Search results for: molecular evolution
Commenced in January 2007
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Edition: International
Paper Count: 3782

Search results for: molecular evolution

122 Seismic Stratigraphy of the First Deposits of the Kribi-Campo Offshore Sub-basin (Gulf of Guinea): Pre-cretaceous Early Marine Incursion and Source Rocks Modeling

Authors: Mike-Franck Mienlam Essi, Joseph Quentin Yene Atangana, Mbida Yem

Abstract:

The Kribi-Campo sub-basin belongs to the southern domain of the Cameroon Atlantic Margin in the Gulf of Guinea. It is the African homologous segment of the Sergipe-Alagoas Basin, located at the northeast side of the Brazil margin. The onset of the seafloor spreading period in the Southwest African Margin in general and the study area particularly remains controversial. Various studies locate this event during the Cretaceous times (Early Aptian to Late Albian), while others suggested that this event occurred during Pre-Cretaceous period (Palaeozoic or Jurassic). This work analyses 02 Cameroon Span seismic lines to re-examine the Early marine incursion period of the study area for a better understanding of the margin evolution. The methodology of analysis in this study is based on the delineation of the first seismic sequence, using the reflector’s terminations tracking and the analysis of its internal reflections associated to the external configuration of the package. The results obtained indicate from the bottom upwards that the first deposits overlie a first seismic horizon (H1) associated to “onlap” terminations at its top and underlie a second horizon which shows “Downlap” terminations at its top (H2). The external configuration of this package features a prograded fill pattern, and it is observed within the depocenter area with discontinuous reflections that pinch out against the basement. From east to west, this sequence shows two seismic facies (SF1 and SF2). SF1 has parallel to subparallel reflections, characterized by high amplitude, and SF2 shows parallel and stratified reflections, characterized by low amplitude. The distribution of these seismic facies reveals a lateral facies variation observed. According to the fundamentals works on seismic stratigraphy and the literature review of the geological context of the study area, particularly, the stratigraphical natures of the identified horizons and seismic facies have been highlighted. The seismic horizons H1 and H2 correspond to Top basement and “Downlap Surface,” respectively. SF1 indicates continental sediments (Sands/Sandstone) and SF2 marine deposits (shales, clays). Then, the prograding configuration observed suggests a marine regression. The correlation of these results with the lithochronostratigraphic chart of Sergipe-Alagoas Basin reveals that the first marine deposits through the study area are dated from Pre-Cretaceous times (Palaeozoic or Jurassic). The first deposits onto the basement represents the end of a cycle of sedimentation. The hypothesis of Mike.F. Mienlam Essi is with the Earth Sciences Department of the Faculty of Science of the University of Yaoundé I, P.O. BOX 812 CAMEROON (e-mail: [email protected]). Joseph.Q. Yene Atangana is with the Earth Sciences Department of the Faculty of Science of the University of Yaoundé I, P.O. BOX 812 CAMEROON (e-mail: [email protected]). Mbida Yem is with the Earth Sciences Department of the Faculty of Science of the University of Yaoundé I, P.O. BOX 812 CAMEROON (e-mail: [email protected]). Cretaceous seafloor spreading through the study area is the onset of another cycle of sedimentation. Furthermore, the presence of marine sediments into the first deposits implies that this package could contain marine source rocks. The spatial tracking of these deposits reveals that they could be found in some onshore parts of the Kribi-Campo area or even in the northern side.

Keywords: cameroon span seismic, early marine incursion, kribi-campo sub-basin, pre-cretaceous period, sergipe-alagoas basin

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121 Histogenesis of the Stomach of Pre-Hatching Quail: A Light and Electron Microscopic Study

Authors: Soha A Soliman, Yasser A Ahmed, Mohamed A Khalaf

Abstract:

Although the enormous literature describing the histology of the stomach of different avian species during the posthatching development, the available literature on the pre-hatching development of quail stomach development is scanty. Thus, the current study was undertaken to provide a careful description of the main histological events during the embryonic development of quail stomach. To achieve this aim, daily histological specimens from the stomach of quail of 4 days post-incubation till the day 17 (few hours before hatching) were examined with light microscopy. The current study showed that the primitive gut tube of the embryonic quail appeared at the 4th day post incubation, and both parts of stomach (proventriculus and gizzard) were similar in structure and composed of endodermal epithelium of pseudostratified type surrounded by undifferentiated mesenchymal tissue. The sequences of the developmental events in the gut tube were preceded in a cranio-caudal pattern. By the 5th day, the endodermal covering of the primitive proventriculus gave rise to sac-like invaginations. The primitive gizzard was distinguished into thick-walled bodies and thin-walled sacs. In the 6th day, the prospective proventricular glandular epithelium became canalized and the muscular layer was developed in the cranial part of the proventriculus, whereas the primitive muscular coat of the gizzard was represented by a layer of condensed mesenchyme. In the 7th day, the proventricular glandular epithelial invaginations increased in depth and number, while, the muscularis mucosa and the muscular layer began to be distinguished. In the 8th day, the myoblasts differentiated into spindle shaped smooth muscle fibers. In the 10th day, branching of the proventricular glands began. The branching continued later on. The surface and the glandular epithelium were transformed into simple columnar type in the 12th day. The epithelial covering of the gizzard gave rise to tubular invaginations lined by simple cuboidal epithelium and the surface epithelium became simple columnar. Canalization of the tubular glands was recognized in the 14th day. In the 15th day, the proventricular surface epithelium invaginated in an concentric manner around a central cavity to form immature secretory units. The central cavity was lined by eosinophilic cells which form the ductal epithelia. The peripheral lamellae were lined by basophilic cells; the undifferentiated oxyntico-peptic cells. Entero-endocrine cells stained positive for silver impregnation in the proventricular glands. The mucosal folding in the gizzard appeared in the 15th day to form the plicae and the sulci. The wall of the proventriculus and gizzard in the 17th day acquired the main histological features of post-hatching birds, but neither the surface nor the ductal epithelium were differentiated to mucous producing cells. The current results shoed be considered in the molecular developmental studies.

Keywords: quail, proventriculus, gizzard, pre-hatching, histology

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120 Characterization and Evaluation of the Dissolution Increase of Molecular Solid Dispersions of Efavirenz

Authors: Leslie Raphael de M. Ferraz, Salvana Priscylla M. Costa, Tarcyla de A. Gomes, Giovanna Christinne R. M. Schver, Cristóvão R. da Silva, Magaly Andreza M. de Lyra, Danilo Augusto F. Fontes, Larissa A. Rolim, Amanda Carla Q. M. Vieira, Miracy M. de Albuquerque, Pedro J. Rolim-Neto

Abstract:

Efavirenz (EFV) is a drug used as first-line treatment of AIDS. However, it has poor aqueous solubility and wettability, presenting problems in the gastrointestinal tract absorption and bioavailability. One of the most promising strategies to improve the solubility is the use of solid dispersions (SD). Therefore, this study aimed to characterize SD EFZ with the polymers: PVP-K30, PVPVA 64 and SOLUPLUS in order to find an optimal formulation to compose a future pharmaceutical product for AIDS therapy. Initially, Physical Mixtures (PM) and SD with the polymers were obtained containing 10, 20, 50 and 80% of drug (w/w) by the solvent method. The best formulation obtained between the SD was selected by in vitro dissolution test. Finally, the drug-carrier system chosen, in all ratios obtained, were analyzed by the following techniques: Differential Scanning Calorimetry (DSC), polarization microscopy, Scanning Electron Microscopy (SEM) and spectrophotometry of absorption in the region of infrared (IR). From the dissolution profiles of EFV, PM and SD, the values of area Under The Curve (AUC) were calculated. The data showed that the AUC of all PM is greater than the isolated EFV, this result is derived from the hydrophilic properties of the polymers thus favoring a decrease in surface tension between the drug and the dissolution medium. In adittion, this ensures an increasing of wettability of the drug. In parallel, it was found that SD whom had higher AUC values, were those who have the greatest amount of polymer (with only 10% drug). As the amount of drug increases, it was noticed that these results either decrease or are statistically similar. The AUC values of the SD using the three different polymers, followed this decreasing order: SD PVPVA 64-EFV 10% > SD PVP-K30-EFV 10% > SD Soluplus®-EFV 10%. The DSC curves of SD’s did not show the characteristic endothermic event of drug melt process, suggesting that the EFV was converted to its amorphous state. The analysis of polarized light microscopy showed significant birefringence of the PM’s, but this was not observed in films of SD’s, thus suggesting the conversion of the drug from the crystalline to the amorphous state. In electron micrographs of all PM, independently of the percentage of the drug, the crystal structure of EFV was clearly detectable. Moreover, electron micrographs of the SD with the two polymers in different ratios investigated, we observed the presence of particles with irregular size and morphology, also occurring an extensive change in the appearance of the polymer, not being possible to differentiate the two components. IR spectra of PM corresponds to the overlapping of polymer and EFV bands indicating thereby that there is no interaction between them, unlike the spectra of all SD that showed complete disappearance of the band related to the axial deformation of the NH group of EFV. Therefore, this study was able to obtain a suitable formulation to overcome the solubility limitations of the EFV, since SD PVPVA 64-EFZ 10% was chosen as the best system in delay crystallization of the prototype, reaching higher levels of super saturation.

Keywords: characterization, dissolution, Efavirenz, solid dispersions

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119 Gold Nanoprobes Assay for the Identification of Foodborn Pathogens Such as Staphylococcus aureus, Listeria monocytogenes and Salmonella enteritis

Authors: D. P. Houhoula, J. Papaparaskevas, S. Konteles, A. Dargenta, A. Farka, C. Spyrou, M. Ziaka, S. Koussisis, E. Charvalos

Abstract:

Objectives: Nanotechnology is providing revolutionary opportunities for the rapid and simple diagnosis of many infectious diseases. Staphylococcus aureus, Listeria monocytogenes and Salmonella enteritis are important human pathogens. Diagnostic assays for bacterial culture and identification are time consuming and laborious. There is an urgent need to develop rapid, sensitive, and inexpensive diagnostic tests. In this study, a gold nanoprobe strategy developed and relies on the colorimetric differentiation of specific DNA sequences based approach on differential aggregation profiles in the presence or absence of specific target hybridization. Method: Gold nanoparticles (AuNPs) were purchased from Nanopartz. They were conjugated with thiolated oligonucleotides specific for the femA gene for the identification of members of Staphylococcus aureus, the mecA gene for the differentiation of Staphylococcus aureus and MRSA Staphylococcus aureus, hly gene encoding the pore-forming cytolysin listeriolysin for the identification of Listeria monocytogenes and the invA sequence for the identification of Salmonella enteritis. DNA isolation from Staphylococcus aureus Listeria monocytogenes and Salmonella enteritis cultures was performed using the commercial kit Nucleospin Tissue (Macherey Nagel). Specifically 20μl of DNA was diluted in 10mMPBS (pH5). After the denaturation of 10min, 20μl of AuNPs was added followed by the annealing step at 58oC. The presence of a complementary target prevents aggregation with the addition of acid and the solution remains pink, whereas in the opposite event it turns to purple. The color could be detected visually and it was confirmed with an absorption spectrum. Results: Specifically, 0.123 μg/μl DNA of St. aureus, L.monocytogenes and Salmonella enteritis was serially diluted from 1:10 to 1:100. Blanks containing PBS buffer instead of DNA were used. The application of the proposed method on isolated bacteria produced positive results with all the species of St. aureus and L. monocytogenes and Salmonella enteritis using the femA, mecA, hly and invA genes respectively. The minimum detection limit of the assay was defined at 0.2 ng/μL of DNA. Below of 0.2 ng/μL of bacterial DNA the solution turned purple after addition of HCl, defining the minimum detection limit of the assay. None of the blank samples was positive. The specificity was 100%. The application of the proposed method produced exactly the same results every time (n = 4) the evaluation was repeated (100% repeatability) using the femA, hly and invA genes. Using the gene mecA for the differentiation of Staphylococcus aureus and MRSA Staphylococcus aureus the method had a repeatability 50%. Conclusion: The proposed method could be used as a highly specific and sensitive screening tool for the detection and differentiation of Staphylococcus aureus Listeria monocytogenes and Salmonella enteritis. The use AuNPs for the colorimetric detection of DNA targets represents an inexpensive and easy-to-perform alternative to common molecular assays. The technology described here, may develop into a platform that could accommodate detection of many bacterial species.

