Search results for: cell-free protein expression

Authors: A. M. Tarola, R. Preti, A. M. Girelli, P. Campana

Abstract:

Goat dairy products are gaining popularity worldwide. In developing countries, but also in many marginal regions of the Mediterranean area, goats represent a great part of the economy and ensure food security. In fact, these small ruminants are able to convert efficiently poor weedy plants and small trees into traditional products of high nutritional quality, showing great resilience to different climatic and environmental conditions. In developed countries, goat milk is appreciated for the presence of health-promoting compounds, bioactive compounds such as conjugated linoleic acids, oligosaccharides, sphingolipids and polyammines. This paper focuses on the recent advances in literature on the nutritional properties of goat milk and on innovative techniques to improve its quality as to become a promising functional food. The environmental sustainability of different methodologies of production has also been examined. Goat milk is valued today as a food of high nutritional value and functional properties as well as small environmental footprint. It is widely consumed in many countries due to high nutritional value, lower allergenic potential, and better digestibility when compared to bovine milk, that makes this product suitable for infants, elderly or sensitive patients. The main differences in chemical composition between a cow and goat milk rely on fat globules that in goat milk are smaller and in fatty acids that present a smaller chain length, while protein, fat, and lactose concentration are comparable. Milk nutritional properties have demonstrated to be strongly influenced by animal diet, genotype, and welfare, but also by season and production systems. Furthermore, there is a growing interest in the dairy industry in goat milk for its relatively high concentration of prebiotics and a good amount of probiotics, which have recently gained importance for their therapeutic potential. Therefore, goat milk is studied as a promising matrix to develop innovative functional foods. In addition to the economic and nutritional value, goat milk is considered a sustainable product for its small environmental footprint, as they require relatively little water and land, and less medical treatments, compared to cow, these characteristics make its production naturally vocated to organic farming. Organic goat milk production has becoming more and more interesting both for farmers and consumers as it can answer to several concerns like environment protection, animal welfare and economical sustainment of rural populations living in marginal lands. These evidences make goat milk an ancient food with novel properties and advantages to be valorized and exploited.

Keywords: goat milk, nutritional quality, bioactive compounds, sustainable production, animal welfare

Procedia PDF Downloads 130

Authors: Tomás Sobrino, Lara García-Varela, Marta Aramburu-Núñez, Mónica Castro, Noemí Gómez-Lado, Mariña Rodríguez-Arrizabalaga, Antía Custodia, Juan Manuel Pías-Peleteiro, José Manuel Aldrey, Daniel Romaus-Sanjurjo, Ángeles Almeida, Pablo Aguiar, Alberto Ouro

Abstract:

Introduction: Alzheimer’s disease (AD) and other tauopathies are primary causes of dementia, causing progressive cognitive deterioration that entails serious repercussions for the patients' performance of daily tasks. Currently, there is no effective approach for the early diagnosis and treatment of AD and tauopathies. This study suggests a theragnostic approach based on the importance of phosphorylated tau protein (p-Tau) in the early pathophysiological processes of AD. We have developed a novel theragnostic monoclonal antibody (mAb) to provide both diagnostic and therapeutic effects. Methods/Results: We have developed a p-Tau mAb, which was doped with deferoxamine for radiolabeling with Zirconium-89 (89Zr) for PET imaging, as well as fluorescence dies for immunofluorescence assays. The p-Tau mAb was evaluated in vitro for toxicity by MTT assay, LDH activity, propidium iodide/Annexin V assay, caspase-3, and mitochondrial membrane potential (MMP) assay in both mouse endothelial cell line (bEnd.3) and cortical primary neurons cell cultures. Importantly, non-toxic effects (up to concentrations of p-Tau mAb greater than 100 ug/mL) were detected. In vivo experiments in the tauopathy model mice (PS19) show that the 89Zr-pTau-mAb and 89Zr-Fragments-pTau-mAb are stable in circulation for up to 10 days without toxic effects. However, only less than 0.2% reached the brain, so further strategies have to be designed for crossing the Brain-Blood-Barrier (BBB). Moreover, an intraparenchymal treatment strategy was carried out. The PS19 mice were operated to implement osmotic pumps (Alzet 1004) at two different times, at 4 and 7 months, to stimulate the controlled release for one month each of the B6 antibody or the IgG1 control antibody. We demonstrated that B6-treated mice maintained their motor and memory abilities significantly compared with IgG1 treatment. In addition, we observed a significant reduction in p-Tau deposits in the brain. Conclusions /Discussion: A theragnostic pTau-mAb was developed. Moreover, we demonstrated that our p-Tau mAb recognizes very-early pathology forms of p-Tau by non-invasive techniques, such as PET. In addition, p-Tau mAb has non-toxic effects, both in vitro and in vivo. Although the p-Tau mAb is stable in circulation, only 0.2% achieve the brain. However, direct intraventricular treatment significantly reduces cognitive impairment in Alzheimer's animal models, as well as the accumulation of toxic p-Tau species.

Keywords: alzheimer's disease, theragnosis, tau, PET, immunotherapy, tauopathies

Procedia PDF Downloads 49

Authors: N. Demertzis, J. L. Bowen

Abstract:

Sepsis is a complex and rapidly evolving condition, resulting from uncontrolled prolonged activation of host immune system due to pathogenic insult. The aim of this study is the development of a multiplex electrochemical sensing platform, capable of detecting both pathogen associated and host immune markers to enable the rapid and definitive diagnosis of sepsis. A combination of aptamers and molecular imprinting approaches have been employed to generate sensing systems for lipopolysaccharide (LPS), c-reactive protein (CRP) and procalcitonin (PCT). Gold working electrodes were mechanically polished and electrochemically cleaned with 0.1 M sulphuric acid using cyclic voltammetry (CV). Following activation, a self-assembled monolayer (SAM) was generated, by incubating the electrodes with a thiolated anti-LPS aptamer / dithiodibutiric acid (DTBA) mixture (1:20). 3-aminophenylboronic acid (3-APBA) in combination with the anti-LPS aptamer was used for the development of the hybrid molecularly imprinted sensor (apta-MIP). Aptasensors, targeting PCT and CRP were also fabricated, following the same approach as in the case of LPS, with mercaptohexanol (MCH) replacing DTBA. In the case of the CRP aptasensor, the SAM was formed following incubation of a 1:1 aptamer: MCH mixture. However, in the case of PCT, the SAM was formed with the aptamer itself, with subsequent backfilling with 1 μM MCH. The binding performance of all systems has been evaluated using electrochemical impedance spectroscopy. The apta-MIP’s polymer thickness is controlled by varying the number of electropolymerisation cycles. In the ideal number of polymerisation cycles, the polymer must cover the electrode surface and create a binding pocket around LPS and its aptamer binding site. Less polymerisation cycles will create a hybrid system which resembles an aptasensor, while more cycles will be able to cover the complex and demonstrate a bulk polymer-like behaviour. Both aptasensor and apta-MIP were challenged with LPS and compared to conventional imprinted (absence of aptamer from the binding site, polymer formed in presence of LPS) and non-imprinted polymers (NIPS, absence of LPS whilst hybrid polymer is formed). A stable LPS aptasensor, capable of detecting down to 5 pg/ml of LPS was generated. The apparent Kd of the system was estimated at 17 pM, with a Bmax of approximately 50 pM. The aptasensor demonstrated high specificity to LPS. The apta-MIP demonstrated superior recognition properties with a limit of detection of 1 fg/ml and a Bmax of 100 pg/ml. The CRP and PCT aptasensors were both able to detect down to 5 pg/ml. Whilst full binding performance is currently being evaluated, there is none of the sensors demonstrate cross-reactivity towards LPS, CRP or PCT. In conclusion, stable aptasensors capable of detecting LPS, PCT and CRP at low concentrations have been generated. The realisation of a multiplex panel such as described herein, will effectively contribute to the rapid, personalised diagnosis of sepsis.

Keywords: aptamer, electrochemical impedance spectroscopy, molecularly imprinted polymers, sepsis

Procedia PDF Downloads 113

Authors: Fruzsina Luca Kézér, Viktor Jurkovich, Ottó Szenci, János Tőzsér, Levente Kovács

Abstract:

Positive interaction between people and animals could have a favorable effect on the welfare and production by reducing stress levels. However, to the repeated contact with humans (e.g. farm staff, veterinarians or herdsmen), animals may respond with escape behavior or avoidance, which both have negative effects on the ease of handling, welfare and may lead to the expression of aggressive behaviors. Rough or aversive handling can impair health and the function of the cardiac autonomic activity due to fear and stress, which also can be determined by certain parameters of heart rate variability (HRV). Although the essential relationships between fear from humans and basal tone of the autonomic nervous system were described by the authors previously, several questions remained unclear in terms of the associations between different coping strategies (behavioral syndromes) of the animals and physiological responsiveness to humans. The main goal of this study was to find out whether human behavior and emotions to the animals have an impact on cardiac function and behavior of animals with different coping styles in response situations. Therefore, in the present study, special (fear, approaching, restraint, novel arena, novel object) tests were performed on healthy, 2-year old heifers (n = 104) differing in coping styles [reactive (passive) vs. proactive (active) coping]. Animals were categorized as reactive or proactive based on the following tests: 1) aggressive behavior at the feeding bunk, 2) avoidance from an approaching person, 3) immobility, and 4) daily activity (number of posture changes). Heart rate, the high frequency (HF) component of HRV as a measure of vagal activity and the ratio between the low frequency (LF) and HF components (LF/HF ratio) as a parameter of sympathetic nervous system activity were calculated for all individual during lying posture (baseline) and for response situations in novel object, novel arena, and unfamiliar person tests (both for 5 min), respectively. The differences between baseline and response were compared between groups. Higher sympathetic (higher heart rates and LF/HF ratios) and lower parasympathetic activity (lower HF) was found for proactive animals in response situations than for reactive (passive) animals either during the novel object, the novel arena and the unfamiliar person test. It suggests that animals with different behavioral traits differ in their immediate autonomic adaptation to novelty and people. Based on our preliminary results, it seems, that the analysis of HRV can help to understand the physiological manifestation of responsiveness to novelty and human presence in dairy cattle with different behavioral syndromes.

