Search results for: engineered physiology
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 423

Search results for: engineered physiology

63 Biofilm Text Classifiers Developed Using Natural Language Processing and Unsupervised Learning Approach

Authors: Kanika Gupta, Ashok Kumar

Abstract:

Biofilms are dense, highly hydrated cell clusters that are irreversibly attached to a substratum, to an interface or to each other, and are embedded in a self-produced gelatinous matrix composed of extracellular polymeric substances. Research in biofilm field has become very significant, as biofilm has shown high mechanical resilience and resistance to antibiotic treatment and constituted as a significant problem in both healthcare and other industry related to microorganisms. The massive information both stated and hidden in the biofilm literature are growing exponentially therefore it is not possible for researchers and practitioners to automatically extract and relate information from different written resources. So, the current work proposes and discusses the use of text mining techniques for the extraction of information from biofilm literature corpora containing 34306 documents. It is very difficult and expensive to obtain annotated material for biomedical literature as the literature is unstructured i.e. free-text. Therefore, we considered unsupervised approach, where no annotated training is necessary and using this approach we developed a system that will classify the text on the basis of growth and development, drug effects, radiation effects, classification and physiology of biofilms. For this, a two-step structure was used where the first step is to extract keywords from the biofilm literature using a metathesaurus and standard natural language processing tools like Rapid Miner_v5.3 and the second step is to discover relations between the genes extracted from the whole set of biofilm literature using pubmed.mineR_v1.0.11. We used unsupervised approach, which is the machine learning task of inferring a function to describe hidden structure from 'unlabeled' data, in the above-extracted datasets to develop classifiers using WinPython-64 bit_v3.5.4.0Qt5 and R studio_v0.99.467 packages which will automatically classify the text by using the mentioned sets. The developed classifiers were tested on a large data set of biofilm literature which showed that the unsupervised approach proposed is promising as well as suited for a semi-automatic labeling of the extracted relations. The entire information was stored in the relational database which was hosted locally on the server. The generated biofilm vocabulary and genes relations will be significant for researchers dealing with biofilm research, making their search easy and efficient as the keywords and genes could be directly mapped with the documents used for database development.

Keywords: biofilms literature, classifiers development, text mining, unsupervised learning approach, unstructured data, relational database

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62 Expression of CASK Antibody in Non-Mucionus Colorectal Adenocarcinoma and Its Relation to Clinicopathological Prognostic Factors

Authors: Reham H. Soliman, Noha Noufal, Howayda AbdelAal

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Calcium/calmodulin-dependent serine protein kinase (CASK) belongs to the membrane-associated guanylate kinase (MAGUK) family and has been proposed as a mediator of cell-cell adhesion and proliferation, which can contribute to tumorogenesis. CASK has been linked as a good prognostic factor with some tumor subtypes, while considered as a poor prognostic marker in others. To our knowledge, no sufficient evidence of CASK role in colorectal cancer is available. The aim of this study is to evaluate the expression of Calcium/calmodulin-dependent serine protein kinase (CASK) in non-mucinous colorectal adenocarcinoma and adenomatous polyps as precursor lesions and assess its prognostic significance. The study included 42 cases of conventional colorectal adenocarcinoma and 15 biopsies of adenomatous polyps with variable degrees of dysplasia. They were reviewed for clinicopathological prognostic factors and stained by CASK; mouse, monoclonal antibody using heat-induced antigen retrieval immunohistochemical techniques. The results showed that CASK protein was significantly overexpressed (p <0.05) in CRC compared with adenoma samples. The CASK protein was overexpressed in the majority of CRC samples with 85.7% of cases showing moderate to strong expression, while 46.7% of adenomas were positive. CASK overexpression was significantly correlated with both TNM stage and grade of differentiation (p <0.05). There was a significantly higher expression in tumor samples with early stages (I/II) rather than advanced stage (III/IV) and with low grade (59.5%) rather than high grade (40.5%). Another interesting finding was found among the adenomas group, where the stronger intensity of staining was observed in samples with high grade dysplasia (33.3%) than those of lower grades (13.3%). In conclusion, this study shows that there is significant overexpression of CASK protein in CRC as well as in adenomas with high grade dysplasia. This indicates that CASK is involved in the process of carcinogenesis and functions as a potential trigger of the adenoma-carcinoma cascade. CASK was significantly overexpressed in early stage and low-grade tumors rather than tumors with advanced stage and higher histological grades. This suggests that CASK protein is a good prognostic factor. We suggest that CASK affects CRC in two different ways derived from its physiology. CASK as part of MAGUK family can stimulate proliferation and through its cell membrane localization and as a mediator of cell-cell adhesion might contribute in tumor confinement and localization.

Keywords: CASK, colorectal cancer, overexpression, prognosis

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61 Obtaining Composite Cotton Fabric by Cyclodextrin Grafting

Authors: U. K. Sahin, N. Erdumlu, C. Saricam, I. Gocek, M. H. Arslan, H. Acikgoz-Tufan, B. Kalav

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Finishing is an important part of fabric processing with which a wide range of features are imparted to greige or colored fabrics for various end-uses. Especially, by the addition or impartation of nano-scaled particles to the fabric structure composite fabrics, a kind of composite materials can be acquired. Composite materials, generally shortened as composites or in other words composition materials, are engineered or naturally occurring materials made from two or more component materials with significantly different physical, mechanical or chemical characteristics remaining separate and distinctive at the macroscopic or microscopic scale within the end product structure. Therefore, the technique finishing which is one of the fundamental methods to be applied on fabrics for obtainment of composite fabrics with many functionalities was used in the current study with the same purpose. However, regardless of the finishing materials applied, the efficient life of finished product on offering desired feature is low, since the durability of finishes on the material is limited. Any increase in durability of these finishes on textiles would enhance the life of use for textiles, which will result in happier users. Therefore, in this study, since higher durability was desired for the finishing materials fixed on the fabrics, nano-scaled hollow structured cyclodextrins were chemically imparted by grafting to the structure of conventional cotton fabrics by the help of finishing technique in order to be fixed permanently. By this way, a processed and functionalized base fabric having potential to be treated in the subsequent processes with many different finishing agents and nanomaterials could be obtained. Henceforth, this fabric can be used as a multi-functional fabric due to the encapturing ability of cyclodextrins to molecules/particles via physical/chemical means. In this study, scoured and rinsed woven bleached plain weave 100% cotton fabrics were utilized because textiles made of cotton are the most demanded textile products in the textile market by the textile consumers in daily life. Cotton fabric samples were immersed in treating baths containing β-cyclodextrin and 1,2,3,4-butanetetracarboxylic acid and to reduce the curing temperature the catalyst sodium hypophosphite monohydrate was used. All impregnated fabric samples were pre-dried. The reaction of grafting was performed in dry state. The treated and cured fabric samples were rinsed with warm distilled water and dried. The samples were dried for 4 h and weighed before and after finishing and rinsing. Stability and durability of β-cyclodextrins on fabric surface against external factors such as washing as well as strength of functionalized fabric in terms of tensile and tear strength were tested. Presence and homogeneity of distribution of β-cyclodextrins on fabric surface were characterized.

Keywords: cotton fabric, cyclodextrine, improved durability, multifunctional composite textile

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60 A Study of the Effect of Early and Late Meal Time on Anthropometric and Biochemical Parameters in Patients of Type 2 Diabetes

Authors: Smriti Rastogi, Narsingh Verma

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Background: A vast body of research exists on the use of oral hypoglycaemic drugs, insulin injections and the like in managing diabetes but no such research exists that has taken into consideration the parameter of time restricted meal intake and its positive effects in managing diabetes. The utility of this project is immense as it offers a solution to the woes of diabetics based on circadian rhythm and normal physiology of the human body. Method: 80 Diabetics, enrolled from the Out Patient Department of Endocrinology, KGMU (King George's Medical University) were randomly divided based on consent to early dinner TRM(time restricted meal) group or not (control group). Follow up was done at six months and 12 months for anthropometric measurement, height, weight, waist-hip ratio, neck size, fasting, postprandial blood sugar, HbA1c, serum urea, serum creatinine, and lipid profile. The patient was given a clear understanding of chronomedicine and how it affects their health. A single intervention was done - the timing of dinner was at or around 7 pm for TRM group. Result: 65% of TRM group and 40 %(non- TRM) had normal HbA1c after 12 months. HbA1c in TRM Group (first visit to second follow up) had a significant p value=0.017. A p value of <0.0001 was observed on comparing the values of blood sugar (fasting) in TRM Group from the first visit and second follow up. The values of blood sugar (postprandial) in TRM Group (first visit and second follow up) showed a p-value <0.0001 (highly significant). Values of the three parameters were non- significant in the control group. Hip size(First Visit to Second Follow Up) TRM Group showed a p-value = 0.0344 (Significant) (Difference between means=2.762 ± 1.261)Detailed results of the above parameters and a few newer ones will be presented at the conference. Conclusion: Time restricted meal intake in diabetics shows promise and is worth exploring further. Time Restricted Meal intake in Type 2 diabetics has a significant effect in controlling and maintaining HbA1c as the reduction in HbA1c value was very significant in the TRM group vs. the control group. Similar highly significant results were obtained in the case of fasting and postprandial values of blood sugar in the TRM group when compared to the control group. The effects of time restricted meal intake in diabetics show promise and are worth exploring further. It is one of the first studies which have been undertaken in Indian diabetics, although the initial data obtained is encouraging yet further research and study are required to corroborate results.

