Search results for: simultaneous tooth cracks

Authors: Tomohiko Utsuki, Kyoka Sato

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In emergency medicine, it is recognized that brain resuscitation is very important for the reduction of mortality rate and neurological sequelae. Especially, the control of brain temperature (BT), intracranial pressure (ICP), and cerebral blood flow (CBF) are most required for stabilizing brain’s physiological state in the treatment for such as brain injury, stroke, and encephalopathy. However, the manual control of BT, ICP, and CBF frequently requires the decision and operation of medical staff, relevant to medication and the setting of therapeutic apparatus. Thus, the integration and the automation of the control of those is very effective for not only improving therapeutic effect but also reducing staff burden and medical cost. For realizing such integration and automation, a mathematical model of brain physiological state is necessary as the controlled object in simulations, because the performance test of a prototype of the control system using patients is not ethically allowed. A model of cerebral blood circulation has already been constructed, which is the most basic part of brain physiological state. Also, a migration model of extracellular fluid in brain has been constructed, however the condition that the total volume of intracranial cavity is almost changeless due to the hardness of cranial bone has not been considered in that model. Therefore, in this research, the dynamic migration model of extracellular fluid in brain was constructed on the consideration of the changelessness of intracranial cavity’s total volume. This model is connectable to the cerebral blood circulation model. The constructed model consists of fourteen compartments, twelve of which corresponds to perfused area of bilateral anterior, middle and posterior cerebral arteries, the others corresponds to cerebral ventricles and subarachnoid space. This model enable to calculate the migration of tissue fluid from capillaries to gray matter and white matter, the flow of tissue fluid between compartments, the production and absorption of cerebrospinal fluid at choroid plexus and arachnoid granulation, and the production of metabolic water. Further, the volume, the colloid concentration, and the tissue pressure of/in each compartment are also calculable by solving 40-dimensional non-linear simultaneous differential equations. In this research, the obtained model was analyzed for its validation under the four condition of a normal adult, an adult with higher cerebral capillary pressure, an adult with lower cerebral capillary pressure, and an adult with lower colloid concentration in cerebral capillary. In the result, calculated fluid flow, tissue volume, colloid concentration, and tissue pressure were all converged to suitable value for the set condition within 60 minutes at a maximum. Also, because these results were not conflict with prior knowledge, it is certain that the model can enough represent physiological state of brain under such limited conditions at least. One of next challenges is to integrate this model and the already constructed cerebral blood circulation model. This modification enable to simulate CBF and ICP more precisely due to calculating the effect of blood pressure change to extracellular fluid migration and that of ICP change to CBF.

Keywords: dynamic model, cerebral extracellular migration, brain resuscitation, automatic control

Procedia PDF Downloads 128

Authors: Lina E. Homman, Alexis C. Edwards, Seung Bin Cho, Danielle M. Dick, Kenneth S. Kendler

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Research supports an association between alcohol problems and internalising symptoms, but the understanding of how the two phenotypes relate to each other is poor. It has been hypothesized that the relationship between the phenotypes is causal; however investigations in regards to direction are inconsistent. Clarity of the relationship between the two phenotypes may be provided by investigating the phenotypes developmental inter-relationships longitudinally. The objective of the study was to investigate a) changes in alcohol problems and internalising symptoms in college students across time and b) the direction of effect of growth between alcohol problems and internalising symptoms from late adolescent to emerging adulthood c) possible gender differences. The present study adds to the knowledge of comorbidity of alcohol problems and internalising symptoms by examining a longitudinal sample of college students and by examining the simultaneous development of the symptoms. A sample of college students is of particular interest as symptoms of both phenotypes often have their onset around this age. A longitudinal sample of college students from a large, urban, public university in the United States was used. Data was collected over a time period of 2 years at 3 time points. Latent growth models were applied to examine growth trajectories. Parallel process growth models were used to assess whether initial level and rate of change of one symptom affected the initial level and rate of change of the second symptom. Possible effects of gender and ethnicity were investigated. Alcohol problems significantly increased over time, whereas internalizing symptoms remained relatively stable. The two phenotypes were significantly correlated in each wave, correlations were stronger among males. Initial level of alcohol problems was significantly positively correlated with initial level of internalising symptoms. Rate of change of alcohol problems positively predicted rate of change of internalising symptoms for females but not for males. Rate of change of internalising symptoms did not predict rate of change of alcohol problems for either gender. Participants of Black and Asian ethnicities indicated significantly lower levels of alcohol problems and a lower increase of internalising symptoms across time, compared to White participants. Participants of Black ethnicity also reported significantly lower levels of internalising symptoms compared to White participants. The present findings provide additional support for a positive relationship between alcohol problems and internalising symptoms in youth. Our findings indicated that both internalising symptoms and alcohol problems increased throughout the sample and that the phenotypes were correlated. The findings mainly implied a bi-directional relationship between the phenotypes in terms of significant associations between initial levels as well as rate of change. No direction of causality was indicated in males but significant results were found in females where alcohol problems acted as the main driver for the comorbidity of alcohol problems and internalising symptoms; alcohol may have more detrimental effects in females than in males. Importantly, our study examined a population-based longitudinal sample of college students, revealing that the observed relationships are not limited to individuals with clinically diagnosed mental health or substance use problems.

Keywords: alcohol, comorbidity, internalising symptoms, longitudinal modelling

Procedia PDF Downloads 323

Authors: Ekaterina A. Savchenko, Elena N. Velichko, Evgenii T. Aksenov

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The development of pathological processes, such as cardiovascular and oncological diseases, are accompanied by changes in molecular parameters in cells, tissues, and serum. The study of the behavior of protein molecules in solutions is of primarily importance for diagnosis of such diseases. Various physical and chemical methods are used to study molecular systems. With the advent of the laser and advances in electronics, optical methods, such as scanning electron microscopy, sedimentation analysis, nephelometry, static and dynamic light scattering, have become the most universal, informative and accurate tools for estimating the parameters of nanoscale objects. The electrophoretic light scattering is the most effective technique. It has a high potential in the study of biological solutions and their properties. This technique allows one to investigate the processes of aggregation and dissociation of different macromolecules and obtain information on their shapes, sizes and molecular weights. Electrophoretic light scattering is an analytical method for registration of the motion of microscopic particles under the influence of an electric field by means of quasi-elastic light scattering in a homogeneous solution with a subsequent registration of the spectral or correlation characteristics of the light scattered from a moving object. We modified the technique by using the regime of total internal reflection with the aim of increasing its sensitivity and reducing the volume of the sample to be investigated, which opens the prospects of automating simultaneous multiparameter measurements. In addition, the method of total internal reflection allows one to study biological fluids on the level of single molecules, which also makes it possible to increase the sensitivity and the informativeness of the results because the data obtained from an individual molecule is not averaged over an ensemble, which is important in the study of bimolecular fluids. To our best knowledge the study of electrophoretic light scattering in the regime of total internal reflection is proposed for the first time, latex microspheres 1 μm in size were used as test objects. In this study, the total internal reflection regime was realized on a quartz prism where the free electrophoresis regime was set. A semiconductor laser with a wavelength of 655 nm was used as a radiation source, and the light scattering signal was registered by a pin-diode. Then the signal from a photodetector was transmitted to a digital oscilloscope and to a computer. The autocorrelation functions and the fast Fourier transform in the regime of Brownian motion and under the action of the field were calculated to obtain the parameters of the object investigated. The main result of the study was the dependence of the autocorrelation function on the concentration of microspheres and the applied field magnitude. The effect of heating became more pronounced with increasing sample concentrations and electric field. The results obtained in our study demonstrated the applicability of the method for the examination of liquid solutions, including biological fluids.

Keywords: light scattering, electrophoretic light scattering, electrophoresis, total internal reflection

Procedia PDF Downloads 187

Authors: Ahlam Said Al Azkawi, Nallusamy Sivakumar, Saif Nasser Al Bahri

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Diverse approaches and pathways are under development with the determination to develop cellulosic biofuels and other bio-products eventually at commercial scale in “bio-refineries”; however, the key challenge is mainly the high level of complexity in processing the feedstock which is complicated and energy consuming. To overcome the complications in utilizing the naturally occurring lignocellulose biomass, using waste paper as a feedstock for bio-production may solve the problem. Besides being abundant and cheap, bioprocessing of waste paper has evolved in response to the public concern from rising landfill cost from shrinking landfill capacity. Cardboard (CB) is one of the major components of municipal solid waste and one of the most important items to recycle. Although 50-70% of cardboard constitute is known to be cellulose and hemicellulose, the presence of lignin around them cause hydrophobic cross-link which physically obstructs the hydrolysis by rendering it resistant to enzymatic cleavage. Therefore, pretreatment is required to disrupt this resistance and to enhance the exposure of the targeted carbohydrates to the hydrolytic enzymes. Several pretreatment approaches have been explored, and the best ones would be those can influence cellulose conversion rates and hydrolytic enzyme performance with minimal or less cost and downstream processes. One of the promising strategies in this field is the application of surfactants, especially non-ionic surfactants. In this study, triton-X 100 was used as surfactants to treat cardboard prior enzymatic hydrolysis and compare it with acid treatment using 0.1% H2SO4. The effect of the surfactant enhancement was evaluated through its effect on hydrolysis rate in respect to time in addition to evaluating the structural changes and modification by scanning electron microscope (SEM) and X-ray diffraction (XRD) and through compositional analysis. Further work was performed to produce ethanol from CB treated with triton-X 100 via separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF). The hydrolysis studies have demonstrated enhancement in saccharification by 35%. After 72 h of hydrolysis, a saccharification rate of 98% was achieved from CB enhanced with triton-X 100, while only 89 of saccharification achieved from acid pre-treated CB. At 120 h, the saccharification % exceeded 100 as reducing sugars continued to increase with time. This enhancement was not supported by any significant changes in the cardboard content as the cellulose, hemicellulose and lignin content remained same after treatment, but obvious structural changes were observed through SEM images. The cellulose fibers were clearly exposed with very less debris and deposits compared to cardboard without triton-X 100. The XRD pattern has also revealed the ability of the surfactant in removing calcium carbonate, a filler found in waste paper known to have negative effect on enzymatic hydrolysis. The cellulose crystallinity without surfactant was 73.18% and reduced to 66.68% rendering it more amorphous and susceptible to enzymatic attack. Triton-X 100 has proved to effectively enhance CB hydrolysis and eventually had positive effect on the ethanol yield via SSF. Treating cardboard with only triton-X 100 was a sufficient treatment to enhance the enzymatic hydrolysis and ethanol production.