Keywords: behavioral syndromes, human-cattle interaction, novel arena test, physiological responsiveness, proactive coping, reactive coping

Procedia PDF Downloads 338

Authors: Sijie Chen, Xiaofang Chen, Juan Wang, Yingying Zhang, Yu Hong, Wanyu Zhuang, Xinxin Huang, Ping Ou, Longsheng Huang

Abstract:

Purpose: Autistic symptom improvement can be observed in children treated with acupuncture, but the mechanism is still being explored. In the present study, we used scalp acupuncture to treat autism rat model, and then their improvement in the abnormal behaviors and specific mechanisms behind were revealed by detecting animal behaviors, analyzing the RNA sequencing of the prefrontal cortex(PFC), and observing the ultrastructure of PFC neurons under the transmission electron microscope. Methods: On gestational day 12.5, Wistar rats were given valproic acid (VPA) by intraperitoneal injection, and their offspring were considered to be reliable rat models of autism. They were randomized to VPA or VPA-acupuncture group (n=8). Offspring of Wistar pregnant rats that were simultaneously injected with saline were randomly selected as the wild-type group (WT). VPA_acupuncture group rats received acupuncture intervention at 23 days of age for 4 weeks, and the other two groups followed without intervention. After the intervention, all experimental rats underwent behavioral tests. Immediately afterward, they were euthanized by cervical dislocation, and their prefrontal cortex was isolated for RNA sequencing and transmission electron microscopy. Results: The main results are as follows: 1. Animal behavioural tests: VPA group rats showed more anxiety-like behaviour and repetitive, stereotyped behaviour than WT group rats. While VPA group rats showed less spatial exploration ability, activity level, social interaction, and social novelty preference than WT group rats. It was gratifying to observe that acupuncture indeed improved these abnormal behaviors of autism rat model. 2. RNA-sequencing: The three groups of rats differed in the expression and enrichment pathways of multiple genes related to synaptic function, neural signal transduction, and circadian rhythm regulation. Our experiments indicated that acupuncture can alleviate the major symptoms of ASD by improving these neurological abnormalities. 3. Under the transmission electron microscopy, several lysosomes and mitochondrial structural abnormalities were observed in the prefrontal neurons of VPA group rats, which were manifested as atrophy of the mitochondrial membran, blurring or disappearance of the mitochondrial cristae, and even vacuolization. Moreover, the number of synapses and synaptic vesicles was relatively small. Conversely, the mitochondrial structure of rats in the WT group and VPA_acupuncture was normal, and the number of synapses and synaptic vesicles was relatively large. Conclusion: Acupuncture effectively improved the abnormal behaviors of autism rat model and the ultrastructure of the PFC neurons, which might worked by improving their abnormal synaptic function, synaptic plasticity and promoting neuronal signal transduction.

Keywords: autism spectrum disorder, acupuncture, animal behavior, RNA sequencing, transmission electron microscope

Procedia PDF Downloads 25

Authors: Elif Erdem, Nalan Kaya, Gonca Ozan, Durrin Ozlem Dabak, Enver Ozan

Abstract:

This study was carried out to determine the histological and biochemical changes in the lungs of the rat pups exposed to tobacco smoke during pregnancy period and to investigate the protective effects of alpha lipoic acid, which is administered during pregnancy, on these changes. In our study, 24 six-week old Spraque-Dawley female rats weighing 160 ± 10 g were used (n:7). Rats were randomly divided into four equal groups: group I (control), group II (tobacco smoke), group III (tobacco smoke + alpha lipoic acid) and group IV (alpha lipoic acid). Rats in the group II, group III were exposed to tobacco smoke twice a day for one hour starting from eight weeks before mating and during pregnancy. In addition to tobacco smoke, 20 mg/kg of alpha lipoic acid was administered via oral gavage to the rats in the group III. Only alpha lipoic acid was administered to the rats in the group IV. Once after the delivery, all administrations were stopped. On the 7 and 21th days, the seven pups of all groups were decapitated. A portion of the lung was taken and stained with HE, PAS and Masson. In addition to immunohistochemical staining of surfactant protein A, vascular endothelial growth factor, caspase-3, TUNEL method was also used to determine apoptosis. Biochemical analyzes were performed with some part of the lung tissue specimens. In the histological evaluations performed under light microscopy, inflammatory cell increase, hemorrhagic areas, edema, interalveolar septal thickening, alveolar numbers decrease, degeneration of some bronchi and bronchial epithelium, epithelial cells that were fallen into the lumen and hyaline membrane formation were observed in tobacco smoke group. These findings were ameliorated in tobacco smoke + ALA group. Hyaline membrane formation was not detected in this group. The TUNEL positive cell numbers a significant increase was detected in the tobacco smoke group, whereas a significant decrease was detected in the tobacco smoke + ALA group. In terms of the immunoreactivity of both SP-A and VEGF, a significant decrease was observed in the tobacco smoke group, and a significant increase was observed in the tobacco smoke + ALA group. Regarding the immunoreactivity of caspase-3, there was a significant increase in the group of tobacco smoke and a significant decrease in the group of tobacco smoke + ALA. The malondialdehyde levels were determined to be significantly increased in the tobacco smoke group, and a significant decreased in the tobacco smoke + ALA. Glutathione and superoxide dismutase enzyme activities showed a significant decrease in the group of tobacco smoke and a significant increase in the tobacco smoke + ALA group. In conclusion, we suggest that the exposure to tobacco smoke during pregnancy leads to morphological, histopathological and functional changes on lung development by causing oxidative damage in lung tissues of neonatal rats and the maternal use of alpha lipoic acid can provide a protective effect on the neonatal lung development against this oxidative stress originating from tobacco smoke.

Keywords: alpha lipoic acid, lung, neonate, tobacco smoke, pregnancy

Procedia PDF Downloads 193

Authors: Delphine Morales, Florian Lombart, Agathe Truchot, Pauline Maire, Pascale Vigneron, Antoine Galmiche, Catherine Lok, Muriel Vayssade

Abstract:

Targeted therapy molecules are used as a first-line treatment for metastatic melanoma with B-Raf mutation. Nevertheless, these molecules can cause side effects to patients and are efficient on 50 to 60 % of them. Indeed, melanoma cell sensitivity to targeted therapy molecules is dependent on tumor microenvironment (cell-cell and cell-extracellular matrix interactions). To better unravel factors modulating cell sensitivity to B-Raf inhibitor, we have developed and compared several melanoma models: from metastatic melanoma cells cultured as monolayer (2D) to a co-culture in a 3D dermal equivalent. Cell response was studied in different melanoma cell lines such as SK-MEL-28 (mutant B-Raf (V600E), sensitive to Vemurafenib), SK-MEL-3 (mutant B-Raf (V600E), resistant to Vemurafenib) and a primary culture of dermal human fibroblasts (HDFn). Assays have initially been performed in a monolayer cell culture (2D), then a second time on a 3D dermal equivalent (dermal human fibroblasts embedded in a collagen gel). All cell lines were treated with Vemurafenib (a B-Raf inhibitor) for 48 hours at various concentrations. Cell sensitivity to treatment was assessed under various aspects: Cell proliferation (cell counting, EdU incorporation, MTS assay), MAPK signaling pathway analysis (Western-Blotting), Apoptosis (TUNEL), Cytokine release (IL-6, IL-1α, HGF, TGF-β, TNF-α) upon Vemurafenib treatment (ELISA) and histology for 3D models. In 2D configuration, the inhibitory effect of Vemurafenib on cell proliferation was confirmed on SK-MEL-28 cells (IC50=0.5 µM), and not on the SK-MEL-3 cell line. No apoptotic signal was detected in SK-MEL-28-treated cells, suggesting a cytostatic effect of the Vemurafenib rather than a cytotoxic one. The inhibition of SK-MEL-28 cell proliferation upon treatment was correlated with a strong expression decrease of phosphorylated proteins involved in the MAPK pathway (ERK, MEK, and AKT/PKB). Vemurafenib (from 5 µM to 10 µM) also slowed down HDFn proliferation, whatever cell culture configuration (monolayer or 3D dermal equivalent). SK-MEL-28 cells cultured in the dermal equivalent were still sensitive to high Vemurafenib concentrations. To better characterize all cell population impacts (melanoma cells, dermal fibroblasts) on Vemurafenib efficacy, cytokine release is being studied in 2D and 3D models. We have successfully developed and validated a relevant 3D model, mimicking cutaneous metastatic melanoma and tumor microenvironment. This 3D melanoma model will become more complex by adding a third cell population, keratinocytes, allowing us to characterize the epidermis influence on the melanoma cell sensitivity to Vemurafenib. In the long run, the establishment of more relevant 3D melanoma models with patients’ cells might be useful for personalized therapy development. The authors would like to thank the Picardie region and the European Regional Development Fund (ERDF) 2014/2020 for the funding of this work and Oise committee of "La ligue contre le cancer".