Keywords: gold nanoparticles, pathogens, nanotechnology, bacteria

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118 Sex Differences in Age-Related AMPK-Sirt1 Axis Alteration in Human Heart

Authors: Maria Luisa Barcena De Arellano, Sofya Pozdniakova, Pavelas Karkacas, Anja Kuhl, Istvan Baczko, Yury Ladilov, Vera Regitz-Zagrosek

Abstract:

Introduction: Aging is associated with deterioration of the physiological function, leading to systemic inflammation and mitochondrial dysfunction that promote the development of cardiovascular diseases. Sex differences in aging-related cardiovascular diseases have been postulated. However, their precise mechanisms remain unclear. In the current study, we aimed to investigate the sex difference in the age-related alteration in Sirt1-AMPK signaling and its relation to the mitochondrial biogenesis and inflammation. Methods: Male and female human non-disease lateral left ventricular wall tissue (young (17–40 years; n= 7 male and 7 female) and old (50–68 years; n= 9 male and 8 female)) were used. qRT-PCR, western blot and immunohistochemistry assays were performed for expression analyses of Sirt1, AMPK, pAMPK, ac-Ku70, TFAM, PGC-1α, Sirt3, SOD2 and catalase. CD68 was used as a marker for macrophages and the ratio of IL-12:IL10 (pro-inflammatory phenotype (high IL-12/low IL-10) and anti-inflammatory phenotype (low IL-12/high IL-10) was used to examine the inflammatory stage in the heart. Results: Sirt1 expression was significantly higher in young females compared to young males, whereas in aged hearts Sirt1 expression was significantly downregulated in females, but not in males. In line with the Sirt1 downregulation in aged females, acetylation of nuclear Ku70, a direct target of Sirt1, in aged female hearts was significantly elevated. The activity of AMPK was significantly decreased in aged individuals, however no sex differences in the AMPK expression or activity were found in young or old individuals. The expression of mitochondrial proteins TOM40, SOD2 and Sirt3 was significantly higher in young females compared to young males, while in aged female hearts SOD2 and TOM40 were downregulated. In addition, the expression of catalase, a key cytosolic and mitochondrial anti-oxidative enzyme was significantly higher in young females and this female sex benefit was lost in aged hearts. In addition, the number of cardiac macrophages was significantly increased in old female, but not in male hearts. Consistently, the pro-inflammatory shift in old females was further confirmed by differences in the IL12/IL10 ratio in young female cardiac tissue in a favour of the anti-inflammatory mediator IL-10 (ratio 1:4) compared to young males (ratio 1:1). The anti-inflammatory environment in the heart was lost in aged females (ratio 1:1). Conclusion: Aging leads to the significant downregulation of Sirt1 expression and elevated acetylation of Ku70 in female, but not in male hearts. Furthermore, a beneficial upregulation of mitochondrial and anti-oxidative proteins in young females is lost with aging. Moreover, the malfunctions in the expression of Sirt1 and mitochondrial proteins in aged female hearts is accompanied by a significant pro-inflammatory shift. The study provides a molecular basis for the increased incidence of cardiovascular diseases in old women.

Keywords: inflammation, mitochondrial dysfunction, aging, Sirt1-AMPK axis

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117 Gut Microbial Dynamics in a Mouse Model of Inflammation-Linked Carcinogenesis as a Result of Diet Supplementation with Specific Mushroom Extracts

Authors: Alvarez M., Chapela M. J., Balboa E., Rubianes D., Sinde E., Fernandez de Ana C., Rodríguez-Blanco A.

Abstract:

The gut microbiota plays an important role as gut inflammation could contribute to colorectal cancer development; however, this role is still not fully understood, and tools able to prevent this progression are yet to be developed. The main objective of this study was to monitor the effects of a mushroom extracts formulation in gut microbial community composition of an Azoxymethane (AOM)/Dextran sodium sulfate (DSS) mice model of inflammation-linked carcinogenesis. For the in vivo study, 41 adult male mice of the C57BL / 6 strain were obtained. 36 of them have been induced in a state of colon carcinogenesis by a single intraperitoneal administration of AOM at a dose of 12.5 mg/kg; the control group animals received instead of the same volume of 0.9% saline. DSS is an extremely toxic polysaccharide sulfate that causes chronic inflammation of the colon mucosa, favoring the appearance of severe colitis and the production of tumors induced by AOM. Induction by AOM/DSS is an interesting platform for chemopreventive intervention studies. This time the model was used to monitor gut microbiota changes as a result of supplementation with a specific mushroom extracts formulation previously shown to have prebiotic activity. The animals have been divided into three groups: (i) Cancer + mushroom extracts formulation experimental group: to which the MicoDigest2.0 mushroom extracts formulation developed by Hifas da Terra S.L has been administered dissolved in drinking water at an estimated concentration of 100 mg / ml. (ii) Control group of animals with Cancer: to which normal water has been administered without any type of treatment. (iii) Control group of healthy animals: these are the animals that have not been induced cancer or have not received any treatment in drinking water. This treatment has been maintained for a period of 3 months, after which the animals were sacrificed to obtain tissues that were subsequently analyzed to verify the effects of the mushroom extract formulation. A microbiological analysis has been carried out to compare the microbial communities present in the intestines of the mice belonging to each of the study groups. For this, the methodology of massive sequencing by molecular analysis of the 16S gene has been used (Ion Torrent technology). Initially, DNA extraction and metagenomics libraries were prepared using the 16S Metagenomics kit, always following the manufacturer's instructions. This kit amplifies 7 of the 9 hypervariable regions of the 16S gene that will then be sequenced. Finally, the data obtained will be compared with a database that makes it possible to determine the degree of similarity of the sequences obtained with a wide range of bacterial genomes. Results obtained showed that, similarly to certain natural compounds preventing colorectal tumorigenesis, a mushroom formulation enriched the Firmicutes and Proteobacteria phyla and depleted Bacteroidetes. Therefore, it was demonstrated that the consumption of the mushroom extracts’ formulation developed could promote the recovery of the microbial balance that is disrupted in the mice model of carcinogenesis. More preclinical and clinical studies are needed to validate this promising approach.

Keywords: carcinogenesis, microbiota, mushroom extracts, inflammation

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116 Profiling of the Cell-Cycle Related Genes in Response to Efavirenz, a Non-Nucleoside Reverse Transcriptase Inhibitor in Human Lung Cancer

Authors: Rahaba Marima, Clement Penny

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The Health-related quality of life (HRQoL) for HIV positive patients has improved since the introduction of the highly active antiretroviral treatment (HAART). However, in the present HAART era, HIV co-morbidities such as lung cancer, a non-AIDS (NAIDS) defining cancer have been documented to be on the rise. Under normal physiological conditions, cells grow, repair and proliferate through the cell-cycle as cellular homeostasis is important in the maintenance and proper regulation of tissues and organs. Contrarily, the deregulation of the cell-cycle is a hallmark of cancer, including lung cancer. The association between lung cancer and the use of HAART components such as Efavirenz (EFV) is poorly understood. This study aimed at elucidating the effects of EFV on the cell-cycle genes’ expression in lung cancer. For this purpose, the human cell-cycle gene array composed of 84 genes was evaluated on both normal lung fibroblasts (MRC-5) cells and adenocarcinoma (A549) lung cells, in response to 13µM EFV or 0.01% vehicle. The ±2 up or down fold change was used as a basis of target selection, with p < 0.05. Additionally, RT-qPCR was done to validate the gene array results. Next, In-silico bio-informatics tools, Search Tool for the Retrieval of Interacting Genes/Proteins (STRING), Reactome, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and Ingenuity Pathway Analysis (IPA) were used for gene/gene interaction studies as well as to map the molecular and biological pathways influenced by the identified targets. Interestingly, the DNA damage response (DDR) pathway genes such as p53, Ataxia telangiectasia mutated and Rad3 related (ATR), Growth arrest and DNA damage inducible alpha (GADD45A), HUS1 checkpoint homolog (HUS1) and Role of radiation (RAD) genes were shown to be upregulated following EFV treatment, as revealed by STRING analysis. Additionally, functional enrichment analysis by the KEGG pathway revealed that most of the differentially expressed gene targets function at the cell-cycle checkpoint such as p21, Aurora kinase B (AURKB) and Mitotic Arrest Deficient-Like 2 (MAD2L2). Core analysis by IPA revealed that p53 downstream targets such as survivin, Bcl2, and cyclin/cyclin dependent kinases (CDKs) complexes are down-regulated, following exposure to EFV. Furthermore, Reactome analysis showed a significant increase in cellular response to stress genes, DNA repair genes, and apoptosis genes, as observed in both normal and cancerous cells. These findings implicate the genotoxic effects of EFV on lung cells, provoking the DDR pathway. Notably, the constitutive expression of this pathway (DDR) often leads to uncontrolled cell proliferation and eventually tumourigenesis, which could be the attribute of HAART components’ (such as EFV) effect on human cancers. Targeting the cell-cycle and its regulation holds a promising therapeutic intervention to the potential HAART associated carcinogenesis, particularly lung cancer.