Keywords: chronomedicine, diabetes, endocrinology, time restricted meal intake

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59 Physical Contact Modulation of Macrophage-Mediated Anti-Inflammatory Response in Osteoimmune Microenvironment by Pollen-Like Nanoparticles

Authors: Qing Zhang, Janak L. Pathak, Macro N. Helder, Richard T. Jaspers, Yin Xiao

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Introduction: Nanomaterial-based bone regeneration is greatly influenced by the immune microenvironment. Tissue-engineered nanomaterials mediate the inflammatory response of macrophages to regulate bone regeneration. Silica nanoparticles have been widely used in tissue engineering-related preclinical studies. However, the effect of topological features on the surface of silica nanoparticles on the immune response of macrophages remains unknown. Purposes: The aims of this research are to compare the influences of normal and pollen-like silica nano-surface topography on macrophage immune responses and to obtain insight into their potential regulatory mechanisms. Method: Macrophages (RAW 264.7 cells) were exposed to mesoporous silica nanoparticles with normal morphology (MSNs) and pollen-like morphology (PMSNs). RNA-seq, RT-qPCR, and LSCM were used to assess the changes in expression levels of immune response-related genes and proteins. SEM and TEM were executed to evaluate the contact and adherence of silica nanoparticles by macrophages. For the assessment of the immunomodulation-mediated osteogenic potential, BMSCs were cultured with conditioned medium (CM) from LPS pre-stimulated macrophage cultures treated with MSNs or PMSNs. Osteoimmunomodulatory potential of MSNs and PMSNs in vivo was tested in a mouse cranial bone osteolysis model. Results: The results of the RNA-seq, RT-qPCR, and LSCM assays showed that PMSNs inhibited the expression of pro-inflammatory genes and proteins in macrophages. SEM images showed distinct macrophage membrane surface binding patterns of MSNs and PMSNs. MSNs were more evenly dispersed across the macrophage cell membrane, while PMSNs were aggregated. PMSNs-induced macrophage anti-inflammatory response was associated with upregulation of the cell surface receptor CD28 and inhibition of ERK phosphorylation. TEM images showed that both MSNs and PMSNs could be phagocytosed by macrophages, and inhibiting nanoparticle phagocytosis did not affect the expression of anti-inflammatory genes and proteins. Moreover, PMSNs-induced conditioned medium from macrophages enhanced BMP-2 expression and osteogenic differentiation mBMSCs. Similarly, PMSNs prevented LPS-induced bone resorption via downregulation of inflammatory reaction. Conclusions: PMSNs can promote bone regeneration by modulating osteoimmunological processes through surface topography. The study offers insights into how surface physical contact cues can modulate the regulation of osteoimmunology and provides a basis for the application of nanoparticles with pollen-like morphology to affect immunomodulation in bone tissue engineering and regeneration.

Keywords: physical contact, osteoimmunology, macrophages, silica nanoparticles, surface morphology, membrane receptor, osteogenesis, inflammation

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58 A Brief Review on the Relationship between Pain and Sociology

Authors: Hanieh Sakha, Nader Nader, Haleh Farzin

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Introduction: Throughout history, pain theories have been supposed by biomedicine, especially regarding its diagnosis and treatment aspects. Therefore, the feeling of pain is not only a personal experience and is affected by social background; therefore, it involves extensive systems of signals. The challenges in emotional and sentimental dimensions of pain originate from scientific medicine (i.e., the dominant theory is also referred to as the specificity theory); however, this theory has accepted some alterations by emerging physiology. Then, Von Frey suggested the theory of cutaneous senses (i.e., Muller’s concept: the common sensation of combined four major skin receptors leading to a proper sensation) 50 years after the specificity theory. The pain pathway was composed of spinothalamic tracts and thalamus with an inhibitory effect on the cortex. Pain is referred to as a series of unique experiences with various reasons and qualities. Despite the gate control theory, the biological aspect overcomes the social aspect. Vrancken provided a more extensive definition of pain and found five approaches: Somatico-technical, dualistic body-oriented, behaviorist, phenomenological, and consciousness approaches. The Western model combined physical, emotional, and existential aspects of the human body. On the other hand, Kotarba felt confused about the basic origins of chronic pain. Freund demonstrated and argued with Durkhemian about the sociological approach to emotions. Lynch provided a piece of evidence about the correlation between cardiovascular disease and emotionally life-threatening occurrences. Helman supposed a distinction between private and public pain. Conclusion: The consideration of the emotional aspect of pain could lead to effective, emotional, and social responses to pain. On the contrary, the theory of embodiment is based on the sociological view of health and illness. Social epidemiology shows an imbalanced distribution of health, illness, and disability among various social groups. The social support and socio-cultural level can result in several types of pain. It means the status of athletes might define their pain experiences. Gender is one of the important contributing factors affecting the type of pain (i.e., females are more likely to seek health services for pain relief.) Chronic non-cancer pain (CNCP) has become a serious public health issue affecting more than 70 million people globally. CNCP is a serious public health issue which is caused by the lack of awareness about chronic pain management among the general population.

Keywords: pain, sociology, sociological, body

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57 DEKA-1 a Dose-Finding Phase 1 Trial: Observing Safety and Biomarkers using DK210 (EGFR) for Inoperable Locally Advanced and/or Metastatic EGFR+ Tumors with Progressive Disease Failing Systemic Therapy

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

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

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

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56 Mechanism of Veneer Colouring for Production of Multilaminar Veneer from Plantation-Grown Eucalyptus Globulus

Authors: Ngoc Nguyen

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There is large plantation of Eucalyptus globulus established which has been grown to produce pulpwood. This resource is not suitable for the production of decorative products, principally due to low grades of wood and “dull” appearance but many trials have been already undertaken for the production of veneer and veneer-based engineered wood products, such as plywood and laminated veneer lumber (LVL). The manufacture of veneer-based products has been recently identified as an unprecedented opportunity to promote higher value utilisation of plantation resources. However, many uncertainties remain regarding the impacts of inferior wood quality of young plantation trees on product recovery and value, and with respect to optimal processing techniques. Moreover, the quality of veneer and veneer-based products is far from optimal as trees are young and have small diameters; and the veneers have the significant colour variation which affects to the added value of final products. Developing production methods which would enhance appearance of low-quality veneer would provide a great potential for the production of high-value wood products such as furniture, joinery, flooring and other appearance products. One of the methods of enhancing appearance of low quality veneer, developed in Italy, involves the production of multilaminar veneer, also named “reconstructed veneer”. An important stage of the multilaminar production is colouring the veneer which can be achieved by dyeing veneer with dyes of different colours depending on the type of appearance products, their design and market demand. Although veneer dyeing technology has been well advanced in Italy, it has been focused on poplar veneer from plantation which wood is characterized by low density, even colour, small amount of defects and high permeability. Conversely, the majority of plantation eucalypts have medium to high density, have a lot of defects, uneven colour and low permeability. Therefore, detailed study is required to develop dyeing methods suitable for colouring eucalypt veneers. Brown reactive dye is used for veneer colouring process. Veneers from sapwood and heartwood of two moisture content levels are used to conduct colouring experiments: green veneer and veneer dried to 12% MC. Prior to dyeing, all samples are treated. Both soaking (dipping) and vacuum pressure methods are used in the study to compare the results and select most efficient method for veneer dyeing. To date, the results of colour measurements by CIELAB colour system showed significant differences in the colour of the undyed veneers produced from heartwood part. The colour became moderately darker with increasing of Sodium chloride, compared to control samples according to the colour measurements. It is difficult to conclude a suitable dye solution used in the experiments at this stage as the variables such as dye concentration, dyeing temperature or dyeing time have not been done. The dye will be used with and without UV absorbent after all trials are completed using optimal parameters in colouring veneers.

Keywords: Eucalyptus globulus, veneer colouring/dyeing, multilaminar veneer, reactive dye

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55 Switchable Lipids: From a Molecular Switch to a pH-Sensitive System for the Drug and Gene Delivery

Authors: Jeanne Leblond, Warren Viricel, Amira Mbarek

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Although several products have reached the market, gene therapeutics are still in their first stages and require optimization. It is possible to improve their lacking efficiency by the use of carefully engineered vectors, able to carry the genetic material through each of the biological barriers they need to cross. In particular, getting inside the cell is a major challenge, because these hydrophilic nucleic acids have to cross the lipid-rich plasmatic and/or endosomal membrane, before being degraded into lysosomes. It takes less than one hour for newly endocytosed liposomes to reach highly acidic lysosomes, meaning that the degradation of the carried gene occurs rapidly, thus limiting the transfection efficiency. We propose to use a new pH-sensitive lipid able to change its conformation upon protonation at endosomal pH values, leading to the disruption of the lipidic bilayer and thus to the fast release of the nucleic acids into the cytosol. It is expected that this new pH-sensitive mechanism promote endosomal escape of the gene, thereby its transfection efficiency. The main challenge of this work was to design a preparation presenting fast-responding lipidic bilayer destabilization properties at endosomal pH 5 while remaining stable at blood pH value and during storage. A series of pH-sensitive lipids able to perform a conformational switch upon acidification were designed and synthesized. Liposomes containing these switchable lipids, as well as co-lipids were prepared and characterized. The liposomes were stable at 4°C and pH 7.4 for several months. Incubation with siRNA led to the full entrapment of nucleic acids as soon as the positive/negative charge ratio was superior to 2. The best liposomal formulation demonstrated a silencing efficiency up to 10% on HeLa cells, very similar to a commercial agent, with a lowest toxicity than the commercial agent. Using flow cytometry and microscopy assays, we demonstrated that drop of pH was required for the transfection efficiency, since bafilomycin blocked the transfection efficiency. Additional evidence was brought by the synthesis of a negative control lipid, which was unable to switch its conformation, and consequently exhibited no transfection ability. Mechanistic studies revealed that the uptake was mediated through endocytosis, by clathrin and caveolae pathways, as reported for previous lipid nanoparticle systems. This potent system was used for the treatment of hypercholesterolemia. The switchable lipids were able to knockdown PCSK9 expression on human hepatocytes (Huh-7). Its efficiency is currently evaluated on in vivo mice model of PCSK9 KO mice. In summary, we designed and optimized a new cationic pH-sensitive lipid for gene delivery. Its transfection efficiency is similar to the best available commercial agent, without the usually associated toxicity. The promising results lead to its use for the treatment of hypercholesterolemia on a mice model. Anticancer applications and pulmonary chronic disease are also currently investigated.