Keywords: cardboard, enhancement, ethanol, hydrolysis, treatment, Triton-X 100

Procedia PDF Downloads 117

Authors: S. Sharma, Gaballa Aqeelah, Tawfig Alghbaili, Ali Elmessmari

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There were abrupt discontinuities in the Brute Stack in the northernmost locations during the acquisition of 2D (2007) and 3D (2021) seismic data in the northwest region of the Ghadames Basin, Libya. In both campaigns, complete fluid circulation loss was seen in these regions during up-hole drilling. Geophysics, sedimentology and shallow subsurface geology were all integrated to look into what was causing the seismic signal to disappear at shallow depths. The Upper Cretaceous Nalut Formation is the near-surface or surface formation in the studied area. It is distinguished by abnormally high resistivity in all the neighboring wells. The Nalut Formation in all the nearby wells from the present study and previous outcrop study suggests lithology of dolomite and chert/flint in nodular or layered forms. There are also reports of karstic caverns, vugs, and thick cracks, which all work together to produce the high resistivity. Four up-hole samples that were analyzed for microfacies revealed a near-coastal to tidal environment. Algal (Chara) infested deposits up to 30 feet thick and monotonous, very porous, are seen in two up-hole sediments; these deposits are interpreted to be scattered, continental algal travertine mounds. Chert/flint, dolomite, and calcite in varying amounts are confirmed by XRD analysis. Regional tracking of the high resistivity of the Nalut Formation, which is thought to be connected to the sea level drop that created the paleokarst layer, is possible. It is abruptly overlain by a blanket marine transgressive deposit caused by rapid sea level rise, which is a regional, relatively high radioactive layer of argillaceous limestone. The examined area's close proximity to the mountainous, E-W trending ridges of northern Libya made it easier for recent freshwater circulation, which later enhanced cavern development and mineralization in the paleokarst layer. Seismic signal loss at shallow depth is caused by extremely heterogeneous mineralogy of pore- filling or lack thereof. Scattering effect of shallow karstic layer on seismic signal has been well documented. Higher velocity inflection points at shallower depths in the northern part and deeper intervals in the southern part, in both cases at Nalut level, demonstrate the layer's influence on the seismic signal. During the Permian-Carboniferous, the Ghadames Basin underwent uplift and extensive erosion, which resulted in this karstic layer of the Nalut Formation uplifted to a shallow depth in the northern part of the studied area weakening the acoustic signal, whereas in the southern part of the 3D acquisition area the Nalut Formation remained at the deeper interval without affecting the seismic signal. Results from actions taken during seismic processing to deal with this signal loss are visible and have improved. This study recommends using denser spacing or dynamite to circumvent the karst layer in a comparable geographic area in order to prevent signal loss at lesser depths.

Keywords: well logging, seismic data acquisition, sesimic data processing, up-holes

Procedia PDF Downloads 53

Authors: Nebojsa Stefanovic, Sasa Milijic, Natasa Danilovic Hristic

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Spatial plan of the special purpose area constitutes a basic tool in the planning of infrastructure corridor of a highway. The aim of the plan is to define the planning basis and provision of spatial conditions for the construction and operation of the highway, as well as for developing other infrastructure systems in the corridor. This paper presents a methodology and approach to the preparation of the Spatial Plan for the special purpose area for the infrastructure corridor of the highway E-80, Section Niš-Merdare in Serbia. The applied methodological approach is based on the combined application of the integrative and participatory method in the decision-making process on the sustainable development of the highway corridor. It was found that, for the planning and management of the infrastructure corridor, a key problem is coordination of spatial and urban planning, strategic environmental assessment and sectoral traffic planning and designing. Through the development of the plan, special attention is focused on increasing the accessibility of the local and regional surrounding, reducing the adverse impacts on the development of settlements and the economy, protection of natural resources, natural and cultural heritage, and the development of other infrastructure systems in the corridor of the highway. As a result of the applied methodology, this paper analyzes the basic features such as coverage, the concept, protected zones, service facilities and objects, the rules of development and construction, etc. Special emphasis is placed to methodology and results of the Strategic Environmental Assessment of the Spatial Plan, and to the importance of protection measures, with the special significance of air and noise protection measures. For evaluation in the Strategic Environmental Assessment, a multicriteria expert evaluation (semi-quantitative method) of planned solutions was used in relation to the set of goals and relevant indicators, based on the basic set of indicators of sustainable development. Evaluation of planned solutions encompassed the significance and size, spatial conditions and probability of the impact of planned solutions on the environment, and the defined goals of strategic assessment. The framework of the implementation of the Spatial Plan is presented, which is determined for the simultaneous elaboration of planning solutions at two levels: the strategic level of the spatial plan and detailed urban plan level. It is also analyzed the relationship of the Spatial Plan to other applicable planning documents for the planning area. The effects of this methodological approach relate to enabling integrated planning of the sustainable development of the infrastructure corridor of the highway and its surrounding area, through coordination of spatial, urban and sectoral traffic planning and design, as well as the participation of all key actors in the adoption and implementation of planned decisions. By the conclusions of the paper, it is pointed to the direction for further research, particularly in terms of harmonizing methodology of planning documentation and preparation of technical-design documentation.

Keywords: corridor, environment, highway, impact, methodology, spatial plan, urban

Procedia PDF Downloads 181

Authors: M. Hosseini, N. Ghorbani Amirabad, M. Zhian

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Seismic design codes accept structural and nonstructural damages after the sever earthquakes (provided that the building is prevented from collapse), so that in many cases demolishing and reconstruction of the building is inevitable, and this is usually very difficult, costly and time consuming. Therefore, designing and constructing of buildings in such a way that they can be easily repaired after earthquakes, even major ones, is quite desired. For this purpose giving the possibility of rocking or see-saw motion to the building structure, partially or as a whole, has been used by some researchers in recent decade .the central support which has a main role in creating the possibility of see-saw motion in the building’s structural system. In this paper, paying more attention to the key role of the central fuse and support, an innovative energy dissipater which can act as the central fuse and support of the building with seesaw motion is introduced, and the process of reaching an optimal geometry for that by using finite element analysis is presented. Several geometric shapes were considered for the proposed central fuse and support. In each case the hysteresis moment rotation behavior of the considered fuse were obtained under simultaneous effect of vertical and horizontal loads, by nonlinear finite element analyses. To find the optimal geometric shape, the maximum plastic strain value in the fuse body was considered as the main parameter. The rotational stiffness of the fuse under the effect of acting moments is another important parameter for finding the optimum shape. The proposed fuse and support can be called Yielding Curved Bars and Clipped Hemisphere Core (YCB&CHC or more briefly YCB) energy dissipater. Based on extensive nonlinear finite element analyses it was found out the using rectangular section for the curved bars gives more reliable results. Then, the YCB energy dissipater with the optimal shape was used in a structural model of a 12 story regular building as its central fuse and support to give it the possibility of seesaw motion, and its seismic responses were compared to those of a the building in the fixed based conditions, subjected to three-components acceleration of several selected earthquakes including Loma Prieta, Northridge, and Park Field. In building with see-saw motion some simple yielding-plate energy dissipaters were also used under circumferential columns.The results indicated that equipping the buildings with central and circumferential fuses result in remarkable reduction of seismic responses of the building, including the base shear, inter story drift, and roof acceleration. In fact by using the proposed technique the plastic deformations are concentrated in the fuses in the lowest story of the building, so that the main body of the building structure remains basically elastic, and therefore, the building can be easily repaired after earthquake.

Keywords: rocking mechanism, see-saw motion, finite element analysis, hysteretic behavior

Procedia PDF Downloads 387

Authors: Alexandra Sargent, Sarah Ferris, Ioannis Theofanous

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The Abbott RealTime High Risk HPV test (RealTime HPV) is one of five assays clinically validated and approved by the English NHS Cervical Screening Programme (CSP) for HPV triage of low grade dyskaryosis and test-of-cure of treated Cervical Intraepithelial Neoplasia. The assay is a highly automated multiplex real-time PCR test for detecting 14 high risk (hr) HPV types, with simultaneous differentiation of HPV 16 and HPV 18 versus non-HPV 16/18 hrHPV. An endogenous internal control ensures sample cellularity, controls extraction efficiency and PCR inhibition. The original cervical specimen collected in SurePath (SP) liquid-based cytology (LBC) medium (BD Diagnostics) and the SP post-gradient cell pellets (SPG) after cytological processing are both CE marked for testing with the RealTime HPV test. During the 2011 NHSCSP validation of new tests only the original aliquot of SP LBC medium was investigated. Residual sample volume left after cytology slide preparation is low and may not always have sufficient volume for repeat HPV testing or for testing of other biomarkers that may be implemented in testing algorithms in the future. The SPG samples, however, have sufficient volumes to carry out additional testing and necessary laboratory validation procedures. This study investigates the correlation of RealTime HPV results of cervical specimens collected in SP LBC medium from women with low grade cytological abnormalities observed with matched pairs of original SP LBC medium and SP post-gradient cell pellets (SPG) after cytology processing. Matched pairs of SP and SPG samples from 750 women with borderline (N = 392) and mild (N = 351) cytology were available for this study. Both specimen types were processed and parallel tested for the presence of hrHPV with RealTime HPV according to the manufacturer´s instructions. HrHPV detection rates and concordance between test results from matched SP and SPGCP pairs were calculated. A total of 743 matched pairs with valid test results on both sample types were available for analysis. An overall-agreement of hrHPV test results of 97.5% (k: 0.95) was found with matched SP/SPG pairs and slightly lower concordance (96.9%; k: 0.94) was observed on 392 pairs from women with borderline cytology compared to 351 pairs from women with mild cytology (98.0%; k: 0.95). Partial typing results were highly concordant in matched SP/SPG pairs for HPV 16 (99.1%), HPV 18 (99.7%) and non-HPV16/18 hrHPV (97.0%), respectively. 19 matched pairs were found with discrepant results: 9 from women with borderline cytology and 4 from women with mild cytology were negative on SPG and positive on SP; 3 from women with borderline cytology and 3 from women with mild cytology were negative on SP and positive on SPG. Excellent correlation of hrHPV DNA test results was found between matched pairs of SP original fluid and post-gradient cell pellets from women with low grade cytological abnormalities tested with the Abbott RealTime High-Risk HPV assay, demonstrating robust performance of the test with both specimen types and reassuring the utility of the assay for cytology triage with both specimen types.