Keywords: 3D human skin model, melanoma, tissue engineering, vemurafenib efficiency

Procedia PDF Downloads 288

Authors: Sandra Christina Gressler

Abstract:

Resting is essentially the physical and mental relaxation. So, can behaviors that go beyond the merely physical relaxation to some extent be understood as a behavior of restoration? Studies on restorative environments emphasize the physical, mental and social benefits that some environments can provide and suggest that activities in natural environments reduce the stress of daily lives, promoting recovery against the daily wear. These studies, though specific in their results, do not unify the different possibilities of restoration. Considering the importance of restorative environments by promoting well-being, this research aims to verify the applicability of the theory on restorative environments in a Brazilian context, inquiring about the environment/behavior of rest. The research sought to achieve its goals by; a) identifying daily ways of how participants interact/connect with nature; b) identifying the resting environments/behavior; c) verifying if rest strategies match the restorative environments suggested by restorative studies; and d) verifying different rest strategies related to time. Workers from different companies in which certain functions require focused attention, and high school students from different schools, participated in this study. An interview was used to collect data and information. The data obtained were compared with studies of attention restoration theory and stress recovery. The collected data were analyzed through the basic descriptive inductive statistics and the use of the software ALCESTE® (Analyse Lexicale par Contexte d'un Ensemble de Segments de Texte). The open questions investigate perception of nature on a daily basis – analysis using ALCESTE; rest periods – daily, weekends and holidays – analysis using ALCESTE with tri-croisé; and resting environments and activities – analysis using a simple descriptive statistics. According to the results, environments with natural characteristics that are compatible with personal desires (physical aspects and distance) and residential environments when they fulfill the characteristics of refuge, safety, and self-expression, characteristics of primary territory, meet the requirements of restoration. Analyzes suggest that the perception of nature has a wide range that goes beyond the objects nearby and possible to be touched, as well as observation and contemplation of details. The restoration processes described in the studies of attention restoration theory occur gradually (hierarchically), starting with being away, following compatibility, fascination, and extent. They are also associated with the time that is available for rest. The relation between rest behaviors and the bio-demographic characteristics of the participants are noted. It reinforces, in studies of restoration, the need to insert not only investigations regarding the physical characteristics of the environment but also behavior, social relationship, subjective reactions, distance and time available. The complexity of the theme indicates the necessity for multimethod studies. Practical contributions provide subsidies for developing strategies to promote the welfare of the population.

Keywords: attention restoration theory, environmental psychology, rest behavior, restorative environments

Procedia PDF Downloads 166

Authors: Mustafa M. Donma, Orkide Donma

Abstract:

Obesity is known to be associated with many clinically important diseases including metabolic syndrome (MetS). Vitamin B₁₂ plays essential roles in fat and protein metabolisms and its cooperation with vitamin B₉ is well-known. The aim of this study is to investigate the possible contributions as well as associations of these micronutrients upon obesity and MetS during childhood. A total of 128 children admitted to Namik Kemal University, Medical Faculty, Department of Pediatrics Outpatient Clinics were included into the scope of this study. The mean age±SEM of 92 morbid obese (MO) children and 36 with MetS were 118.3±3.8 months and 129.5±6.4 months, respectively (p > 0.05). The study was approved by Namık Kemal University, Medical Faculty Ethics Committee. Written informed consent forms were obtained from the parents. Demographic features and anthropometric measurements were recorded. WHO BMI-for age percentiles were used. The values above 99 percentile were defined as MO. Components of MetS [waist circumference (WC), fasting blood glucose (FBG), triacylglycerol (TRG), high density lipoprotein cholesterol (HDL-Chol), systolic pressure (SP), diastolic pressure (DP)] were determined. Routine laboratory tests were performed. Serum vitamin B₁₂ concentrations were measured using electrochemiluminescence immunoassay. Vitamin B₉ was analyzed by an immunoassay analyzer. Values for vitamin B₁₂ < 148 pmol/L, 148-221 pmol/L, > 221 pmol/L were accepted as low, borderline and normal, respectively. Vitamin B₉ levels ≤ 4 mcg/L defined deficiency state. Statistical evaluations were performed by SPSSx Version 16.0. p≤0.05 was accepted as statistical significance level. Statistically higher body mass index (BMI), WC, hip circumference (C) and neck C were calculated in MetS group compared to children with MO. No difference was noted for head C. All MetS components differed between the groups (SP, DP p < 0.001; WC, FBG, TRG p < 0.01; HDL-Chol p < 0.05). Significantly decreased vitamin B₉ and vitamin B₁₂ levels were detected (p < 0.05) in children with MetS. In both groups percentage of folate deficiency was 5.5%. No cases were below < 148 pmol/L. However, in MO group 14.3% and in MetS group 22.2% of the cases were of borderline status. In MO group B₁₂ levels were negatively correlated with BMI, WC, hip C and head C, but not with neck C. WC, hip C, head C and neck C were all negatively correlated with HDL-Chol. None of these correlations were observed in the group of children with MetS. Strong positive correlation between FBG and insulin as well as strong negative correlation between TRG and HDL-Chol detected in MO children were lost in MetS group. Deficiency state end-products of both B₉ and B₁₂ may interfere with the expected profiles of MetS components. In this study, the alterations in MetS components affected vitamin B₁₂ metabolism and also its associations with anthropometric body measurements. Further increases in vitamin B₁₂ and vitamin B₉ deficiency in MetS associated with the increased vitamin B₁₂ as well as vitamin B₉ deficiency metabolites may add to MetS parameters.

Keywords: children, cobalamin, folate, metabolic syndrome, obesity

Procedia PDF Downloads 174

Authors: Vaidehi Raipat

Abstract:

A city is an expression of coexistence between its users and built environment. The way in which its spaces are animated signify the quality of this coexistence. Urban sustainability is the ability of a city to respond efficiently towards its people, culture, environment, visual image, history, visions and identity. The quality of built environment determines the quality of our lifestyles, but poor ability of the built environment to adapt and sustain itself through the changes leads to degradation of cities. Ranchi was created in November 2000, as the capital of the newly formed state Jharkhand, located on eastern side of India. Before this Ranchi was known as summer capital of Bihar and was a little larger than a town in terms of development. But since then it has been vigorously expanding in size, infrastructure as well as population. This sudden expansion has created a stress on existing built environment. The large forest covers, agricultural land, diverse culture and pleasant climatic conditions have degraded and decreased to a large extent. Narrow roads and old buildings are unable to bear the load of the changing requirements, fast improving technology and growing population. The built environment has hence been rendered unsustainable and unadaptable through fastidious changes of present era. Some of the common hazards that can be easily spotted in the built environment are half-finished built forms, pedestrians and vehicles moving on the same part of the road. Unpaved areas on street edges. Over-sized, bright and randomly placed hoardings. Negligible trees or green spaces. The old buildings have been poorly maintained and the new ones are being constructed over them. Roads are too narrow to cater to the increasing traffic, both pedestrian and vehicular. The streets have a large variety of activities taking place on them, but haphazardly. Trees are being cut down for road widening and new constructions. There is no space for greenery in the commercial as well as old residential areas. The old infrastructure is deteriorating because of poor maintenance and the economic limitations. Pseudo understanding of functionality as well as aesthetics drive the new infrastructure. It is hence necessary to evaluate the extent of sustainability of existing built environment of the city and create or regenerate the existing built environment into a more sustainable and adaptable one. For this purpose, research titled “Sustainability of the Built Environment of Ranchi District” has been carried out. In this research the condition of the built environment of Ranchi are explored so as to figure out the problems and shortcomings existing in the city and provide for design strategies that can make the existing built-environment sustainable. The built environment of Ranchi that include its outdoor spaces like streets, parks, other open areas, its built forms as well as its users, has been analyzed in terms of various urban design parameters. Based on which strategies have been suggested to make the city environmentally, socially, culturally and economically sustainable.

Keywords: adaptable, built-environment, sustainability, urban

Procedia PDF Downloads 217

Authors: Sudipta Garai

Abstract:

Films as a cultural, social practice remains a dominant space for creation and destruction of ideologies and practices which make the sociological viewing, analysis, and interpretation of the same a complex affair. It remains the doorway between the interpretations and understanding of the writer/director and the reader/viewer. India, being a multi-linguistic culture, the media plays a much intriguing role than that of newspaper, books, stories, novels or any other medium of expression. Known to be the largest democracy, the State seem to guarantee and safeguard people’s choices and life of dignity through its Fundamental Rights and Directives. However, the laws contradict themselves when it comes to IPC 377 criminalizing anything except penovaginal sexual intercourse restricting alternative sexual preferences and practices questioning its sense of ‘democracy.' In this context, the issue of homosexuality came up in bits and pieces through various representations in ‘popular’ cinema mostly with sudden references of mockery and laughter where the explicit narratives of ‘queer’ seemed missing. Rituparno Ghosh, an eminent film maker of Bengal, came up as the ‘queer’ face in Kolkata specifically through his ‘queer’ trilogy (Memories in March, 2010; Arekti Premer Golpo, 2010; Chitrangada: A Crowning Wish, 2012) coming out of his own closet and speaking about his own sexual choices not only through the explicit narratives in films but also in person which made these films an important point of departure in Bengali film history. A sociological reading of these films through a discourse analysis is being done with the critical questions of ‘choice,' ’freedom,' ‘love and marriage’ and most importantly the ‘change.' This study not only focuses on the films and its analysis of content but also to engage with its audience, queer and not in order to extend beyond the art form into the actual vulnerabilities of life and experiences through informal interviews, focused group discussions and engaging with the real life narratives. A research of this kind is always looked upon as a medium of change hoping for a better world wiping away the discrimination and ‘shame’ the ‘queer’ faces in their everyday life, but a social science research is limited but its ‘time’ and academic boundary where the hope of change might be initiated but not fulfilled. The experiences and reflections of the ‘queer’ not only redefined the narratives of the films but also me as a researcher. The perspectives of the ‘hetero-normative’ informants gave a broader picture of the study and the socio-cultural complications that are intrigued with the ideas of resistance and change. The issues on subjectivity, power, and position couldn’t be wiped out in a study of this kind as both politics and aesthetics become integrated with each other in the creation of any art form be it films or a study of research.