Keywords: cell-cycle, DNA damage response, Efavirenz, lung cancer

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115 An E-Maintenance IoT Sensor Node Designed for Fleets of Diverse Heavy-Duty Vehicles

Authors: George Charkoftakis, Panagiotis Liosatos, Nicolas-Alexander Tatlas, Dimitrios Goustouridis, Stelios M. Potirakis

Abstract:

E-maintenance is a relatively new concept, generally referring to maintenance management by monitoring assets over the Internet. One of the key links in the chain of an e-maintenance system is data acquisition and transmission. Specifically for the case of a fleet of heavy-duty vehicles, where the main challenge is the diversity of the vehicles and vehicle-embedded self-diagnostic/reporting technologies, the design of the data acquisition and transmission unit is a demanding task. This clear if one takes into account that a heavy-vehicles fleet assortment may range from vehicles with only a limited number of analog sensors monitored by dashboard light indicators and gauges to vehicles with plethora of sensors monitored by a vehicle computer producing digital reporting. The present work proposes an adaptable internet of things (IoT) sensor node that is capable of addressing this challenge. The proposed sensor node architecture is based on the increasingly popular single-board computer – expansion boards approach. In the proposed solution, the expansion boards undertake the tasks of position identification by means of a global navigation satellite system (GNSS), cellular connectivity by means of 3G/long-term evolution (LTE) modem, connectivity to on-board diagnostics (OBD), and connectivity to analog and digital sensors by means of a novel design of expansion board. Specifically, the later provides eight analog plus three digital sensor channels, as well as one on-board temperature / relative humidity sensor. The specific device offers a number of adaptability features based on appropriate zero-ohm resistor placement and appropriate value selection for limited number of passive components. For example, although in the standard configuration four voltage analog channels with constant voltage sources for the power supply of the corresponding sensors are available, up to two of these voltage channels can be converted to provide power to the connected sensors by means of corresponding constant current source circuits, whereas all parameters of analog sensor power supply and matching circuits are fully configurable offering the advantage of covering a wide variety of industrial sensors. Note that a key feature of the proposed sensor node, ensuring the reliable operation of the connected sensors, is the appropriate supply of external power to the connected sensors and their proper matching to the IoT sensor node. In standard mode, the IoT sensor node communicates to the data center through 3G/LTE, transmitting all digital/digitized sensor data, IoT device identity, and position. Moreover, the proposed IoT sensor node offers WiFi connectivity to mobile devices (smartphones, tablets) equipped with an appropriate application for the manual registration of vehicle- and driver-specific information, and these data are also forwarded to the data center. All control and communication tasks of the IoT sensor node are performed by dedicated firmware. It is programmed with a high-level language (Python) on top of a modern operating system (Linux). Acknowledgment: This research has been co-financed by the European Union and Greek national funds through the Operational Program Competitiveness, Entrepreneurship, and Innovation, under the call RESEARCH—CREATE—INNOVATE (project code: T1EDK- 01359, IntelligentLogger).

Keywords: IoT sensor nodes, e-maintenance, single-board computers, sensor expansion boards, on-board diagnostics

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114 Previously Undescribed Cardiac Abnormalities in Two Unrelated Autistic Males with Causative Variants in CHD8

Authors: Mariia A. Parfenenko, Ilya S. Dantsev, Sergei V. Bochenkov, Natalia V. Vinogradova, Olga S. Groznova, Victoria Yu. Voinova

Abstract:

Introduction: Autism is the most common neurodevelopmental disorder. Autism is characterized by difficulties in social interaction and adherence to stereotypic behavioral patterns and frequently co-occurs with epilepsy, intellectual disabilities, connective tissue disorders, and other conditions. CHD8 codes for chromodomain-helicase-DNA-binding protein 8 - a chromatin remodeler that regulates cellular proliferation and neurodevelopment in embryogenesis. CHD8 is one of the genes most frequently involved in autism. Patients and methods: 2 unrelated male patients, P3 and P12, aged 3 and 12 years old, underwent whole genome sequencing, which determined that they both had different likely pathogenic variants, both previously undescribed in literature. Sanger sequencing later determined that P12 inherited the variant from his affected mother. Results: P3 and P12 presented with autism, a developmental delay, ataxia, sleep disorders, overgrowth, and macrocephaly, as well as other clinical features typically present in patients with causative variants in CHD8. The mother of P12 also has autistic traits, as well as ataxia, hypotonia, sleep disorders, and other symptoms. However, P3 and P12 also have different cardiac abnormalities. P3 had signs of a repolarization disorder: a flattened T wave in the III and aVF derivations and a negative T wave in the V1-V2 derivations. He also had structural valve anomalies with associated regurgitation, local contractility impairment of the left ventricular, and diastolic dysfunction of the right ventricle. Meanwhile, P12 had Wolff-Parkinson-White syndrome and underwent radiofrequency ablation at the age of 2 years. At the time of observation, P12 had mild sinus arrhythmia and an incomplete right bundle branch block, as well as arterial hypertension. Discussion: Cardiac abnormalities were not previously reported in patients with causative variants in CHD8. The underlying mechanism for the formation of those abnormalities is currently unknown. However, the two hypotheses are either a disordered interaction with CHD7 – another chromodomain remodeler known to be directly involved in the cardiophenotype of CHARGE syndrome – a rare condition characterized by coloboma, heart defects and growth abnormalities, or the disrupted functioning of CHD8 as an A-Kinase Anchoring Protein, which are known to modulate cardiac function. Conclusion: We observed 2 unrelated autistic males with likely pathogenic variants in CHD8 that presented with typical symptoms of CHD8-related neurodevelopmental disorder, as well as cardiac abnormalities. Cardiac abnormalities have, until now, been considered uncharacteristic for patients with causative variants in CHD8. Further accumulation of data, including experimental evidence of the involvement of CHD8 in heart formation, will elucidate the mechanism underlying the cardiophenotype of those patients. Acknowledgements: Molecular genetic testing of the patients was made possible by the Charity Fund for medical and social genetic aid projects «Life Genome.»

Keywords: autism spectrum disorders, chromodomain-helicase-DNA-binding protein 8, neurodevelopmental disorder, cardio phenotype

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113 Bio-Detoxification of Mycotoxins by Lactic Acid Bacteria from Different Food Matrices

Authors: António Inês, Ana Guimarães, José Maria, Vânia Laranjo, Armando Venâncio, Luís Abrunhosa

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Lactic acid bacteria (LAB) play a key role in the biopreservation of a wide range of fermented food products, such as yogurt, cheese, fermented milks, meat, fish, vegetables (sauerkraut, olives and pickles), certain beer brands, wines and silage, allowing their safe consumption, which gave to these bacteria a GRAS (Generally Recognised as Safe) status. Besides that, the use of LAB in food and feed is a promising strategy to reduce the exposure to dietary mycotoxins, improving their shelf life and reducing health risks, given the unique mycotoxin decontaminating characteristic of some LAB. Mycotoxins present carcinogenic, mutagenic, teratogenic, neurotoxic and immunosuppressive effects over animals and Humans, being the most important ochratoxin A (OTA), aflatoxins (AFB1), trichothecenes, zearalenone (ZEA), fumonisin (FUM) and patulin. In a previous work of our group it was observed OTA biodegradation by some strains of Pediococcus parvulus isolated from Douro wines. So, the aim of this study was to enlarge the screening of the biodetoxification over more mycotoxins besides OTA, including AFB1, and ZEA. This ability was checked in a collection of LAB isolated from vegetable (wine, olives, fruits and silage) and animal (milk and dairy products, sausages) sources. All LAB strains were characterized phenotypically (Gram, catalase) and genotypically. Molecular characterisation of all LAB strains was performed using genomic fingerprinting by MSP-PCR with (GTG)5 and csM13 primers. The identification of the isolates was confirmed by 16S rDNA sequencing. To study the ability of LAB strains to degrade OTA, AFB1 and ZEA, a MRS broth medium was supplemented with 2.0 μg/mL of each mycotoxin. For each strain, 2 mL of MRS supplemented with the mycotoxins was inoculated in triplicate with 109 CFU/mL. The culture media and bacterial cells were extracted by the addition of an equal volume of acetonitrile/methanol/acetic acid (78:20:2 v/v/v) to the culture tubes. A 2 mL sample was then collected and filtered into a clean 2 mL vial using PP filters with 0.45 μm pores. The samples were preserved at 4 °C until HPLC analysis. Among LAB tested, 10 strains isolated from milk were able to eliminate AFB1, belonging to Lactobacillus casei (7), Lb. paracasei (1), Lb. plantarum (1) and 1 to Leuconostoc mesenteroides. Two strains of Enterococcus faecium and one of Ec. faecalis from sausage eliminated ZEA. Concerning to strains of vegetal origin, one Lb. plantarum isolated from elderberry fruit, one Lb. buchnerii and one Lb. parafarraginis both isolated from silage eliminated ZEA. Other 2 strains of Lb. plantarum from silage were able to degrade both ZEA and OTA, and 1 Lb. buchnerii showed activity over AFB1. These enzymatic activities were also verified genotypically through specific gene PCR and posteriorly confirmed by sequencing analysis. In conclusion, due the ability of some strains of LAB isolated from different sources to eliminate OTA, AFB1 and ZEA one can recognize their potential biotechnological application to reduce the health hazards associated with these mycotoxins. They may be suitable as silage inoculants or as feed additives or even in food industry.

Keywords: bio-detoxification, lactic acid bacteria, mycotoxins, food and feed

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112 Nurturing Minds, Shaping Futures: A Reflective Journey of 32 Years as a Teacher Educator

Authors: Mary Isobelle Mullaney

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The maxim "an unexamined life is not worth living," attributed to Socrates, prompts a contemplative reflection spanning over 32 years as a teacher educator in the Republic of Ireland. Taking time to contemplate the changes that have occurred and the current landscape provides valuable insights into the dynamic terrain of teacher preparation. The reflective journey traverses the impacts of global and societal shifts, responding to challenges, embracing advancements, and navigating the delicate balance between responsiveness to the world and the active shaping of it. The transformative events of the COVID-19 pandemic spotlighted the indispensable role of teachers in Ireland, reinforcing the critical nature of education for the well-being of pupils. Research solidifies the understanding that teachers matter and so it is worth exploring the pivotal role of the teacher educator. This reflective piece examines the changes in teacher education and explores the juxtapositions that have emerged in response to three decades of profound change. The attractiveness of teaching as a career is juxtaposed against the reality of the demands of the job, with conditions for public servants in Ireland undergoing a shift. High-level strategic discussions about increasing teacher numbers now contrast with a previous oversupply. The delicate balance between the imperative to increase enrolment (getting "bums on seats") and the gatekeeper role of teacher educators is explored, raising questions about maintaining high standards amid changing student profiles. Another poignant dichotomy involves the high demand for teachers versus the hurdles candidates face in becoming teachers. The rising cost and duration of teacher education courses raise concerns about attracting quality candidates. The perceived attractiveness of teaching as a career contends with the reality of increased demands on educators. One notable juxtaposition centres around the rapid evolution of Irish initial teacher education versus the potential risk of change overload. The Teaching Council of Ireland has spearheaded considerable changes, raising questions about the timing and evaluation of these changes. This reflection contemplates the vision of a professional teaching council versus its evolving reality and the challenges posed by the value placed on school placement in teacher preparation. The juxtapositions extend to the classroom, where theory may not seamlessly align with the lived experience. Inconsistencies between college expectations and the classroom reality prompt reflection on the effectiveness of teacher preparation programs. Addressing the changing demographic landscape of society and schools, there is a persistent incongruity between the diversity of Irish society and the profile of second-level teachers. As education undergoes a digital revolution, the enduring philosophies of education confront technological advances. This reflection highlights the tension between established practices and contemporary demands, acknowledging the irreplaceable value of face-to-face interaction while integrating technology into teacher training programs. In conclusion, this reflective journey encapsulates the intricate web of juxtapositions in Irish Initial Teacher Education. It emphasises the enduring commitment to fostering education, recognising the profound influence educators wield, and acknowledging the challenges and gratifications inherent in shaping the minds and futures of generations to come.