Keywords: Abbott realtime test, HPV, SurePath liquid based cytology, surepath post-gradient cell pellet

Procedia PDF Downloads 229

Authors: Budur Almutairi

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Introduction: There is an alarming increase in the number of diabetic patients in Saudi Arabia during the last twenty years. The World Health Organization (WHO) reports that the country ranks seventh in the world for the rate of diabetes. It is also estimated that around 7 million of the population are diabetic and almost around 3 million have pre-diabetes. The prevalence is more in urban area than in rural and more in women than in men and it is closely associated with the parallel rise in obesity rates. Diabetes is found to be contributing to the increasing mortality, morbidity and vascular complications and becoming a significant cause of medical complications and even death. The trends shown by the numbers are worrying as the prevalence is steadily doubling every two decades and particularly in Saudi Arabia, this could soon reach 50% in those over 50 years of age. The economic growth and prosperity have shown notable changes in the lifestyle of the people. Most importantly, along with an increased consumption of fast foods and sugar-rich carbonated soft drinks, eating habits became less healthy and the level of physical activity is decreased. The simultaneous technological advancement and the introduction of new mechanical devices like, elevators, escalators, remotes and vehicles pushed people to a situation of leading a more sedentary life. This study is attempting to evaluate the success of the campaign that introduced through popular social media in the country. Methodology: The Ministry of Health (MoH) has initiated a novel method of campaign activity to generate discussion among public about diabetes. There were mythical monsters introduced through popular social media with disguised messages about the condition of diabetes has generated widespread discussions about the disease among the general public. The characters that started appearing in social media About 600 retweets of the original post was testimonial for the success of the Twitter campaign. The second most successful form of campaign was a video that adopted a very popular approach of using Dark Comedy in which, the diabetes was represented through a twisted negative character that talks about his meticulous plans of how he is going to take the common people into his clutches. This fictional character gained more popularity when introduced into twitter and people started interacting with him raising various questions and challenging his anti-social activities. Major findings: The video generated more than 3,200,000 views ranking 9th in You Tube’s most popular video in Saudi Arabia and was shared 7000 times in a single week. Also, the hashtag got over 4,500,000impressions and over one million visits. Conclusion: Diabetes mellitus in Saudi Arabia is emerging as an epidemic of massive proportions, threatening to negate the benefits of modernization and economic revival. It is highly possible that healthy practices connected with the prevention and management of DM can easily be implemented in a manner that does not conflict with the cultural milieu of Saudi Arabia.

Keywords: campaign, diabetes, Saudi, social media

Procedia PDF Downloads 107

Authors: Eliska Smidova, Petr Kabele

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This paper describes non-linear finite element simulation of laminated veneer lumber (LVL) under tensile and shear loads that induce cracking along fibers. For this purpose, we use 2D homogeneous orthotropic constitutive model of tensile and shear fracture in timber that has been recently developed and implemented into ATENA® finite element software by the authors. The model captures (i) material orthotropy for small deformations in both linear and non-linear range, (ii) elastic behavior until anisotropic failure criterion is fulfilled, (iii) inelastic behavior after failure criterion is satisfied, (iv) different post-failure response for cracks along and across the grain, (v) unloading/reloading behavior. The post-cracking response is treated by fixed smeared crack model where Reinhardt-Hordijk function is used. The model requires in total 14 input parameters that can be obtained from standard tests, off-axis test results and iterative numerical simulation of compact tension (CT) or compact tension-shear (CTS) test. New engineered timber composites, such as laminated veneer lumber (LVL), offer improved structural parameters compared to sawn timber. LVL is manufactured by laminating 3 mm thick wood veneers aligned in one direction using water-resistant adhesives (e.g. polyurethane). Thus, 3 main grain directions, namely longitudinal (L), tangential (T), and radial (R), are observed within the layered LVL product. The core of this work consists in 3 numerical simulations of experiments where Radiata Pine LVL and Yellow Poplar LVL were involved. The first analysis deals with calibration and validation of the proposed model through off-axis tensile test (at a load-grain angle of 0°, 10°, 45°, and 90°) and CTS test (at a load-grain angle of 30°, 60°, and 90°), both of which were conducted for Radiata Pine LVL. The second finite element simulation reproduces load-CMOD curve of compact tension (CT) test of Yellow Poplar with the aim of obtaining cohesive law parameters to be used as an input in the third finite element analysis. That is four point bending test of small-size arch of 780 mm span that is made of Yellow Poplar LVL. The arch is designed with a through crack between two middle layers in the crown. Curved laminated beams are exposed to high radial tensile stress compared to timber strength in radial tension in the crown area. Let us note that in this case the latter parameter stands for tensile strength in perpendicular direction with respect to the grain. Standard tests deliver most of the relevant input data whereas traction-separation law for crack along the grain can be obtained partly by inverse analysis of compact tension (CT) test or compact tension-shear test (CTS). The initial crack was modeled as a narrow gap separating two layers in the middle the arch crown. Calculated load-deflection curve is in good agreement with the experimental ones. Furthermore, crack pattern given by numerical simulation coincides with the most important observed crack paths.

Keywords: compact tension (CT) test, compact tension shear (CTS) test, fixed smeared crack model, four point bending test, laminated arch, laminated veneer lumber LVL, off-axis test, orthotropic elasticity, orthotropic fracture criterion, Radiata Pine LVL, traction-separation law, yellow poplar LVL, 2D constitutive model

Procedia PDF Downloads 256

Authors: Tianfang Zhang, Luxi He, Zhengbin He, Songlin Yi

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Moso bamboo is a common non-wood forest resource in Asia that is widely used in construction, furniture, and other fields. Influenced by the heterogeneous structure and various hygroscopic groups of bamboo, the deformation occurs as moisture absorption and desorption when the environment temperature and humidity conditions change. Thermal modification is a well-established commercial technology for improving the dimensional stability of bamboo. However, the higher energy consumption and carbon emissions limit its further development. Previous studies have indicated that inorganic salt-assisted thermal modification could lead to significant reductions in moisture absorption and energy consumption. Represented by metal chlorides, it could show Lewis acid properties when dissolved in water, generating metal ion ligand complexes. In addition, ultrasonic treatment, as an efficient and environmentally friendly physical treatment method, improved the accessibility of pretreatment chemical impregnation agents and intensified mass and heat transfer during reactions. To save energy and reduce deformation, this study elucidates the influence of zinc chloride-ultrasonic treatment on the physicochemical properties of heat-treated bamboo, and the details of the bamboo deformation mechanism with Lewis acid are explained. Three sets of parameters (inorganic salt concentration, ultrasonic frequency and heat treatment temperature) were designed, and an optimized process was proposed to clarify this scientific question, that is: 5% (w/w) zinc chloride solution, 40 kHz ultrasonic waves and heat treatment at 160 °C. The samples were characterized by different means to analyze changes in their macroscopic features, pore structure, chemical structure and chemical composition. The results suggested that the maximum weight loss rate was reduced by at least 19.75%. The maximum thermal degradation peak of hemicellulose was significantly shifted forward. The hygroscopicity was reduced by 10.15%, the relative crystallinity was increased by 4.4%, the surface contact angle was increased by 25.2%, and the color change was increased by 23.60 in the optimal condition. From the electron microscope observation, the treated surface became rougher, and cracks appeared in some weaker areas, accelerating starch loss and removing granular attachments around the pits. By ion diffusion, zinc ions diffused into hemicellulose and a partial amorphous region of cellulose. Parts of the cell wall structure were subjected to swelling and degradation, leading to the broken state of parenchyma cells. From the Raman spectrum, compared to conventional thermal modifications, hemicellulose thermal degradation and lignin migration is promoted by Lewis acid under dilute acid-thermal condition. As shown in this work, the combined Lewis acid and ultrasonic pretreatment as an environmentally friendly, safe, and efficient physic-chemical combined pretreatment method improved the dimensional stability of Moso bamboo and lowered the thermal degradation conditions. This method has great potential for development in the field of bamboo heat treatment, and it might provide some guidance for making dark bamboo flooring.

Keywords: Moso bamboo, Lewis acid, ultrasound, heat treatment

Procedia PDF Downloads 42

Authors: J. Lino Alves, L. Lopes

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It is not new that industrial design projects are a result of cooperative work from different areas of knowledge. However, in the academic teaching of Industrial Design and Mechanical Engineering courses, it is not recurrent that those competences are mixed before the professional life arrives. This abstract intends to describe two semester experiences carried out by two professors - a mechanical engineer and an industrial designer - in the last two academic years, for which they created mixed teams of Industrial Design and Mechanical Engineering (UPorto University). The two experiences differ in several factors; the main one is related to the challenges of online education, a constraint that affected the second experience. In the first year, even before foreseeing the effects that the pandemic would reconfigure the education system, a partnership with the Education Service of Águas do Porto was established. The purpose of the exercise was the project development of a game that could be an interaction element oriented to potentiate a positive experience and as an educational contribution to the children. In the second year, already foreseeing that the teaching experience would be carried out online, it was decided to design an open briefing, which allowed the groups to choose among three themes: a hand scale game using additive manufacturing; a modular system for ventilated facade using a parametric design basis; or, a modular system for vertical gardens. In methodological terms, besides the weekly follow-up, with the simultaneous support of the two professors, a group self-evaluation was requested; and a form to be filled individually to evaluate other groups. One of the first conclusions is related to the briefing format. Industrial Design students seem comfortable working on an open briefing that allows them to draw the project on a conceptual basis created for that purpose; on the other hand, Mechanical Engineering students were uncomfortable and insecure in the initial phase due to the absence of concrete, closed "order." In other words, it is not recurrent for Mechanical Engineering students that the creative component is stimulated, seemingly leaving them reserved to the technical solution and execution, depriving them of the co-creation phase during the conceptual construction of the project's own brief. Another fact that was registered is related to the leadership positions in the groups, which alternated according to the state of development of the project: design students took the lead during the ideation/concept phase, while mechanical engineering ones took a greater lead during the intermediate development process, namely in the definition of constructive solutions, mass/volume calculations, manufacturing, and material resistance. Designers' competences were again more evident and assumed in the final phase, especially in communication skills, as well as in simulations in the context of use. However, at some moments, it was visible the capacity for quite balanced leadership between engineering and design, in a constant debate centered on the human factor of the project - evidenced in the final solution, in the compromise and balance between technical constraints, functionality, usability, and aesthetics.