Keywords: cinema, alternative sexualities, narratives, sexual choices, state and society

Procedia PDF Downloads 343

Authors: Drago Bratkovic, Curtis Gravance, David Ketteridge, Ravi Krishnan, Michael Imperiale

Abstract:

The mucopolysaccharidoses (MPSs) are a group of lysosomal storage diseases that have a common defect in the catabolism of glycosaminoglycans (GAGs). MPS I is the most common of the MPS diseases. Manifestations of MPS I include coarsening of facial features, corneal clouding, developmental delay, short stature, skeletal manifestations, hearing loss, cardiac valve disease, hepatosplenomegaly, and umbilical and inguinal hernias. Treatments for MPS I restore or activate the missing or deficient enzyme in the case of enzyme replacement therapy (ERT) and haematopoietic stem cell transplantation (HSCT). Pentosan polysulfate sodium (PPS) is a potential treatment to improve bone and joint manifestations of MPS I. The mechanisms of action of PPS that are relevant to the treatment of MPS I are the ability to: (i) Reduce systemic and accumulated GAG, (ii) Reduce inflammatory effects via the inhibition of NF-kB, resulting in the reduction in pro-inflammatory mediators. (iii) Reduce the expression of the pain mediator nerve growth factor in osteocytes from degenerating joints. (iv) Inhibit the cartilage degrading enzymes related to joint dysfunction in MPS I. PPS is being evaluated as an adjunctive therapy to ERT and/or HSCT in an open-label, single-centre, phase 2 study. Patients are ≥ 5 years of age with a diagnosis of MPS I and previously received HSCT and/or ERT. Three white, female, patients with MPS I-Hurler, ages 14, 15, and 19 years, and one, white male patient aged 15 years are enrolled. All were diagnosed at ≤2 years of age. All patients received HSCT ≤ 6 months after diagnosis. Two of the patients were treated with ERT prior to HSCT, and 1 patient received ERT commencing 3 months prior to HSCT. Two patients received 0.75mg/kg and 2 patients received 1.5mg/kg of PPS. PPS was well tolerated at doses of 0.75 and 1.5 mg/kg to 47 weeks of continuous dosing. Of the 19 adverse events (AEs), 2 were related to PPS. One AE was moderate (pre-syncope) and 1 was mild (injection site bruising), experienced in the same patient. All AEs were reported as mild or moderate. There have been no SAEs. One subject experienced a COVID-19 infection and PPS was interrupted. The MPS I signature GAG fragments, sulfated disaccharide and UA-HNAc S, tended to decrease in 3 patients from baseline through Week 25. Week 25 GAG data are pending for the 4th patient. Overall, most biomarkers (inflammatory, cartilage degeneration, and bone turnover) evaluated in the 3 patients with 25-week assessments have indicated either no change or a reduction in levels compared to baseline. In 3 patients, there was a trend toward improvement in the 2MWT from baseline to Week 48 with > 100% increase in 1 patient (01-201). In the 3 patients that had Week 48 assessments, patients and proxies reported improvement in PGIC, including “worthwhile difference” (n=1), or “made all the difference” (n=2).

Keywords: MPS I, pentosan polysulfate sodium, clinical study, 2MWT, QoL

Procedia PDF Downloads 98

Authors: Omnia Y. Shawky, Asmaa M. Megahed, Alaa E. ElKomy, A. E. Abd-El-Hamid, Y. A. Attia

Abstract:

This study was carried out to elevate the broilers physiological response against heat stress and reduce this impact by adding vitamin C (VC), vitamin E (VE) alone/or with organic zinc (Zn) to chicks’ rations. A total of 192, 26-day-old Arbor Acers male chicks were randomly divided into equal 8 groups (4 replicates for each). All experimental groups were treated as follow: Group 2 was served as a heat stress control that reared at 37ºC with relative humidity 53 ± 8% for 6 hours/day for three successive days/week and fed the basal diet only. Groups 3-8 were heat stressed in a like manner to group 2 and fed basal diet inclusion 200mg VC (group 3), 200mg VE (group 4), 200mg VC+200mg VE (group 5), 200mg VC+30mg Zn (group 6), 200mg VE+30mg Zn (group 7) and 200mg VC+200mg VE+30mg Zn (group 8) /kg feed, while Group 1 was served as a positive control that reared on a neutral temperature (NT) (approximately 21ºC) and fed the basal diet only. Respiration rate and rectal temperature were boosted of HS chicks (80.8 breath/min and 41.97ºC) compared to NT group (60.12 breath/min and 40.9ºC), while, adding VC alone and with VE or Zn resulted in decrease these measurements. Heat stress had a significantly negative effect on chicks body weight gain, feed consumption and feed conversion ratio compared to the NT group, this harmful effect could be overcome by adding VC and VE individually or with Zn. Chicks exposed to heat stress showed slightly increase hemoglobin concentration compared to NT group, while, adding VC, VE individually or with Zn alleviated this effect. Plasma glucose concentration was significantly increased in HS group than the NT group, but adding VC, VE individually or with Zn resulted in a reduction plasma glucose level, which it was still higher than the NT group. Heat stress caused an increase in plasma total lipids and cholesterol concentration compared to the NT group and inclusion VC or VE alone or with Zn was not able to reduce this effect. The increased liver enzymes activities (AST and ALT) that observed in HS group compared to NT group were removed by adding VC and VE individually or with Zn. As well, exposure of broiler chicks to heat stress resulted in a slightly decrease in plasma total antioxidant capacity level (TAC) superoxide dismutase and catalase enzymes activities, while inclusion VC and VE individually or with Zn in chicks rations caused an increased in these measurements. Broiler chicks that exposed to HS revealed a significant increase in heat shock protein (Hsp 70) compared to the NT group, while, adding VC or VE individually or with Zn resulted in a significant decrease in Hsp70 than the HS group and VE alone or with VC had the greatest effect. In conclusion, it could be overcome the harmful and the negative effect of heat stress on broiler chicks’ productive performance and physiological status by inclusion VC (200mg) or VE (200mg) individual or in a combination with organic zinc (30 mg) in chicks’ rations.

Keywords: heat stress, broiler, vitamin C, vitamin E, organic zinc

Procedia PDF Downloads 187

Authors: Tomasz Lewinski

Abstract:

The right to know becomes gradually recognised as an increasing number of states adopts national legislations regarding access to state-held information. Laws differ from each other in the scope of the right to information (hereinafter: RTI). In all regimes of RTI, there are exceptions from the general notion of the right. States’ authorities too often use exceptions to justify refusals to requests for state-held information. This paper sets out how states hamper RTI basing on the notion of exception and by not providing an effective procedure that could redress unlawful denials. This paper bases on two selected examples of RTI incorporation into the national legal regime, United Kingdom, and South Africa. It succinctly outlines the international standard given in Article 19 of the International Covenant on Civil and Political Rights (hereinafter: ICCPR) and its influence on the RTI in selected countries. It shortly demonstrates as a background to further analysis the Human Rights Committee’s jurisprudence and standards articulated by successive Special Rapporteurs on freedom of opinion and expression. Subsequently, it presents a brief comparison of these standards with the regional standards, namely the African Charter on Human and Peoples' Rights and the European Convention on Human Rights. It critically discusses the regimes of exceptions in RTI legislations in respective national laws. It shows how excessive these regimes are, what implications they have for the transparency in general. Also, the objective is to divide exceptions enumerated in legislations of selected states in relation to exceptions provided in Article 19 of the ICCPR. Basing on the established division of exceptions by its natures, it compares both regimes of exceptions related to the principle of national security. That is to compare jurisprudence of domestic courts, and overview practices of states’ authorities applied to RTI requests. The paper evaluates remedies available in legislations, including contexts of the length and costs of the subsequent proceedings. This provides a general assessment of the given mechanisms and present potential risks of its ineffectiveness. The paper relies on examination of the national legislations, comments of the credible non-governmental organisations (e.g. The Public's Right to Know Principles on Freedom of Information Legislation by the Article 19, The Tshwane Principles on National Security and the Right to Information), academics and also the research of the relevant judgements delivered by domestic and international courts. Conclusion assesses whether selected countries’ legislations go in line with international law and trends, whether the jurisprudence of the regional courts provide appropriate benchmarks for national courts to address RTI issues effectively. Furthermore, it identifies the largest disadvantages of current legislations and to what outcomes it leads in domestic courts jurisprudences. In the end, it provides recommendations and policy arguments for states to improve transparency and support local organisations in their endeavours to establish more transparent states and societies.