Keywords: Irish post primary teaching, juxtapositions, reflection, teacher education

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111 Photo-Fenton Degradation of Organic Compounds by Iron(II)-Embedded Composites

Authors: Marius Sebastian Secula, Andreea Vajda, Benoit Cagnon, Ioan Mamaliga

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One of the most important classes of pollutants is represented by dyes. The synthetic character and complex molecular structure make them more stable and difficult to be biodegraded in water. The treatment of wastewaters containing dyes in order to separate/degrade dyes is of major importance. Various techniques have been employed to remove and/or degrade dyes in water. Advanced oxidation processes (AOPs) are known as among the most efficient ones towards dye degradation. The aim of this work is to investigate the efficiency of a cheap Iron-impregnated activated carbon Fenton-like catalyst in order to degrade organic compounds in aqueous solutions. In the presented study an anionic dye, Indigo Carmine, is considered as a model pollutant. Various AOPs are evaluated for the degradation of Indigo Carmine to establish the effect of the prepared catalyst. It was found that the Iron(II)-embedded activated carbon composite enhances significantly the degradation process of Indigo Carmine. Using the wet impregnation procedure, 5 g of L27 AC material were contacted with Fe(II) solutions of FeSO4 precursor at a theoretical iron content in the resulted composite of 1 %. The L27 AC was impregnated for 3h at 45°C, then filtered, washed several times with water and ethanol and dried at 55 °C for 24 h. Thermogravimetric analysis, Fourier transform infrared, X-ray diffraction, and transmission electron microscopy were employed to investigate the structural, textural, and micromorphology of the catalyst. Total iron content in the obtained composites and iron leakage were determined by spectrophotometric method using phenantroline. Photo-catalytic tests were performed using an UV - Consulting Peschl Laboratory Reactor System. UV light irradiation tests were carried out to determine the performance of the prepared Iron-impregnated composite towards the degradation of Indigo Carmine in aqueous solution using different conditions (17 W UV lamps, with and without in-situ generation of O3; different concentrations of H2O2, different initial concentrations of Indigo Carmine, different values of pH, different doses of NH4-OH enhancer). The photocatalytic tests were performed after the adsorption equilibrium has been established. The obtained results emphasize an enhancement of Indigo Carmine degradation in case of the heterogeneous photo-Fenton process conducted with an O3 generating UV lamp in the presence of hydrogen peroxide. The investigated process obeys the pseudo-first order kinetics. The photo-Fenton degradation of IC was tested at different values of initial concentration. The obtained results emphasize an enhancement of Indigo Carmine degradation in case of the heterogeneous photo-Fenton process conducted with an O3 generating UV lamp in the presence of hydrogen peroxide. Acknowledgments: This work was supported by a grant of the Romanian National Authority for Scientific Research and Innovation, CNCS - UEFISCDI, project number PN-II-RU-TE-2014-4-0405.

Keywords: photodegradation, heterogeneous Fenton, anionic dye, carbonaceous composite, screening factorial design

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110 Tangible Losses, Intangible Traumas: Re-envisioning Recovery Following the Lytton Creek Fire 2021 through Place Attachment Lens

Authors: Tugba Altin

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In an era marked by pronounced climate change consequences, communities are observed to confront traumatic events that yield both tangible and intangible repercussions. Such events not only cause discernible damage to the landscape but also deeply affect the intangible aspects, including emotional distress and disruptions to cultural landscapes. The Lytton Creek Fire of 2021 serves as a case in point. Beyond the visible destruction, the less overt but profoundly impactful disturbance to place attachment (PA) is scrutinized. PA, representing the emotional and cognitive bonds individuals establish with their environments, is crucial for understanding how such events impact cultural identity and connection to the land. The study underscores the significance of addressing both tangible and intangible traumas for holistic community recovery. As communities renegotiate their affiliations with altered environments, the cultural landscape emerges as instrumental in shaping place-based identities. This renewed understanding is pivotal for reshaping adaptation planning. The research advocates for adaptation strategies rooted in the lived experiences and testimonies of the affected populations. By incorporating both the tangible and intangible facets of trauma, planning efforts are suggested to be more culturally attuned and emotionally insightful, fostering true resonance with the affected communities. Through such a comprehensive lens, this study contributes enriching the climate change discourse, emphasizing the intertwined nature of tangible recovery and the imperative of emotional and cultural healing after environmental disasters. Following the pronounced aftermath of the Lytton Creek Fire in 2021, research aims to deeply understand its impact on place attachment (PA), encompassing the emotional and cognitive bonds individuals form with their environments. The interpretive phenomenological approach, enriched by a hermeneutic framework, is adopted, emphasizing the experiences of the Lytton community and co-researchers. Phenomenology informed the understanding of 'place' as the focal point of attachment, providing insights into its formation and evolution after traumatic events. Data collection departs from conventional methods. Instead of traditional interviews, walking audio sessions and photo elicitation methods are utilized. These allow co-researchers to immerse themselves in the environment, re-experience, and articulate memories and feelings in real-time. Walking audio facilitates reflections on spatial narratives post-trauma, while photo voices captured intangible emotions, enabling the visualization of place-based experiences. The analysis is collaborative, ensuring the co-researchers' experiences and interpretations are central. Emphasizing their agency in knowledge production, the process is rigorous, facilitated by the harmonious blend of interpretive phenomenology and hermeneutic insights. The findings underscore the need for adaptation and recovery efforts to address emotional traumas alongside tangible damages. By exploring PA post-disaster, the research not only fills a significant gap but advocates for an inclusive approach to community recovery. Furthermore, the participatory methodologies employed challenge traditional research paradigms, heralding potential shifts in qualitative research norms.

Keywords: wildfire recovery, place attachment, trauma recovery, cultural landscape, visual methodologies

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109 Post Harvest Fungi Diversity and Level of Aflatoxin Contamination in Stored Maize: Cases of Kitui, Nakuru and Trans-Nzoia Counties in Kenya

Authors: Gachara Grace, Kebira Anthony, Harvey Jagger, Wainaina James

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Aflatoxin contamination of maize in Africa poses a major threat to food security and the health of many African people. In Kenya, aflatoxin contamination of maize is high due to the environmental, agricultural and socio-economic factors. Many studies have been conducted to understand the scope of the problem, especially at pre-harvest level. This research was carried out to gather scientific information on the fungi population, diversity and aflatoxin level during the post-harvest period. The study was conducted in three geographical locations of; Kitui, Kitale and Nakuru. Samples were collected from storage structures of farmers and transported to the Biosciences eastern and central Africa (BecA), International Livestock and Research Institute (ILRI) hub laboratories. Mycoflora was recovered using the direct plating method. A total of five fungal genera (Aspergillus, Penicillium, Fusarium, Rhizopus and Bssyochlamys spp.) were isolated from the stored maize samples. The most common fungal species that were isolated from the three study sites included A. flavus at 82.03% followed by A.niger and F.solani at 49% and 26% respectively. The aflatoxin producing fungi A. flavus was recovered in 82.03% of the samples. Aflatoxin levels were analysed on both the maize samples and in vitro. Most of the A. flavus isolates recorded a high level of aflatoxin when they were analysed for presence of aflatoxin B1 using ELISA. In Kitui, all the samples (100%) had aflatoxin levels above 10ppb with a total aflatoxin mean of 219.2ppb. In Kitale, only 3 samples (n=39) had their aflatoxin levels less than 10ppb while in Nakuru, the total aflatoxin mean level of this region was 239.7ppb. When individual samples were analysed using Vicam fluorometer method, aflatoxin analysis revealed that most of the samples (58.4%) had been contaminated. The means were significantly different (p=0.00<0.05) in all the three locations. Genetic relationships of A. flavus isolates were determined using 13 Simple Sequence Repeats (SSRs) markers. The results were used to generate a phylogenetic tree using DARwin5 software program. A total of 5 distinct clusters were revealed among the genotypes. The isolates appeared to cluster separately according to the geographical locations. Principal Coordinates Analysis (PCoA) of the genetic distances among the 91 A. flavus isolates explained over 50.3% of the total variation when two coordinates were used to cluster the isolates. Analysis of Molecular Variance (AMOVA) showed a high variation of 87% within populations and 13% among populations. This research has shown that A. flavus is the main fungal species infecting maize grains in Kenya. The influence of aflatoxins on human populations in Kenya demonstrates a clear need for tools to manage contamination of locally produced maize. Food basket surveys for aflatoxin contamination should be conducted on a regular basis. This would assist in obtaining reliable data on aflatoxin incidence in different food crops. This would go a long way in defining control strategies for this menace.

Keywords: aflatoxin, Aspergillus flavus, genotyping, Kenya

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108 Carbon Nanotubes (CNTs) as Multiplex Surface Enhanced Raman Scattering Sensing Platforms

Authors: Pola Goldberg Oppenheimer, Stephan Hofmann, Sumeet Mahajan

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Owing to its fingerprint molecular specificity and high sensitivity, surface-enhanced Raman scattering (SERS) is an established analytical tool for chemical and biological sensing capable of single-molecule detection. A strong Raman signal can be generated from SERS-active platforms given the analyte is within the enhanced plasmon field generated near a noble-metal nanostructured substrate. The key requirement for generating strong plasmon resonances to provide this electromagnetic enhancement is an appropriate metal surface roughness. Controlling nanoscale features for generating these regions of high electromagnetic enhancement, the so-called SERS ‘hot-spots’, is still a challenge. Significant advances have been made in SERS research, with wide-ranging techniques to generate substrates with tunable size and shape of the nanoscale roughness features. Nevertheless, the development and application of SERS has been inhibited by the irreproducibility and complexity of fabrication routes. The ability to generate straightforward, cost-effective, multiplex-able and addressable SERS substrates with high enhancements is of profound interest for miniaturised sensing devices. Carbon nanotubes (CNTs) have been concurrently, a topic of extensive research however, their applications for plasmonics has been only recently beginning to gain interest. CNTs can provide low-cost, large-active-area patternable substrates which, coupled with appropriate functionalization capable to provide advanced SERS-platforms. Herein, advanced methods to generate CNT-based SERS active detection platforms will be discussed. First, a novel electrohydrodynamic (EHD) lithographic technique will be introduced for patterning CNT-polymer composites, providing a straightforward, single-step approach for generating high-fidelity sub-micron-sized nanocomposite structures within which anisotropic CNTs are vertically aligned. The created structures are readily fine-tuned, which is an important requirement for optimizing SERS to obtain the highest enhancements with each of the EHD-CNTs individual structural units functioning as an isolated sensor. Further, gold-functionalized VACNTFs are fabricated as SERS micro-platforms. The dependence on the VACNTs’ diameters and density play an important role in the Raman signal strength, thus highlighting the importance of structural parameters, previously overlooked in designing and fabricating optimized CNTs-based SERS nanoprobes. VACNTs forests patterned into predesigned pillar structures are further utilized for multiplex detection of bio-analytes. Since CNTs exhibit electrical conductivity and unique adsorption properties, these are further harnessed in the development of novel chemical and bio-sensing platforms.

