thus, isolation and characterization of probiotic bacteria from same fish host is very much necessary. In this study, attempts were made to isolate potent probiotic lactic acid bacteria (LAB) from intestinal microflora of rohu fish. Twenty-five experimental rohu fishes (mean weight 400 ± 20gm, mean standard length 20 ± 3cm) were used in the study to collect fish gut after dissection in a sterile condition. A total of 150 tentative LAB isolates from selective agar media (de Man-Rogosa-Sharpe (MRS)) were screened for their antimicrobial activity against Aeromonas hydrophila and Microccocus leuteus. A total of 17 isolates, identified as Lactobacillus plantarum and Lactococcus lactis, identified by biochemical tests and PCR amplification and sequencing of 16S rRNA gene fragment, displayed promising antimicrobial activity against both the pathogens. Two isolates from each species (FLB1, FLB2 from L. plantarum; and FLC1, FLC2 from L. lactis) were subjected to downstream probiotic potential characterization. These isolates were compared in vitro for their hemolytic activity, acid and bile tolerance for growth kinetics, auto-aggregation, cell-surface hydrophobicity against xylene, and chloroform, tolerance to phenol, cell adhesion, and safety parameters (by intraperitoneal and intramuscular injections). None of the tested isolates showed any hemolytic activity indicating their potential safety. Moreover, these isolates were tolerant to 0.3% bile (75-82% survival), phenol stress (96-99% survival) with 100% viability at pH 3 over a period of 3 h. Antibiotic sensitivity test revealed that all the tested LAB isolates were resistant to vancomycin, gentamicin, streptomycin, and erythromycin and sensitive to Erythromycin, Chloramphenicol, Ampicillin, Trimethoprim, and Nitrofurantoin. Tetracycline resistance was found in L. plantarum (FLB1 and FLB2 isolates), whereas L. lactis were susceptible to it. Intramuscular and intraperitoneal challenges to fingerlings of rohu fish (5 ± 1gm weight) with FLB1 showed no pathogenicity and occurrence of disease symptoms in fishes over an observation period of 7 days. The results revealed FLB1 as a potential probiotic candidate for aquaculture application among other isolates.

Keywords: aquaculture, Lactobacillus plantarum, Lactococcus lactis, probiotics

Procedia PDF Downloads 107

Authors: Kavitha Madhu, Karthik K. Srinivasan, R. Sivanandan

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Indian traffic can be considered as mixed and heterogeneous due to the presence of various types of vehicles that operate with weak lane discipline. Consequently, vehicles can position themselves anywhere in the traffic stream depending on availability of gaps. The choice of lateral positioning is an important component in representing and characterizing mixed traffic. The field data provides evidence that the trajectory of vehicles in Indian urban roads have significantly varying longitudinal and lateral components. Further, the notion of headway which is widely used for homogeneous traffic simulation is not well defined in conditions lacking lane discipline. From field data it is clear that following is not strict as in homogeneous and lane disciplined conditions and neighbouring vehicles ahead of a given vehicle and those adjacent to it could also influence the subject vehicles choice of position, speed and acceleration. Given these empirical features, the suitability of using headway distributions to characterize mixed traffic in Indian cities is questionable, and needs to be modified appropriately. To address these issues, this paper attempts to analyze the time gap distribution between consecutive vehicles (in a time-sense) crossing a section of roadway. More specifically, to characterize the complex interactions noted above, the influence of composition, manoeuvre types, and lateral placement characteristics on time gap distribution is quantified in this paper. The developed model is used for evaluating various performance measures such as link speed, midblock delay and intersection delay which further helps to characterise the vehicular fuel consumption and emission on urban roads of India. Identifying and analyzing exact interactions between various classes of vehicles in the traffic stream is essential for increasing the accuracy and realism of microscopic traffic flow modelling. In this regard, this study aims to develop and analyze time gap distribution models and quantify it by lead lag pair, manoeuvre type and lateral position characteristics in heterogeneous non-lane based traffic. Once the modelling scheme is developed, this can be used for estimating the vehicle kilometres travelled for the entire traffic system which helps to determine the vehicular fuel consumption and emission. The approach to this objective involves: data collection, statistical modelling and parameter estimation, simulation using calibrated time-gap distribution and its validation, empirical analysis of simulation result and associated traffic flow parameters, and application to analyze illustrative traffic policies. In particular, video graphic methods are used for data extraction from urban mid-block sections in Chennai, where the data comprises of vehicle type, vehicle position (both longitudinal and lateral), speed and time gap. Statistical tests are carried out to compare the simulated data with the actual data and the model performance is evaluated. The effect of integration of above mentioned factors in vehicle generation is studied by comparing the performance measures like density, speed, flow, capacity, area occupancy etc under various traffic conditions and policies. The implications of the quantified distributions and simulation model for estimating the PCU (Passenger Car Units), capacity and level of service of the system are also discussed.

Keywords: lateral movement, mixed traffic condition, simulation modeling, vehicle following models

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Authors: Sarah Nauwelaerts, Koen De Cremer, Alfred Bernard, Meredith Verlooy, Kristel Heremans, Natalia Bustos Sierra, Katrien Tersago, Tim Nawrot, Jordy Vercauteren, Christophe Stroobants, Sigrid C. J. De Keersmaecker, Nancy Roosens

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Projected climate changes could lead to exacerbation of respiratory disorders associated with reduced air quality. Air pollution and climate changes influence each other through complex interactions. The poor air quality in urban and rural areas includes high levels of particulate matter (PM), ozone (O3) and nitrogen oxides (NOx), representing a major threat to public health and especially for the most vulnerable population strata, and especially young children. In this study, we aim to develop generic standardized policy supporting tools and methods that allow evaluating in future follow-up larger scale epidemiological studies the risks of the combined short-term effects of O3 and PM on the cardiorespiratory system of children. We will use non-invasive indicators of airway damage/inflammation and of genetic or epigenetic variations by using urine or saliva as alternative to blood samples. Therefore, a multi-phase field study will be organized in order to assess the sensitivity and applicability of these tests in large cohorts of children during episodes of air pollution. A first test phase was planned in March 2018, not yet taking into account ‘critical’ pollution periods. Working with non-invasive samples, choosing the right set-up for the field work and the volunteer selection were parameters to consider, as they significantly influence the feasibility of this type of study. During this test phase, the selection of the volunteers was done in collaboration with medical doctors from the Centre for Student Assistance (CLB), by choosing a class of pre-pubertal children of 9-11 years old in a primary school in Flemish Brabant, Belgium. A questionnaire, collecting information on the health and background of children and an informed consent document were drawn up for the parents as well as a simplified cartoon-version of this document for the children. A detailed study protocol was established, giving clear information on the study objectives, the recruitment, the sample types, the medical examinations to be performed, the strategy to ensure anonymity, and finally on the sample processing. Furthermore, the protocol describes how this field study will be conducted in relation with the prevision and monitoring of air pollutants for the future phases. Potential protein, genetic and epigenetic biomarkers reflecting the respiratory function and the levels of air pollution will be measured in the collected samples using unconventional technologies. The test phase results will be used to address the most important bottlenecks before proceeding to the following phases of the study where the combined effect of O3 and PM during pollution peaks will be examined. This feasibility study will allow identifying possible bottlenecks and providing missing scientific knowledge, necessary for the preparation, implementation and evaluation of federal policies/strategies, based on the most appropriate epidemiological studies on the health effects of air pollution. The research leading to these results has been funded by the Belgian Science Policy Office through contract No.: BR/165/PI/PMOLLUGENIX-V2.