Keywords: liposomes, siRNA, pH-sensitive, molecular switch

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54 Piled Critical Size Bone-Biomimetic and Biominerizable Nanocomposites: Formation of Bioreactor-Induced Stem Cell Gradients under Perfusion and Compression

Authors: W. Baumgartner, M. Welti, N. Hild, S. C. Hess, W. J. Stark, G. Meier Bürgisser, P. Giovanoli, J. Buschmann

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Perfusion bioreactors are used to solve problems in tissue engineering in terms of sufficient nutrient and oxygen supply. Such problems especially occur in critical size grafts because vascularization is often too slow after implantation ending up in necrotic cores. Biominerizable and biocompatible nanocomposite materials are attractive and suitable scaffold materials for bone tissue engineering because they offer mineral components in organic carriers – mimicking natural bone tissue. In addition, human adipose derived stem cells (ASCs) can potentially be used to increase bone healing as they are capable of differentiating towards osteoblasts or endothelial cells among others. In the present study, electrospun nanocomposite disks of poly-lactic-co-glycolic acid and amorphous calcium phosphate nanoparticles (PLGA/a-CaP) were seeded with human ASCs and eight disks were stacked in a bioreactor running with normal culture medium (no differentiation supplements). Under continuous perfusion and uniaxial cyclic compression, load-displacement curves as a function of time were assessed. Stiffness and energy dissipation were recorded. Moreover, stem cell densities in the layers of the piled scaffold were determined as well as their morphologies and differentiation status (endothelial cell differentiation, chondrogenesis and osteogenesis). While the stiffness of the cell free constructs increased over time caused by the transformation of the a-CaP nanoparticles into flake-like apatite, ASC-seeded constructs showed a constant stiffness. Stem cell density gradients were histologically determined with a linear increase in the flow direction from the bottom to the top of the 3.5 mm high pile (r2 > 0.95). Cell morphology was influenced by the flow rate, with stem cells getting more roundish at higher flow rates. Less than 1 % osteogenesis was found upon osteopontin immunostaining at the end of the experiment (9 days), while no endothelial cell differentiation and no chondrogenesis was triggered under these conditions. All ASCs had mainly remained in their original pluripotent status within this time frame. In summary, we have fabricated a critical size bone graft based on a biominerizable bone-biomimetic nanocomposite with preserved stiffness when seeded with human ASCs. The special feature of this bone graft was that ASC densities inside the piled construct varied with a linear gradient, which is a good starting point for tissue engineering interfaces such as bone-cartilage where the bone tissue is cell rich while the cartilage exhibits low cell densities. As such, this tissue-engineered graft may act as a bone-cartilage interface after the corresponding differentiation of the ASCs.

Keywords: bioreactor, bone, cartilage, nanocomposite, stem cell gradient

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53 Functional Switching of Serratia marcescens Transcriptional Regulator from Activator to Inhibitor of Quorum Sensing by Exogenous Addition

Authors: Norihiro Kato, Yuriko Takayama

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Some gram-negative bacteria enable the simultaneous activation of gene expression involved in N-acylhomoserine lactone (AHL) dependent cell-to-cell communication system. Such regulatory system for the bacterial group behavior is termed as quorum sensing (QS) because a diffusible AHL signal can accumulate around the cell during the increase of the cell density and trigger activation of the sequential QS process. By blocking the QS, the expression of diverse genes related to infection, antibiotic production, and biofilm formation is inhibited. Conditioning of QS by regulation of the DNA-receptor-AHL interaction is a potential target for enhancing host defenses against pathogenicity. We focused on engineered application of transcriptional regulator SpnR produced in opportunistic human pathogen Serratia marcescens. The SpnR can interact with AHL signals at an N-terminal domain and also with a promoter region of a QS target gene at a C-terminal domain. As the initial process of the QS activation, the SpnR forms a complex with the AHL to enhance the expression of pig cluster; the SpnR normally acts as an activator for the expression of the QS-dependent gene. In this research, we attempt to artificially control QS by changing the role of SpnR. The QS-dependent prodigiosin production is expected to inhibit by externally added SpnR in the culture broth of AS-1 strain because the AHL concentration was kept below the threshold by AHL-SpnR complex formation. Maltose-binding protein (MBP)-tagged SpnR (MBP-SpnR) was overexpressed in Escherichia coli and purified using an affinity chromatography equipped with an amylose resin column. The specific interaction between AHL and MBP-SpnR was demonstrated by quartz crystal microbalance (QCM) sensor. AHL with amino end-group was coupled with COOH-terminated self-assembled monolayer prepared on a gold electrode of 27-MHz quartz crystal sensor using water-soluble carbodiimide. After the injection of MBP-SpnR into a cup-type sensor cell filled with the buffer solution, time course of resonant frequency change (ΔFs) was determined. A decrease of ΔFs clearly showed the uptake of MBP-SpnR onto the AHL-immobilized electrode. Furthermore, no binding affinity was observed after the heat-inactivation of MBP-SpnR at 80ºC. These results suggest that MBP-SpnR possesses a specific affinity for AHL. MBP-SpnR was added to the culture medium as an AHL trap to study inhibitory effects on intracellularly accumulated prodigiosin. With approximately 2 µM MBP-SpnR, the amount of prodigiosin induced was half that of the control without any additives. In conclusion, the function of SpnR could be switched by adding it to the cell culture. Exogenously added MBP-SpnR possesses high affinity for AHL derived from cells and acts as an inhibitor of AHL-mediated QS.

Keywords: intracellular signaling, microbial biotechnology, quorum sensing, transcriptional regulator

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52 Automatic Identification and Classification of Contaminated Biodegradable Plastics using Machine Learning Algorithms and Hyperspectral Imaging Technology

Authors: Nutcha Taneepanichskul, Helen C. Hailes, Mark Miodownik

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Plastic waste has emerged as a critical global environmental challenge, primarily driven by the prevalent use of conventional plastics derived from petrochemical refining and manufacturing processes in modern packaging. While these plastics serve vital functions, their persistence in the environment post-disposal poses significant threats to ecosystems. Addressing this issue necessitates approaches, one of which involves the development of biodegradable plastics designed to degrade under controlled conditions, such as industrial composting facilities. It is imperative to note that compostable plastics are engineered for degradation within specific environments and are not suited for uncontrolled settings, including natural landscapes and aquatic ecosystems. The full benefits of compostable packaging are realized when subjected to industrial composting, preventing environmental contamination and waste stream pollution. Therefore, effective sorting technologies are essential to enhance composting rates for these materials and diminish the risk of contaminating recycling streams. In this study, it leverage hyperspectral imaging technology (HSI) coupled with advanced machine learning algorithms to accurately identify various types of plastics, encompassing conventional variants like Polyethylene terephthalate (PET), Polypropylene (PP), Low density polyethylene (LDPE), High density polyethylene (HDPE) and biodegradable alternatives such as Polybutylene adipate terephthalate (PBAT), Polylactic acid (PLA), and Polyhydroxyalkanoates (PHA). The dataset is partitioned into three subsets: a training dataset comprising uncontaminated conventional and biodegradable plastics, a validation dataset encompassing contaminated plastics of both types, and a testing dataset featuring real-world packaging items in both pristine and contaminated states. Five distinct machine learning algorithms, namely Partial Least Squares Discriminant Analysis (PLS-DA), Support Vector Machine (SVM), Convolutional Neural Network (CNN), Logistic Regression, and Decision Tree Algorithm, were developed and evaluated for their classification performance. Remarkably, the Logistic Regression and CNN model exhibited the most promising outcomes, achieving a perfect accuracy rate of 100% for the training and validation datasets. Notably, the testing dataset yielded an accuracy exceeding 80%. The successful implementation of this sorting technology within recycling and composting facilities holds the potential to significantly elevate recycling and composting rates. As a result, the envisioned circular economy for plastics can be established, thereby offering a viable solution to mitigate plastic pollution.