Keywords: education, industrial design, mechanical engineering, teaching ethodologies

Procedia PDF Downloads 153

Authors: Ugo Galvanetto, Pirto G. Pavan, Mirco Zaccariotto

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The discipline of Computer-integrated surgery (CIS) will provide equipment able to improve the efficiency of healthcare systems and, which is more important, clinical results. Surgeons and machines will cooperate in new ways that will extend surgeons’ ability to train, plan and carry out surgery. Patient specific CIS of the brain requires several steps: 1 - Fast generation of brain models. Based on image recognition of MR images and equipped with artificial intelligence, image recognition techniques should differentiate among all brain tissues and segment them. After that, automatic mesh generation should create the mathematical model of the brain in which the various tissues (white matter, grey matter, cerebrospinal fluid …) are clearly located in the correct positions. 2 – Reliable and fast simulation of the surgical process. Computational mechanics will be the crucial aspect of the entire procedure. New algorithms will be used to simulate the mechanical behaviour of cutting through cerebral tissues. 3 – Real time provision of visual and haptic feedback A sophisticated human-machine interface based on ergonomics and psychology will provide the feedback to the surgeon. The present work will address in particular point 2. Modelling the cutting of soft tissue in a structure as complex as the human brain is an extremely challenging problem in computational mechanics. The finite element method (FEM), that accurately represents complex geometries and accounts for material and geometrical nonlinearities, is the most used computational tool to simulate the mechanical response of soft tissues. However, the main drawback of FEM lies in the mechanics theory on which it is based, classical continuum Mechanics, which assumes matter is a continuum with no discontinuity. FEM must resort to complex tools such as pre-defined cohesive zones, external phase-field variables, and demanding remeshing techniques to include discontinuities. However, all approaches to equip FEM computational methods with the capability to describe material separation, such as interface elements with cohesive zone models, X-FEM, element erosion, phase-field, have some drawbacks that make them unsuitable for surgery simulation. Interface elements require a-priori knowledge of crack paths. The use of XFEM in 3D is cumbersome. Element erosion does not conserve mass. The Phase Field approach adopts a diffusive crack model instead of describing true tissue separation typical of surgical procedures. Modelling discontinuities, so difficult when using computational approaches based on classical continuum Mechanics, is instead easy for novel computational methods based on Peridynamics (PD). PD is a non-local theory of mechanics formulated with no use of spatial derivatives. Its governing equations are valid at points or surfaces of discontinuity, and it is, therefore especially suited to describe crack propagation and fragmentation problems. Moreover, PD does not require any criterium to decide the direction of crack propagation or the conditions for crack branching or coalescence; in the PD-based computational methods, cracks develop spontaneously in the way which is the most convenient from an energy point of view. Therefore, in PD computational methods, crack propagation in 3D is as easy as it is in 2D, with a remarkable advantage with respect to all other computational techniques.

Keywords: computational mechanics, peridynamics, finite element, biomechanics

Procedia PDF Downloads 52

Authors: Jatziri Y. Moreno-Martinez, Arturo Galvan, Xavier Chavez Cardenas, Hiram Arroyo

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Several methods have been utilized to study the prediction of cracking of concrete structural under loading. The finite element analysis is an alternative that shows good results. The aim of this work was the numerical study of the width crack in reinforced concrete beams with dapped ends, these are frequently found in bridge girders and precast concrete construction. Properly restricting cracking is an important aspect of the design in dapped ends, it has been observed that the cracks that exceed the allowable widths are unacceptable in an aggressive environment for reinforcing steel. For simulating the crack width, the discrete crack approach was considered by means of a Cohesive Zone (CZM) Model using a function to represent the crack opening. Two cases of dapped-end were constructed and tested in the laboratory of Structures and Materials of Engineering Institute of UNAM. The first case considers a reinforcement based on hangers as well as on vertical and horizontal ring, the second case considers 50% of the vertical stirrups in the dapped end to the main part of the beam were replaced by an equivalent area (vertically projected) of diagonal bars under. The loading protocol consisted on applying symmetrical loading to reach the service load. The models were performed using the software package ANSYS v. 16.2. The concrete structure was modeled using three-dimensional solid elements SOLID65 capable of cracking in tension and crushing in compression. Drucker-Prager yield surface was used to include the plastic deformations. The reinforcement was introduced with smeared approach. Interface delamination was modeled by traditional fracture mechanics methods such as the nodal release technique adopting softening relationships between tractions and the separations, which in turn introduce a critical fracture energy that is also the energy required to break apart the interface surfaces. This technique is called CZM. The interface surfaces of the materials are represented by a contact elements Surface-to-Surface (CONTA173) with bonded (initial contact). The Mode I dominated bilinear CZM model assumes that the separation of the material interface is dominated by the displacement jump normal to the interface. Furthermore, the opening crack was taken into consideration according to the maximum normal contact stress, the contact gap at the completion of debonding, and the maximum equivalent tangential contact stress. The contact elements were placed in the crack re-entrant corner. To validate the proposed approach, the results obtained with the previous procedure are compared with experimental test. A good correlation between the experimental and numerical Load-Displacement curves was presented, the numerical models also allowed to obtain the load-crack width curves. In these two cases, the proposed model confirms the capability of predicting the maximum crack width, with an error of ± 30 %. Finally, the orientation of the crack is a fundamental for the prediction of crack width. The results regarding the crack width can be considered as good from the practical point view. Load-Displacement curve of the test and the location of the crack were able to obtain favorable results.

Keywords: cohesive zone model, dapped-end beams, discrete crack approach, finite element analysis

Procedia PDF Downloads 140

Authors: Omar Nouri, Wafa Mnejja, Fatma Dhouib, Syrine Zouari, Wicem Siala, Ilhem Charfeddine, Afef Khanfir, Leila Farhat, Nejla Fourati, Jamel Daoud

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Purpose and Objective: Intensity modulated radiation (IMRT) technique, associated with induction chemotherapy (IC) and/or concomitant chemotherapy (CC), is actually the recommended treatment modality for nasopharyngeal carcinomas (NPC). The aim of this study was to evaluate the therapeutic results and the patterns of relapse with this treatment protocol. Material and methods: A retrospective monocentric study of 145 patients with NPC treated between June 2016 and July 2021. All patients received IMRT with integrated simultaneous boost (SIB) of 33 daily fractions at a dose of 69.96 Gy for high-risk volume, 60 Gy for intermediate risk volume and 54 Gy for low-risk volume. The high-risk volume dose was 66.5 Gy in children. Survival analysis was performed according to the Kaplan-Meier method, and the Log-rank test was used to compare factors that may influence survival. Results: Median age was 48 years (11-80) with a sex ratio of 2.9. One hundred-twenty tumors (82.7%) were classified as stages III-IV according to the 2017 UICC TNM classification. Ten patients (6.9%) were metastatic at diagnosis. One hundred-thirty-five patient (93.1%) received IC, 104 of which (77%) were TPF-based (taxanes, cisplatin and 5 fluoro-uracil). One hundred-thirty-eight patient (95.2%) received CC, mostly cisplatin in 134 cases (97%). After a median follow-up of 50 months [22-82], 46 patients (31.7%) had a relapse: 12 (8.2%) experienced local and/or regional relapse after a median of 18 months [6-43], 29 (20%) experienced distant relapse after a median of 9 months [2-24] and 5 patients (3.4%) had both. Thirty-five patients (24.1%) died, including 5 (3.4%) from a cause other than their cancer. Three-year overall survival (OS), cancer specific survival, disease free survival, metastasis free survival and loco-regional free survival were respectively 78.1%, 81.3%, 67.8%, 74.5% and 88.1%. Anatomo-clinic factors predicting OS were age > 50 years (88.7 vs. 70.5%; p=0.004), diabetes history (81.2 vs. 66.7%; p=0.027), UICC N classification (100 vs. 95 vs. 77.5 vs. 68.8% respectively for N0, N1, N2 and N3; p=0.008), the practice of a lymph node biopsy (84.2 vs. 57%; p=0.05), and UICC TNM stages III-IV (93.8 vs. 73.6% respectively for stage I-II vs. III-IV; p=0.044). Therapeutic factors predicting OS were a number of CC courses (less than 4 courses: 65.8 vs. 86%; p=0.03, less than 5 courses: 71.5 vs. 89%; p=0.041), a weight loss > 10% during treatment (84.1 vs. 60.9%; p=0.021) and a total cumulative cisplatin dose, including IC and CC, < 380 mg/m² (64.4 vs. 87.6%; p=0.003). Radiotherapy delay and total duration did not significantly affect OS. No grade 3-4 late side effects were noted in the evaluable 127 patients (87.6%). The most common toxicity was dry mouth which was grade 2 in 47 cases (37%) and grade 1 in 55 cases (43.3%).Conclusion: IMRT for nasopharyngeal carcinoma granted a high loco-regional control rate for patients during the last five years. However, distant relapses remain frequent and conditionate the prognosis. We identified many anatomo-clinic and therapeutic prognosis factors. Therefore, high-risk patients require a more aggressive therapeutic approach, such as radiotherapy dose escalation or adding adjuvant chemotherapy.