Keywords: air pollution, biomarkers, children, field study, feasibility study, non-invasive

Procedia PDF Downloads 142

Authors: Maria Karnachoriti, Ellas Spyratou, Dimitrios Lykidis, Maria Lambropoulou, Yiannis S. Raptis, Ioannis Seimenis, Efstathios P. Efstathopoulos, Athanassios G. Kontos

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Colorectal cancer is the third most common cancer diagnosed in Europe, according to the latest incidence data provided by the World Health Organization (WHO), and early diagnosis has proved to be the key in reducing cancer-related mortality. In cases where surgical interventions are required for cancer treatment, the accurate discrimination between healthy and cancerous tissues is critical for the postoperative care of the patient. The current study focuses on the ex vivo handling of surgically excised colorectal specimens and the acquisition of their spectral fingerprints using Raman spectroscopy. Acquired data were analyzed in an effort to discriminate, in microscopic scale, between healthy and malignant margins. Raman spectroscopy is a spectroscopic technique with high detection sensitivity and spatial resolution of few micrometers. The spectral fingerprint which is produced during laser-tissue interaction is unique and characterizes the biostructure and its inflammatory or cancer state. Numerous published studies have demonstrated the potential of the technique as a tool for the discrimination between healthy and malignant tissues/cells either ex vivo or in vivo. However, the handling of the excised human specimens and the Raman measurement conditions remain challenging, unavoidably affecting measurement reliability and repeatability, as well as the technique’s overall accuracy and sensitivity. Therefore, tissue handling has to be optimized and standardized to ensure preservation of cell integrity and hydration level. Various strategies have been implemented in the past, including the use of balanced salt solutions, small humidifiers or pump-reservoir-pipette systems. In the current study, human colorectal specimens of 10X5 mm were collected from 5 patients up to now who underwent open surgery for colorectal cancer. A novel, non-toxic zinc-based fixative (Z7) was used for tissue preservation. Z7 demonstrates excellent protein preservation and protection against tissue autolysis. Micro-Raman spectra were recorded with a Renishaw Invia spectrometer from successive random 2 micrometers spots upon excitation at 785 nm to decrease fluorescent background and secure avoidance of tissue photodegradation. A temperature-controlled approach was adopted to stabilize the tissue at 2 °C, thus minimizing dehydration effects and consequent focus drift during measurement. A broad spectral range, 500-3200 cm-1,was covered with five consecutive full scans that lasted for 20 minutes in total. The average spectra were used for least square fitting analysis of the Raman modes.Subtle Raman differences were observed between normal and cancerous colorectal tissues mainly in the intensities of the 1556 cm-1 and 1628 cm-1 Raman modes which correspond to v(C=C) vibrations in porphyrins, as well as in the range of 2800-3000 cm-1 due to CH2 stretching of lipids and CH3 stretching of proteins. Raman spectra evaluation was supported by histological findings from twin specimens. This study demonstrates that Raman spectroscopy may constitute a promising tool for real-time verification of clear margins in colorectal cancer open surgery.

Keywords: colorectal cancer, Raman spectroscopy, malignant margins, spectral fingerprints

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Authors: Hasmik V. Zanginyan, Gayane S. Ghazaryan, Laura M. Hovsepyan

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Experimental autoimmune encephalomyelitis (EAE) is an inflammatory demyelinating disease of the central nervous system that is induced in laboratory animals by developing an immune response against myelin epitopes. The typical clinical course is ascending palsy, which correlates with inflammation and tissue damage in the thoracolumbar spinal cord, although the optic nerves and brain (especially the subpial white matter and brainstem) are also often affected. With multiple sclerosis, there is a violation of lipid metabolism in myelin. When membrane lipids (glycosphingolipids, phospholipids) are disturbed, metabolites not only play a structural role in membranes but are also sources of secondary mediators that transmit multiple cellular signals. The purpose of this study was to investigate the effect of ganglioside as a therapeutic agent in experimental multiple sclerosis. The biological activity of a ganglioside-containing medicinal preparation (Cronassial) was evaluated in an experimental model of multiple sclerosis in laboratory animals. An experimental model of multiple sclerosis in rats was obtained by immunization with myelin basic protein (MBP), as well as homogenization of the spinal cord or brain. EAE was induced by administering a mixture of an encephalitogenic mixture (EGM) with Complete Freund’s Adjuvant. Mitochondrial fraction was isolated in a medium containing 0,25 M saccharose and 0, 01 M tris buffer, pH - 7,4, by a method of differential centrifugation on a K-24 centrifuge. Glutathione peroxidase activity was assessed by reduction reactions of hydrogen peroxide (H₂O₂) and lipid hydroperoxides (ROOH) in the presence of GSH. LPO activity was assessed by the amount of malondialdehyde (MDA) in the total homogenate and mitochondrial fraction of the spinal cord and brain of control and experimental autoimmune encephalomyelitis rats. MDA was assessed by a reaction with Thiobarbituric acid. For statistical data analysis on PNP, SPSS (Statistical Package for Social Science) package was used. The nature of the distribution of the obtained data was determined by the Kolmogorov-Smirnov criterion. The comparative analysis was performed using a nonparametric Mann-Whitney test. The differences were statistically significant when р ≤ 0,05 or р ≤ 0,01. Correlational analysis was conducted using a nonparametric Spearman test. In the work, refrigeratory centrifuge, spectrophotometer LKB Biochrom ULTROSPECII (Sweden), pH-meter PL-600 mrc (Israel), guanosine, and ATP (Sigma). The study of the process of lipid peroxidation in the total homogenate of the brain and spinal cord in experimental animals revealed an increase in the content of malonic dialdehyde. When applied, Cronassial observed normalization of lipid peroxidation processes. Reactive oxygen species, causing lipid peroxidation processes, can be toxic both for neurons and for oligodendrocytes that form myelin, causing a violation of their lipid composition. The high content of lipids in the brain and the uniqueness of their structure determines the nature of the development of LPO processes. The lipid layer of cellular and intracellular membranes performs two main functions -barrier and matrix (structural). Damage to the barrier leads to dysregulation of intracellular processes and severe disorders of cellular functions.