Keywords: biodegradable plastics, sorting technology, hyperspectral imaging technology, machine learning algorithms

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51 Temporal Estimation of Hydrodynamic Parameter Variability in Constructed Wetlands

Authors: Mohammad Moezzibadi, Isabelle Charpentier, Adrien Wanko, Robert Mosé

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The calibration of hydrodynamic parameters for subsurface constructed wetlands (CWs) is a sensitive process since highly non-linear equations are involved in unsaturated flow modeling. CW systems are engineered systems designed to favour natural treatment processes involving wetland vegetation, soil, and their microbial flora. Their significant efficiency at reducing the ecological impact of urban runoff has been recently proved in the field. Numerical flow modeling in a vertical variably saturated CW is here carried out by implementing the Richards model by means of a mixed hybrid finite element method (MHFEM), particularly well adapted to the simulation of heterogeneous media, and the van Genuchten-Mualem parametrization. For validation purposes, MHFEM results were compared to those of HYDRUS (a software based on a finite element discretization). As van Genuchten-Mualem soil hydrodynamic parameters depend on water content, their estimation is subject to considerable experimental and numerical studies. In particular, the sensitivity analysis performed with respect to the van Genuchten-Mualem parameters reveals a predominant influence of the shape parameters α, n and the saturated conductivity of the filter on the piezometric heads, during saturation and desaturation. Modeling issues arise when the soil reaches oven-dry conditions. A particular attention should also be brought to boundary condition modeling (surface ponding or evaporation) to be able to tackle different sequences of rainfall-runoff events. For proper parameter identification, large field datasets would be needed. As these are usually not available, notably due to the randomness of the storm events, we thus propose a simple, robust and low-cost numerical method for the inverse modeling of the soil hydrodynamic properties. Among the methods, the variational data assimilation technique introduced by Le Dimet and Talagrand is applied. To that end, a variational data assimilation technique is implemented by applying automatic differentiation (AD) to augment computer codes with derivative computations. Note that very little effort is needed to obtain the differentiated code using the on-line Tapenade AD engine. Field data are collected for a three-layered CW located in Strasbourg (Alsace, France) at the water edge of the urban water stream Ostwaldergraben, during several months. Identification experiments are conducted by comparing measured and computed piezometric head by means of the least square objective function. The temporal variability of hydrodynamic parameter is then assessed and analyzed.

Keywords: automatic differentiation, constructed wetland, inverse method, mixed hybrid FEM, sensitivity analysis

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50 The Impact of Housing Design on the Health and Well-Being of Populations: A Case-Study of Middle-Class Families in the Metropolitan Region of Port-Au-Prince, Haiti

Authors: A. L. Verret, N. Prince, Y. Jerome, A. Bras

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The effects of housing design on the health and well-being of populations are quite intangible. In fact, healthy housing parameters are generally difficult to establish scientifically. It is often unclear the direction of a cause-and-effect relationship between health variables and housing. However, the lack of clear and definite measurements does not entail the absence of relationship between housing, health, and well-being. Research has thus been conducted. It has mostly aimed the physical rather than the psychological or social well-being of a population, given the difficulties to establish cause-effect relationships because of the subjectivity of the psychological symptoms and of the challenge in determining the influence of other factors. That said, a strong relationship has been exposed between light and physiology. Both the nervous and endocrine systems, amongst others, are affected by different wavelengths of natural light within a building. Daylight in the workplace is indeed associated to decreased absenteeism, errors and product defects, fatigue, eyestrain, increased productivity and positive attitude. Similar associations can also be made to residential housing. Lower levels of sunlight within the home have been proven to result in impaired cognition in depressed participants of a cross-sectional case study. Moreover, minimum space (area and volume) has been linked to healthy housing and quality of life, resulting in norms and regulations for such parameters for home constructions. As a matter of fact, it is estimated that people spend the two-thirds of their lives within the home and its immediate environment. Therefore, it is possible to deduct that the health and well-being of the occupants are potentially at risk in an unhealthy housing situation. While the impact of architecture on health and well-being is acknowledged and considered somewhat crucial in various countries of the north and the south, this issue is barely raised in Haiti. In fact, little importance is given to architecture for many reasons (lack of information, lack of means, societal reflex, poverty…). However, the middle-class is known for its residential strategies and trajectories in search of better-quality homes and environments. For this reason, it would be pertinent to use this group and its strategies and trajectories to isolate the impact of housing design on the overall health and well-being. This research aims to analyze the impact of housing architecture on the health and well-being of middle-class families in the metropolitan region of Port-au-Prince. It is a case study which uses semi-structured interviews and observations as research methods. Although at an early stage, this research anticipates that homes affect their occupants both psychologically and physiologically, and consequently, public policies and the population should take into account the architectural design in the planning and construction of housing and, furthermore, cities.

Keywords: architectural design, health and well-being, middle-class housing, Port-au-Prince, Haiti

Procedia PDF Downloads 136
49 Cardiac Biosignal and Adaptation in Confined Nuclear Submarine Patrol

Authors: B. Lefranc, C. Aufauvre-Poupon, C. Martin-Krumm, M. Trousselard

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Isolated and confined environments (ICE) present several challenges which may adversely affect human’s psychology and physiology. Submariners in Sub-Surface Ballistic Nuclear (SSBN) mission exposed to these environmental constraints must be able to perform complex tasks as part of their normal duties, as well as during crisis periods when emergency actions are required or imminent. The operational and environmental constraints they face contribute to challenge human adaptability. The impact of such a constrained environment has yet to be explored. Establishing a knowledge framework is a determining factor, particularly in view of the next long space travels. Ensuring that the crews are maintained in optimal operational conditions is a real challenge because the success of the mission depends on them. This study focused on the evaluation of the impact of stress on mental health and sensory degradation of submariners during a mission on SSBN using cardiac biosignal (heart rate variability, HRV) clustering. This is a pragmatic exploratory study of a prospective cohort included 19 submariner volunteers. HRV was recorded at baseline to classify by clustering the submariners according to their stress level based on parasympathetic (Pa) activity. Impacts of high Pa (HPa) versus low Pa (LPa) level at baseline were assessed on emotional state and sensory perception (interoception and exteroception) as a cardiac biosignal during the patrol and at a recovery time one month after. Whatever the time, no significant difference was found in mental health between groups. There are significant differences in the interoceptive, exteroceptive and physiological functioning during the patrol and at recovery time. To sum up, compared to the LPa group, the HPa maintains a higher level in psychosensory functioning during the patrol and at recovery but exhibits a decrease in Pa level. The HPa group has less adaptable HRV characteristics, less unpredictability and flexibility of cardiac biosignals while the LPa group increases them during the patrol and at recovery time. This dissociation between psychosensory and physiological adaptation suggests two treatment modalities for ICE environments. To our best knowledge, our results are the first to highlight the impact of physiological differences in the HRV profile on the adaptability of submariners. Further studies are needed to evaluate the negative emotional and cognitive effects of ICEs based on the cardiac profile. Artificial intelligence offers a promising future for maintaining high level of operational conditions. These future perspectives will not only allow submariners to be better prepared, but also to design feasible countermeasures that will help support analog environments that bring us closer to a trip to Mars.

Keywords: adaptation, exteroception, HRV, ICE, interoception, SSBN

Procedia PDF Downloads 181
48 Development and Characterization of Topical 5-Fluorouracil Solid Lipid Nanoparticles for the Effective Treatment of Non-Melanoma Skin Cancer

Authors: Sudhir Kumar, V. R. Sinha

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Background: The topical and systemic toxicity associated with present nonmelanoma skin cancer (NMSC) treatment therapy using 5-Fluorouracil (5-FU) make it necessary to develop a novel delivery system having lesser toxicity and better control over drug release. Solid lipid nanoparticles offer many advantages like: controlled and localized release of entrapped actives, nontoxicity, and better tolerance. Aim:-To investigate safety and efficacy of 5-FU loaded solid lipid nanoparticles as a topical delivery system for the treatment of nonmelanoma skin cancer. Method: Topical solid lipid nanoparticles of 5-FU were prepared using Compritol 888 ATO (Glyceryl behenate) as lipid component and pluronic F68 (Poloxamer 188), Tween 80 (Polysorbate 80), Tyloxapol (4-(1,1,3,3-Tetramethylbutyl) phenol polymer with formaldehyde and oxirane) as surfactants. The SLNs were prepared with emulsification method. Different formulation parameters viz. type and ratio of surfactant, ratio of lipid and ratio of surfactant:lipid were investigated on particle size and drug entrapment efficiency. Results: Characterization of SLNs like–Transmission Electron Microscopy (TEM), Differential Scannig calorimetry (DSC), Fourier transform infrared spectroscopy (FTIR), Particle size determination, Polydispersity index, Entrapment efficiency, Drug loading, ex vivo skin permeation and skin retention studies, skin irritation and histopathology studies were performed. TEM results showed that shape of SLNs was spherical with size range 200-500nm. Higher encapsulation efficiency was obtained for batches having higher concentration of surfactant and lipid. It was found maximum 64.3% for SLN-6 batch with size of 400.1±9.22 nm and PDI 0.221±0.031. Optimized SLN batches and marketed 5-FU cream were compared for flux across rat skin and skin drug retention. The lesser flux and higher skin retention was obtained for SLN formulation in comparison to topical 5-FU cream, which ensures less systemic toxicity and better control of drug release across skin. Chronic skin irritation studies lacks serious erythema or inflammation and histopathology studies showed no significant change in physiology of epidermal layers of rat skin. So, these studies suggest that the optimized SLN formulation is efficient then marketed cream and safer for long term NMSC treatment regimens. Conclusion: Topical and systemic toxicity associated with long-term use of 5-FU, in the treatment of NMSC, can be minimized with its controlled release with significant drug retention with minimal flux across skin. The study may provide a better alternate for effective NMSC treatment.