Keywords: therapeutic results, prognostic factors, intensity-modulated radiotherapy, nasopharyngeal carcinoma

Procedia PDF Downloads 41

Authors: A. V. Harutyunyan

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The investigation results of rocks at high pressures and temperatures have revealed the intervals of changes of seismic waves and density, as well as some processes taking place in rocks. In the serpentinized rocks, as a consequence of dehydration, abrupt changes in seismic waves and density have been recorded. Hydrogen-bearing components are released which combine with carbon-bearing components. As a result, hydrocarbons formed. The investigated samples are smelted. Then, geofluids and hydrocarbons migrate into the upper horizons of the Earth crust by the deep faults. Then their differentiation and accumulation in the jointed rocks of the faults and in the layers with collecting properties takes place. Under the majority of the hydrocarbon deposits, at a certain depth, magmatic centers and deep faults are recorded. The investigation results of the serpentinized rocks with numerous geological-geophysical factual data allow understanding that hydrocarbons are mainly formed in both the offshore part of the ocean and at different depths of the continental crust. Experiments have also shown that the dehydration of the serpentinized rocks is accompanied by an explosion with the instantaneous increase in pressure and temperature and smelting the studied rocks. According to numerous publications, hydrocarbons and diamonds are formed in the upper part of the mantle, at the depths of 200-400km, and as a consequence of geodynamic processes, they rise to the upper horizons of the Earth crust through narrow channels. However, the genesis of metamorphogenic diamonds and the diamonds found in the lava streams formed within the Earth crust, remains unclear. As at dehydration, super high pressures and temperatures arise. It is assumed that diamond crystals are formed from carbon containing components present in the dehydration zone. It can be assumed that besides the explosion at dehydration, secondary explosions of the released hydrogen take place. The process is naturally accompanied by seismic phenomena, causing earthquakes of different magnitudes on the surface. As for the diamondiferous kimberlites, it is well-known that the majority of them are located within the ancient shield and platforms not obligatorily connected with the deep faults. The kimberlites are formed at the shallow location of dehydrated masses in the Earth crust. Kimberlites are younger in respect of containing ancient rocks containing serpentinized bazites and ultrbazites of relicts of the paleooceanic crust. Sometimes, diamonds containing water and hydrocarbons showing their simultaneous genesis are found. So, the geofluids, hydrocarbons and diamonds, according to the new concept put forward, are formed simultaneously from serpentinized rocks as a consequence of their dehydration at different depths of the Earth crust. Based on the concept proposed by us, we suggest discussing the following: -Genesis of gigantic hydrocarbon deposits located in the offshore area of oceans (North American, Mexican Gulf, Cuanza-Kamerunian, East Brazilian etc.) as well as in the continental parts of different mainlands (Kanadian-Arctic Caspian, East Siberian etc.) - Genesis of metamorphogenic diamonds and diamonds in the lava streams (Guinea-Liberian, Kokchetav, Kanadian, Kamchatka-Tolbachinian, etc.).

Keywords: dehydration, diamonds, hydrocarbons, serpentinites

Procedia PDF Downloads 316

Authors: Amrei Voelkner, Nils Peters, Thomas Mannheim

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The worldwide exponential growth of the population and the simultaneous increasing food production requires the strategic realization of sustainable and improved cultivation systems to ensure the fertility of arable land and to guarantee the food supply for the whole world. To fulfill this target, large quantities of fertilizers have to be applied to the field, but the long-term environmental impacts remain uncertain. Thus, a combined system would be necessary to increase the nutrient availability for plants while reducing nutrient losses (e.g. NO3- by leaching) to the environment. To enhance the nutrient efficiency, polymer coated fertilizer with a controlled release behavior have been developed. This kind of fertilizer ensures a delayed release of nutrients to synchronize the nutrient supply with the demand of different crops. In the last decades, research focused primarily on semi-permeable polyurethane coatings, which remain in the soil for a long period after the complete solvation of the fertilizer core. Within the implementation of the new European Regulation Directive the replacement of non-degradable synthetic polymers by degradable coatings is necessary. It was, therefore, the objective of this study to develop a total biodegradable polymer (to CO2 and H2O) coating according to ISO 17556 and to compare the retarding effect of the biodegradable coatings with commercially available non-degradable products. To investigate the effect of ten selected coated urea fertilizer on the yield of annual ryegrass and maize, the fresh and dry mass, the percentage of total nitrogen and main nutrients were analyzed in greenhouse experiments in sixfold replications using near-infrared spectroscopy. For the experiments, a homogenized and air-dried loamy sand (Cambic Luvisol) was equipped with a basic fertilization of P, K, Mg and S. To investigate the effect of nitrogen level increase, three levels (80%, 100%, 120%) were established, whereas the impact of CRF granules was determined using a N-level of 100%. Additionally, leaching of NO3- from pots planted with annual ryegrass was examined to evaluate the retention capacity of urea by the polymer coating. For this, leachate from Kick-Brauckmann-Pots was collected daily and analyzed for total nitrogen, NO3- and NH4+ in twofold repetition once a week using near-infrared spectroscopy. We summarize from the results that the coated fertilizer have a clear impact on the yield of annual ryegrass and maize. Compared to the control, an increase of fresh and dry mass could be recognized. Partially, the non-degradable coatings showed a retarding effect for a longer period, which was however reflected by a lower fresh and dry mass. It was ascertained that the percentage of leached-out nitrate could be reduced markedly. As a conclusion, it could be pointed out that the impact of coated fertilizer of all polymer types might contribute to a reduction of negative environmental impacts in addition to their fertilizing effect.

Keywords: biodegradable polymers, coating, enhanced efficiency fertilizers, nitrate leaching

Procedia PDF Downloads 249

Authors: Gerd Wilsch, Tobias Guenther, Tobias Voelker

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In view of the ageing of vital infrastructure facilities, a reliable condition assessment of concrete structures is becoming of increasing interest for asset owners to plan timely and appropriate maintenance and repair interventions. For concrete structures, reinforcement corrosion induced by penetrating chlorides is the dominant deterioration mechanism affecting the serviceability and, eventually, structural performance. The determination of the quantitative chloride ingress is required not only to provide valuable information on the present condition of a structure, but the data obtained can also be used for the prediction of its future development and associated risks. At present, wet chemical analysis of ground concrete samples by a laboratory is the most common test procedure for the determination of the chloride content. As the chloride content is expressed by the mass of the binder, the analysis should involve determination of both the amount of binder and the amount of chloride contained in a concrete sample. This procedure is laborious, time-consuming, and costly. The chloride profile obtained is based on depth intervals of 10 mm. LIBS is an economically viable alternative providing chloride contents at depth intervals of 1 mm or less. It provides two-dimensional maps of quantitative element distributions and can locate spots of higher concentrations like in a crack. The results are correlated directly to the mass of the binder, and it can be applied on-site to deliver instantaneous results for the evaluation of the structure. Examples for the application of the method in the laboratory for the investigation of diffusion and migration of chlorides, sulfates, and alkalis are presented. An example for the visualization of the Li transport in concrete is also shown. These examples show the potential of the method for a fast, reliable, and automated two-dimensional investigation of transport processes. Due to the better spatial resolution, more accurate input parameters for model calculations are determined. By the simultaneous detection of elements such as carbon, chlorine, sodium, and potassium, the mutual influence of the different processes can be determined in only one measurement. Furthermore, the application of a mobile LIBS system in a parking garage is demonstrated. It uses a diode-pumped low energy laser (3 mJ, 1.5 ns, 100 Hz) and a compact NIR spectrometer. A portable scanner allows a two-dimensional quantitative element mapping. Results show the quantitative chloride analysis on wall and floor surfaces. To determine the 2-D distribution of harmful elements (Cl, C), concrete cores were drilled, split, and analyzed directly on-site. Results obtained were compared and verified with laboratory measurements. The results presented show that the LIBS method is a valuable addition to the standard procedures - the wet chemical analysis of ground concrete samples. Currently, work is underway to develop a technical code of practice for the application of the method for the determination of chloride concentration in concrete.

Keywords: chemical analysis, concrete, LIBS, spectroscopy

Procedia PDF Downloads 88

Authors: J. Gabrielczyk, J. Kluitmann, T. Dammeyer, H. J. Jördening

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High-level expression of proteins in bacteria often causes production of insoluble protein aggregates, called inclusion bodies (IB). They contain mainly one type of protein and offer an easy and efficient way to get purified protein. On the other hand, proteins in IB are normally devoid of function and therefore need a special treatment to become active. Most refolding techniques aim at diluting the solubilizing chaotropic agents. Unfortunately, optimal refolding conditions have to be found empirically for every protein. For large-scale applications, a simple refolding process with high yields and high final enzyme concentrations is still missing. The constructed plasmid pASK-IBA63b containing the sequence of fructosyltransferase (FTF, EC 2.4.1.162) from Bacillus subtilis NCIMB 11871 was transformed into E. coli BL21 (DE3) Rosetta. The bacterium was cultivated in a fed-batch bioreactor. The produced FTF was obtained mainly as IB. For refolding experiments, five different amounts of IBs were solubilized in urea buffer with protein concentration of 0.2-8.5 g/L. Solubilizates were refolded with batch or continuous dialysis. The refolding yield was determined by measuring the protein concentration of the clear supernatant before and after the dialysis. Particle size was measured by dynamic light scattering. We tested the solubilization properties of fructosyltransferase IBs. The particle size measurements revealed that the solubilization of the aggregates is achieved at urea concentration of 5M or higher and confirmed by absorption spectroscopy. All results confirm previous investigations that refolding yields are dependent upon initial protein concentration. In batch dialysis, the yields dropped from 67% to 12% and 72% to 19% for continuous dialysis, in relation to initial concentrations from 0.2 to 8.5 g/L. Often used additives such as sucrose and glycerol had no effect on refolding yields. Buffer screening indicated a significant increase in activity but also temperature stability of FTF with citrate/phosphate buffer. By adding citrate to the dialysis buffer, we were able to increase the refolding yields to 82-47% in batch and 90-74% in the continuous process. Further experiments showed that in general, higher ionic strength of buffers had major impact on refolding yields; doubling the buffer concentration increased the yields up to threefold. Finally, we achieved corresponding high refolding yields by reducing the chamber volume by 75% and the amount of buffer needed. The refolded enzyme had an optimal activity of 12.5±0.3 x104 units/g. However, detailed experiments with native FTF revealed a reaggregation of the molecules and loss in specific activity depending on the enzyme concentration and particle size. For that reason, we actually focus on developing a process of simultaneous enzyme refolding and immobilization. The results of this study show a new approach in finding optimal refolding conditions for inclusion bodies at high concentrations. Straightforward buffer screening and increase of the ionic strength can optimize the refolding yield of the target protein by 400%. Gentle removal of chaotrope with continuous dialysis increases the yields by an additional 65%, independent of the refolding buffer applied. In general time is the crucial parameter for successful refolding of solubilized proteins.