Keywords: experimental autoimmune encephalomyelitis, multiple sclerosis, neuroinflammation, therapy

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Authors: Andrew Read, Alice Parry, Kate Woods

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Since the advent of the SARS-CoV-2 pandemic, the search for medications that can reduce mortality and morbidity has been a global research priority. Several multi-center trials have recently demonstrated improved mortality associated with the use of Tocilizumab, an interleukin-6 receptor antagonist, in patients with severe SARS-CoV-2 pneumonia. Initial data supported the administration in patients requiring respiratory support (non-invasive or invasive ventilation), but more recent data has shown benefit in all hypoxic patients. At the height of the second wave of COVID-19 infections in London, our hospital introduced the use of Tocilizumab for patients with severe COVID-19. Tocilizumab is licensed for use in chronic inflammatory conditions and has been associated with an increased risk of severe bacterial and fungal infections, as well as reactivation of chronic viral infections (e.g., hepatitis B). It is a specialist drug that suppresses the formation of C-reactive protein (CRP) for 6 – 12 weeks. It is not widely used by the general medical community. We aimed to assess Tocilizumab use in our hospital and to implement changes to the protocol as required to ensure administration was safe and appropriate. A retrospective study design was used to assess prescriptions over an initial 3-week period in both intensive care and on the medical wards. This amounted to a total of 13 patients. The initial data collection identified four key areas of concern: adherence to national and local inclusion & exclusion criteria; a collection of appropriate screening blood prior to administration; documentation of informed consent or best interest decision and documentation of Tocilizumab administration on patient discharge information, to alert future healthcare providers that typical measures of inflammation and infection, such as CRP, are unreliable for up to 3-months. Data were collected from electronic notes, blood results and observation charts, and cross referenced with pharmacy data. Initial results showed that all four key areas were completed in approximately 50% of cases. Of particular concern was adherence to exclusion criteria, such as current evidence of bacterial infection, and ensuring the correct screening blood was sent to exclude infections such as hepatitis. To remedy this and improve patient safety, the initial data was presented to relevant healthcare professionals. Subsequently, three interventions were introduced and education on each provided to hospital staff. An electronic ‘order set’ collating the appropriate screening blood was created simplifying the screening process. Pre-formed electronic documentation which can be inserted into the notes was created to provide a framework for consent discussions and reduce the time needed for junior doctors to complete this task. Additionally, a ‘Tocilizumab’ administration card was created and administered via pharmacy. This was distributed to each patient on discharge to ensure future healthcare professionals were aware of the potential effects of Tocilizumab administration, including suppression of CRP. Following these changes, repeat data collection over two months illustrated that each of the 4 safety aspects was met with a 100% success rate in every patient. Although this demonstrates good progress and effective interventions the challenge will be to maintain this progress. The audit data collection is ongoing

Keywords: education, patient safety , SARS-CoV-2, Tocilizumab

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Authors: Priyanka Mokashi, Aparna Khanna, Nancy Pandita

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Diabetes mellitus is a chronic metabolic disorder which will be the 7th leading cause of death by 2030. The current line of treatment for the diabetes mellitus is oral antidiabetic drugs (biguanides, sulfonylureas, meglitinides, thiazolidinediones and alpha-glycosidase inhibitors) and insulin therapy depending upon the type 1 or type 2 diabetes mellitus. But, these treatments have their disadvantages, ranging from the developing of resistance to the drugs and adverse effects caused by them. Alternative to these synthetic agents, natural products provides a new insight for the development of more efficient and safe drugs due to their therapeutic values. Enicostema littorale blume (A. Raynal) is a traditional Indian plant belongs to the Gentianaceae family. It is widely distributed in Asia, Africa, and South America. There are few reports on Swrtiamarin, major component of this plant for its antidiabetic activity. However, the antidiabetic activity of flavonoids from E. littorale and their mechanism of action have not yet been elucidated. Flavonoids have a positive relationship with disease prevention and can act on various molecular targets and regulate different signaling pathways in pancreatic β-cells, adipocytes, hepatocytes and skeletal myofibers. They may exert beneficial effects in diabetes by (i) improving hyperglycemia through regulation of glucose metabolism in hepatocytes; (ii) enhancing insulin secretion and reducing apoptosis and promoting proliferation of pancreatic β-cells; (iii) increasing glucose uptake in hepatocytes, skeletal muscle and white adipose tissue (iv) reducing insulin resistance, inflammation and oxidative stress. Therefore, we have isolated four flavonoid rich fractions, Fraction A (FA), Fraction B (FB), Fraction C (FC), Fraction D (FD) from crude alcoholic hot (AH) extract from E. littorale, identified by LC/MS. Total eight flavonoids were identified on the basis of fragmentation pattern. Flavonoid FA showed the presence of swertisin, isovitexin, and saponarin; FB showed genkwanin, quercetin, isovitexin, FC showed apigenin, swertisin, quercetin, 5-O-glucosylswertisin and 5-O-glucosylisoswertisin whereas FD showed the presence of swertisin. Further, these fractions were assessed for their antidiabetic activity on stimulating glucose uptake in insulin-resistant HepG2 cell line model (IR/HepG2). The results showed that FD containing C-glycoside Swertisin has significantly increased the glucose uptake rate of IR/HepG2 cells at the concentration of 10 µg/ml as compared to positive control Metformin (0.5mM) which was determined by glucose oxidase- peroxidase method. It has been reported that enhancement of glucose uptake of cells occurs due the translocation of Glut4 vesicles to cell membrane through IR/IRS1/AKT pathway. Therefore, we have studied expressions of three genes IRS1, AKT and Glut4 by real-time PCR to evaluate whether they follow the same pathway or not. It was seen that the glucose uptake rate has increased in FD treated IR/HepG2 cells due to the activation of insulin receptor substrate-1 (IRS1) followed by protein kinase B (AKT) through phosphoinositide 3-kinase (PI3K) leading to translocation of Glut 4 vesicles to cell membrane, thereby enhancing glucose uptake and insulin sensitivity of insulin resistant HepG2 cells. Hence, the up-regulation indicated the mechanism of action through which FD (Swertisin) acts as antidiabetic candidate in the treatment of type 2 diabetes mellitus.