Keywords: 5-FU, topical formulation, solid lipid nanoparticles, non melanoma skin cancer

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47 Engineering Topology of Photonic Systems for Sustainable Molecular Structure: Autopoiesis Systems

Authors: Moustafa Osman Mohammed

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This paper introduces topological order in descried social systems starting with the original concept of autopoiesis by biologists and scientists, including the modification of general systems based on socialized medicine. Topological order is important in describing the physical systems for exploiting optical systems and improving photonic devices. The stats of topological order have some interesting properties of topological degeneracy and fractional statistics that reveal the entanglement origin of topological order, etc. Topological ideas in photonics form exciting developments in solid-state materials, that being; insulating in the bulk, conducting electricity on their surface without dissipation or back-scattering, even in the presence of large impurities. A specific type of autopoiesis system is interrelated to the main categories amongst existing groups of the ecological phenomena interaction social and medical sciences. The hypothesis, nevertheless, has a nonlinear interaction with its natural environment 'interactional cycle' for exchange photon energy with molecules without changes in topology. The engineering topology of a biosensor is based on the excitation boundary of surface electromagnetic waves in photonic band gap multilayer films. The device operation is similar to surface Plasmonic biosensors in which a photonic band gap film replaces metal film as the medium when surface electromagnetic waves are excited. The use of photonic band gap film offers sharper surface wave resonance leading to the potential of greatly enhanced sensitivity. So, the properties of the photonic band gap material are engineered to operate a sensor at any wavelength and conduct a surface wave resonance that ranges up to 470 nm. The wavelength is not generally accessible with surface Plasmon sensing. Lastly, the photonic band gap films have robust mechanical functions that offer new substrates for surface chemistry to understand the molecular design structure and create sensing chips surface with different concentrations of DNA sequences in the solution to observe and track the surface mode resonance under the influences of processes that take place in the spectroscopic environment. These processes led to the development of several advanced analytical technologies: which are; automated, real-time, reliable, reproducible, and cost-effective. This results in faster and more accurate monitoring and detection of biomolecules on refractive index sensing, antibody-antigen reactions with a DNA or protein binding. Ultimately, the controversial aspect of molecular frictional properties is adjusted to each other in order to form unique spatial structure and dynamics of biological molecules for providing the environment mutual contribution in investigation of changes due to the pathogenic archival architecture of cell clusters.

Keywords: autopoiesis, photonics systems, quantum topology, molecular structure, biosensing

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46 Interplay of Material and Cycle Design in a Vacuum-Temperature Swing Adsorption Process for Biogas Upgrading

Authors: Federico Capra, Emanuele Martelli, Matteo Gazzani, Marco Mazzotti, Maurizio Notaro

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Natural gas is a major energy source in the current global economy, contributing to roughly 21% of the total primary energy consumption. Production of natural gas starting from renewable energy sources is key to limit the related CO2 emissions, especially for those sectors that heavily rely on natural gas use. In this context, biomethane produced via biogas upgrading represents a good candidate for partial substitution of fossil natural gas. The upgrading process of biogas to biomethane consists in (i) the removal of pollutants and impurities (e.g. H2S, siloxanes, ammonia, water), and (ii) the separation of carbon dioxide from methane. Focusing on the CO2 removal process, several technologies can be considered: chemical or physical absorption with solvents (e.g. water, amines), membranes, adsorption-based systems (PSA). However, none emerged as the leading technology, because of (i) the heterogeneity in plant size, ii) the heterogeneity in biogas composition, which is strongly related to the feedstock type (animal manure, sewage treatment, landfill products), (iii) the case-sensitive optimal tradeoff between purity and recovery of biomethane, and iv) the destination of the produced biomethane (grid injection, CHP applications, transportation sector). With this contribution, we explore the use of a technology for biogas upgrading and we compare the resulting performance with benchmark technologies. The proposed technology makes use of a chemical sorbent, which is engineered by RSE and consists of Di-Ethanol-Amine deposited on a solid support made of γ-Alumina, to chemically adsorb the CO2 contained in the gas. The material is packed into fixed beds that cyclically undergo adsorption and regeneration steps. CO2 is adsorbed at low temperature and ambient pressure (or slightly above) while the regeneration is carried out by pulling vacuum and increasing the temperature of the bed (vacuum-temperature swing adsorption - VTSA). Dynamic adsorption tests were performed by RSE and were used to tune the mathematical model of the process, including material and transport parameters (i.e. Langmuir isotherms data and heat and mass transport). Based on this set of data, an optimal VTSA cycle was designed. The results enabled a better understanding of the interplay between material and cycle tuning. As exemplary application, the upgrading of biogas for grid injection, produced by an anaerobic digester (60-70% CO2, 30-40% CH4), for an equivalent size of 1 MWel was selected. A plant configuration is proposed to maximize heat recovery and minimize the energy consumption of the process. The resulting performances are very promising compared to benchmark solutions, which make the VTSA configuration a valuable alternative for biomethane production starting from biogas.

Keywords: biogas upgrading, biogas upgrading energetic cost, CO2 adsorption, VTSA process modelling

Procedia PDF Downloads 273
45 Selection and Preparation of High Performance, Natural and Cost-Effective Hydrogel as a Bio-Ink for 3D Bio-Printing and Organ on Chip Applications

Authors: Rawan Ashraf, Ahmed E. Gomaa, Gehan Safwat, Ayman Diab

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Background: Three-dimensional (3D) bio-printing has become a versatile and powerful method for generating a variety of biological constructs, including bone or extracellular matrix scaffolds endo- or epithelial, muscle tissue, as well as organoids. Aim of the study: Fabricate a low cost DIY 3D bio-printer to produce 3D bio-printed products such as anti-microbial packaging or multi-organs on chips. We demonstrate the alignment between two types of 3D printer technology (3D Bio-printer and DLP) on Multi-organ-on-a-chip (multi-OoC) devices fabrication. Methods: First, Design and Fabrication of the Syringe Unit for Modification of an Off-the-Shelf 3D Printer, then Preparation of Hydrogel based on natural polymers Sodium Alginate and Gelatin, followed by acquisition of the cell suspension, then modeling the desired 3D structure. Preparation for 3D printing, then Cell-free and cell-laden hydrogels went through the printing process at room temperature under sterile conditions and finally post printing curing process and studying the printed structure regards physical and chemical characteristics. The hard scaffold of the Organ on chip devices was designed and fabricated using the DLP-3D printer, following similar approaches as the Microfluidics system fabrication. Results: The fabricated Bio-Ink was based onHydrogel polymer mix of sodium alginate and gelatin 15% to 0.5%, respectively. Later the 3D printing process was conducted using a higher percentage of alginate-based hydrogels because of it viscosity and the controllable crosslinking, unlike the thermal crosslinking of Gelatin. The hydrogels were colored to simulate the representation of two types of cells. The adaption of the hard scaffold, whether for the Microfluidics system or the hard-tissues, has been acquired by the DLP 3D printers with fabricated natural bioactive essential oils that contain antimicrobial activity, followed by printing in Situ three complex layers of soft-hydrogel as a cell-free Bio-Ink to simulate the real-life tissue engineering process. The final product was a proof of concept for a rapid 3D cell culturing approaches that uses an engineered hard scaffold along with soft-tissues, thus, several applications were offered as products of the current prototype, including the Organ-On-Chip as a successful integration between DLP and 3D bioprinter. Conclusion: Multiple designs for the organ-on-a-chip (multi-OoC) devices have been acquired in our study with main focus on the low cost fabrication of such technology and the potential to revolutionize human health research and development. We describe circumstances in which multi-organ models are useful after briefly examining the requirement for full multi-organ models with a systemic component. Following that, we took a look at the current multi-OoC platforms, such as integrated body-on-a-chip devices and modular techniques that use linked organ-specific modules.

Keywords: 3d bio-printer, hydrogel, multi-organ on chip, bio-inks

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44 Sensitivity Improvement of Optical Ring Resonator for Strain Analysis with the Direction of Strain Recognition Possibility

Authors: Tayebeh Sahraeibelverdi, Ahmad Shirazi Hadi Veladi, Mazdak Radmalekshah

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Optical sensors became attractive due to preciseness, low power consumption, and intrinsic electromagnetic interference-free characteristic. Among the waveguide optical sensors, cavity-based ones attended for the high Q-factor. Micro ring resonators as a potential platform have been investigated for various applications as biosensors to pressure sensors thanks to their sensitive ring structure responding to any small change in the refractive index. Furthermore, these small micron size structures can come in an array, bringing the opportunity to have any of the resonance in a specific wavelength and be addressed in this way. Another exciting application is applying a strain to the ring and making them an optical strain gauge where the traditional ones are based on the piezoelectric material. Making them in arrays needs electrical wiring and about fifty times bigger in size. Any physical element that impacts the waveguide cross-section, Waveguide elastic-optic property change, or ring circumference can play a role. In comparison, ring size change has a larger effect than others. Here an engineered ring structure is investigated to study the strain effect on the ring resonance wavelength shift and its potential for more sensitive strain devices. At the same time, these devices can measure any strain by mounting on the surface of interest. The idea is to change the" O" shape ring to a "C" shape ring with a small opening starting from 2π/360 or one degree. We used the Mode solution of Lumbrical software to investigate the effect of changing the ring's opening and the shift induced by applied strain. The designed ring radius is a three Micron silicon on isolator ring which can be fabricated by standard complementary metal-oxide-semiconductor (CMOS) micromachining. The measured wavelength shifts from1-degree opening of the ring to a 6-degree opening have been investigated. Opening the ring for 1-degree affects the ring's quality factor from 3000 to 300, showing an order of magnitude Q-factor reduction. Assuming a strain making the ring-opening from 1 degree to 6 degrees, our simulation results showing negligible Q-factor reduction from 300 to 280. A ring resonator quality factor can reach up to 108 where an order of magnitude reduction is negligible. The resonance wavelength shift showed a blue shift and was obtained to be 1581, 1579,1578,1575nm for 1-, 2-, 4- and 6-degree ring-opening, respectively. This design can find the direction of the strain-induced by applying the opening on different parts of the ring. Moreover, by addressing the specified wavelength, we can precisely find the direction. We can open a significant opportunity to find cracks and any surface mechanical property very specifically and precisely. This idea can be implemented on polymer ring resonators while they can come with a flexible substrate and can be very sensitive to any strain making the two ends of the ring in the slit part come closer or further.