Keywords: carbon nanotubes (CNTs), EHD patterning, SERS, vertically aligned carbon nanotube forests (VACNTF)

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107 A Parallel Cellular Automaton Model of Tumor Growth for Multicore and GPU Programming

Authors: Manuel I. Capel, Antonio Tomeu, Alberto Salguero

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Tumor growth from a transformed cancer-cell up to a clinically apparent mass spans through a range of spatial and temporal magnitudes. Through computer simulations, Cellular Automata (CA) can accurately describe the complexity of the development of tumors. Tumor development prognosis can now be made -without making patients undergo through annoying medical examinations or painful invasive procedures- if we develop appropriate CA-based software tools. In silico testing mainly refers to Computational Biology research studies of application to clinical actions in Medicine. To establish sound computer-based models of cellular behavior, certainly reduces costs and saves precious time with respect to carrying out experiments in vitro at labs or in vivo with living cells and organisms. These aim to produce scientifically relevant results compared to traditional in vitro testing, which is slow, expensive, and does not generally have acceptable reproducibility under the same conditions. For speeding up computer simulations of cellular models, specific literature shows recent proposals based on the CA approach that include advanced techniques, such the clever use of supporting efficient data structures when modeling with deterministic stochastic cellular automata. Multiparadigm and multiscale simulation of tumor dynamics is just beginning to be developed by the concerned research community. The use of stochastic cellular automata (SCA), whose parallel programming implementations are open to yield a high computational performance, are of much interest to be explored up to their computational limits. There have been some approaches based on optimizations to advance in multiparadigm models of tumor growth, which mainly pursuit to improve performance of these models through efficient memory accesses guarantee, or considering the dynamic evolution of the memory space (grids, trees,…) that holds crucial data in simulations. In our opinion, the different optimizations mentioned above are not decisive enough to achieve the high performance computing power that cell-behavior simulation programs actually need. The possibility of using multicore and GPU parallelism as a promising multiplatform and framework to develop new programming techniques to speed-up the computation time of simulations is just starting to be explored in the few last years. This paper presents a model that incorporates parallel processing, identifying the synchronization necessary for speeding up tumor growth simulations implemented in Java and C++ programming environments. The speed up improvement that specific parallel syntactic constructs, such as executors (thread pools) in Java, are studied. The new tumor growth parallel model is proved using implementations with Java and C++ languages on two different platforms: chipset Intel core i-X and a HPC cluster of processors at our university. The parallelization of Polesczuk and Enderling model (normally used by researchers in mathematical oncology) proposed here is analyzed with respect to performance gain. We intend to apply the model and overall parallelization technique presented here to solid tumors of specific affiliation such as prostate, breast, or colon. Our final objective is to set up a multiparadigm model capable of modelling angiogenesis, or the growth inhibition induced by chemotaxis, as well as the effect of therapies based on the presence of cytotoxic/cytostatic drugs.

Keywords: cellular automaton, tumor growth model, simulation, multicore and manycore programming, parallel programming, high performance computing, speed up

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106 Internet of Assets: A Blockchain-Inspired Academic Program

Authors: Benjamin Arazi

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Blockchain is the technology behind cryptocurrencies like Bitcoin. It revolutionizes the meaning of trust in the sense of offering total reliability without relying on any central entity that controls or supervises the system. The Wall Street Journal states: “Blockchain Marks the Next Step in the Internet’s Evolution”. Blockchain was listed as #1 in Linkedin – The Learning Blog “most in-demand hard skills needed in 2020”. As stated there: “Blockchain’s novel way to store, validate, authorize, and move data across the internet has evolved to securely store and send any digital asset”. GSMA, a leading Telco organization of mobile communications operators, declared that “Blockchain has the potential to be for value what the Internet has been for information”. Motivated by these seminal observations, this paper presents the foundations of a Blockchain-based “Internet of Assets” academic program that joins under one roof leading application areas that are characterized by the transfer of assets over communication lines. Two such areas, which are pillars of our economy, are Fintech – Financial Technology and mobile communications services. The next application in line is Healthcare. These challenges are met based on available extensive professional literature. Blockchain-based assets communication is based on extending the principle of Bitcoin, starting with the basic question: If digital money that travels across the universe can ‘prove its own validity’, can this principle be applied to digital content. A groundbreaking positive answer here led to the concept of “smart contract” and consequently to DLT - Distributed Ledger Technology, where the word ‘distributed’ relates to the non-existence of reliable central entities or trusted third parties. The terms Blockchain and DLT are frequently used interchangeably in various application areas. The World Bank Group compiled comprehensive reports, analyzing the contribution of DLT/Blockchain to Fintech. The European Central Bank and Bank of Japan are engaged in Project Stella, “Balancing confidentiality and auditability in a distributed ledger environment”. 130 DLT/Blockchain focused Fintech startups are now operating in Switzerland. Blockchain impact on mobile communications services is treated in detail by leading organizations. The TM Forum is a global industry association in the telecom industry, with over 850 member companies, mainly mobile operators, that generate US$2 trillion in revenue and serve five billion customers across 180 countries. From their perspective: “Blockchain is considered one of the digital economy’s most disruptive technologies”. Samples of Blockchain contributions to Fintech (taken from a World Bank document): Decentralization and disintermediation; Greater transparency and easier auditability; Automation & programmability; Immutability & verifiability; Gains in speed and efficiency; Cost reductions; Enhanced cyber security resilience. Samples of Blockchain contributions to the Telco industry. Establishing identity verification; Record of transactions for easy cost settlement; Automatic triggering of roaming contract which enables near-instantaneous charging and reduction in roaming fraud; Decentralized roaming agreements; Settling accounts per costs incurred in accordance with agreement tariffs. This clearly demonstrates an academic education structure where fundamental technologies are studied in classes together with these two application areas. Advanced courses, treating specific implementations then follow separately. All are under the roof of “Internet of Assets”.

Keywords: blockchain, education, financial technology, mobile telecommunications services

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105 Evolution of Plio/Pleistocene Sedimentary Processes in Patraikos Gulf, Offshore Western Greece

Authors: E. K. Tripsanas, D. Spanos, I. Oikonomopoulos, K. Stathopoulou, A. S. Abdelsamad, A. Pagoulatos

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Patraikos Gulf is located offshore western Greece, and it is limited to the west by the Zante, Cephalonia, and Lefkas islands. The Plio/Pleistocene sequence is characterized by two depocenters, the east and west Patraikos basins separated from each other by a prominent sill. This study is based on the Plio/Pleistocene seismic stratigraphy analysis of a newly acquired 3D PSDM (Pre-Stack depth migration) seismic survey in the west Patraikos Basin and few 2D seismic profiles throughout the entire Patraikos Gulf. The eastern Patraikos Basin, although completely buried today with water depths less than 100 m, it was a deep basin during Pliocene ( > 2 km of Pliocene-Pleistocene sediments) and appears to have gathered most of Achelous River discharges. The west Patraikos Gulf was shallower ( < 1300 m of Pliocene-Pleistocene sediments) and characterized by a hummocky relief due to thrust-belt tectonics and Miocene to Pleistocene halokinetic processes. The transition from Pliocene to Miocene is expressed by a widespread erosional unconformity with evidence of fluvial drainage patterns. This indicates that west Patraikos Basin was aerially exposed during the Messinian Salinity Crisis. Continuous to semi-continuous, parallel reflections in the lower, early- to mid-Pliocene seismic packet provides evidence that the re-connection of the Mediterranean Sea with the Atlantic Ocean during Zanclean resulted in the flooding of the west Patraikos basin and the domination of hemipelagic sedimentation interrupted by occasional gravity flows. This is evident in amplitude and semblance horizon slices, which clearly show the presence of long-running, meandering submarine channels sourced from the southeast (northwest Peloponnese) and north. The long-running nature of the submarine channels suggests mobile efficient turbidity currents, probably due to the participation of a sufficient amount of clay minerals in their suspended load. The upper seismic section in the study area mainly consists of several successions of clinoforms, interpreted as progradational delta complexes of Achelous River. This sudden change from marine to shallow marine sedimentary processes is attributed to climatic changes and eustatic perturbations since late Pliocene onwards (~ 2.6 Ma) and/or a switch of Achelous River from the east Patraikos Basin to the west Patraikos Basin. The deltaic seismic unit consists of four delta complexes. The first two complexes result in the infill of topographic depressions and smoothing of an initial hummocky bathymetry. The distribution of the upper two delta complexes is controlled by compensational stacking. Amplitude and semblance horizon slices depict the development of several almost straight and short (a few km long) distributary submarine channels at the delta slopes and proximal prodeltaic plains with lobate sand-sheet deposits at their mouths. Such channels are interpreted to result from low-efficiency turbidity currents with low content in clay minerals. Such a differentiation in the nature of the gravity flows is attributed to the switch of the sediment supply from clay-rich sediments derived from the draining of flysch formations of the Ionian and Gavrovo zones, to the draining of poor in clay minerals carbonate formations of Gavrovo zone through the Achelous River.

Keywords: sequence stratigraphy, basin analysis, river deltas, submarine channels

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104 Effect of Endurance Training on Serum Chemerin Levels and Lipid Profile of Plasma in Obese Women

Authors: A. Moghadasein, M. Ghasemi, S. Fazelifar

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Aim: Chemerin is a novel adipokine that play an important role in regulating lipid metabolism and abiogenesis. Chemerin is dependent on autocrine and paracrine signals for the differentiation and maturation of fat cells; it also regulates glucose uptake in fat cells and stimulates lipolysis. It has been reported that in adipocytes, chemerin enhances the insulin-stimulated glucose and causes the phosphorylation of tyrosine in Insulin receptor substrate. According to the studies, Chemerin may increase insulin sensitivity in adipose tissue and is largely associated with Body mass index, triglycerides, and blood pressure in those with normal glucose tolerance. There is limited information available regarding the effect of exercise training on serum chemerin concentrations. The purpose of this study was to investigate the effect of endurance training on serum chemerin levels and lipids of plasma in overweight women. Methodology: This study was a quasi-experimental research with a pre-post test design. After required examination and verification of high pressure by the physician, 22 obese subjects (age: 35.64±5.55 yr, weight: 75.62±9.30 kg, body mass index: 32.4±1.6 kg/m2) were randomly assigned to aerobic training (n= 12) and control (n= 12) groups. Participants completed a questionnaire indicating the lack of sports history during the past six months, the lack of anti-hypertension drugs use, hormone therapy, cardiovascular problems, and complete stoppage of menstrual cycle. Aerobic training was performed 3 times weekly for 8 weeks. Resting levels of chemerin plasma, metabolic parameters were measured prior to and after the intervention. The control group did not participate in any training program. In this study, ethical considerations included the complete description of the objectives to the study participants, ensuring the confidentiality of their information. Kolmogorov-Smirnov and Levin test were used for determining the normal distribution of data and homogeneity of variances, respectively. Analyze of variance with repeated measure were used to investigate the changes in the intra-group and the differences in inter-group of variables. Statistical operations were performed using SPSS 16 and the significance level of the tests was considered at P < 0.05. Results: After an 8 week aerobic training, levels of chemerin plasma were significantly decreased in aerobic trained group when compared with their control groups (p < 0.05).Concurrently, levels of HDL-c were significantly decreased (p < 0.05) whereas, levels of cholesterol, TG and LDL-c, showed no significant changes (p > 0.05). No significant correlations between chemerin levels and weight loss were observed in subjects with overweight women. Conclusion: The present study demonstrated, 8 weeks aerobic training, reduced serum chemerin concentrations in overweight women. Whereas, aerobic training exercise programmers affected the lipid profile response of obese subjects differently. However further research is warranted in order to unravel the molecular mechanism for the range of responses and the role of serum chemerin.