Keywords: E. littorale, glucose transporter, glucose uptake rate, insulin resistance

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Authors: Jun Byung Joon, Seo Seok Hyun, Lee Seo Young

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As carbon dioxide emissions increase due to rapid industrialization and reckless development, abnormal climates such as floods and droughts are occurring. In order to respond to such climate change, the use of existing fossil fuels is reduced, and the proportion of eco-friendly renewable energy is gradually increasing. Korea is an energy resource-poor country that depends on imports for 93% of its total energy. As the global energy supply chain instability experienced due to the Russia-Ukraine crisis increases, countries around the world are resetting energy policies to minimize energy dependence and strengthen security. Seawater thermal energy is a renewable energy that replaces the existing air heat energy. It uses the characteristic of having a higher specific heat than air to cool and heat main spaces of buildings to increase heat transfer efficiency and minimize power consumption to generate electricity using fossil fuels, and Carbon dioxide emissions can be minimized. In addition, the effect on the marine environment is very small by using only the temperature characteristics of seawater in a limited way. K-water carried out a demonstration project of supplying cooling and heating energy to spaces such as the central control room and presentation room in the management building by acquiring the heat source of seawater circulated through the power plant's waterway by using the characteristics of the tidal power plant. Compared to the East Sea and the South Sea, the main system was designed in consideration of the large tidal difference, small temperature difference, and low-temperature characteristics, and its performance was verified through operation during the demonstration period. In addition, facility improvements were made for major deficiencies to strengthen monitoring functions, provide user convenience, and improve facility soundness. To spread these achievements, the basic concept was to expand the seawater heating and cooling system with a scale of 200 USRT at the Tidal Culture Center. With the operational experience of the demonstration system, it will be possible to establish an optimal seawater heat cooling and heating system suitable for the characteristics of the west coast ocean. Through this, it is possible to reduce operating costs by KRW 33,31 million per year compared to air heat, and through industry-university-research joint research, it is possible to localize major equipment and materials and develop key element technologies to revitalize the seawater heat business and to advance into overseas markets. The government's efforts are needed to expand the seawater heating and cooling system. Seawater thermal energy utilizes only the thermal energy of infinite seawater. Seawater thermal energy has less impact on the environment than river water thermal energy, except for environmental pollution factors such as bottom dredging, excavation, and sand or stone extraction. Therefore, it is necessary to increase the sense of speed in project promotion by innovatively simplifying unnecessary licensing/permission procedures. In addition, support should be provided to secure business feasibility by dramatically exempting the usage fee of public waters to actively encourage development in the private sector.

Keywords: seawater thermal energy, marine energy, tidal power plant, energy consumption

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Authors: Eman Ghoneim

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The Arabian deserts include some of the driest areas on Earth. Yet, its landforms reserved a record of past wet climates. During humid phases, the desert was green and contained permanent rivers, inland deltas and lakes. Some of their water would have seeped and replenished the groundwater aquifers. When the wet periods came to an end, several thousand years ago, the entire region transformed into an extended band of desert and its original fluvial surface was totally covered by windblown sand. In this work, radar and thermal infrared images were used to reveal numerous hidden surface/subsurface features. Radar long wavelength has the unique ability to penetrate surface dry sands and uncover buried subsurface terrain. Thermal infrared also proven to be capable of spotting cooler moist areas particularly in hot dry surfaces. Integrating Radarsat images and GIS revealed several previously unknown paleoriver and lake basins in the region. One of these systems, known as the Kufrah, is the largest yet identified river basin in the Eastern Sahara. This river basin, which straddles the border between Egypt and Libya, flowed north parallel to the adjacent Nile River with an extensive drainage area of 235,500 km2 and massive valley width of 30 km in some parts. This river was most probably served as a spillway for an overflow from Megalake Chad to the Mediterranean Sea and, thus, may have acted as a natural water corridor used by human ancestors to migrate northward across the Sahara. The Gilf-Kebir is another large paleoriver system located just east of Kufrah and emanates from the Gilf Plateau in Egypt. Both river systems terminate with vast inland deltas at the southern margin of the Great Sand Sea. The trends of their distributary channels indicate that both rivers drained to a topographic depression that was periodically occupied by a massive lake. During dry climates, the lake dried up and roofed by sand deposits, which is today forming the Great Sand Sea. The enormity of the lake basin provides explanation as to why continuous extraction of groundwater in this area is possible. A similar lake basin, delimited by former shorelines, was detected by radar space data just across the border of Sudan. This lake, called the Northern Darfur Megalake, has a massive size of 30,750 km2. These former lakes and rivers could potentially hold vast reservoirs of groundwater, oil and natural gas at depth. Similar to radar data, thermal infrared images were proven to be useful in detecting potential locations of subsurface water accumulation in desert regions. Analysis of both Aster and daily MODIS thermal channels reveal several subsurface cool moist patches in the sandy desert of the Arabian Peninsula. Analysis indicated that such evaporative cooling anomalies were resulted from the subsurface transmission of the Monsoonal rainfall from the mountains to the adjacent plain. Drilling a number of wells in several locations proved the presence of productive water aquifers confirming the validity of the used data and the adopted approaches for water exploration in dry regions.

Keywords: radarsat, SRTM, MODIS, thermal infrared, near-surface water, ancient rivers, desert, Sahara, Arabian peninsula

Procedia PDF Downloads 216

Authors: Chia-Pei Chen, Yu-Ju Chen, Yu-Juei Hsu

Abstract:

The prevalence and incidence of end-stage renal disease in Taiwan ranks the highest in the world. According to the statistical survey of the Ministry of Health and Welfare in 2019, kidney disease is the ninth leading cause of death in Taiwan. It leads to autonomic dysfunction, inflammatory response and symptom distress, and further increases the damage to the structure and function of the kidneys, leading to increased demand for renal replacement therapy and risks of cardiovascular disease, which also has medical costs for the society. If we can intervene in a feasible manual to effectively regulate the autonomic nerve function of CKD patients, reduce the inflammatory response and symptom distress. To prolong the progression of the disease, it will be the main goal of caring for CKD patients. This study aims to test the effect of heart rate variability biofeedback (HRVBF) on improving autonomic nerve function (Heart Rate Variability, HRV), inflammatory response (Interleukin-6 [IL-6], C reaction protein [CRP] ), symptom distress (Piper fatigue scale, Pittsburgh Sleep Quality Index [PSQI], and Beck Depression Inventory-II [BDI-II] ) in patients with chronic kidney disease. This study was experimental research, with a convenience sampling. Participants were recruited from the nephrology clinic at a medical center in northern Taiwan. With signed informed consent, participants were randomly assigned to the HRVBF or control group by using the Excel BINOMDIST function. The HRVBF group received four weekly hospital-based HRVBF training, and 8 weeks of home-based self-practice was done with StressEraser. The control group received usual care. We followed all participants for 3 months, in which we repeatedly measured their autonomic nerve function (HRV), inflammatory response (IL-6, CRP), and symptom distress (Piper fatigue scale, PSQI, and BDI-II) on their first day of study participation (baselines), 1 month, and 3 months after the intervention to test the effects of HRVBF. The results were analyzed by SPSS version 23.0 statistical software. The data of demographics, HRV, IL-6, CRP, Piper fatigue scale, PSQI, and BDI-II were analyzed by descriptive statistics. To test for differences between and within groups in all outcome variables, it was used by paired sample t-test, independent sample t-test, Wilcoxon Signed-Rank test and Mann-Whitney U test. Results: Thirty-four patients with chronic kidney disease were enrolled, but three of them were lost to follow-up. The remaining 31 patients completed the study, including 15 in the HRVBF group and 16 in the control group. The characteristics of the two groups were not significantly different. The four-week hospital-based HRVBF training combined with eight-week home-based self-practice can effectively enhance the parasympathetic nerve performance for patients with chronic kidney disease, which may against the disease-related parasympathetic nerve inhibition. In the inflammatory response, IL-6 and CRP in the HRVBF group could not achieve significant improvement when compared with the control group. Self-reported fatigue and depression significantly decreased in the HRVBF group, but they still failed to achieve a significant difference between the two groups. HRVBF has no significant effect on improving the sleep quality for CKD patients.

Keywords: heart rate variability biofeedback, autonomic nerve function, inflammatory response, symptom distress, chronic kidney disease

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Authors: Chung-Ming Lo, Chung-Chien Lee

Abstract:

In many countries, the lifetime prevalence of shoulder pain is up to 70%. In America, the health care system spends 7 billion per year about the healthy issues of shoulder pain. With respect to the origin, up to 70% of shoulder pain is attributed to rotator cuff lesions This study proposed a computer-aided diagnosis (CAD) system to assist radiologists classifying rotator cuff lesions with less operator dependence. Quantitative features were extracted from the shoulder ultrasound images acquired using an ALOKA alpha-6 US scanner (Hitachi-Aloka Medical, Tokyo, Japan) with linear array probe (scan width: 36mm) ranging from 5 to 13 MHz. During examination, the postures of the examined patients are standard sitting position and are followed by the regular routine. After acquisition, the shoulder US images were drawn out from the scanner and stored as 8-bit images with pixel value ranging from 0 to 255. Upon the sonographic appearance, the boundary of each lesion was delineated by a physician to indicate the specific pattern for analysis. The three lesion categories for classification were composed of 20 cases of tendon inflammation, 18 cases of calcific tendonitis, and 18 cases of supraspinatus tear. For each lesion, second-order statistics were quantified in the feature extraction. The second-order statistics were the texture features describing the correlations between adjacent pixels in a lesion. Because echogenicity patterns were expressed via grey-scale. The grey-scale co-occurrence matrixes with four angles of adjacent pixels were used. The texture metrics included the mean and standard deviation of energy, entropy, correlation, inverse different moment, inertia, cluster shade, cluster prominence, and Haralick correlation. Then, the quantitative features were combined in a multinomial logistic regression classifier to generate a prediction model of rotator cuff lesions. Multinomial logistic regression classifier is widely used in the classification of more than two categories such as the three lesion types used in this study. In the classifier, backward elimination was used to select a feature subset which is the most relevant. They were selected from the trained classifier with the lowest error rate. Leave-one-out cross-validation was used to evaluate the performance of the classifier. Each case was left out of the total cases and used to test the trained result by the remaining cases. According to the physician’s assessment, the performance of the proposed CAD system was shown by the accuracy. As a result, the proposed system achieved an accuracy of 86%. A CAD system based on the statistical texture features to interpret echogenicity values in shoulder musculoskeletal ultrasound was established to generate a prediction model for rotator cuff lesions. Clinically, it is difficult to distinguish some kinds of rotator cuff lesions, especially partial-thickness tear of rotator cuff. The shoulder orthopaedic surgeon and musculoskeletal radiologist reported greater diagnostic test accuracy than general radiologist or ultrasonographers based on the available literature. Consequently, the proposed CAD system which was developed according to the experiment of the shoulder orthopaedic surgeon can provide reliable suggestions to general radiologists or ultrasonographers. More quantitative features related to the specific patterns of different lesion types would be investigated in the further study to improve the prediction.

Keywords: shoulder ultrasound, rotator cuff lesions, texture, computer-aided diagnosis

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Authors: Cátia Gonçalves, Teresa Nunes, Inês Pina, Ana Vicente, C. Alves, Felix Charvet, Daniel Neves, A. Matos

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The production of charcoal relies on rudimentary technologies using traditional brick kilns. Charcoal is produced under pyrolysis conditions: breaking down the chemical structure of biomass under high temperature in the absence of air. The amount of the pyrolysis products (charcoal, pyroligneous extract, and flue gas) depends on various parameters, including temperature, time, pressure, kiln design, and wood characteristics like the moisture content. This activity is recognized for its inefficiency and high pollution levels, but it is poorly characterized. This activity is widely distributed and is a vital economic activity in certain regions of Portugal, playing a relevant role in the management of woody residues. The location of the units establishes the biomass used for charcoal production. The Portalegre district, in the Alto Alentejo region (Portugal), is a good example, essentially with rural characteristics, with a predominant farming, agricultural, and forestry profile, and with a significant charcoal production activity. In this district, a recent inventory identifies almost 50 charcoal production units, equivalent to more than 450 kilns, of which 80% appear to be in operation. A field campaign was designed with the objective of determining the composition of the emissions released during a charcoal production cycle. A total of 30 samples of particulate matter and 20 gas samples in Tedlar bags were collected. Particulate and gas samplings were performed in parallel, 2 in the morning and 2 in the afternoon, alternating the inlet heads (PM₁₀ and PM₂.₅), in the particulate sampler. The gas and particulate samples were collected in the plume as close as the emission chimney point. The biomass (dry basis) used in the carbonization process was a mixture of cork oak (77 wt.%), holm oak (7 wt.%), stumps (11 wt.%), and charred wood (5 wt.%) from previous carbonization processes. A cylindrical batch kiln (80 m³) with 4.5 m diameter and 5 m of height was used in this study. The composition of the gases was determined by gas chromatography, while the particulate samples (PM₁₀, PM₂.₅) were subjected to different analytical techniques (thermo-optical transmission technique, ion chromatography, HPAE-PAD, and GC-MS after solvent extraction) after prior gravimetric determination, to study their organic and inorganic constituents. The charcoal production cycle presents widely varying operating conditions, which will be reflected in the composition of gases and particles produced and emitted throughout the process. The concentration of PM₁₀ and PM₂.₅ in the plume was calculated, ranging between 0.003 and 0.293 g m⁻³, and 0.004 and 0.292 g m⁻³, respectively. Total carbon, inorganic ions, and sugars account, in average, for PM10 and PM₂.₅, 65 % and 56 %, 2.8 % and 2.3 %, 1.27 %, and 1.21 %, respectively. The organic fraction studied until now includes more than 30 aliphatic compounds and 20 PAHs. The emission factors of particulate matter to produce charcoal in the traditional kiln were 33 g/kg (wooddb) and 27 g/kg (wooddb) for PM₁₀ and PM₂.₅, respectively. With the data obtained in this study, it is possible to fill the lack of information about the environmental impact of the traditional charcoal production in Portugal. Acknowledgment: Authors thanks to FCT – Portuguese Science Foundation, I.P. and to Ministry of Science, Technology and Higher Education of Portugal for financial support within the scope of the project CHARCLEAN (PCIF/GVB/0179/2017) and CESAM (UIDP/50017/2020 + UIDB/50017/2020).