Keywords: optical ring resonator, strain gauge, strain sensor, surface mechanical property analysis

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43 A Holistic View of Microbial Community Dynamics during a Toxic Harmful Algal Bloom

Authors: Shi-Bo Feng, Sheng-Jie Zhang, Jin Zhou

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The relationship between microbial diversity and algal bloom has received considerable attention for decades. Microbes undoubtedly affect annual bloom events and impact the physiology of both partners, as well as shape ecosystem diversity. However, knowledge about interactions and network correlations among broader-spectrum microbes that lead to the dynamics in a complete bloom cycle are limited. In this study, pyrosequencing and network approaches simultaneously assessed the associate patterns among bacteria, archaea, and microeukaryotes in surface water and sediments in response to a natural dinoflagellate (Alexandrium sp.) bloom. In surface water, among the bacterial community, Gamma-Proteobacteria and Bacteroidetes dominated in the initial bloom stage, while Alpha-Proteobacteria, Cyanobacteria, and Actinobacteria become the most abundant taxa during the post-stage. In the archaea biosphere, it clustered predominantly with Methanogenic members in the early pre-bloom period while the majority of species identified in the later-bloom stage were ammonia-oxidizing archaea and Halobacteriales. In eukaryotes, dinoflagellate (Alexandrium sp.) was dominated in the onset stage, whereas multiply species (such as microzooplankton, diatom, green algae, and rotifera) coexistence in bloom collapse stag. In sediments, the microbial species biomass and richness are much higher than the water body. Only Flavobacteriales and Rhodobacterales showed a slight response to bloom stages. Unlike the bacteria, there are small fluctuations of archaeal and eukaryotic structure in the sediment. The network analyses among the inter-specific associations show that bacteria (Alteromonadaceae, Oceanospirillaceae, Cryomorphaceae, and Piscirickettsiaceae) and some zooplankton (Mediophyceae, Mamiellophyceae, Dictyochophyceae and Trebouxiophyceae) have a stronger impact on the structuring of phytoplankton communities than archaeal effects. The changes in population were also significantly shaped by water temperature and substrate availability (N & P resources). The results suggest that clades are specialized at different time-periods and that the pre-bloom succession was mainly a bottom-up controlled, and late-bloom period was controlled by top-down patterns. Additionally, phytoplankton and prokaryotic communities correlated better with each other, which indicate interactions among microorganisms are critical in controlling plankton dynamics and fates. Our results supplied a wider view (temporal and spatial scales) to understand the microbial ecological responses and their network association during algal blooming. It gives us a potential multidisciplinary explanation for algal-microbe interaction and helps us beyond the traditional view linked to patterns of algal bloom initiation, development, decline, and biogeochemistry.

Keywords: microbial community, harmful algal bloom, ecological process, network

Procedia PDF Downloads 111
42 Bio-Functionalized Silk Nanofibers for Peripheral Nerve Regeneration

Authors: Kayla Belanger, Pascale Vigneron, Guy Schlatter, Bernard Devauchelle, Christophe Egles

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A severe injury to a peripheral nerve leads to its degeneration and the loss of sensory and motor function. To this day, there still lacks a more effective alternative to the autograft which has long been considered the gold standard for nerve repair. In order to overcome the numerous drawbacks of the autograft, tissue engineered biomaterials may be effective alternatives. Silk fibroin is a favorable biomaterial due to its many advantageous properties such as its biocompatibility, its biodegradability, and its robust mechanical properties. In this study, bio-mimicking multi-channeled nerve guidance conduits made of aligned nanofibers achieved by electrospinning were functionalized with signaling biomolecules and were tested in vitro and in vivo for nerve regeneration support. Silk fibroin (SF) extracted directly from silkworm cocoons was put in solution at a concentration of 10wt%. Poly(ethylene oxide) (PEO) was added to the resulting SF solution to increase solution viscosity and the following three electrospinning solutions were made: (1) SF/PEO solution, (2) SF/PEO solution with nerve growth factor and ciliary neurotrophic factor, and (3) SF/PEO solution with nerve growth factor and neurotrophin-3. Each of these solutions was electrospun into a multi-layer architecture to obtain mechanically optimized aligned nanofibrous mats. For in vitro studies, aligned fibers were treated to induce β-sheet formation and thoroughly rinsed to eliminate presence of PEO. Each material was tested using rat embryo neuron cultures to evaluate neurite extension and the interaction with bio-functionalized or non-functionalized aligned fibers. For in vivo studies, the mats were rolled into 5mm long multi-, micro-channeled conduits then treated and thoroughly rinsed. The conduits were each subsequently implanted between a severed rat sciatic nerve. The effectiveness of nerve repair over a period of 8 months was extensively evaluated by cross-referencing electrophysiological, histological, and movement analysis results to comprehensively evaluate the progression of nerve repair. In vitro results show a more favorable interaction between growing neurons and bio-functionalized silk fibers compared to pure silk fibers. Neurites can also be seen having extended unidirectionally along the alignment of the nanofibers which confirms a guidance factor for the electrospun material. The in vivo study has produced positive results for the regeneration of the sciatic nerve over the length of the study, showing contrasts between the bio-functionalized material and the non-functionalized material along with comparisons to the experimental control. Nerve regeneration has been evaluated not only by histological analysis, but also by electrophysiological assessment and motion analysis of two separate natural movements. By studying these three components in parallel, the most comprehensive evaluation of nerve repair for the conduit designs can be made which can, therefore, more accurately depict their overall effectiveness. This work was supported by La Région Picardie and FEDER.

Keywords: electrospinning, nerve guidance conduit, peripheral nerve regeneration, silk fibroin

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41 A Brief Review on Doping in Sports and Performance-Enhancing Drugs

Authors: Zahra Mohajer, Afsaneh Soltani

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Doping is a major issue in competitive sports and is favored by vast groups of athletes. The feeling of being higher-ranking than others and gaining fame has caused many athletes to misuse drugs. The definition of doping is to use prohibited substances and/or methods that help physical or mental performances or both. Doping counts as the illegal use of chemical substances or drugs, excessive amounts of physiological substances to increase the performance at or out of competition or even the use of inappropriate medications to treat an injury to gain the ability to participate in a competition. The International Olympic Committee (IOC) and World Anti-Doping Agency (WADA) have forbidden these substances to ensure fair and equal competition and also the health of the competitors. As of 2004 WADA has published an international list of illegal substances used for doping, which is updated annually. In the process of the Genome Project scientists have gained the ability to treat numerous diseases by gene therapy, which may result in bodily performance increase and therefore a potential opportunity to misuse by some athletes. Gene doping is defined as the non-therapeutic direct and indirect genetic modifications using genetic materials that can improve the performances in sports events. Biosynthetic drugs are a form of indirect genetic engineering. The method can be performed in three ways such as injecting the DNA directly into the muscle, inserting the genetically engineered cells, or transferring the DNA using a virus as a vector. Erythropoietin is a hormone majorly released by the kidney and in small amounts by the liver. Its function is to stimulate the erythropoiesis and therefore the more production of red blood cells (RBC) which causes an increase in Hemoglobin (Hb). During this process, the oxygen delivery to muscles will increase, which will improve athletic performance and postpone exhaustion. There are ways to increase the oxygen transferred to muscles such as blood transfusion, stimulating the production of red blood cells by using Erythropoietin (EPO), and also using allosteric effectors of Hemoglobin. EPO can either be injected as a protein or can be inserted into the cells as the gene which encodes EPO. Adeno-associated viruses have been employed to deliver the EPO gene to the cells. Employing the genes that naturally exist in the human body such as the EPO gene can reduce the risk of detecting gene doping. The first research about blood doping was conducted in 1947. The study has shown that an increase in hematocrit (HCT) up to 55% following homologous transfusion makes it more unchallenging for the body to perform the exercise at the altitude. Thereafter athletes’ attraction to blood infusion escalated. Also, a study has demonstrated that by reinfusing their own blood 4 weeks after being drawn, three men have shown a rise in Hb level which improved the oxygen uptake, and a delay in exhaustion. The list of performance-enhancing drugs is published by WADA annually and includes the following drugs: anabolic agents, hormones, Beta-2 agonists, Beta-blockers, Diuretics, Stimulants, narcotics, cannabinoids, and corticosteroids.