Keywords: access to information, freedom of information, national security, right to know, transparency

Procedia PDF Downloads 195

Authors: Joseph Falzon, Margaret Nelson

Abstract:

Background: Regeneration and re-use of abandoned historic buildings present a continuous challenge for policy makers and stakeholders in the tourism and leisure industry. Obsolete historic buildings provide great potential for tourism and leisure accommodation, presenting unique heritage experiences to travellers and host communities. Contemporary demands in the hospitality industry continuously require higher standards, some of which are in conflict with heritage conservation principles. Objective: The aim of this research paper is to critically discuss regeneration policies with stakeholders of the tourism and leisure industry and to examine current practices in policy development and the resultant impact of policies on the Maltese tourism and leisure industry. Research Design: Six semi-structured interviews with stakeholders involved in the tourism and leisure industry participated in the research. A number of measures were taken to reduce bias and thus improve trustworthiness. Clear statements of the purpose of the research study were provided at the start of each interview to reduce expectancy bias. The interviews were semi-structured to minimise interviewer bias. Interviewees were allowed to expand and elaborate as necessary, with only necessary probing questions, to allow free expression of opinion and practices. Interview guide was submitted to participants at least two weeks before the interview to allow participants to prepare for the interview and prevent recall bias during the interview as much as possible. Interview questions and probes contained both positive and negative aspects to prevent interviewer bias. Policy documents were available during the interview to prevent recall bias. Interview recordings were transcribed ‘intelligent’ verbatim. Analysis was carried out using thematic analysis with the coding frame developed independently by two researchers. All phases of the study were governed by research ethics. Findings: Findings were grouped in main themes: financing of regeneration, governance, legislation and policies. Other key issues included value of historic buildings and approaches for regeneration. Whist regeneration of historic buildings was noted, participants discussed a number of barriers that hindered regeneration. Stakeholders identified gaps in policies and gaps at policy implementation stages. European Union funding policies facilitated regeneration initiatives but funding criteria based on economic deliverables presented the intangible heritage gap. Stakeholders identified niche markets for heritage tourism accommodation. Lack of research-based policies was also identified. Conclusion: Potential of regeneration is hindered by inadequate legal framework that supports contemporary needs of the tourism industry. Policies should be developed by active stakeholder participation. Adequate funding schemes have to support the tangible and intangible components of the built heritage.

Keywords: governance, historic buildings, policy, tourism

Procedia PDF Downloads 218

Authors: Jeanne Leblond, Warren Viricel, Amira Mbarek

Abstract:

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

Procedia PDF Downloads 188

Authors: Smita Lahkar

Abstract:

Sattriya, the youngest of the eight principal Classical Indian dance traditions, has undergone too many changes and modifications to arrive at its present stage of performing art form extracting itself from age-old religious confinement. Although some of the other traditions have been revived in the recent past, Sattriya has a living tradition since its inception in the 15th century by Srimanta Sankardeva, the great Vaishnavite saint, poet, playwright, lyricist, painter, singer and dancer of Assam, a primary north-eastern state of India. This living dance tradition from the Sattras, the Vaishnavite monasteries, has been practiced for over five hundred years by celibate male monks, as a powerful medium for propagating the Vaishnava religious faith. Sankardeva realised the potential of the vocalised word integrated with the visual image as a powerful medium of expression and communication. So he used this principal medium for propagating his newly found message of devotion among the people of his time. Earlier, Sattriya was performed by male monks alone in monasteries (Sattras) as a part of daily rituals. The females were not even allowed to learn this art form. But, in present time, Sattriya has come out from the Sattras to proscenium stage, performed mostly by female as well as few male dancers also. The technique of performing movements, costumes, ornaments, music and style of performance too have experienced too many changes and modifications. For example, earlier and even today in Sattra, the ‘Pataka’ hand gesture is depicted in conformity with the original context (religious) of creation of the dance form. But, today stage-performers prefer the instructions of the scripture ‘Srihastamuktavali’ and depict the ‘Pataka’ in a sophisticated manner affecting decontextualisation to a certain extent. This adds aesthetic beauty to the dance form as an art distancing it from its context of being a vehicle for propagating Vaishnava religion. The Sattriya dance today stands at the crossroads of past and future, tradition and modernity, devotion and display, spirituality and secularism. The traditional exponents trained under the tutelage of Sattra maestros and imbibing a devotionally inspired rigour of the religion, try to retain the traditional nuances; while the young artists being trained outside the monasteries are more interested in taking up the discipline purely from the perspective of ‘performing arts’ bereft of the philosophy of religion or its sacred associations. Hence, this paper will be an endeavor to establish the hypothesis that the Sattriya, whose origin was for propagating Vaishnava faith, has now entered the world of performing arts with highly aesthetical components. And as a transformed art form, Sattriya may be expected to carve a niche in world dance arena. This will be done with the help of historical evidences, observations from the recorded past and expert rendezvous.

Keywords: dance, performing art, religion, Sattriya

Procedia PDF Downloads 200

Authors: Jesús Horacio, Alfredo Ollero, Víctor Bouzas-Blanco, Augusto Pérez-Alberti

Abstract:

Rivers have traditionally been classified, assessed and managed in terms of hydrological, chemical and / or biological criteria. Geomorphological classifications had in the past a secondary role, although proposals like River Styles Framework, Catchment Baseline Survey or Stroud Rural Sustainable Drainage Project did incorporate geomorphology for management decision-making. In recent years many studies have been attracted to the geomorphological component. The geomorphological processes and their associated forms determine the structure of a river system. Understanding these processes and forms is a critical component of the sustainable rehabilitation of aquatic ecosystems. The fluvial auto-classification approach suggests that a river is a self-built natural system, with processes and forms designed to effectively preserve their ecological function (hydrologic, sedimentological and biological regime). Fluvial systems are formed by a wide range of elements with multiple non-linear interactions on different spatial and temporal scales. Besides, the fluvial auto-classification concept is built using data from the river itself, so that each classification developed is peculiar to the river studied. The variables used in the classification are specific stream power and mean grain size. A discriminant analysis showed that these variables are the best characterized processes and forms. The statistical technique applied allows to get an individual discriminant equation for each geomorphological type. The geomorphological classification was developed using sites with high naturalness. Each site is a control point of high ecological and geomorphological quality. The changes in the conditions of the control points will be quickly recognizable, and easy to apply a right management measures to recover the geomorphological type. The study focused on Galicia (NW Spain) and the mapping was made analyzing 122 control points (sites) distributed over eight river basins. In sum, this study provides a method for fluvial geomorphological classification that works as an open and flexible tool underlying the fluvial auto-classification concept. The hydrographic mapping is the visual expression of the results, such that each river has a particular map according to its geomorphological characteristics. Each geomorphological type is represented by a particular type of hydraulic geometry (channel width, width-depth ratio, hydraulic radius, etc.). An alteration of this geometry is indicative of a geomorphological disturbance (whether natural or anthropogenic). Hydrographic mapping is also dynamic because its meaning changes if there is a modification in the specific stream power and/or the mean grain size, that is, in the value of their equations. The researcher has to check annually some of the control points. This procedure allows to monitor the geomorphology quality of the rivers and to see if there are any alterations. The maps are useful to researchers and managers, especially for conservation work and river restoration.

Keywords: fluvial auto-classification concept, mapping, geomorphology, river

Procedia PDF Downloads 354

Authors: Ming-Chien Su, Yi-Zih Chen, Nien-Hsin Kao, Hideaki Shibata

Abstract:

The major component of the atmosphere is nitrogen, yet atmospheric nitrogen has limited availability for biological use. Human activities have produced different types of nitrogen related compounds such as nitrogen oxides from combustion, nitrogen fertilizers from farming, and the nitrogen compounds from waste and wastewater, all of which have impacted the environment. Many studies have indicated the N-footprint is dominated by food, followed by housing, transportation, and goods and services sectors. To solve the impact issues from agricultural land, nitrogen cycle research is one of the key solutions. The study site is located in Hualien County, Taiwan, a major rice and food production area of Taiwan. Importantly, environmentally friendly farming has been promoted for years, and an environmental indicator system has been established by previous authors based on the concept of resilience capacity index (RCI) and environmental performance index (EPI). Nitrogen management is required for food production, as excess N causes environmental pollution. Therefore it is very important to develop a roadmap of the nitrogen footprint, and to integrate it with environmental indicators. The key focus of the study thus addresses (1) understanding the environmental impact caused by the nitrogen cycle of food products and (2) uncovering the trend of the N-footprint of agricultural products in Hualien, Taiwan. The N-footprint model was applied, which included both crops and energy consumption in the area. All data were adapted from government statistics databases and crosschecked for consistency before modeling. The actions involved with agricultural production were evaluated and analyzed for nitrogen loss to the environment, as well as measuring the impacts to humans and the environment. The results showed that rice makes up the largest share of agricultural production by weight, at 80%. The dominant meat production is pork (52%) and poultry (40%); fish and seafood were at similar levels to pork production. The average per capita food consumption in Taiwan is 2643.38 kcal capita−1 d−1, primarily from rice (430.58 kcal), meats (184.93 kcal) and wheat (ca. 356.44 kcal). The average protein uptake is 87.34 g capita−1 d−1, and 51% is mainly from meat, milk, and eggs. The preliminary results showed that the nitrogen footprint of food production is 34 kg N per capita per year, congruent with the results of Shibata et al. (2014) for Japan. These results provide a better understanding of the nitrogen demand and loss in the environment, and the roadmap can furthermore support the establishment of nitrogen policy and strategy. Additionally, the results serve to develop a roadmap of the nitrogen cycle of an environmentally friendly farming area, thus illuminating the nitrogen demand and loss of such areas.