Keywords: brick kilns, charcoal, emission factors, PAHs, total carbon

Procedia PDF Downloads 106

Authors: Sudeshna Chandra, Christian Gäbler, Christian Schliebe, Heinrich Lang

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Highly branched dendrimers provide structural homogeneity, controlled composition, comparable size to biomolecules, internal porosity and multiple functional groups for conjugating reactions. Electro-active dendrimers containing multiple redox units have generated great interest in their use as electrode modifiers for development of biosensors. The electron transfer between the redox-active dendrimers and the biomolecules play a key role in developing a biosensor. Ferrocenes have multiple and electrochemically equivalent redox units that can act as electron “pool” in a system. The ferrocenyl-terminated polyamidoamine dendrimer is capable of transferring multiple numbers of electrons under the same applied potential. Therefore, they can be used for dual purposes: one in building a film over the electrode for immunosensors and the other for immobilizing biomolecules for sensing. Electrochemical immunosensor, thus developed, exhibit fast and sensitive analysis, inexpensive and involve no prior sample pre-treatment. Electrochemical amperometric immunosensors are even more promising because they can achieve a very low detection limit with high sensitivity. Detection of the cancer biomarkers at an early stage can provide crucial information for foundational research of life science, clinical diagnosis and prevention of disease. Elevated concentration of biomarkers in body fluid is an early indication of some type of cancerous disease and among all the biomarkers, IgG is the most common and extensively used clinical cancer biomarkers. We present an IgG (=immunoglobulin) electrochemical immunosensor using a newly synthesized redox-active ferrocenyl dendrimer of generation 2 (G2Fc) as glassy carbon electrode material for immobilizing the antibody. The electrochemical performance of the modified electrodes was assessed in both aqueous and non-aqueous media using varying scan rates to elucidate the reaction mechanism. The potential shift was found to be higher in an aqueous electrolyte due to presence of more H-bond which reduced the electrostatic attraction within the amido groups of the dendrimers. The cyclic voltammetric studies of the G2Fc-modified GCE in 0.1 M PBS solution of pH 7.2 showed a pair of well-defined redox peaks. The peak current decreased significantly with the immobilization of the anti-goat IgG. After the immunosensor is blocked with BSA, a further decrease in the peak current was observed due to the attachment of the protein BSA to the immunosensor. A significant decrease in the current signal of the BSA/anti-IgG/G2Fc/GCE was observed upon immobilizing IgG which may be due to the formation of immune-conjugates that blocks the tunneling of mass and electron transfer. The current signal was found to be directly related to the amount of IgG captured on the electrode surface. With increase in the concentration of IgG, there is a formation of an increasing amount of immune-conjugates that decreased the peak current. The incubation time and concentration of the antibody was optimized for better analytical performance of the immunosensor. The developed amperometric immunosensor is sensitive to IgG concentration as low as 2 ng/mL. Tailoring of redox-active dendrimers provides enhanced electroactivity to the system and enlarges the sensor surface for binding the antibodies. It may be assumed that both electron transfer and diffusion contribute to the signal transformation between the dendrimers and the antibody.

Keywords: ferrocenyl dendrimers, electrochemical immunosensors, immunoglobulin, amperometry

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Authors: Lacramioara Ochiuz, Aurelia Vasile, Iulian Stoleriu, Cristina Ghiciuc, Maria Ignat

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Topical administration of chemotherapeutic agents (eg. carmustine, bexarotene, mechlorethamine etc.) in local treatment of cutaneous T-cell lymphoma (CTCL) is accompanied by multiple side effects, such as contact hypersensitivity, pruritus, skin atrophy or even secondary malignancies. A known method of reducing the side effects of anticancer agent is the development of modified drug release systems using drug incapsulation in biocompatible nanoporous inorganic matrices, such as mesoporous MCM-41 silica. Mesoporous MCM-41 silica is characterized by large specific surface, high pore volume, uniform porosity, and stable dispersion in aqueous medium, excellent biocompatibility, in vivo biodegradability and capacity to be functionalized with different organic groups. Therefore, MCM-41 is an attractive candidate for a wide range of biomedical applications, such as controlled drug release, bone regeneration, protein immobilization, enzymes, etc. The main advantage of this material lies in its ability to host a large amount of the active substance in uniform pore system with adjustable size in a mesoscopic range. Silanol groups allow surface controlled functionalization leading to control of drug loading and release. This study shows (I) the amino-grafting optimization of mesoporous MCM-41 silica matrix by means of co-condensation during synthesis and post-synthesis using APTES (3-aminopropyltriethoxysilane); (ii) loading the therapeutic agent (carmustine) obtaining a modified drug release systems; (iii) determining the profile of in vitro carmustine release from these systems; (iv) assessment of carmustine release kinetics by fitting on four mathematical models. Obtained powders have been described in terms of structure, texture, morphology thermogravimetric analysis. The concentration of the therapeutic agent in the dissolution medium has been determined by HPLC method. In vitro dissolution tests have been done using cell Enhancer in a 12 hours interval. Analysis of carmustine release kinetics from mesoporous systems was made by fitting to zero-order model, first-order model Higuchi model and Korsmeyer-Peppas model, respectively. Results showed that both types of highly ordered mesoporous silica (amino grafted by co-condensation process or post-synthesis) are thermally stable in aqueous medium. In what regards the degree of loading and efficiency of loading with the therapeutic agent, there has been noticed an increase of around 10% in case of co-condensation method application. This result shows that direct co-condensation leads to even distribution of amino groups on the pore walls while in case of post-synthesis grafting many amino groups are concentrated near the pore opening and/or on external surface. In vitro dissolution tests showed an extended carmustine release (more than 86% m/m) both from systems based on silica functionalized directly by co-condensation and after synthesis. Assessment of carmustine release kinetics revealed a release through diffusion from all studied systems as a result of fitting to Higuchi model. The results of this study proved that amino-functionalized mesoporous silica may be used as a matrix for optimizing the anti-cancer topical therapy by loading carmustine and developing prolonged-release systems.