Keywords: doping, PEDs, sports, WADA

Procedia PDF Downloads 105
40 Trauma Scores and Outcome Prediction After Chest Trauma

Authors: Mohamed Abo El Nasr, Mohamed Shoeib, Abdelhamid Abdelkhalik, Amro Serag

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Background: Early assessment of severity of chest trauma, either blunt or penetrating is of critical importance in prediction of patient outcome. Different trauma scoring systems are widely available and are based on anatomical or physiological parameters to expect patient morbidity or mortality. Up till now, there is no ideal, universally accepted trauma score that could be applied in all trauma centers and is suitable for assessment of severity of chest trauma patients. Aim: Our aim was to compare various trauma scoring systems regarding their predictability of morbidity and mortality in chest trauma patients. Patients and Methods: This study was a prospective study including 400 patients with chest trauma who were managed at Tanta University Emergency Hospital, Egypt during a period of 2 years (March 2014 until March 2016). The patients were divided into 2 groups according to the mode of trauma: blunt or penetrating. The collected data included age, sex, hemodynamic status on admission, intrathoracic injuries, and associated extra-thoracic injuries. The patients outcome including mortality, need of thoracotomy, need for ICU admission, need for mechanical ventilation, length of hospital stay and the development of acute respiratory distress syndrome were also recorded. The relevant data were used to calculate the following trauma scores: 1. Anatomical scores including abbreviated injury scale (AIS), Injury severity score (ISS), New injury severity score (NISS) and Chest wall injury scale (CWIS). 2. Physiological scores including revised trauma score (RTS), Acute physiology and chronic health evaluation II (APACHE II) score. 3. Combined score including Trauma and injury severity score (TRISS ) and 4. Chest-Specific score Thoracic trauma severity score (TTSS). All these scores were analyzed statistically to detect their sensitivity, specificity and compared regarding their predictive power of mortality and morbidity in blunt and penetrating chest trauma patients. Results: The incidence of mortality was 3.75% (15/400). Eleven patients (11/230) died in blunt chest trauma group, while (4/170) patients died in penetrating trauma group. The mortality rate increased more than three folds to reach 13% (13/100) in patients with severe chest trauma (ISS of >16). The physiological scores APACHE II and RTS had the highest predictive value for mortality in both blunt and penetrating chest injuries. The physiological score APACHE II followed by the combined score TRISS were more predictive for intensive care admission in penetrating injuries while RTS was more predictive in blunt trauma. Also, RTS had a higher predictive value for expectation of need for mechanical ventilation followed by the combined score TRISS. APACHE II score was more predictive for the need of thoracotomy in penetrating injuries and the Chest-Specific score TTSS was higher in blunt injuries. The anatomical score ISS and TTSS score were more predictive for prolonged hospital stay in penetrating and blunt injuries respectively. Conclusion: Trauma scores including physiological parameters have a higher predictive power for mortality in both blunt and penetrating chest trauma. They are more suitable for assessment of injury severity and prediction of patients outcome.

Keywords: chest trauma, trauma scores, blunt injuries, penetrating injuries

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39 Effect of Rolling Shear Modulus and Geometric Make up on the Out-Of-Plane Bending Performance of Cross-Laminated Timber Panel

Authors: Md Tanvir Rahman, Mahbube Subhani, Mahmud Ashraf, Paul Kremer

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Cross-laminated timber (CLT) is made from layers of timber boards orthogonally oriented in the thickness direction, and due to this, CLT can withstand bi-axial bending in contrast with most other engineered wood products such as laminated veneer lumber (LVL) and glued laminated timber (GLT). Wood is cylindrically anisotropic in nature and is characterized by significantly lower elastic modulus and shear modulus in the planes perpendicular to the fibre direction, and is therefore classified as orthotropic material and is thus characterized by 9 elastic constants which are three elastic modulus in longitudinal direction, tangential direction and radial direction, three shear modulus in longitudinal tangential plane, longitudinal radial plane and radial tangential plane and three Poisson’s ratio. For simplification, timber materials are generally assumed to be transversely isotropic, reducing the number of elastic properties characterizing it to 5, where the longitudinal plane and radial planes are assumed to be planes of symmetry. The validity of this assumption was investigated through numerical modelling of CLT with both orthotropic mechanical properties and transversely isotropic material properties for three softwood species, which are Norway spruce, Douglas fir, Radiata pine, and three hardwood species, namely Victorian ash, Beech wood, and Aspen subjected to uniformly distributed loading under simply supported boundary condition. It was concluded that assuming the timber to be transversely isotropic results in a negligible error in the order of 1 percent. It was also observed that along with longitudinal elastic modulus, ratio of longitudinal shear modulus (GL) and rolling shear modulus (GR) has a significant effect on a deflection for CLT panels of lower span to depth ratio. For softwoods such as Norway spruce and Radiata pine, the ratio of longitudinal shear modulus, GL to rolling shear modulus GR is reported to be in the order of 12 to 15 times in literature. This results in shear flexibility in transverse layers leading to increased deflection under out-of-plane loading. The rolling shear modulus of hardwoods has been found to be significantly higher than those of softwoods, where the ratio between longitudinal shear modulus to rolling shear modulus as low as 4. This has resulted in a significant rise in research into the manufacturing of CLT from entirely from hardwood, as well as from a combination of softwood and hardwoods. The commonly used beam theory to analyze the performance of CLT panels under out-of-plane loads are the Shear analogy method, Gamma method, and k-method. The shear analogy method has been found to be the most effective method where shear deformation is significant. The effect of the ratio of longitudinal shear modulus and rolling shear modulus of cross-layer on the deflection of CLT under uniformly distributed load with respect to its length to depth ratio was investigated using shear analogy method. It was observed that shear deflection is reduced significantly as the ratio of the shear modulus of the longitudinal layer and rolling shear modulus of cross-layer decreases. This indicates that there is significant room for improvement of the bending performance of CLT through developing hybrid CLT from a mix of softwood and hardwood.

Keywords: rolling shear modulus, shear deflection, ratio of shear modulus and rolling shear modulus, timber

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38 Surface Acoustic Waves Nebulisation of Liposomes Manufactured in situ for Pulmonary Drug Delivery

Authors: X. King, E. Nazarzadeh, J. Reboud, J. Cooper

Abstract:

Pulmonary diseases, such as asthma, are generally treated by the inhalation of aerosols that has the advantage of reducing the off-target (e.g., toxicity) effects associated with systemic delivery in blood. Effective respiratory drug delivery requires a droplet size distribution between 1 and 5 µm. Inhalation of aerosols with wide droplet size distribution, out of this range, results in deposition of drug in not-targeted area of the respiratory tract, introducing undesired side effects on the patient. In order to solely deliver the drug in the lower branches of the lungs and release it in a targeted manner, a control mechanism to produce the aerosolized droplets is required. To regulate the drug release and to facilitate the uptake from cells, drugs are often encapsulated into protective liposomes. However, a multistep process is required for their formation, often performed at the formulation step, therefore limiting the range of available drugs or their shelf life. Using surface acoustic waves (SAWs), a pulmonary drug delivery platform was produced, which enabled the formation of defined size aerosols and the formation of liposomes in situ. SAWs are mechanical waves, propagating along the surface of a piezoelectric substrate. They were generated using an interdigital transducer on lithium niobate with an excitation frequency of 9.6 MHz at a power of 1W. Disposable silicon superstrates were etched using photolithography and dry etch processes to create an array of cylindrical through-holes with different diameters and pitches. Superstrates were coupled with the SAW substrate through water-based gel. As the SAW propagates on the superstrate, it enables nebulisation of a lipid solution deposited onto it. The cylindrical cavities restricted the formation of large drops in the aerosol, while at the same time unilamellar liposomes were created. SAW formed liposomes showed a higher monodispersity compared to the control sample, as well as displayed, a faster production rate. To test the aerosol’s size, dynamic light scattering and laser diffraction methods were used, both showing the size control of the aerosolised particles. The use of silicon superstate with cavity size of 100-200 µm, produced an aerosol with a mean droplet size within the optimum range for pulmonary drug delivery, containing the liposomes in which the medicine could be loaded. Additionally, analysis of liposomes with Cryo-TEM showed formation of vesicles with narrow size distribution between 80-100 nm and optimal morphology in order to be used for drug delivery. Encapsulation of nucleic acids in liposomes through the developed SAW platform was also investigated. In vitro delivery of siRNA and DNA Luciferase were achieved using A549 cell line, lung carcinoma from human. In conclusion, SAW pulmonary drug delivery platform was engineered, in order to combine multiple time consuming steps (formation of liposomes, drug loading, nebulisation) into a unique platform with the aim of specifically delivering the medicament in a targeted area, reducing the drug’s side effects.