Keywords: agriculture productions, energy consumption, environmental indicator, nitrogen footprint

Procedia PDF Downloads 288

Authors: Lucas Derks, Christine Beenhakker, Michiel Brandt, Gert Arts, Ruud van Langeveld

Abstract:

Background: The method Depression in Awareness Space (DAS) is a psychotherapeutic intervention technique based on the principles of spatial cognition and imagination therapy with spatial components. The basic assumptions are: mental space is the primary organizing principle in the mind, and all psychological issues can be treated by first locating and by next relocating the conceptualizations involved. The most clinical experience was gathered over the last 20 years in the area of social issues (with the social panorama model). The latter work led to the conclusion that a mental object (image) gains emotional impact when it is placed more central, closer and higher in the visual field – and vice versa. Changing the locations of mental objects in space thus alters the (socio-) emotional meaning of the relationships. The experience of depression seems always associated with darkness. Psychologists tend to see the link between depression and darkness as a metaphor. However, clinical practice hints to the existence of more literal forms of darkness. Aims: The aim of the method Depression in Awareness Space is to reduce the distress of clients with depression in the clinical counseling practice, as a reliable alternative method of psychological therapy for the treatment of depression. The method Depression in Awareness Space aims at making dark areas smaller, lighter and more transparent in order to identify the problem or the cause of the depression which lies behind the darkness. It was hypothesized that the darkness is a subjective side-effect of the neurological process of repression. After reducing the dark clouds the real problem behind the depression becomes more visible, allowing the client to work on it and in that way reduce their feelings of depression. This makes repression of the issue obsolete. Results: Clients could easily get into their 'sadness' when asked to do so and finding the location of the dark zones proved pretty easy as well. In a recent pilot study with five participants with mild depressive symptoms (measured on two different scales and tested against an untreated control group with similar symptoms), the first results were also very promising. If the mental spatial approach to depression can be proven to be really effective, this would be very good news. The Society of Mental Space Psychology is now looking for sponsoring of an up scaled experiment. Conclusions: For spatial cognition and the research into spatial psychological phenomena, the discovery of dark areas can be a step forward. Beside out of pure scientific interest, it is great to know that this discovery has a clinical implication: when darkness can be connected to depression. Also, darkness seems to be more than metaphorical expression. Progress can be monitored over measurement tools that quantify the level of depressive symptoms and by reviewing the areas of darkness.

Keywords: depression, spatial cognition, spatial imagery, social panorama

Procedia PDF Downloads 151

Authors: Rafael Santos, Clara Pimenta Do Vale, Barbara Bogoni, Poul Henning Kirkegaard

Abstract:

In the last decades, the relevance of hand drawing has decreased in the scope of architectural education. However, some schools continue to recognize its decisive role, not only in the architectural design teaching, but in the whole of architectural training. With this paper it is intended to present the results of a research developed on the following problem: the practise of hand drawing as a premier form of representation in architectural design teaching. The research had as its object the educational model of the Faculty of Architecture of the University of Porto (FAUP) and was led by three main objectives: to identify the circumstance that promoted hand drawing as a form of representation in FAUP's model; to characterize the types of hand drawing and their role in that model; to determine the particularities of hand drawing as a premier form of representation in architectural design teaching. Methodologically, the research was conducted according to a qualitative embedded single-case study design. The object – i.e., the educational model – was approached in FAUP case considering its Context and three embedded unities of analysis: the educational Purposes, Principles and Practices. In order to guide the procedures of data collection and analysis, a Matrix for the Characterization (MCC) was developed. As a methodological tool, the MCC allowed to relate the three embedded unities of analysis with the three main sources of evidence where the object manifests itself: the professors, expressing how the model is Assumed; the architectural design classes, expressing how the model is Achieved; and the students, expressing how the model is Acquired. The main research methods used were the naturalistic and participatory observation, in-person-interview and documentary and bibliographic review. The results reveal that the educational model of FAUP – following the model of the former Porto School – was largely due to the methodological foundations created with the hand drawing teaching-learning processes. In the absence of a culture of explicit theoretical elaboration or systematic research, hand drawing was the support for the continuity of the school, an expression of a unified thought about what should be the reflection and practice of architecture. As a form of representation, hand drawing plays a transversal role in the entire educational model, since its purposes are not limited to the conception of architectural design – it is also a means for perception, analysis and synthesis. Regarding the architectural design teaching, there seems to be an understanding of three complementary dimensions of didactics: the instrumental, methodological and propositional dimension. At FAUP, hand drawing is recognized as the common denominator among these dimensions, according to the idea of "globality of drawing". It is expected that the knowledge base developed in this research may have three main contributions: to contribute to the maintenance and valorisation of FAUP’s model; through the precise description of the methodological procedures, to contribute by transferability to similar studies; through the critical and objective framework of the problem underlying the hand drawing in architectural design teaching, to contribute to the broader discussion concerning the contemporary challenges on architectural education.

Keywords: architectural design teaching, architectural education, forms of representation, hand drawing

Procedia PDF Downloads 110

Authors: H. Mayton, X. Yan, A. G. Taylor

Abstract:

Infested tomato seeds were used to investigate the influence of Xanthomonas euvesicatoria on germination and seed to seedling transmission in a controlled environment and greenhouse assays in an effort to develop effective seed treatments and characterize seed borne transmission of bacterial leaf spot of tomato. Bacterial leaf spot of tomato, caused by four distinct Xanthomonas species, X. euvesicatoria, X. gardneri, X. perforans, and X. vesicatoria, is a serious disease worldwide. In the United States, disease prevention is expensive for commercial growers in warm, humid regions of the country, and crop losses can be devastating. In this study, four different infested tomato seed lots were extracted from tomato fruits infected with bacterial leaf spot from a field in New York State in 2017 that had been inoculated with X. euvesicatoria. In addition, vacuum infiltration at 61 kilopascals for 1, 5, 10, and 15 minutes and seed soaking for 5, 10, 15, and 30 minutes with different bacterial concentrations were used to artificially infest seed in the laboratory. For controlled environment assays, infested tomato seeds from the field and laboratory were placed othe n moistened blue blotter in square plastic boxes (10 cm x 10 cm) and incubated at 20/30 ˚C with an 8/16 hour light cycle, respectively. Infested tomato seeds from the field and laboratory were also planted in small plastic trays in soil (peat-lite medium) and placed in the greenhouse with 24/18 ˚C day and night temperatures, respectively, with a 14-hour photoperiod. Seed germination was assessed after eight days in the laboratory and 14 days in the greenhouse. Polymerase chain reaction (PCR) using the hrpB7 primers (RST65 [5’- GTCGTCGTTACGGCAAGGTGGTG-3’] and RST69 [5’-TCGCCCAGCGTCATCAGGCCATC-3’]) was performed to confirm presence or absence of the bacterial pathogen in seed lots collected from the field and in germinating seedlings in all experiments. For infested seed lots from the field, germination was lowest (84%) in the seed lot with the highest level of bacterial infestation (55%) and ranged from 84-98%. No adverse effect on germination was observed from artificially infested seeds for any bacterial concentration and method of infiltration when compared to a non-infested control. Germination in laboratory assays for artificially infested seeds ranged from 82-100%. In controlled environment assays, 2.5 % were PCR positive for the pathogen, and in the greenhouse assays, no infected seedlings were detected. From these experiments, X. euvesicatoria does not appear to adversely influence germination. The lowest rate of germination from field collected seed may be due to contamination with multiple pathogens and saprophytic organisms as no effect of artificial bacterial seed infestation in the laboratory on germination was observed. No evidence of systemic movement from seed to seedling was observed in the greenhouse assays; however, in the controlled environment assays, some seedlings were PCR positive. Additional experiments are underway with green fluorescent protein-expressing isolates to further characterize seed to seedling transmission of the bacterial leaf spot pathogen in tomato.

Keywords: bacterial leaf spot, seed germination, tomato, Xanthomonas euvesicatoria

Procedia PDF Downloads 118

Authors: Bibha Kumari, Nigel P. Brunton, Dilip K. Rai, Brijesh K. Tiwari

Abstract:

Edible mushrooms possess numerous functional components like homo- and hetero- β-glucans [β(1→3), β(1→4) and β(1→6) glucosidic linkages], chitins, ergosterols, bioactive polysaccharides and peptides imparting health beneficial properties to mushrooms. Some of the proven biological activities of mushroom extracts are antioxidant, antimicrobial, immunomodulatory, cholesterol lowering activity by inhibiting a key cholesterol metabolism enzyme i.e. 3-hydroxy-3-methyl-glutaryl CoA reductase (HMGCR), angiotensin I-converting enzyme (ACE) inhibition. Application of novel extraction technologies like pulsed electric field (PEF) and high power ultrasound offers clean, green, faster and efficient extraction alternatives with enhanced and good quality extracts. Sequential PEF followed by ultrasound assisted extraction (UAE) were applied to recover bioactive enriched fractions from industrial white button mushroom (Agaricus bisporus) stalk waste using environmentally friendly and GRAS solvents i.e. water and water/ethanol combinations. The PEF treatment was carried out at 60% output voltage, 2 Hz frequency for 500 pulses of 20 microseconds pulse width, using KCl salt solution of 0.6 mS/cm conductivity by the placing 35g of chopped fresh mushroom stalks and 25g of salt solution in the 4x4x4cm3 treatment chamber. Sequential UAE was carried out on the PEF pre-treated samples using ultrasonic-water-bath (USB) of three frequencies (25 KHz, 35 KHz and 45 KHz) for various treatment times (15-120 min) at 80°C. Individual treatment using either PEF or UAE were also investigation to compare the effect of each treatment along with the combined effect on the recovery and bioactivity of the crude extracts. The freeze dried mushroom stalk powder was characterised for proximate compositional parameters (dry weight basis) showing 64.11% total carbohydrate, 19.12% total protein, 7.21% total fat, 31.2% total dietary fiber, 7.9% chitin (as glucosamine equivalent) and 1.02% β-glucan content. The total phenolic contents (TPC) were determined by the Folin-Ciocalteu procedure and expressed as gallic-acid-equivalents (GAE). The antioxidant properties were ascertained using DPPH and FRAP assays and expressed as trolox-equivalents (TE). HMGCR activity and molecular mass of β-glucans will be measured using the commercial HMG-CoA Reductase Assay kit (Sigma-Aldrich) and size exclusion chromatography (HPLC-SEC), respectively. Effects of PEF, UAE and their combination on the antioxidant capacity, HMGCR inhibition and β-glucans content will be presented.