Keywords: dialysis, inclusion body, refolding, solubilization

Procedia PDF Downloads 275

Authors: Sandhya Parameswaran, Prajakta Dhuri

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Despite great improvements in health care, the world oral health report states that dental problems still persist, particularly among underprivileged groups in both developing and developed countries. Dental caries and periodontal diseases are identified as the most important oral health problems globally. Acidic foods and beverages can affect natural teeth, and chronic exposure often leads to the development of dental erosion, abrasion, and decay. In recent years, there has been an increased interest toward essential oils. These are secondary metabolites and possess antibacterial, antifungal and antioxidant properties. Essential oils are volatile and chemically unstable in the presence of air, light, moisture and high temperature. Hence many novel methods like a liposomal encapsulation of oils have been introduced to enhance the stability and bioavailability. This research paper focuses on two essential oils, clove and cinnamon oil. Clove oil was obtained from Syzygium aromaticum Linn using clavengers apparatus. It contains eugenol and β caryophyllene. Cinnamon oil, from the barks of Cinnamomum cassia, contains cinnamaldehyde, The objective of the current research was to develop a liposomal carrier system containing clove and cinnamon oil and study their synergistic activity against dental pathogens when formulated as a gel. Methodology: The essential oil were first tested for their antimicrobial activity against dental pathogens, Lactobacillus acidophillus (MTCC No. 10307, MRS broth) and Streptococcus Mutans (MTCC No .890, Brain Heart Infusion agar). The oils were analysed by UV spectroscopy for eugenol and cinnamaldehyde content. Standard eugenol was linear between 5ppm to 25ppm at 282nm and standard cinnamaldehde from 1ppm to 5pmm at 284nm. The concentration of eugenol in clove oil was found to be 62.65 % w/w, and that of cinnamaldehyde was found to be 5.15%s w/w. The oils were then formulated into liposomes. Liposomes were prepared by thin film hydration method using Phospholipid, Cholesterol, and other oils dissolved in a chloroform methanol (3:1) mixture. The organic solvent was evaporated in a rotary evaporator above lipid transition temperature. The film was hydrated with phosphate buffer (pH 5.5).The various batches of liposomes were characterized and compared for their size, loading rate, encapsulation efficiency and morphology. The prepared liposomes when evaluated for entrapment efficiency showed 65% entrapment for clove and 85% for cinnamon oil. They were also tested for their antimicrobial activity against dental pathogens and their synergistic activity studied. Based on the activity and the entrapment efficiency the amount of liposomes required to prepare 1gm of the gel was calculated. The gel was prepared using a simple ointment base and contained 0.56% of cinnamon and clove liposomes. A simultaneous method of analysis for eugenol and cinnamaldehyde.was then developed using HPLC. The prepared gels were then studied for their stability as per ICH guidelines. Conclusion: It was found that liposomes exhibited spherical shaped vesicles and protected the essential oil from degradation. Liposomes, therefore, constitute a suitable system for encapsulation of volatile, unstable essential oil constituents.

Keywords: cinnamon oil, clove oil, dental caries, liposomes

Procedia PDF Downloads 161

Authors: Andrey Cherdantsev, Mikhail Cherdantsev, Sergey Isaenkov, Dmitriy Markovich

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In annular gas-liquid flow, liquid flows as a film along pipe walls sheared by high-velocity gas stream. Film surface is covered by large-scale disturbance waves which affect pressure drop and heat transfer in the system and are necessary for entrainment of liquid droplets from film surface into the core of gas stream. Disturbance waves are a highly complex and their properties are affected by numerous parameters. One of such aspects is flow development, i.e., change of flow properties with the distance from the inlet. In the present work, this question is studied using brightness-based laser-induced fluorescence (BBLIF) technique. This method enables one to perform simultaneous measurements of local film thickness in large number of points with high sampling frequency. In the present experiments first 50 cm of upward and downward annular flow in a vertical pipe of 11.7 mm i.d. is studied with temporal resolution of 10 kHz and spatial resolution of 0.5 mm. Thus, spatiotemporal evolution of film surface can be investigated, including scenarios of formation, acceleration and coalescence of disturbance waves. The behaviour of disturbance waves' velocity depending on phases flow rates and downstream distance was investigated. Besides measuring the waves properties, the goal of the work was to investigate the interrelation between disturbance waves properties and integral characteristics of the flow such as interfacial shear stress and flow rate of dispersed phase. In particular, it was shown that the initial acceleration of disturbance waves, defined by the value of shear stress, linearly decays with downstream distance. This lack of acceleration which may even lead to deceleration is related to liquid entrainment. Flow rate of disperse phase linearly grows with downstream distance. During entrainment events, liquid is extracted directly from disturbance waves, reducing their mass, area of interaction to the gas shear and, hence, velocity. Passing frequency of disturbance waves at each downstream position was measured automatically with a new algorithm of identification of characteristic lines of individual disturbance waves. Scenarios of coalescence of individual disturbance waves were identified. Transition from initial high-frequency Kelvin-Helmholtz waves appearing at the inlet to highly nonlinear disturbance waves with lower frequency was studied near the inlet using 3D realisation of BBLIF method in the same cylindrical channel and in a rectangular duct with cross-section of 5 mm by 50 mm. It was shown that the initial waves are generally two-dimensional but are promptly broken into localised three-dimensional wavelets. Coalescence of these wavelets leads to formation of quasi two-dimensional disturbance waves. Using cross-correlation analysis, loss and restoration of two-dimensionality of film surface with downstream distance were studied quantitatively. It was shown that all the processes occur closer to the inlet at higher gas velocities.

Keywords: annular flow, disturbance waves, entrainment, flow development

Procedia PDF Downloads 229

Authors: Joao Ferreira Do Rosario, Maria De Lurdes Calisto, Ana Teresa Machado, Nuno Gustavo, Rui Gonçalves

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The tourism sector is increasingly important to the economic performance of countries and a relevant theme to academic research, increasing the importance of understanding how and why tourists evaluate tourism locations. The city of Lisbon is currently a tourist destination of excellence in the European and world-wide panorama, registering a significant growth of the economic weight of its tourist activities in the Gross Added Value of the region. Although there is research on the feedback of those who visit tourist sites and different methodologies for studying tourist sites have been applied, this research seeks to be innovative in the objective of obtaining insights on the competitiveness in terms of attractiveness of the city of Lisbon as a tourist destination, based the feedback of tourists in the Facebook pages of the most visited museums and monuments of Lisbon, an interpretation that is relevant in the development of strategies of tourist attraction. The intangible dimension of the tourism offer, due to its unique condition of simultaneous production and consumption, makes eWOM particularly relevant. The testimony of consumers is thus a decisive factor in the decision-making and buying process in tourism. Online social networks are one of the most used platforms for tourists to evaluate the attractiveness's points of a tourism destination (e.g. cultural and historical heritage), with this user-generated feedback enabling relevant information about the customer-tourists. This information is related to the tourist experience representing the true voice of the customer. Furthermore, this voice perceived by others as genuine, opposite to marketing messages, may have a powerful word-of-mouth influence on other potential tourists. The relevance of online reviews sharing, however, becomes particularly complex, considering social media users’ different profiles or the possible and different sources of information available, as well as their associated reputation associated with each source. In the light of these trends, our research focuses on the tourists’ feedback on Facebook pages of the most visited museums and monuments of Lisbon that contribute to its attractiveness as a tourism destination. Sentiment Analysis is the methodology selected for this research, using public available information in the online context, which was deemed as an appropriate non-participatory observation method. Data will be collected from two museums (Museu dos Coches and Museu de Arte Antiga) and three monuments ((Mosteiro dos Jerónimos, Torre de Belém and Panteão Nacional) Facebook pages during a period of one year. The research results will help in the evaluation of the considered places by the tourists, their contribution to the city attractiveness and present insights helpful for the management decisions regarding this museums and monuments. The results of this study will also contribute to a better knowledge of the tourism sector, namely the identification of attributes in the evaluation and choice of the city of Lisbon as a tourist destination. Further research will evaluate the Lisbon attraction points for tourists in different categories beyond museums and monuments, will also evaluate the tourist feedback from other sources like TripAdvisor and apply the same methodology in other cities and country regions.

Keywords: Lisbon tourism, opinion mining, sentiment analysis, tourism location attractiveness evaluation

Procedia PDF Downloads 204

Authors: Ruth Goldschmidt, Vyacheslav Kalchenko, Lilah Agemy, Rachel Elmoalem, Avigdor Scherz

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Vascular Targeted Photodynamic therapy (VTP) is a new modality for selective cancer treatment that leads to the complete tumor ablation. A photosensitizer, a bacteriochlorophyll derivative in our case, is first administered to the patient and followed by the illumination of the tumor area, by a near-IR laser for its photoactivation. The photoactivated drug releases reactive oxygen species (ROS) in the circulation, which reacts with blood cells and the endothelium leading to the occlusion of the blood vasculature. If the blood vessels are only partially closed, the tumor may recover, and cancer cells could survive. On the other hand, excessive treatment may lead to toxicity of healthy tissues nearby. Simultaneous VTP monitoring and image processing independent of the photoexcitation laser has not yet been reported, to our knowledge. Here we present a method for blood flow monitoring, using a real-time laser speckle imaging (RTLSI) in the tumor during VTP. We have synthesized over the years a library of bacteriochlorophyll derivatives, among them WST11 and STL-6014. Both are water soluble derivatives that are retained in the blood vasculature through their partial binding to HSA. WST11 has been approved in Mexico for VTP treatment of prostate cancer at a certain drug dose, and time/intensity of illumination. Application to other bacteriochlorophyll derivatives or other cancers may require different treatment parameters (such as light/drug administration). VTP parameters for STL-6014 are still under study. This new derivative mainly differs from WST11 by its lack of the central Palladium, and its conjugation to an Arg-Gly-Asp (RGD) sequence. RGD is a tumor-specific ligand that is used for targeting the necrotic tumor domains through its affinity to αVβ3 integrin receptors. This enables the study of cell-targeted VTP. We developed a special RTLSI module, based on Labview software environment for data processing. The new module enables to acquire raw laser speckle images and calculate the values of the laser temporal statistics of time-integrated speckles in real time, without additional off-line processing. Using RTLSI, we could monitor the tumor’s blood flow following VTP in a CT26 colon carcinoma ear model. VTP with WST11 induced an immediate slow down of the blood flow within the tumor and a complete final flow arrest, after some sporadic reperfusions. If the irradiation continued further, the blood flow stopped also in the blood vessels of the surrounding healthy tissue. This emphasizes the significance of light dose control. Using our RTLSI system, we could prevent any additional healthy tissue damage by controlling the illumination time and restrict blood flow arrest within the tumor only. In addition, we found that VTP with STL-6014 was the most effective when the photoactivation was conducted 4h post-injection, in terms of tumor ablation success in-vivo and blood vessel flow arrest. In conclusion, RTSLI application should allow to optimize VTP efficacy vs. toxicity in both the preclinical and clinical arenas.