Keywords: acoustics, drug delivery, liposomes, surface acoustic waves

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37 Criticality of Socio-Cultural Factors in Public Policy: A Study of Reproductive Health Care in Rural West Bengal

Authors: Arindam Roy

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Public policy is an intriguing terrain, which involves complex interplay of administrative, social political and economic components. There is hardly any fit-for all formulation of public policy as Lindbloom has aptly categorized it as a science of muddling through. In fact, policies are both temporally and contextually determined as one the proponents of policy sciences Harold D Lasswell has underscored it in his ‘contextual-configurative analysis’ as early as 1950s. Though, a lot of theoretical efforts have been made to make sense of this intricate dynamics of policy making, at the end of the day the applied area of public policy negates any such uniform, planned and systematic formulation. However, our policy makers seem to have learnt very little of that. Until recently, policy making was deemed as an absolutely specialized exercise to be conducted by a cadre of professionally trained seasoned mandarin. Attributes like homogeneity, impartiality, efficiency, and neutrality were considered as the watchwords of delivering common goods. Citizen or clientele was conceptualized as universal political or economic construct, to be taken care of uniformly. Moreover, policy makers usually have the proclivity to put anything into straightjacket, and to ignore the nuances therein. Hence, least attention has been given to the ground level reality, especially the socio-cultural milieu where the policy is supposed to be applied. Consequently, a substantial amount of public money goes in vain as the intended beneficiaries remain indifferent to the delivery of public policies. The present paper in the light of Reproductive Health Care policy in rural West Bengal has tried to underscore the criticality of socio-cultural factors in public health delivery. Indian health sector has traversed a long way. From a near non-existent at the time of independence, the Indian state has gradually built a country-wide network of health infrastructure. Yet it has to make a major breakthrough in terms of coverage and penetration of the health services in the rural areas. Several factors are held responsible for such state of things. These include lack of proper infrastructure, medicine, communication, ambulatory services, doctors, nursing services and trained birth attendants. Policy makers have underlined the importance of supply side in policy formulation and implementation. The successive policy documents concerning health delivery bear the testimony of it. The present paper seeks to interrogate the supply-side oriented explanations for the failure of the delivery of health services. Instead, it identified demand side to find out the answer. The state-led and bureaucratically engineered public health measures fail to engender demands as these measures mostly ignore socio-cultural nuances of health and well-being. Hence, the hiatus between supply side and demand side leads to huge wastage of revenue as health infrastructure, medicine and instruments remain unutilized in most cases. Therefore, taking proper cognizance of these factors could have streamlined the delivery of public health.

Keywords: context, policy, socio-cultural factor, uniformity

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36 Biomaterials Solutions to Medical Problems: A Technical Review

Authors: Ashish Thakur

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This technical paper was written in view of focusing the biomaterials and its various applications in modern industries. Author tires to elaborate not only the medical, infect plenty of application in other industries. The scope of the research area covers the wide range of physical, biological and chemical sciences that underpin the design of biomaterials and the clinical disciplines in which they are used. A biomaterial is now defined as a substance that has been engineered to take a form which, alone or as part of a complex system, is used to direct, by control of interactions with components of living systems, the course of any therapeutic or diagnostic procedure. Biomaterials are invariably in contact with living tissues. Thus, interactions between the surface of a synthetic material and biological environment must be well understood. This paper reviews the benefits and challenges associated with surface modification of the metals in biomedical applications. The paper also elaborates how the surface characteristics of metallic biomaterials, such as surface chemistry, topography, surface charge, and wettability, influence the protein adsorption and subsequent cell behavior in terms of adhesion, proliferation, and differentiation at the biomaterial–tissue interface. The chapter also highlights various techniques required for surface modification and coating of metallic biomaterials, including physicochemical and biochemical surface treatments and calcium phosphate and oxide coatings. In this review, the attention is focused on the biomaterial-associated infections, from which the need for anti-infective biomaterials originates. Biomaterial-associated infections differ markedly for epidemiology, aetiology and severity, depending mainly on the anatomic site, on the time of biomaterial application, and on the depth of the tissues harbouring the prosthesis. Here, the diversity and complexity of the different scenarios where medical devices are currently utilised are explored, providing an overview of the emblematic applicative fields and of the requirements for anti-infective biomaterials. In addition to this, chapter introduces nanomedicine and the use of both natural and synthetic polymeric biomaterials, focuses on specific current polymeric nanomedicine applications and research, and concludes with the challenges of nanomedicine research. Infection is currently regarded as the most severe and devastating complication associated to the use of biomaterials. Osteoporosis is a worldwide disease with a very high prevalence in humans older than 50. The main clinical consequences are bone fractures, which often lead to patient disability or even death. A number of commercial biomaterials are currently used to treat osteoporotic bone fractures, but most of these have not been specifically designed for that purpose. Many drug- or cell-loaded biomaterials have been proposed in research laboratories, but very few have received approval for commercial use. Polymeric nanomaterial-based therapeutics plays a key role in the field of medicine in treatment areas such as drug delivery, tissue engineering, cancer, diabetes, and neurodegenerative diseases. Advantages in the use of polymers over other materials for nanomedicine include increased functionality, design flexibility, improved processability, and, in some cases, biocompatibility.

Keywords: nanomedicine, tissue, infections, biomaterials

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35 Numerical Study of Leisure Home Chassis under Various Loads by Using Finite Element Analysis

Authors: Asem Alhnity, Nicholas Pickett

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The leisure home industry is experiencing an increase in sales due to the rise in popularity of staycations. However, there is also a demand for improvements in thermal and structural behaviour from customers. Existing standards and codes of practice outline the requirements for leisure home design. However, there is a lack of expertise in applying Finite Element Analysis (FEA) to complex structures in this industry. As a result, manufacturers rely on standardized design approaches, which often lead to excessively engineered or inadequately designed products. This study aims to address this issue by investigating the impact of the habitation structure on chassis performance in leisure homes. The aim of this research is to comprehensively analyse the impact of the habitation structure on chassis performance in leisure homes. By employing FEA on the entire unit, including both the habitation structure and the chassis, this study seeks to develop a novel framework for designing and analysing leisure homes. The objectives include material reduction, enhancing structural stability, resolving existing design issues, and developing innovative modular and wooden chassis designs. The methodology used in this research is quantitative in nature. The study utilizes FEA to analyse the performance of leisure home chassis under various loads. The analysis procedures involve running the FEA simulations on the numerical model of the leisure home chassis. Different load scenarios are applied to assess the stress and deflection performance of the chassis under various conditions. FEA is a numerical method that allows for accurate analysis of complex systems. The research utilizes flexible mesh sizing to calculate small deflections around doors and windows, with large meshes used for macro deflections. This approach aims to minimize run-time while providing meaningful stresses and deflections. Moreover, it aims to investigate the limitations and drawbacks of the popular approach of applying FEA only to the chassis and replacing the habitation structure with a distributed load. The findings of this study indicate that the popular approach of applying FEA only to the chassis and replacing the habitation structure with a distributed load overlooks the strengthening generated from the habitation structure. By employing FEA on the entire unit, it is possible to optimize stress and deflection performance while achieving material reduction and enhanced structural stability. The study also introduces innovative modular and wooden chassis designs, which show promising weight reduction compared to the existing heavily fabricated lattice chassis. In conclusion, this research provides valuable insights into the impact of the habitation structure on chassis performance in leisure homes. By employing FEA on the entire unit, the study demonstrates the importance of considering the strengthening generated from the habitation structure in chassis design. The research findings contribute to advancements in material reduction, structural stability, and overall performance optimization. The novel framework developed in this study promotes sustainability, cost-efficiency, and innovation in leisure home design.

Keywords: static homes, caravans, motor homes, holiday homes, finite element analysis (FEA)

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34 A Biophysical Study of the Dynamic Properties of Glucagon Granules in α Cells by Imaging-Derived Mean Square Displacement and Single Particle Tracking Approaches

Authors: Samuele Ghignoli, Valentina de Lorenzi, Gianmarco Ferri, Stefano Luin, Francesco Cardarelli

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Insulin and glucagon are the two essential hormones for maintaining proper blood glucose homeostasis, which is disrupted in Diabetes. A constantly growing research interest has been focused on the study of the subcellular structures involved in hormone secretion, namely insulin- and glucagon-containing granules, and on the mechanisms regulating their behaviour. Yet, while several successful attempts were reported describing the dynamic properties of insulin granules, little is known about their counterparts in α cells, the glucagon-containing granules. To fill this gap, we used αTC1 clone 9 cells as a model of α cells and ZIGIR as a fluorescent Zinc chelator for granule labelling. We started by using spatiotemporal fluorescence correlation spectroscopy in the form of imaging-derived mean square displacement (iMSD) analysis. This afforded quantitative information on the average dynamical and structural properties of glucagon granules having insulin granules as a benchmark. Interestingly, the iMSD sensitivity to average granule size allowed us to confirm that glucagon granules are smaller than insulin ones (~1.4 folds, further validated by STORM imaging). To investigate possible heterogeneities in granule dynamic properties, we moved from correlation spectroscopy to single particle tracking (SPT). We developed a MATLAB script to localize and track single granules with high spatial resolution. This enabled us to classify the glucagon granules, based on their dynamic properties, as ‘blocked’ (i.e., trajectories corresponding to immobile granules), ‘confined/diffusive’ (i.e., trajectories corresponding to slowly moving granules in a defined region of the cell), or ‘drifted’ (i.e., trajectories corresponding to fast-moving granules). In cell-culturing control conditions, results show this average distribution: 32.9 ± 9.3% blocked, 59.6 ± 9.3% conf/diff, and 7.4 ± 3.2% drifted. This benchmarking provided us with a foundation for investigating selected experimental conditions of interest, such as the glucagon-granule relationship with the cytoskeleton. For instance, if Nocodazole (10 μM) is used for microtubule depolymerization, the percentage of drifted motion collapses to 3.5 ± 1.7% while immobile granules increase to 56.0 ± 10.7% (remaining 40.4 ± 10.2% of conf/diff). This result confirms the clear link between glucagon-granule motion and cytoskeleton structures, a first step towards understanding the intracellular behaviour of this subcellular compartment. The information collected might now serve to support future investigations on glucagon granules in physiology and disease. Acknowledgment: This work has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement No 866127, project CAPTUR3D).

Keywords: glucagon granules, single particle tracking, correlation spectroscopy, ZIGIR

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