Keywords: chemerin, aerobic training, lipid profile, obese women

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103 Preparation of Biodegradable Methacrylic Nanoparticles by Semicontinuous Heterophase Polymerization for Drugs Loading: The Case of Acetylsalicylic Acid

Authors: J. Roberto Lopez, Hened Saade, Graciela Morales, Javier Enriquez, Raul G. Lopez

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Implementation of systems based on nanostructures for drug delivery applications have taken relevance in recent studies focused on biomedical applications. Although there are several nanostructures as drugs carriers, the use of polymeric nanoparticles (PNP) has been widely studied for this purpose, however, the main issue for these nanostructures is the size control below 50 nm with a narrow distribution size, due to they must go through different physiological barriers and avoid to be filtered by kidneys (< 10 nm) or the spleen (> 100 nm). Thus, considering these and other factors, it can be mentioned that drug-loaded nanostructures with sizes varying between 10 and 50 nm are preferred in the development and study of PNP/drugs systems. In this sense, the Semicontinuous Heterophase Polymerization (SHP) offers the possibility to obtain PNP in the desired size range. Considering the above explained, methacrylic copolymer nanoparticles were obtained under SHP. The reactions were carried out in a jacketed glass reactor with the required quantities of water, ammonium persulfate as initiator, sodium dodecyl sulfate/sodium dioctyl sulfosuccinate as surfactants, methyl methacrylate and methacrylic acid as monomers with molar ratio of 2/1, respectively. The monomer solution was dosed dropwise during reaction at 70 °C with a mechanical stirring of 650 rpm. Nanoparticles of poly(methyl methacrylate-co-methacrylic acid) were loaded with acetylsalicylic acid (ASA, aspirin) by a chemical adsorption technique. The purified latex was put in contact with a solution of ASA in dichloromethane (DCM) at 0.1, 0.2, 0.4 or 0.6 wt-%, at 35°C during 12 hours. According to the boiling point of DCM, as well as DCM and water densities, the loading process is completed when the whole DCM is evaporated. The hydrodynamic diameter was measured after polymerization by quasi-elastic light scattering and transmission electron microscopy, before and after loading procedures with ASA. The quantitative and qualitative analyses of PNP loaded with ASA were measured by infrared spectroscopy, differential scattering calorimetry and thermogravimetric analysis. Also, the molar mass distributions of polymers were determined in a gel permeation chromatograph apparatus. The load capacity and efficiency were determined by gravimetric analysis. The hydrodynamic diameter results for methacrylic PNP without ASA showed a narrow distribution with an average particle size around 10 nm and a composition methyl methacrylate/methacrylic acid molar ratio equal to 2/1, same composition of Eudragit S100, which is a commercial compound widely used as excipient. Moreover, the latex was stabilized in a relative high solids content (around 11 %), a monomer conversion almost 95 % and a number molecular weight around 400 Kg/mol. The average particle size in the PNP/aspirin systems fluctuated between 18 and 24 nm depending on the initial percentage of aspirin in the loading process, being the drug content as high as 24 % with an efficiency loading of 36 %. These average sizes results have not been reported in the literature, thus, the methacrylic nanoparticles here reported are capable to be loaded with a considerable amount of ASA and be used as a drug carrier.

Keywords: aspirin, biocompatibility, biodegradable, Eudragit S100, methacrylic nanoparticles

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102 Mixed Monolayer and PEG Linker Approaches to Creating Multifunctional Gold Nanoparticles

Authors: D. Dixon, J. Nicol, J. A. Coulter, E. Harrison

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The ease with which they can be functionalized, combined with their excellent biocompatibility, make gold nanoparticles (AuNPs) ideal candidates for various applications in nanomedicine. Indeed several promising treatments are currently undergoing human clinical trials (CYT-6091 and Auroshell). A successful nanoparticle treatment must first evade the immune system, then accumulate within the target tissue, before enter the diseased cells and delivering the payload. In order to create a clinically relevant drug delivery system, contrast agent or radiosensitizer, it is generally necessary to functionalize the AuNP surface with multiple groups; e.g. Polyethylene Glycol (PEG) for enhanced stability, targeting groups such as antibodies, peptides for enhanced internalization, and therapeutic agents. Creating and characterizing the biological response of such complex systems remains a challenge. The two commonly used methods to attach multiple groups to the surface of AuNPs are the creation of a mixed monolayer, or by binding groups to the AuNP surface using a bi-functional PEG linker. While some excellent in-vitro and animal results have been reported for both approaches further work is necessary to directly compare the two methods. In this study AuNPs capped with both PEG and a Receptor Mediated Endocytosis (RME) peptide were prepared using both mixed monolayer and PEG linker approaches. The PEG linker used was SH-PEG-SGA which has a thiol at one end for AuNP attachment, and an NHS ester at the other to bind to the peptide. The work builds upon previous studies carried out at the University of Ulster which have investigated AuNP synthesis, the influence of PEG on stability in a range of media and investigated intracellular payload release. 18-19nm citrate capped AuNPs were prepared using the Turkevich method via the sodium citrate reduction of boiling 0.01wt% Chloroauric acid. To produce PEG capped AuNPs, the required amount of PEG-SH (5000Mw) or SH-PEG-SGA (3000Mw Jenkem Technologies) was added, and the solution stirred overnight at room temperature. The RME (sequence: CKKKKKKSEDEYPYVPN, Biomatik) co-functionalised samples were prepared by adding the required amount of peptide to the PEG capped samples and stirring overnight. The appropriate amounts of PEG-SH and RME peptide were added to the AuNP to produce a mixed monolayer consisting of approximately 50% PEG and 50% RME. The PEG linker samples were first fully capped with bi-functional PEG before being capped with RME peptide. An increase in diameter from 18-19mm for the ‘as synthesized’ AuNPs to 40-42nm after PEG capping was observed via DLS. The presence of PEG and RME peptide on both the mixed monolayer and PEG linker co-functionalized samples was confirmed by both FTIR and TGA. Bi-functional PEG linkers allow the entire AuNP surface to be capped with PEG, enabling in-vitro stability to be achieved using a lower molecular weight PEG. The approach also allows the entire outer surface to be coated with peptide or other biologically active groups, whilst also offering the promise of enhanced biological availability. The effect of mixed monolayer versus PEG linker attachment on both stability and non-specific protein corona interactions was also studied.

Keywords: nanomedicine, gold nanoparticles, PEG, biocompatibility

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101 Ionophore-Based Materials for Selective Optical Sensing of Iron(III)

Authors: Natalia Lukasik, Ewa Wagner-Wysiecka

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Development of selective, fast-responsive, and economical sensors for diverse ions detection and determination is one of the most extensively studied areas due to its importance in the field of clinical, environmental and industrial analysis. Among chemical sensors, vast popularity has gained ionophore-based optical sensors, where the generated analytical signal is a consequence of the molecular recognition of ion by the ionophore. Change of color occurring during host-guest interactions allows for quantitative analysis and for 'naked-eye' detection without the need of using sophisticated equipment. An example of application of such sensors is colorimetric detection of iron(III) cations. Iron as one of the most significant trace elements plays roles in many biochemical processes. For these reasons, the development of reliable, fast, and selective methods of iron ions determination is highly demanded. Taking all mentioned above into account a chromogenic amide derivative of 3,4-dihydroxybenzoic acid was synthesized, and its ability to iron(III) recognition was tested. To the best of authors knowledge (according to chemical abstracts) the obtained ligand has not been described in the literature so far. The catechol moiety was introduced to the ligand structure in order to mimic the action of naturally occurring siderophores-iron(III)-selective receptors. The ligand–ion interactions were studied using spectroscopic methods: UV-Vis spectrophotometry and infrared spectroscopy. The spectrophotometric measurements revealed that the amide exhibits affinity to iron(III) in dimethyl sulfoxide and fully aqueous solution, what is manifested by the change of color from yellow to green. Incorporation of the tested amide into a polymeric matrix (cellulose triacetate) ensured effective recognition of iron(III) at pH 3 with the detection limit 1.58×10⁻⁵ M. For the obtained sensor material parameters like linear response range, response time, selectivity, and possibility of regeneration were determined. In order to evaluate the effect of the size of the sensing material on iron(III) detection nanospheres (in the form of nanoemulsion) containing the tested amide were also prepared. According to DLS (dynamic light scattering) measurements, the size of the nanospheres is 308.02 ± 0.67 nm. Work parameters of the nanospheres were determined and compared with cellulose triacetate-based material. Additionally, for fast, qualitative experiments the test strips were prepared by adsorption of the amide solution on a glass microfiber material. Visual limit of detection of iron(III) at pH 3 by the test strips was estimated at the level 10⁻⁴ M. In conclusion, reported here amide derived from 3,4- dihydroxybenzoic acid proved to be an effective candidate for optical sensing of iron(III) in fully aqueous solutions. N. L. kindly acknowledges financial support from National Science Centre Poland the grant no. 2017/01/X/ST4/01680. Authors thank for financial support from Gdansk University of Technology grant no. 032406.

Keywords: ion-selective optode, iron(III) recognition, nanospheres, optical sensor

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100 Functionalizing Gold Nanostars with Ninhydrin as Vehicle Molecule for Biomedical Applications

Authors: Swati Mishra

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In recent years, there has been an explosion in Gold NanoParticle (GNP) research, with a rapid increase in publications in diverse fields, including imaging, bioengineering, and molecular biology. GNPs exhibit unique physicochemical properties, including surface plasmon resonance (SPR) and bind amine and thiol groups, allowing surface modification and use in biomedical applications. Nanoparticle functionalization is the subject of intense research at present, with rapid progress being made towards developing biocompatible, multi-functional particles. In the present study, the photochemical method has been done to functionalize various-shaped GNPs like nanostars by the molecules like ninhydrin. Ninhydrin is bactericidal, virucidal, fungicidal, antigen-antibody reactive, and used in fingerprint technology in forensics. The GNPs functionalized with ninhydrin efficiently will bind to the amino acids on the target protein, which is of eminent importance during the pandemic, especially where long-term treatments of COVID- 19 bring many side effects of the drugs. The photochemical method is adopted as it provides low thermal load, selective reactivity, selective activation, and controlled radiation in time, space, and energy. The GNPs exhibit their characteristic spectrum, but a distinctly blue or redshift in the peak will be observed after UV irradiation, ensuring efficient ninhydrin binding. Now, the bound ninhydrin in the GNP carrier, upon chemically reacting with any amino acid, will lead to the formation of Rhumann purple. A common method of GNP production includes citrate reduction of Au [III] derivatives such as aurochloric acid (HAuCl4) in water to Au [0] through a one-step synthesis of size-tunable GNPs. The following reagents are prepared to validate the approach. Reagent A solution 1 is0.0175 grams ninhydrin in 5 ml Millipore water Reagent B 30 µl of HAuCl₄.3H₂O in 3 ml of solution 1 Reagent C 1 µl of gold nanostars in 3 ml of solution 1 Reagent D 6 µl of cetrimonium bromide (CTAB) in 3 ml of solution1 ReagentE 1 µl of gold nanostars in 3 ml of ethanol ReagentF 30 µl of HAuCl₄.₃H₂O in 3 ml of ethanol ReagentG 30 µl of HAuCl₄.₃H₂O in 3 ml of solution 2 ReagentH solution 2 is0.0087 grams ninhydrin in 5 ml Millipore water ReagentI 30 µl of HAuCl₄.₃H₂O in 3 ml of water The reagents were irradiated at 254 nm for 15 minutes, followed by their UV Visible spectroscopy. The wavelength was selected based on the one reported for excitation of a similar molecule Pthalimide. It was observed that the solution B and G deviate around 600 nm, while C peaks distinctively at 567.25 nm and 983.9 nm. Though it is tough to say about the chemical reaction happening, butATR-FTIR of reagents will ensure that ninhydrin is not forming Rhumann purple in the absence of amino acids. Therefore, these experiments, we achieved the functionalization of gold nanostars with ninhydrin corroborated by the deviation in the spectrum obtained in a mixture of GNPs and ninhydrin irradiated with UV light. It prepares them as a carrier molecule totake up amino acids for targeted delivery or germicidal action.