Keywords: blood vessel occlusion, cancer treatment, photodynamic therapy, real time imaging

Procedia PDF Downloads 197

Authors: Jing Zhang, Yvonne Reinwald, Nick Poulson, Alicia El Haj, Chung See, Mike Somekh, Melissa Mather

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Biomedical imaging of native tissue using light offers the potential to obtain excellent structural and functional information in a non-invasive manner with good temporal resolution. Image contrast can be derived from intrinsic absorption, fluorescence, or scatter, or through the use of extrinsic contrast. A major challenge in applying optical microscopy to in vivo tissue imaging is the effects of light attenuation which limits light penetration depth and achievable imaging resolution. Recently Selective Plane Illumination Microscopy (SPIM) has been used to map the 3D distribution of fluorophores dispersed in biological structures. In this approach, a focused sheet of light is used to illuminate the sample from the side to excite fluorophores within the sample of interest. Images are formed based on detection of fluorescence emission orthogonal to the illumination axis. By scanning the sample along the detection axis and acquiring a stack of images, 3D volumes can be obtained. The combination of rapid image acquisition speeds with the low photon dose to samples optical sectioning provides SPIM is an attractive approach for imaging biological samples in 3D. To date all implementations of SPIM rely on the use of fluorescence reporters be that endogenous or exogenous. This approach has the disadvantage that in the case of exogenous probes the specimens are altered from their native stage rendering them unsuitable for in vivo studies and in general fluorescence emission is weak and transient. Here we present for the first time to our knowledge a label-free implementation of SPIM that has downstream applications in the clinical setting. The experimental set up used in this work incorporates both label-free and fluorescent illumination arms in addition to a high specification camera that can be partitioned for simultaneous imaging of both fluorescent emission and scattered light from intrinsic sources of optical contrast in the sample being studied. This work first involved calibration of the imaging system and validation of the label-free method with well characterised fluorescent microbeads embedded in agarose gel. 3D constructs of mammalian cells cultured in agarose gel with varying cell concentrations were then imaged. A time course study to track cell proliferation in the 3D construct was also carried out and finally a native tissue sample was imaged. For each sample multiple images were obtained by scanning the sample along the axis of detection and 3D maps reconstructed. The results obtained validated label-free SPIM as a viable approach for imaging cells in a 3D gel construct and native tissue. This technique has the potential use in a near-patient environment that can provide results quickly and be implemented in an easy to use manner to provide more information with improved spatial resolution and depth penetration than current approaches.

Keywords: bioimaging, optics, selective plane illumination microscopy, tissue imaging

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Authors: Maria De Los Angeles Rodriguez Cadena

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This paper discusses how the fictional versions of women of the past contribute to advance today’s ideas of social justice, personal freedom and emancipation as well as to highlight the creative challenge of constructing people and events on fictional narratives on television that incorporate multiple and simultaneous layers of meaning and complexity. This project builds on existing scholarship on audiovisual texts by exploring an influential but under-studied director. In two Mexican television series, Patricia Arriaga Jordan, an award-winning television producer, scriptwriter and director, constructs the life of two outstanding women that have played an influential role in national history and captured Mexican’s popular imagination for generations: Sor Juana Inés de la Cruz, and Malinche. Malinche (2018) tells the story of an extraordinary indigenous woman, Malintzin, during the Spanish Conquest (1511-1550) that is considered to have played a key role in the fall of the Aztec empire by acting as translator, negotiator and cultural mediator for the Spanish conquerors. Juana Ines (2016) portrays Sor Juana, a poet, essayist, playwright, theologian, philosopher, nun, of XVII century colonial Mexico, one of the brightest minds of her time, and now recognized as the first feminist of the Americas who wrote on the rights of women to an education, religious authority and feminist advocacy. Both series, as fictional narratives that recreate defining historical periods, specific events and relevant characters in the History of Mexico can be read as an example of what is called texts of cultural memory. A cultural memory text is a narrative that bonds the concepts of history, identity and belonging, and that is realized and disseminated through symbolic systems such as written documents, visual images, and dramatic representation. Cultural memory, through its narratives of historical fiction, emphasizes memory processes (historiography) and its implications and artifacts (cultural memory) mainly through the medial frameworks of remembering, which are the medial process by which memories (narratives, documents) participate in public knowledge and become collective memory. Historical fiction on television not only creates a portrayal of the past related to the real lives of protagonists, but it also significantly contributes to understand the past as an ever-evolving entity that highlights both, the necessary connection with the present as part of a developing sense of collective identity and belonging, as well as the relevance of the medium in which the past is represented and that ultimately supports the process of historical awareness. Through the emblematic recreation of national heroines and historical events in the unique context of historical drama on television, those texts constitute a venue where concepts of the past and the traditionally established ideas about history and heroines are highlighted, questioned and transformed.

Keywords: cultural memory, historical fiction, Mexico, television, women directors

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Authors: Morello Sara, Pederiva Sabina, Bianchi Manila, Martucci Francesca, Marchis Daniela, Decastelli Lucia

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Vegetables represent an integral part of a healthy diet due to their valuable nutritional properties and the growth in consumer demand in recent years is particularly remarkable for a diet rich in vitamins and micronutrients. However, plant-based products are involved in several food outbreaks connected to various sources of contamination and quite often, bacteria responsible for side effects showed high resistance to antibiotics. The abuse of antibiotics can be one of the main mechanisms responsible for increasing antibiotic resistance (AR). Plants grown for food use can be contaminated directly by spraying antibiotics on crops or indirectly by treatments with antibiotics due to the use of manure, which may contain both antibiotics and genes of antibiotic resistance (ARG). Antibiotic residues could represent a potential way of human health risk due to exposure through the consumption of plant-based foods. The presence of antibiotic-resistant bacteria might pose a particular risk to consumers. The present work aims to investigate through a multidisciplinary approach the occurrence of ARG by means of a biomolecular approach (PCR) and the prevalence of antibiotic residues using a multi residues LC-MS/MS method, both in different plant-based products. During the period from July 2020 to October 2021, a total of 74 plant samples (33 lettuces and 41 tomatoes) were collected from 57 farms located throughout the Piedmont area, and18 out of 74 samples (11 lettuces and 7 tomatoes) were selected to LC-MS/MS analyses. DNA extracted (ExtractME, Blirt, Poland) from plants used on crops and isolated bacteria were analyzed with 6 sets of end-point multiplex PCR (Qiagen, Germany) to detect the presence of resistance genes of the main antibiotic families, such as tet genes (tetracyclines), bla (β-lactams) and mcr (colistin). Simultaneous detection of 43 molecules of antibiotics belonging to 10 different classes (tetracyclines, sulphonamides, quinolones, penicillins, amphenicols, macrolides, pleuromotilines, lincosamides, diaminopyrimidines) was performed using Exion LC system AB SCIEX coupled to a triple quadrupole mass spectrometer QTRAP 5500 from AB SCIEX. The PCR assays showed the presence of ARG in 57% (n=42): tetB (4.8%; n=2), tetA (9.5%; n=4), tetE (2.4%; n=1), tetL (12%; n=5), tetM (26%; n=11), blaSHV (21.5%; n=9), blaTEM (4.8%; n =2) and blaCTX-M (19%; n=8). In none of the analyzed samples was the mcr gene responsible for colistin resistance detected. Results obtained from LC-MS/MS analyses showed that none of the tested antibiotics appear to exceed the LOQ (100 ppb). Data obtained confirmed the presence of bacterial populations containing antibiotic resistance determinants such as tet gene (tetracycline) and bla genes (beta-lactams), widely used in human medicine, which can join the food chain and represent a risk for consumers, especially with raw products. The presence of traces of antibiotic residues in vegetables, in concentration below the LOQ of the LC-MS/MS method applied, cannot be excluded. In conclusion, traces of antibiotic residues could be a health risk to the consumer due to potential involvement in the spread of AR. PCR represents a useful and effective approach to characterize and monitor AR carried by bacteria from the entire food chain.

Keywords: plant-based products, ARG, PCR, antibiotic residues

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Authors: V. Giacometti, R. Mirabelli, V. Patera, D. Pinci, A. Sarti, A. Sciubba, G. Traini, M. Marafini

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The particle therapy (PT) is a very modern technique of non invasive radiotherapy mainly devoted to the treatment of tumours untreatable with surgery or conventional radiotherapy, because localised closely to organ at risk (OaR). Nowadays, PT is available in about 55 centres in the word and only the 20\% of them are able to treat with carbon ion beam. However, the efficiency of the ion-beam treatments is so impressive that many new centres are in construction. The interest in this powerful technology lies to the main characteristic of PT: the high irradiation precision and conformity of the dose released to the tumour with the simultaneous preservation of the adjacent healthy tissue. However, the beam interactions with the patient produce a large component of secondary particles whose additional dose has to be taken into account during the definition of the treatment planning. Despite, the largest fraction of the dose is released to the tumour volume, a non-negligible amount is deposed in other body regions, mainly due to the scattering and nuclear interactions of the neutrons within the patient body. One of the main concerns in PT treatments is the possible occurrence of secondary malignant neoplasm (SMN). While SMNs can be developed up to decades after the treatments, their incidence impacts directly life quality of the cancer survivors, in particular in pediatric patients. Dedicated Treatment Planning Systems (TPS) are used to predict the normal tissue toxicity including the risk of late complications induced by the additional dose released by secondary neutrons. However, no precise measurement of secondary neutrons flux is available, as well as their energy and angular distributions: an accurate characterization is needed in order to improve TPS and reduce safety margins. The project MONDO (MOnitor for Neutron Dose in hadrOntherapy) is devoted to the construction of a secondary neutron tracker tailored to the characterization of that secondary neutron component. The detector, based on the tracking of the recoil protons produced in double-elastic scattering interactions, is a matrix of thin scintillating fibres, arranged in layer x-y oriented. The final size of the object is 10 x 10 x 20 cm3 (squared 250µm scint. fibres, double cladding). The readout of the fibres is carried out with a dedicated SPAD Array Sensor (SBAM) realised in CMOS technology by FBK (Fondazione Bruno Kessler). The detector is under development as well as the SBAM sensor and it is expected to be fully constructed for the end of the year. MONDO will make data tacking campaigns at the TIFPA Proton Therapy Center of Trento, at the CNAO (Pavia) and at HIT (Heidelberg) with carbon ion in order to characterize the neutron component and predict the additional dose delivered on the patients with much more precision and to drastically reduce the actual safety margins. Preliminary measurements with charged particles beams and MonteCarlo FLUKA simulation will be presented.