Keywords: carmustine, silica, controlled, release

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Authors: Seyedeh Nasim Mirbahari, Taha Azad, Mehdi Totonchi

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Oncolytic viruses, which only replicate in tumor cells, are being extensively studied for their use in cancer therapy. A particular virus known as the vaccinia virus, a member of the poxvirus family, has demonstrated oncolytic abilities glioma. Treating Glioma with traditional methods such as chemotherapy and radiotherapy is quite challenging. Even though oncolytic viruses have shown immense potential in cancer treatment, their effectiveness in glioblastoma treatment is still low. Therefore, there is a need to improve and optimize immunotherapies for better results. In this study, we have designed oncoVV-TT, which can more effectively target tumor cells while minimizing replication in normal cells by replacing the thymidine kinase gene with a luc-p2a-GFP gene expression cassette. Human glioblastoma cell line U251 MG, rat glioblastoma cell line C6, and non-tumor cell line HFF were plated at 105 cells in a 12-well plates in 2 mL of DMEM-F2 medium with 10% FBS added to each well. Then incubated at 37°C. After 16 hours, the cells were treated with oncoVV-TT at an MOI of 0.01, 0.1 and left in the incubator for a further 24, 48, 72 and 96 hours. Viral replication assay, fluorescence imaging and viability tests, including trypan blue and crystal violet, were conducted to evaluate the cytotoxic effect of oncoVV-TT. The finding shows that oncoVV-TT had significantly higher cytotoxic activity and proliferation rates in tumor cells in a dose and time-dependent manner, with the strongest effect observed in U251 MG. To conclude, oncoVV-TT has the potential to be a promising oncolytic virus for cancer treatment, with a more cytotoxic effect in human glioblastoma cells versus rat glioma cells. To assess the effectiveness of vaccinia virus-mediated viral therapy, we have tested U251mg and C6 tumor cell lines taken from human and rat gliomas, respectively. The study evaluated oncoVV-TT's ability to replicate and lyse cells and analyzed the survival rates of the tested cell lines when treated with different doses of oncoVV-TT. Additionally, we compared the sensitivity of human and mouse glioma cell lines to the oncolytic vaccinia virus. All experiments regarding viruses were conducted under biosafety level 2. We engineered a Vaccinia-based oncolytic virus called oncoVV-TT to replicate specifically in tumor cells. To propagate the oncoVV-TT virus, HeLa cells (5 × 104/well) were plated in 24-well plates and incubated overnight to attach to the bottom of the wells. Subsequently, 10 MOI virus was added. After 48 h, cells were harvested by scraping, and viruses were collected by 3 sequential freezing and thawing cycles followed by removal of cell debris by centrifugation (1500 rpm, 5 min). The supernatant was stored at −80 ◦C for the following experiments. To measure the replication of the virus in Hela, cells (5 × 104/well) were plated in 24-well plates and incubated overnight to attach to the bottom of the wells. Subsequently, 5 MOI virus or equal dilution of PBS was added. At the treatment time of 0 h, 24 h, 48 h, 72 h and 96 h, the viral titers were determined under the fluorescence microscope (BZ-X700; Keyence, Osaka, Japan). Fluorescence intensity was quantified using the imagej software according to the manufacturer’s protocol. For the isolation of single-virus clones, HeLa cells seeded in six-well plates (5×105 cells/well). After 24 h (100% confluent), the cells were infected with a 10-fold dilution series of TianTan green fluorescent protein (GFP)virus and incubated for 4 h. To examine the cytotoxic effect of oncoVV-TT virus ofn U251mg and C6 cell, trypan blue and crystal violet assay was used.

Keywords: oncolytic virus, immune therapy, glioma, vaccinia virus

Procedia PDF Downloads 50

Authors: Omaima A. Sharaf, Wafaa M. Abd El-Rahim, Hassan Moawad, Michael J. Sadowsky

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Textile industry is one of the important industries in the worldwide. It is known that the eco-friendly industrial and agricultural activities are significant for socio-economic stability of all countries. The absence of appropriate industrial waste water treatments is essential barrier for sustainable development in food and agricultural sectors especially in developing country like Egypt. Thus, the development of enzymatic bioremediation technology for textile dye removal will enhance the collaboration between scientists who develop the technology and industry where this technology will be implemented towards the safe disposal of the textile dye wastes. Highly efficient microorganisms are of most importance in developing and using highly effective biological treatment processes. Bacterial degradation of azo dyes is generally initiated by an enzymatic step that involves cleavage of azo linkages, usually with the aid of an azoreductase as electron donor. Thus, expanding the spectrum of microorganisms with high enzymatic activities as azoreductases and discovering novel azo-dye degrading enzymes, with enhanced stability and superior catalytic properties, are necessary for many environmental and industrial applications. Consequently, the use of molecular tools has become increasingly integrated into the understanding of enzyme properties and characterization. Researchers have utilized a gene cloning and expression methods as a tool to produce recombinant protein for decolorizing dyes more efficiently. Thus, presumptive evidence for the presence of genes encoding azoreductases in the genomes of selected local, and most potent azo-degrading strains were obtained by using specific oligonucleotides primers. These potent strains have been isolated from textile industrial wastewater in Egypt and identified using 16S rRNA sequence analysis as 'Lysinibacillus macrolidesB8, Brevibacillus parabrevisB11, Bacillus coagulansB7, and B. cereusB5'. PCR products of two full length genes designated as (AZO1;621bp and AZO2;534bp) were detected. BLASTx results indicated that AZO1 gene was corresponding to predicted azoreductase from of Bacillus sp. ABP14, complete genome, multispecies azoreductase [Bacillus], It was submitted to the gene bank by an accession no., BankIt2085371 AZO1 MG923210 (621bp; 207 amino acids). AZO1 was generated from the DNA of our identified strains Lysinibacillus macrolidesB8. On the other hand, AZO2 gene was corresponding to a predicted azoreductase from Bacillus cereus strain S2-8. Gene bank accession no. was BankIt2085839 AZO2 MG932081 (534bp;178 amino acids) and it was amplified from our Bacillus coagulansB7. Both genes were successfully cloned into pCR2.1TOPO (Invitrogen) and in pET28b+ vectors, then they transformed into E. coli DH5α and BL21(DE3) cells for heterologous expression studies. Our recombinant azoreductases (AZO1&AZO2) exhibited potential enzyme activity and efficiently decolorized an azo dye (Direct violet). They exhibited pH stability between 6 and 8 with optimum temperature up to 60°C and 37 °C after induction by 1mM and 1.5mM IPTG, for both AZO1 &AZO2, respectively. These results suggested that further optimization and purification of these recombinant proteins by using different heterologous expression systems will give great potential for the sustainable utilization of these recombinant enzymes in several industrial applications especially in wastewater treatments.