Keywords: β-glucan, mushroom stalks, pulsed electric field (PEF), ultrasound assisted extraction (UAE)

Procedia PDF Downloads 273

Authors: Parvesh Singh, Vipan Kumar, Vishu Mehra

Abstract:

Quinoline-4-ones represent an important class of heterocyclic scaffolds that have attracted significant interest due to their various biological and pharmacological activities. This heterocyclic unit also constitutes an integral component in drugs used for the treatment of neurodegenerative diseases, sleep disorders and in antibiotics viz. norfloxacin and ciprofloxacin. The synthetic accessibility and possibility of fictionalization at varied positions in quinoline-4-ones exemplifies an elegant platform for the designing of combinatorial libraries of functionally enriched scaffolds with a range of pharmacological profles. They are also considered to be attractive precursors for the synthesis of medicinally imperative molecules such as non-steroidal androgen receptor antagonists, antimalarial drug Chloroquine and martinellines with antibacterial activity. 2-Aryl-2,3-dihydroquinolin-4(1H)-ones are present in many natural and non-natural compounds and are considered to be the aza-analogs of favanones. The β-lactam class of antibiotics is generally recognized to be a cornerstone of human health care due to the unparalleled clinical efficacy and safety of this type of antibacterial compound. In addition to their biological relevance as potential antibiotics, β-lactams have also acquired a prominent place in organic chemistry as synthons and provide highly efficient routes to a variety of non-protein amino acids, such as oligopeptides, peptidomimetics, nitrogen-heterocycles, as well as biologically active natural and unnatural products of medicinal interest such as indolizidine alkaloids, paclitaxel, docetaxel, taxoids, cyptophycins, lankacidins, etc. A straight forward route toward the synthesis of quinoline-4-ones via the triflic acid assisted Fries rearrangement of N-aryl-βlactams has been reported by Tepe and co-workers. The ring expansion observed in this case was solely attributed to the inherent ring strain in β-lactam ring because -lactam failed to undergo rearrangement under reaction conditions. Theabovementioned protocol has been recently extended by our group for the synthesis of benzo[b]-azocinon-6-ones via a tandem Michael addition–Fries rearrangement of sorbyl anilides as well as for the single-pot synthesis of 2-aryl-quinolin-4(3H)-ones through the Fries rearrangement of 3-dienyl-βlactams. In continuation with our synthetic endeavours with the β-lactam ring and in view of the lack of convenient approaches for the synthesis of C-3 functionalized quinolin-4(1H)-ones, the present work describes the single-pot synthesis of C-3 functionalized quinolin-4(1H)-ones via the trific acid promoted Fries rearrangement of C-3 vinyl/isopropenyl substituted β-lactams. In addition, DFT calculations and MD simulations were performed to investigate the stability profles of synthetic compounds.

Keywords: dihydroquinoline, fries rearrangement, azetidin-2-ones, quinoline-4-ones

Procedia PDF Downloads 232

Authors: Evgeny A. Ermakov, Lyudmila P. Smirnova, Valentina N. Buneva

Abstract:

Schizophrenia associated with dysregulation of neurotransmitter processes in the central nervous system and disturbances in the humoral immune system resulting in the formation of antibodies (Abs) to the various components of the nervous tissue. Abs to different neuronal receptors and DNA were detected in the blood of patients with schizophrenia. Abs hydrolyzing DNA were detected in pool of polyclonal autoantibodies in autoimmune and infectious diseases, such catalytic Abs were named abzymes. It is believed that DNA-hydrolyzing abzymes are cytotoxic, cause nuclear DNA fragmentation and induce cell death by apoptosis. Abzymes with DNAase activity are interesting because of the mechanism of formation and the possibility of use as diagnostic markers. Therefore, in our work we have set following goals: to determine the level anti-DNA Abs in the serum of patients with schizophrenia and to study DNA-hydrolyzing activity of IgG of patients with schizophrenia. Materials and methods: In our study there were included 41 patients with a verified diagnosis of paranoid or simple schizophrenia and 24 healthy donors. Electrophoretically and immunologically homogeneous IgGs were obtained by sequential affinity chromatography of the serum proteins on protein G-Sepharose and gel filtration. The levels of anti-DNA Abs were determined using ELISA. DNA-hydrolyzing activity was detected as the level of supercoiled pBluescript DNA transition in circular and linear forms, the hydrolysis products were analyzed by agarose electrophoresis followed by ethidium bromide stain. To correspond the registered catalytic activity directly to the antibodies we carried out a number of strict criteria: electrophoretic homogeneity of the antibodies, gel filtration (acid shock analysis) and in situ activity. Statistical analysis was performed in ‘Statistica 9.0’ using the non-parametric Mann-Whitney test. Results: The sera of approximately 30% of schizophrenia patients displayed a higher level of Abs interacting with single-stranded (ssDNA) and double-stranded DNA (dsDNA) compared with healthy donors. The average level of Abs interacting with ssDNA was only 1.1-fold lower than that for interacting with dsDNA. IgG of patient with schizophrenia were shown to possess DNA hydrolyzing activity. Using affinity chromatography, electrophoretic analysis of isolated IgG homogeneity, gel filtration in acid shock conditions and in situ DNAse activity analysis we proved that the observed activity is intrinsic property of studied antibodies. We have shown that the relative DNAase activity of IgG in patients with schizophrenia averaged 55.4±32.5%, IgG of healthy donors showed much lower activity (average of 9.1±6.5%). It should be noted that DNAase activity of IgG in patients with schizophrenia with a negative symptoms was significantly higher (73.3±23.8%), than in patients with positive symptoms (43.3±33.1%). Conclusion: Anti-DNA Abs of patients with schizophrenia not only bind DNA, but quite efficiently hydrolyze the substrate. The data show a correlation with the level of DNase activity and leading symptoms of patients with schizophrenia.

Keywords: anti-DNA antibodies, abzymes, DNA hydrolysis, schizophrenia

Procedia PDF Downloads 308

Authors: D. Cole, S. Mahootian, P. Medlock

Abstract:

This research contributes to existing knowledge of the complexities surrounding women’s relationship to beauty standards by examining their lived experiences. While there is much academic work on the effects of culturally imposed and largely unattainable beauty standards, the arguments tend to fall into two paradigms. On the one hand is the radical feminist perspective that argues that women are subjected to absolute oppression within the patriarchal system in which beauty standards have been constructed. This position advocates for a complete restructuring of social institutions to liberate women from all types of oppression. On the other hand, there are liberal feminist arguments that focus on choice, arguing that women’s agency in how to present themselves is empowerment. These arguments center around what women do within the patriarchal system in order to liberate themselves. However, there is very little research on the lived experiences of women negotiating these two realms: the complex negotiation between the pressure to adhere to cultural beauty standards and the agency of self-expression and empowerment. By exploring beauty standards through the intersection of societal messages (including macro-level processes such as social media and advertising as well as smaller-scale interactions such as families and peers) and lived experiences, this study seeks to provide a nuanced understanding of how women navigate and negotiate their own presentation and sense of self-identity. Current research sees a rise in incidents of body dysmorphia, depression and anxiety since the advent of social media. Approximately 91% of women are unhappy with their bodies and resort to dieting to achieve their ideal body shape, but only 5% of women naturally possess the body type often portrayed by Americans in movies and media. It is, therefore, crucial we begin talking about the processes that are affecting self-image and mental health. A question that arises is that, given these negative effects, why do companies continue to advertise and target women with standards that very few could possibly attain? One obvious answer is that keeping beauty standards largely unattainable enables the beauty and fashion industries to make large profits by promising products and procedures that will bring one up to “standard”. The creation of dissatisfaction for some is profit for others. This research utilizes qualitative methods: interviews, questionnaires, and focus groups to investigate women’s relationships to beauty standards and empowerment. To this end, we reached out to potential participants through a video campaign on social media: short clips on Instagram, Facebook, and TikTok and a longer clip on YouTube inviting users to take part in the study. Participants are asked to react to images, videos, and other beauty-related texts. The findings of this research have implications for policy development, advocacy and interventions aimed at promoting healthy inclusivity and empowerment of women.