Keywords: secondary neutrons, particle therapy, tracking detector, elastic scattering

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Authors: Alena Semeradtova, Marcel Stofik, Lucie Mareckova, Petr Maly, Ondrej Stanek, Jan Maly

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Recently, novel strategies based on so-called molecular evolution were shown to be effective for the production of various peptide ligand libraries with high affinities to molecular targets of interest comparable or even better than monoclonal antibodies. The major advantage of these peptide scaffolds is mainly their prevailing low molecular weight and simple structure. This study describes a new high-affinity binding molecules based immunesensor using a simple optical system for human serum albumin (HSA) detection as a model molecule. We present a comparison of two variants of recombinant binders based on albumin binding domain of the protein G (ABD) performed on micropatterned glass chip. Binding domains may be tailored to any specific target of interest by molecular evolution. Micropatterened glass chips were prepared using UV-photolithography on chromium sputtered glasses. Glass surface was modified by (3-aminopropyl)trietoxysilane and biotin-PEG-acid using EDC/NHS chemistry. Two variants of high-affinity binding molecules were used to detect target molecule. Firstly, a variant is based on ABD domain fused with TolA chain. This molecule is in vivo biotinylated and each molecule contains one molecule of biotin and one ABD domain. Secondly, the variant is ABD domain based on streptavidin molecule and contains four gaps for biotin and four ABD domains. These high-affinity molecules were immobilized to the chip surface via biotin-streptavidin chemistry. To eliminate nonspecific binding 1% bovine serum albumin (BSA) or 6% fetal bovine serum (FBS) were used in every step. For both variants range of measured concentrations of fluorescently labelled HSA was 0 – 30 µg/ml. As a control, we performed a simultaneous assay without high-affinity binding molecules. Fluorescent signal was measured using inverse fluorescent microscope Olympus IX 70 with COOL LED pE 4000 as a light source, related filters, and camera Retiga 2000R as a detector. The fluorescent signal from non-modified areas was substracted from the signal of the fluorescent areas. Results were presented in graphs showing the dependence of measured grayscale value on the log-scale of HSA concentration. For the TolA variant the limit of detection (LOD) of the optical immunosensor proposed in this study is calculated to be 0,20 µg/ml for HSA detection in 1% BSA and 0,24 µg/ml in 6% FBS. In the case of streptavidin-based molecule, it was 0,04 µg/ml and 0,07 µg/ml respectively. The dynamical range of the immunosensor was possible to estimate just in the case of TolA variant and it was calculated to be 0,49 – 3,75 µg/ml and 0,73-1,88 µg/ml respectively. In the case of the streptavidin-based the variant we didn´t reach the surface saturation even with the 480 ug/ml concentration and the upper value of dynamical range was not estimated. Lower value was calculated to be 0,14 µg/ml and 0,17 µg/ml respectively. Based on the obtained results, it´s clear that both variants are useful for creating the bio-recognizing layer on immunosensors. For this particular system, it is obvious that the variant based on streptavidin molecule is more useful for biosensing on glass planar surfaces. Immunosensors based on this variant would exhibit better limit of detection and wide dynamical range.

Keywords: high affinity binding molecules, human serum albumin, optical immunosensor, protein G, UV-photolitography

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Authors: Andreas Latz, Daniela Heinz, Fatima Hashemi, Melek Baygül

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Introduction: Feline leukemia virus (FeLV), feline immunodeficiency virus (FIV), and feline coronavirus (FCoV) are serious infectious diseases affecting cats worldwide. Transmission of these viruses occurs primarily through close contact with infected cats (via saliva, nasal secretions, faeces, etc.). FeLV, FIV, and FCoV infections can occur in combination and are expressed in similar clinical symptoms. Diagnosis can therefore be challenging: Symptoms are variable and often non-specific. Sick cats show very similar clinical symptoms: apathy, anorexia, fever, immunodeficiency syndrome, anemia, etc. Sample volume for small companion animals for diagnostic purposes can be challenging to collect. In addition, multiplex diagnosis of diseases can contribute to an easier, cheaper, and faster workflow in the lab as well as to the better differential diagnosis of diseases. For this reason, we wanted to develop a new diagnostic tool that utilizes less sample volume, reagents, and consumables than multiplesingleplex ELISA assays Methods: The Multiplier from Dynextechonogies (USA) has been used as platform to develop a Multiplex diagnostic tool for the detection of antibodies against FIV and FCoV/FIP and antigens for FeLV. Multiplex diagnostics. The Dynex®Multiplier®is a fully automated chemiluminescence immunoassay analyzer that significantly simplifies laboratory workflow. The Multiplier®ease-of-use reduces pre-analytical steps by combining the power of efficiently multiplexing multiple assays with the simplicity of automated microplate processing. Plastic beads have been coated with antigens for FIV and FCoV/FIP, as well as antibodies for FeLV. Feline blood samples are incubated with the beads. Read out of results is performed via chemiluminescence Results: Bead coating was optimized for each individual antigen or capture antibody and then combined in the multiplex diagnostic tool. HRP: Antibody conjugates for FIV and FCoV antibodies, as well as detection antibodies for FeLV antigen, have been adjusted and mixed. 3 individual prototyple batches of the assay have been produced. We analyzed for each disease 50 well defined positive and negative samples. Results show an excellent diagnostic performance of the simultaneous detection of antibodies or antigens against these feline diseases in a fully automated system. A 100% concordance with singleplex methods like ELISA or IFA can be observed. Intra- and Inter-Assays showed a high precision of the test with CV values below 10% for each individual bead. Accelerated stability testing indicate a shelf life of at least 1 year. Conclusion: The new tool can be used for multiplex diagnostics of the most important feline infectious diseases. Only a very small sample volume is required. Fully automation results in a very convenient and fast method for diagnosing animal diseases.With its large specimen capacity to process over 576 samples per 8-hours shift and provide up to 3,456 results, very high laboratory productivity and reagent savings can be achieved.

Keywords: Multiplex, FIV, FeLV, FCoV, FIP

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Authors: Kunal Garg, Louise Theusen Hermansan, Kanoktip Puttaraska, Oliver Hendricks, Heidi Pirttinen, Leona Gilbert

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The Germ Theory (one infectious determinant is equal to one disease) has unarguably evolved our capability to diagnose and treat infectious diseases over the years. Nevertheless, the advent of technology, climate change, and volatile human behavior has brought about drastic changes in our environment, leading us to question the relevance of the Germ Theory in our day, i.e. will vector-borne disease (VBD) sufferers produce multiple immune responses when tested for multiple microbes? Vector diseased patients producing multiple immune responses to different microbes would evidently suggest human polymicrobial infections (HPI). Ongoing diagnostic tools are exceedingly unequipped with the current research findings that would aid in diagnosing patients for polymicrobial infections. This shortcoming has caused misdiagnosis at very high rates, consequently diminishing the patient’s quality of life due to inadequate treatment. Equipped with the state-of-art scientific knowledge, PolyScan intends to address the pitfalls in current VBD diagnostics. PolyScan is a multiplex and multifunctional enzyme linked Immunosorbent assay (ELISA) platform that can test for numerous VBD microbes and allow simultaneous screening for multiple types of antibodies. To validate PolyScan, Lyme Borreliosis (LB) and spondyloarthritis (SpA) patient groups (n = 54 each) were tested for Borrelia burgdorferi, Borrelia burgdorferi Round Body (RB), Borrelia afzelii, Borrelia garinii, and Ehrlichia chaffeensis against IgM and IgG antibodies. LB serum samples were obtained from Germany and SpA serum samples were obtained from Denmark under relevant ethical approvals. The SpA group represented chronic LB stage because reactive arthritis (SpA subtype) in the form of Lyme arthritis links to LB. It was hypothesized that patients from both the groups will produce multiple immune responses that as a consequence would evidently suggest HPI. It was also hypothesized that the multiple immune response proportion in SpA patient group would be significantly larger when compared to the LB patient group across both antibodies. It was observed that 26% LB patients and 57% SpA patients produced multiple immune responses in contrast to 33% LB patients and 30% SpA patients that produced solitary immune responses when tested against IgM. Similarly, 52% LB patients and an astounding 73% SpA patients produced multiple immune responses in contrast to 30% LB patients and 8% SpA patients that produced solitary immune responses when tested against IgG. Interestingly, IgM immune dysfunction in both the patient groups was also recorded. Atypically, 6% of the unresponsive 18% LB with IgG antibody was recorded producing multiple immune responses with the IgM antibody. Similarly, 12% of the unresponsive 19% SpA with IgG antibody was recorded producing multiple immune responses with the IgM antibody. Thus, results not only supported hypothesis but also suggested that IgM may atypically prevail longer than IgG. The PolyScan concept will aid clinicians to detect patients for early, persistent, late, polymicrobial, & immune dysfunction conditions linked to different VBD. PolyScan provides a paradigm shift for the VBD diagnostic industry to follow that will drastically shorten patient’s time to receive adequate treatment.

Keywords: diagnostics, immune dysfunction, polymicrobial, TICK-TAG

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