Keywords: azoreductases, decolorization, enzyme activity, gene cloning and expression

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Authors: Aishwarya Venkat, Anastasia Marshak, Ryan B. Simpson, Elena N. Naumova

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Livestock ownership is simultaneously linked to improved nutritional status through increased availability of animal-source protein, and increased risk of enteric infections through higher exposure to contaminated water sources. Agrarian and agro-pastoral households, especially those with cattle, goats, and sheep, are highly dependent on seasonally various environmental conditions, which directly impact nutrition and health. This study explores global spatiotemporally explicit evidence regarding the relationship between livestock ownership, enteric infections, and malnutrition. Seasonal and cyclical fluctuations, as well as mediating effects, are further examined to elucidate health and nutrition outcomes of individual and communal livestock ownership. The US Agency for International Development’s Demographic and Health Surveys (DHS) and the United Nations International Children's Emergency Fund’s Multi-Indicator Cluster Surveys (MICS) provide valuable sources of household-level information on anthropometry, asset ownership, and disease outcomes. These data are especially important in data-sparse regions, where surveys may only be conducted in the aftermath of emergencies. Child-level disease history, anthropometry, and household-level asset ownership information have been collected since DHS-V (2003-present) and MICS-III (2005-present). This analysis combines over 15 years of survey data from DHS and MICS to study 2,466,257 children under age five from 82 countries. Subnational (administrative level 1) measures of diarrhea prevalence, mean livestock ownership by type, mean and median anthropometric measures (height for age, weight for age, and weight for height) were investigated. Effects of several environmental, market, community, and household-level determinants were studied. Such covariates included precipitation, temperature, vegetation, the market price of staple cereals and animal source proteins, conflict events, livelihood zones, wealth indices and access to water, sanitation, hygiene, and public health services. Children aged 0 – 6 months, 6 months – 2 years, and 2 – 5 years of age were compared separately. All observations were standardized to interview day of year, and administrative units were harmonized for consistent comparisons over time. Geographically weighted regressions were constructed for each outcome and subnational unit. Preliminary results demonstrate the importance of accounting for seasonality in concurrent assessments of malnutrition and enteric infections. Household assets, including livestock, often determine the intensity of these outcomes. In many regions, livestock ownership affects seasonal fluxes in malnutrition and enteric infections, which are also directly affected by environmental and local factors. Regression analysis demonstrates the spatiotemporal variability in nutrition outcomes due to a variety of causal factors. This analysis presents a synthesis of evidence from global survey data on the interrelationship between enteric infections, malnutrition, and livestock. These results provide a starting point for locally appropriate interventions designed to address this nexus in a timely manner and simultaneously improve health, nutrition, and livelihoods.

Keywords: diarrhea, enteric infections, households, livestock, malnutrition, seasonality

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Authors: Monika Dmitrzak-Weglarz, Aleksandra Rajewska-Rager, Maria Skibinska, Natalia Lepczynska, Piotr Sibilski, Joanna Pawlak, Pawel Kapelski, Joanna Hauser

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Epidermal growth factor (EGF) is a well-known neurotrophic factor that involves in neuronal growth and synaptic plasticity. The proteomic research provided in order to identify novel candidate biological markers for mood disorders focused on elevated EGF serum level in patients during depression episode. However, the EGF association with mood disorder spectrum among adolescents and young adults has not been studied extensively. In this study, we aim to investigate the serum levels of EGF in adolescents and young adults during hypo/manic, depressive episodes and in remission compared to healthy control group. In our study, we involved 80 patients aged 12-24 years in 2-year follow-up study with a primary diagnosis of mood disorder spectrum, and 35 healthy volunteers matched by age and gender. Diagnoses were established according to DSM-IV-TR criteria using structured clinical interviews: K-SADS for child and adolescents, and SCID for young adults. Clinical and biological evaluations were made at baseline and euthymic mood (at 3th or 6th month of treatment and after 1 and 2 years). The Young Mania Rating Scale and Hamilton Rating Scale for Depression were used for assessment. The study protocols were approved by the relevant ethics committee. Serum protein concentration was determined by Enzyme-Linked Immunosorbent Assays (ELISA) method. Human EGF (cat. no DY 236) DuoSet ELISA kit was used (R&D Systems). Serum EGF levels were analysed with following variables: age, age under 18 and above 18 years old, sex, family history of affective disorders, drug-free vs. medicated. Shapiro-Wilk test was used to test the normality of the data. The homogeneity of variance was calculated with Levene’s test. EGF levels showed non-normal distribution and the homogeneity of variance was violated. Non-parametric tests: Mann-Whitney U test, Kruskall-Wallis ANOVA, Friedman’s ANOVA, Wilcoxon signed rank test, Spearman correlation coefficient was applied in the analyses The statistical significance level was set at p<0.05. Elevated EGF level at baseline (p=0.001) and at month 24 (p=0.02) was detected in study subjects compared with controls. Increased EGF level in women at month 12 (p=0.02) compared to men in study group have been observed. Using Wilcoxon signed rank test differences in EGF levels were detected: decrease from baseline to month 3 (p=0.014) and increase comparing: month 3 vs. 24 (p=0.013); month 6 vs. 12 (p=0.021) and vs. 24 (p=0.008). EGF level at baseline was negatively correlated with depression and mania occurrence at 24 months. EGF level at 24 months was positively correlated with depression and mania occurrence at 12 months. No other correlations of EGF levels with clinical and demographical variables have been detected. The findings of the present study indicate that EGF serum level is significantly elevated in the study group of patients compared to the controls. We also observed fluctuations in EGF levels during two years of disease observation. EGF seems to be useful as an early marker for prediction of diagnosis, course of illness and treatment response in young patients during first episode od mood disorders, which requires further investigation. Grant was founded by National Science Center in Poland no 2011/03/D/NZ5/06146.

Keywords: biological marker, epidermal growth factor, mood disorders, prediction

Procedia PDF Downloads 164