Keywords: women, beauty, consumerism, social media

Procedia PDF Downloads 32

Authors: Doni Dermawan

Abstract:

Modern sports competition as a consequence of the increase in the value of the business and entertainment in the field of sport has been demanding athletes to always have excellent physical endurance performance. Physical exercise is done in a long time, and intensive may pose a risk of muscle tissue damage caused by the increase of the enzyme creatine kinase. Branched Chain Amino Acids (BCAA) is an essential amino acid that is composed of leucine, isoleucine, and valine which serves to maintain muscle tissue, keeping the immune system, and prevent further loss of coordination and muscle pain. Pea (Pisum sativum L.) is a kind of leguminous plants that are rich in Branched Chain Amino Acids (BCAA) where every one gram of protein pea contains 82.7 mg of leucine; 56.3 mg isoleucine; and 56.0 mg of valine. This research aims to develop Branched Chain Amino Acids (BCAA) from pea extract is applied in dosage forms Gel PVP Kinesio Tape technology using Ultrasound-assisted Extraction. The method used in the writing of this paper is the Cochrane Collaboration Review that includes literature studies, testing the quality of the study, the characteristics of the data collection, analysis, interpretation of results, and clinical trials as well as recommendations for further research. Extraction of BCAA in pea done using ultrasound-assisted extraction technology with optimization variables includes the type of solvent extraction (NaOH 0.1%), temperature (20-250C), time (15-30 minutes) power (80 watt) and ultrasonic frequency (35 KHz). The advantages of this extraction method are the level of penetration of the solvent into the membrane of the cell is high and can increase the transfer period so that the BCAA substance separation process more efficient. BCAA extraction results are then applied to the polymer PVP (Polyvinylpyrrolidone) Gel powder composed of PVP K30 and K100 HPMC dissolved in 10 mL of water-methanol (1: 1) v / v. Preparations Kinesio Tape Gel PVP is the BCAA in the gel are absorbed into the muscle tissue, and joints through tensile force then provides stimulation to the muscle circulation with variable pressure so that the muscle can increase the biomechanical movement and prevent damage to the muscle enzyme creatine kinase. Analysis and evaluation of test preparation include interaction, thickness, weight uniformity, humidity, water vapor permeability, the levels of the active substance, content uniformity, percentage elongation, stability testing, release profile, permeation in vitro and in vivo skin irritation testing.

Keywords: branched chain amino acid, BCAA, Kinesio tape, pea, PVP gel, ultrasound-assisted extraction

Procedia PDF Downloads 270

Authors: Daniel O. Kleinsman

Abstract:

The 2019 terrorist attack on two masjidain in Christchurch, New Zealand, was a shocking demonstration of the harm that can be caused by hate crime. As legal and governmental responses to the attack struggle to provide effective remedies to its victims, restorative justice has emerged as a tool that can assist, in terms of both meeting victims’ needs and discharging the obligations of the state under the International Covenant on Civil and Political Rights (ICCPR), arts 2(3), 26, 27. Restorative justice is a model that emphasizes the repair of harm caused or revealed by unjust behavior. It also prioritises the facilitation of dialogue, the restoration of equitable relationships, and the prevention of future harm. Returning to the case study, in the remarks of the sentencing judge, the terrorist’s actions were described as a hate crime of vicious malevolence that the Court was required to decisively reject, as anathema to the values of acceptance, tolerance and mutual respect upon which New Zealand’s inclusive society is based and which the country strives to maintain. This was one of the reasons for which the terrorist received a life sentence with no possibility of parole. However, in the report of the Royal Commission of Inquiry into the Attack, it was found that victims felt the attack occurred within the context of widespread racism, discrimination and Islamophobia, where hostile behaviors, including hate-based threats and attacks, were rarely recorded, analysed or acted on. It was also found that the Government had inappropriately concentrated intelligence resources on the risk of ‘Islamist’ terrorism and had failed to adequately respond to concerns raised about threats against the Muslim community. In this light, the remarks of the sentencing judge can be seen to reflect a criminal justice system that, in the absence of other remedies, denies systemic accountability and renders hate crime an isolated incident rather than an expression of more widespread discrimination and hate to be holistically addressed. One of the recommendations of the Royal Commission was to explore with victims the desirability and design of restorative justice processes. This presents an opportunity for victims to meet with state representatives and pursue effective remedies (ICCPR art 2(3)) not only for the harm caused by the terrorist but the harm revealed by a system that has exposed the minority Muslim community in New Zealand to hate in all forms, including but not limited to violent extremism. In this sense, restorative justice can also assist the state in discharging its wider obligations to protect all persons from discrimination (art 26) and allow ethnic and religious minorities to enjoy their own culture and profess and practice their own religion (art 27). It can also help give effect to the law and its purpose as a remedy to hate crime, as expressed in this case study by the sentencing judge.

Keywords: hate crime, restorative justice, minorities, victims' rights

Procedia PDF Downloads 91

Authors: Alaa G. M. Osman, Abd-El –Baset M. Abd El Reheem, Khaled Y. Abouelfadl, Usama M. Mahmoud, Mohsen A. Moustafa

Abstract:

This study aimed to explore new sites of biomarker research and to establish the use of blood parameters in wild fish populations. Four hundred and twenty fish samples were collected from six sites along the whole course of the river Nile, Egypt. The mean values of erythrocytes, thrombocytes, hemoglobin concentration, hematocrit value, and mean corpuscular volume were significantly lower in the blood of Nile tilapia and African catfish collected from downstream (contaminated) compared to upstream sites. In contrast, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration in the peripheral blood of both fish species significantly increased from upstream to downstream river Nile. The leukocytes count was significantly decreased in contaminated sites compared to upstream area. Hematological variables in the peripheral blood of Oreochromis niloticus niloticus and Clarias gariepinus exhibited significant (p<0.05) correlation with nearly all the detected chemical and physical parameters along the Nile course. In the present study, lower cellular and nuclear areas and cellular and nuclear shape factor were recorded in the erythrocytes of fish collected from downstream compared to those caught from upstream sites. This was confirmed by higher immature ratios of red cells in the blood of fish sampled from downstream river Nile. Karyorrhetic and enucleated erythrocytes were significantly correlated with physiochemical parameters in water samples collected from the same sites is being higher in the blood of fish collected from downstream sites. To see if there was any correlation between fish altered physiological fitness and environmental stress, we measured serum biochemical variables namely; total protein, cholesterol, triglycerides, calcium, chlorides, alkaline phosphatase activity (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), uric acid activity, creatinine, and serum glucose. The level of all the selected biochemical variables in the blood of O. niloticus niloticus and C. gariepinus were recorded to be significantly higher (p<0.05) in downstream sites. According to the present results, nearly all the detected haematological and blood biochemical variables are suitable indicators of contaminant exposure in O. niloticus niloticus and C. gariepinus. Also the detected erythrocytes malformations in blood collected from Nile tilapia and African catfish were proven to be suitable for bio-monitoring aquatic pollution. The results revealed species-specific differences in sensitivities, suggesting that Nile tilapia may serve as a more sensitive test species compared to African catfish.

Keywords: biomarkers, water pollution, blood parameters, river nile, african catfish, nile tilapia

Procedia PDF Downloads 277

Authors: M. Nerantzaki, I. Koliakou, D. Bikiaris

Abstract:

This study evaluates the hypothesis that the incorporation of fibrous hydroxyapatite nanoparticles (nHA) with high crystallinity and high aspect ratio, synthesized by hydrothermal method, into Poly(butylene succinate) (PBSu), improves the bioactivity of the aliphatic polyester and affects new bone growth inhibiting resorption and enhancing bone formation. Hydroxyapatite nanorods were synthesized using a simple hydrothermal procedure. First, the HPO42- -containing solution was added drop-wise into the Ca2+-containing solution, while the molar ratio of Ca/P was adjusted at 1.67. The HA precursor was then treated hydrothermally at 200°C for 72 h. The resulting powder was characterized using XRD, FT-IR, TEM, and EDXA. Afterwards, PBSu nanocomposites containing 2.5wt% (nHA) were prepared by in situ polymerization technique for the first time and were examined as potential scaffolds for bone engineering applications. For comparison purposes composites containing either 2.5wt% micro-Bioglass (mBG) or 2.5wt% mBG-nHA were prepared and studied, too. The composite scaffolds were characterized using SEM, FTIR, and XRD. Mechanical testing (Instron 3344) and Contact Angle measurements were also carried out. Enzymatic degradation was studied in an aqueous solution containing a mixture of R. Oryzae and P. Cepacia lipases at 37°C and pH=7.2. In vitro biomineralization test was performed by immersing all samples in simulated body fluid (SBF) for 21 days. Biocompatibility was assessed using rat Adipose Stem Cells (rASCs), genetically modified by nucleofection with DNA encoding SB100x transposase and pT2-Venus-neo transposon expression plasmids in order to attain fluorescence images. Cell proliferation and viability of cells on the scaffolds were evaluated using fluoresce microscopy and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide) assay. Finally, osteogenic differentiation was assessed by staining rASCs with alizarine red using cetylpyridinium chloride (CPC) method. TEM image of the fibrous HAp nanoparticles, synthesized in the present study clearly showed the fibrous morphology of the synthesized powder. The addition of nHA decreased significantly the contact angle of the samples, indicating that the materials become more hydrophilic and hence they absorb more water and subsequently degrade more rapidly. In vitro biomineralization test confirmed that all samples were bioactive as mineral deposits were detected by X-ray diffractometry after incubation in SBF. Metabolic activity of rASCs on all PBSu composites was high and increased from day 1 of culture to day 14. On day 28 metabolic activity of rASCs cultured on samples enriched with bioceramics was significantly decreased due to possible differentiation of rASCs to osteoblasts. Staining rASCs with alizarin red after 28 days in culture confirmed our initial hypothesis as the presence of calcium was detected, suggesting osteogenic differentiation of rACS on PBSu/nHAp/mBG 2.5% and PBSu/mBG 2.5% composite scaffolds.

Keywords: biomaterials, hydroxyapatite nanorods, poly(butylene succinate), scaffolds

Procedia PDF Downloads 293