Keywords: gold nanostars, ninhydrin, photochemical method, UV visible specgtroscopy

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99 Branding Capability Developed from Country-Specific and Firm-Specific Resources for Internationalizing Small and Medium Enterprises

Authors: Hsing-Hua Stella Chang, Mong-Ching Lin, Cher-Min Fong

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There has recently been a notable rise in the number of emerging-market industrial small and medium-sized enterprises (SMEs) that have managed to upgrade their operations. Evolving from original equipment manufacturing (OEM) into value-added original or own brand manufacturing (OBM) in such firms represents a specific process of internationalization. The OEM-OBM upgrade requires development of a firm’s own brand. In this respect, the extant literature points out that emerging-market industrial marketers (latecomers) have developed some marketing capabilities, of which branding has been identified as one of the most important. In specific, an industrial non-brand marketer (OEM) marks the division of labor between manufacturing and branding (as part of marketing). In light of this discussion, this research argues that branding capability plays a critical role in supporting the evolution of manufacture upgrade. This is because a smooth transformation from OEM to OBM entails the establishment of strong brands through which branding capability is developed. Accordingly, branding capability can be exemplified as a series of processes and practices in relation to mobilizing branding resources and orchestrating branding activities, which will result in the establishment of business relationships, greater acceptance of business partners (channels, suppliers), and increased industrial brand equity in the firm as key resource advantages). For the study purpose, Taiwan was chosen as the research context, representing a typical case that exemplifies the industrial development path of more-established emerging markets, namely, transformation from OEM to OBM. This research adopted a two-phase research design comprising exploratory (a qualitative study) and confirmatory approaches (a survey study) The findings show that: Country-specific advantage is positively related to branding capability for internationalizing SMEs. Firm-specific advantage is positively related to branding capability for internationalizing SMEs. Hsing-Hua Stella Chang is Assistant Professor with National Taichung University of Education, International Master of Business Administration, (Yingcai Campus) No.227, Minsheng Rd., West Dist., Taichung City 40359, Taiwan, R.O.C. (phone: 886-22183612; e-mail: [email protected]). Mong-Ching Lin is PhD candidate with National Sun Yat-Sen University, Department of Business Management, 70 Lien-hai Rd., Kaohsiung 804, Taiwan, R.O.C. (e-mail: [email protected]). Cher-Min Fong is Full Professor with National Sun Yat-Sen University, Department of Business Management, 70 Lien-hai Rd., Kaohsiung 804, Taiwan, R.O.C. (e-mail: [email protected]). Branding capability is positively related to international performance for internationalizing SMEs. This study presents a pioneering effort to distinguish industrial brand marketers from non-brand marketers in exploring the role of branding capability in the internationalizing small and medium-sized industrial brand marketers from emerging markets. Specifically, when industrial non-brand marketers (OEMs) enter into a more advanced stage of internationalization (i.e., OBM), they must overcome disadvantages (liabilities of smallness, foreignness, outsidership) that do not apply in the case of incumbent developed-country MNEs with leading brands. Such critical differences mark the urgency and significance of distinguishing industrial brand marketers from non-brand marketers on issues relating to their value-adding branding and marketing practices in international markets. This research thus makes important contributions to the international marketing, industrial branding, and SME internationalization literature.

Keywords: brand marketers, branding capability, emerging markets, SME internationalization

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98 Assessment of Cellular Metabolites and Impedance for Early Diagnosis of Oral Cancer among Habitual Smokers

Authors: Ripon Sarkar, Kabita Chaterjee, Ananya Barui

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Smoking is one of the leading causes of oral cancer. Cigarette smoke affects various cellular parameters and alters molecular metabolism of cells. Epithelial cells losses their cytoskeleton structure, membrane integrity, cellular polarity that subsequently initiates the process of epithelial cells to mesenchymal transition due to long exposure of cigarette smoking. It changes the normal cellular metabolic activity which induces oxidative stress and enhances the reactive oxygen spices (ROS) formation. Excessive ROS and associated oxidative stress are considered to be a driving force in alteration in cellular phenotypes, polarity distribution and mitochondrial metabolism. Noninvasive assessment of such parameters plays essential role in development of routine screening system for early diagnosis of oral cancer. Electrical cell-substrate impedance sensing (ECIS) is one of such method applied for detection of cellular membrane impedance which can be correlated to cell membrane integrity. Present study intends to explore the alteration in cellular impedance along with the expression of cellular polarity molecules and cytoskeleton distributions in oral epithelial cells of habitual smokers and to correlate the outcome to that of clinically diagnosed oral leukoplakia and oral squamous cell carcinoma patients. Total 80 subjects were categorized into four study groups: nonsmoker (NS), cigarette smoker (CS), oral leukoplakia (OLPK) and oral squamous cell carcinoma (OSCC). Cytoskeleton distribution was analyzed by staining of actin filament and generation of ROS was measured using assay kit using standard protocol. Cell impedance was measured through ECIS method at different frequencies. Expression of E-cadherin and protease-activated receptor (PAR) proteins were observed through immune-fluorescence method. Distribution of actin filament is well organized in NS group however; distribution pattern was grossly varied in CS, OLPK and OSCC. Generation of ROS was low in NS which subsequently increased towards OSCC. Expressions of E-cadherin and change in cellular electrical impedance in different study groups indicated the hallmark of cancer progression from NS to OSCC. Expressions of E-cadherin, PAR protein, and cell impedance were decreased from NS to CS and farther OSCC. Generally, the oral epithelial cells exhibit apico-basal polarity however with cancer progression these cells lose their characteristic polarity distribution. In this study expression of polarity molecule and ECIS observation indicates such altered pattern of polarity among smoker group. Overall the present study monitored the alterations in intracellular ROS generation and cell metabolic function, membrane integrity in oral epithelial cells in cigarette smokers. Present study thus has clinical significance, and it may help in developing a noninvasive technique for early diagnosis of oral cancer amongst susceptible individuals.

Keywords: cigarette smoking, early oral cancer detection, electric cell-substrate impedance sensing, noninvasive screening

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97 New Hybrid Process for Converting Small Structural Parts from Metal to CFRP

Authors: Yannick Willemin

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Carbon fibre-reinforced plastic (CFRP) offers outstanding value. However, like all materials, CFRP also has its challenges. Many forming processes are largely manual and hard to automate, making it challenging to control repeatability and reproducibility (R&R); they generate significant scrap and are too slow for high-series production; fibre costs are relatively high and subject to supply and cost fluctuations; the supply chain is fragmented; many forms of CFRP are not recyclable, and many materials have yet to be fully characterized for accurate simulation; shelf life and outlife limitations add cost; continuous-fibre forms have design limitations; many materials are brittle; and small and/or thick parts are costly to produce and difficult to automate. A majority of small structural parts are metal due to high CFRP fabrication costs for the small-size class. The fact that CFRP manufacturing processes that produce the highest performance parts also tend to be the slowest and least automated is another reason CFRP parts are generally higher in cost than comparably performing metal parts, which are easier to produce. Fortunately, business is in the midst of a major manufacturing evolution—Industry 4.0— one technology seeing rapid growth is additive manufacturing/3D printing, thanks to new processes and materials, plus an ability to harness Industry 4.0 tools. No longer limited to just prototype parts, metal-additive technologies are used to produce tooling and mold components for high-volume manufacturing, and polymer-additive technologies can incorporate fibres to produce true composites and be used to produce end-use parts with high aesthetics, unmatched complexity, mass customization opportunities, and high mechanical performance. A new hybrid manufacturing process combines the best capabilities of additive—high complexity, low energy usage and waste, 100% traceability, faster to market—and post-consolidation—tight tolerances, high R&R, established materials, and supply chains—technologies. The platform was developed by Zürich-based 9T Labs AG and is called Additive Fusion Technology (AFT). It consists of a design software offering the possibility to determine optimal fibre layup, then exports files back to check predicted performance—plus two pieces of equipment: a 3d-printer—which lays up (near)-net-shape preforms using neat thermoplastic filaments and slit, roll-formed unidirectional carbon fibre-reinforced thermoplastic tapes—and a post-consolidation module—which consolidates then shapes preforms into final parts using a compact compression press fitted with a heating unit and matched metal molds. Matrices—currently including PEKK, PEEK, PA12, and PPS, although nearly any high-quality commercial thermoplastic tapes and filaments can be used—are matched between filaments and tapes to assure excellent bonding. Since thermoplastics are used exclusively, larger assemblies can be produced by bonding or welding together smaller components, and end-of-life parts can be recycled. By combining compression molding with 3D printing, higher part quality with very-low voids and excellent surface finish on A and B sides can be produced. Tight tolerances (min. section thickness=1.5mm, min. section height=0.6mm, min. fibre radius=1.5mm) with high R&R can be cost-competitively held in production volumes of 100 to 10,000 parts/year on a single set of machines.

Keywords: additive manufacturing, composites, thermoplastic, hybrid manufacturing

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96 Enhanced Functional Production of a Crucial Biomolecule Human Serum Albumin in Escherichia coli

Authors: Ashima Sharma

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Human Serum Albumin (HSA)- one of the most demanded therapeutic proteins with immense biotechnological applications- is a large multidomain protein containing 17 disulfide bonds. The current source of HSA is human blood plasma which is a limited and unsafe source. Thus, there exists an indispensable need to promote non-animal derived recombinant HSA (rHSA) production. Escherichia coli is one of the most convenient hosts which had contributed to the production of more than 30% of the FDA approved recombinant pharmaceuticals. It grows rapidly and reaches high cell density using inexpensive and simple substrates. E. coli derived recombinant products have more economic potential as fermentation processes are cheaper compared to the other expression hosts. The major bottleneck in exploiting E. coli as a host for a disulfide-rich multidomain protein is the formation of aggregates of overexpressed protein. The majority of the expressed HSA forms inclusion bodies (more than 90% of the total expressed rHSA) in the E. coli cytosol. Recovery of functional rHSA from inclusion bodies is not preferred because it is difficult to obtain a large multidomain disulfide bond rich protein like rHSA in its functional native form. Purification is tedious, time-consuming, laborious and expensive. Because of such limitations, the E. coli host system was neglected for rHSA production for the past few decades despite its numerous advantages. In the present work, we have exploited the capabilities of E. coli as a host for the enhanced functional production of rHSA (~60% of the total expressed rHSA in the soluble fraction). Parameters like intracellular environment, temperature, induction type, duration of induction, cell lysis conditions etc. which play an important role in enhancing the level of production of the desired protein in its native form in vivo have been optimized. We have studied the effect of assistance of different types of exogenously employed chaperone systems on the functional expression of rHSA in the E. coli host system. Different aspects of cell growth parameters during the production of rHSA in presence and absence of molecular chaperones in E. coli have also been studied. Upon overcoming the difficulties to produce functional rHSA in E. coli, it has been possible to produce significant levels of functional protein through engineering the biological system of protein folding in the cell, the E. coli-derived rHSA has been purified to homogeneity. Its detailed physicochemical characterization has been performed by monitoring its conformational properties, secondary and tertiary structure elements, surface properties, ligand binding properties, stability issues etc. These parameters of the recombinant protein have been compared with the naturally occurring protein from the human source. The outcome of the comparison reveals that the recombinant protein resembles exactly the same as the natural one. Hence, we propose that the E. coli-derived rHSA is an ideal biosimilar for human blood plasma-derived serum albumin. Therefore, in the present study, we have introduced and promoted the E. coli- derived rHSA as an alternative to the preparation from a human source, pHSA.

Keywords: recombinant human serum albumin, Escherichia coli, biosimilar, chaperone assisted protein folding

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95 Investigation of Processing Conditions on Rheological Features of Emulsion Gels and Oleogels Stabilized by Biopolymers

Authors: M. Sarraf, J. E. Moros, M. C. Sánchez

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Oleogels are self-standing systems that are able to trap edible liquid oil into a tridimensional network and also help to use less fat by forming crystallization oleogelators. There are different ways to generate oleogelation and oil structuring, including direct dispersion, structured biphasic systems, oil sorption, and indirect method (emulsion-template). The selection of processing conditions as well as the composition of the oleogels is essential to obtain a stable oleogel with characteristics suitable for its purpose. In this sense, one of the ingredients widely used in food products to produce oleogels and emulsions is polysaccharides. Basil seed gum (BSG), with the scientific name Ocimum basilicum, is a new native polysaccharide with high viscosity and pseudoplastic behavior because of its high molecular weight in the food industry. Also, proteins can stabilize oil in water due to the presence of amino and carboxyl moieties that result in surface activity. Whey proteins are widely used in the food industry due to available, cheap ingredients, nutritional and functional characteristics such as emulsifier and a gelling agent, thickening, and water-binding capacity. In general, the interaction of protein and polysaccharides has a significant effect on the food structures and their stability, like the texture of dairy products, by controlling the interactions in macromolecular systems. Using edible oleogels as oil structuring helps for targeted delivery of a component trapped in a structural network. Therefore, the development of efficient oleogel is essential in the food industry. A complete understanding of the important points, such as the ratio oil phase, processing conditions, and concentrations of biopolymers that affect the formation and stability of the emulsion, can result in crucial information in the production of a suitable oleogel. In this research, the effects of oil concentration and pressure used in the manufacture of the emulsion prior to obtaining the oleogel have been evaluated through the analysis of droplet size and rheological properties of obtained emulsions and oleogels. The results show that the emulsion prepared in the high-pressure homogenizer (HPH) at higher pressure values has smaller droplet sizes and a higher uniformity in the size distribution curve. On the other hand, in relation to the rheological characteristics of the emulsions and oleogels obtained, the predominantly elastic character of the systems must be noted, as they present values of the storage modulus higher than those of losses, also showing an important plateau zone, typical of structured systems. In the same way, if steady-state viscous flow tests have been analyzed on both emulsions and oleogels, the result is that, once again, the pressure used in the homogenizer is an important factor for obtaining emulsions with adequate droplet size and the subsequent oleogel. Thus, various routes for trapping oil inside a biopolymer matrix with adjustable mechanical properties could be applied for the creation of the three-dimensional network in order to the oil absorption and creating oleogel.

Keywords: basil seed gum, particle size, viscoelastic properties, whey protein

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94 MusicTherapy for Actors: An Exploratory Study Applied to Students from University Theatre Faculty

Authors: Adriana De Serio, Adrian Korek

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Aims: This experiential research work presents a Group-MusicTherapy-Theatre-Plan (MusThePlan) the authors have carried out to support the actors. The MusicTherapy gives rise to individual psychophysical feedback and influences the emotional centres of the brain and the subconsciousness. Therefore, the authors underline the effectiveness of the preventive, educational, and training goals of the MusThePlan to lead theatre students and actors to deal with anxiety and to overcome psychophysical weaknesses, shyness, emotional stress in stage performances, to increase flexibility, awareness of one's identity and resources for a positive self-development and psychophysical health, to develop and strengthen social bonds, increasing a network of subjects working for social inclusion and reduction of stigma. Materials-Methods: Thirty students from the University Theatre Faculty participated in weekly music therapy sessions for two months; each session lasted 120 minutes. MusThePlan: Each session began with a free group rhythmic-sonorous-musical-production by body-percussion, voice-canto, instruments, to stimulate communication. Then, a synchronized-structured bodily-rhythmic-sonorous-musical production also involved acting, dances, movements of hands and arms, hearing, and more sensorial perceptions and speech to balance motor skills and the muscular tone. Each student could be the director-leader of the group indicating a story to inspire the group's musical production. The third step involved the students in rhythmic speech and singing drills and in vocal exercises focusing on the musical pitch to improve the intonation and on the diction to improve the articulation and lead up it to an increased intelligibility. At the end of each musictherapy session and of the two months, the Musictherapy Assessment Document was drawn up by analysis of observation protocols and two Indices by the authors: Patient-Environment-Music-Index (time to - tn) to estimate the behavior evolution, Somatic Pattern Index to monitor subject’s eye and mouth and limb motility, perspiration, before, during and after musictherapy sessions. Results: After the first month, the students (non musicians) learned to play percussion instruments and formed a musical band that played classical/modern music on the percussion instruments with the musictherapist/pianist/conductor in a public concert. At the end of the second month, the students performed a public musical theatre show, acting, dancing, singing, and playing percussion instruments. The students highlighted the importance of the playful aspects of the group musical production in order to achieve emotional contact and harmony within the group. The students said they had improved kinetic and vocal and all the skills useful for acting activity and the nourishment of the bodily and emotional balance. Conclusions: The MusThePlan makes use of some specific MusicTherapy methodological models, techniques, and strategies useful for the actors. The MusThePlan can destroy the individual "mask" and can be useful when the verbal language is unable to undermine the defense mechanisms of the subject. The MusThePlan improves actor’s psychophysical activation, motivation, gratification, knowledge of one's own possibilities, and the quality of life. Therefore, the MusThePlan could be useful to carry out targeted interventions for the actors with characteristics of repeatability, objectivity, and predictability of results. Furthermore, it would be useful to plan a University course/master in “MusicTherapy for the Theatre”.

Keywords: musictherapy, sonorous-musical energy, quality of life, theatre

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93 Primary and Secondary Big Bangs Theory of Creation of Universe

Authors: Shyam Sunder Gupta

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For creation of Universe, theory of Big Bang , from Singularity is most acceptable theory, but has limitations as it does not answer ; how Singularity gets created and what causes Big Bang ?Further , Universe is composed of 95% Dark Energy and Dark Matter and balance 5% is visible part of Universe , but no explanation . Recently, it has been reported that there could be very large number of Universes, but only , a stipulation. This research which is based on Bhagvat Puran, a Vedic Scripture answers all questions. There is a Unique Energy Field which is eternal and infinite. The carrier Particles of Unique Energy are Paramanus; God Particles. Paramanus are Fundamental Particles and combination of these particles create bigger particles from which Universe gets created. For creation to initiate, Unique Energy gets represented in three phases; Positive Male Energy, Neutral Energy(creates Eternal Time)and Negative Female Energy. Positive Male Energy further expands in three forms of Creative Energies (CE1,CE2andCE3)and 16 principles get created, namely, Energy of Activation , Energy of Action, Energy of Darkness, Pradhan ( Equilibrium state of three energies ) , Prakriti(Non-equilibrium state of three energies, creating modes of Activation, Action and Darkness),Mahat-tattva ( consists of three modes , dominant in Mode of Darkness), Time, Energy of Consciousness, Ego Energy(consists of three modes , very strongly dominated by Mode of Darkness),Energy of Intellect, Mind Energy , Sky( creates Space and Sound Energy),Air(creates gaseous substances), Fire( creates different forms of energies like thermal, light, electrical etc.), Water( creates liquid substances)and Earth(creates solid substances). CE1 Energy creates Infinite number of Singularities from seven principles, Pradhan , Mahat-tattva, Sky , Air, Fire, Water and Earth . CE1 Energy gets divided as CE2 and enters along with other 9 principles , in each of Singularity and Primary Big Bang takes and infinite number of Universes get created. Each Universe has seven coverings of 7 principles and each layer is 10 times thicker than previous layer. By Energy CE2 , space in Universe under the coverings is divided in two parts , upper part and lower part. Upper part is occupied by Dark Energy which is created from Mode of Darkness in Ego Energy which keeps getting converted in Dark Matter and forms Invisible part of Universe. In the lower part , process of evolution gets initiated and seeds of 24 elements , Consciousness , Ego, Intellect, Mind, 5 Fundamental Elements( space, Air, Fire, Water Earth, which create non-living matter ),5 senses which receive inputs( eyes, nose, ears, tongue , skin), 5 Working Senses (Smell, Taste, Sight, Touch and Hearing);5 elements of Action( Organs of procreation , excretion, locomotion , speech and acquisition ), get created . In EC2 Energy, Singularity gets created which gets exploded by force of Energy of Action ,and Secondary Big Bang takes place and Visible Universe gets created in the shape of Bud of Flower Lotus . Within the Visible part of Universe, a small part gets created , Phenomenal Universe. Diameter of Sun and planetary system ,at the time of formation ,is 6.4 billion km, which is close to reported value . There are 5 different orbits , with reference to our Solar System. Moon around earth takes one month,, earth around sun one year, sun around Milk way one cosmic year(322.58 million years), Milky way around Universe 4.32 billion years and universe around center of universe 311.04 trillion years. Universe creation is a cyclic process with cycle time of 622.08 trillion years.In summary, Universe consists of 4 parts; covering of 7 layers, Dark Energy and Dark Matter, Visible and Phenomenal universe.

Keywords: big bang, creation, dark energy, dark matter, singularity, universe

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