Search results for: substrate specificity

Authors: Yves Mann Elate Lea Mbassi, Marie Solange Evehe Bebandoue, Wilfred Fon Mbacham

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Improving the catalytic performance of enzymes has been a long-standing theme of analytical biochemistry research. Induction of peroxidase activity by metals is a common reaction in higher plants. We thought that this increase in peroxidase activity may be due, on the one hand, to the stimulation of the gene expression of these enzymes but also to a modification of their chemical reactivity following the binding of some metal ions on their active site. We tested the effect of some metal salts (MgCl₂, MnCl₂, ZnCl₂, CaCl₂ and CuSO₄) on the activity and thermostability of peroxidase A6, a thermostable peroxidase that we discovered and purified in a previous study. The chromogenic substrate used was 3,3′,5,5′-tetramethylbenzidine. Of all the metals tested for their effect on A6, only magnesium and copper had a significant effect on the activity of the enzyme at room temperature. The Mann-Whitney test shows a slight inhibitory effect of activity by the magnesium salt (P = 0.043), while the activity of the enzyme is 5 times higher in the presence of the copper salt (P = 0.002). Moreover, the thermostability of peroxidase A6 is increased when calcium and copper salts are present. The activity in the presence of CaCl₂ is 8 times higher than the residual activity of the enzyme alone after incubation at 80°C for 10 min and 35 times higher in the presence of CuSO4 under the same conditions. In addition, manganese and zinc salts slightly reduce the thermostability of the enzyme. The activity and structural stability of peroxidase A6 can clearly be activated by Cu₂+, which therefore enhance the oxidation of 3,3′,5,5′-tetramethylbenzidine, which was used in this study as a chromogenic substrate. Ca₂+ likely has a more stabilizing function for the catalytic site.

Keywords: peroxidase activity, copper ions, calcium ions, thermostability

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Authors: Di Luo, Maria Buchberger, Anja Pickhard

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Objectives: The purpose of this study was to assess and compare the diagnostic accuracy of four common morphological approaches, including sonography, computed tomography (CT), positron emission tomography/computed tomography (PET/CT), and magnetic resonance imaging (MRI) for the evaluation of cervical lymph node metastases in head and neck squamous cell carcinoma (HNSCC) patients. Material and Methods: Included in this retrospective study were 26 patients diagnosed with HNSCC between 2010 and 2011 who all underwent sonography, CT, PET/CT, and MRI imaging before neck dissection. Morphological data were compared to the corresponding histopathological results. Statistical analysis was performed with SPSS statistic software (version 26.0), calculating sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy for detection of cervical lymph node metastases. Results: The 5-year survival rate of the patient collective was 55.5%.Risk factors for survival included initial primary tumor stage, initial lymph node stage, initial metastasis status, and therapeutic approaches. Cox regression showed initial metastasis status(HR 8.671, 95%CI 1.316-57.123, p=0.025) and therapeutic approaches(HR 6.699, 95%CI 1.746-25.700, p=0.006)to be independent predictive risk factors for survival. Sensitivity was highest for MRI (96% compared to 85% for sonography and 89% for CT and PET/CT). Specificity was comparable with 95 % for CT and 98 % for sonography and PET/CT, but only 68% for MRI. While the MRI showed the least PPV (34%) compared to all other methods (85% for sonography,75% for CT, and 86% for PET/CT), the NPV was comparable in all methods(98-99%). The overall accuracy of cervical lymph node metastases detection was comparable for sonography, CT, and PET/CT with 96%,97%,94%, respectively, while MRI had only 72% accuracy. Conclusion: Since the initial status of metastasis is an independent predictive risk factor for patients’ survival, efficient detection is crucial to plan adequate therapeutic approaches. Sonography, CT, and PET/CT have better diagnostic accuracy than MRI for the evaluation of cervical lymph node metastases in HNSCC patients.

Keywords: cervical lymph node metastases, diagnostic accuracy, head and neck squamous carcinoma, risk factors, survival

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Authors: Ahmed El-Sabbagh, Heba Mansour, Magdy El-Hedeny

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This study provided a taphonomic alteration and paleoecology of Cenomanian oysters from the Musabaa Salama area, south western Sinai, Egypt. Three oyster zones can be recognized in the studied area, a lower one of Amphidonte (Ceratostreon) flabellatum (lower-middle Cenomanian), a middle zone of Ilymatogyra (Afrogyra) africana (upper Cenomanian) and an upper one of Exogyra (Costagyra) olisiponensis (upper Cenomanian). Taphonomic features including disarticulation, fragmentation, encrustation and bioerosion were subjected to multivariate statistical analyses. The analyses showed that the distributions of the identified ichnospecies were greatly similar within the identified oyster zones in the Musabaa Salama section. With rare exceptions, Entobia cretacea, Gastrochaenolites torpedo and Maeandropolydora decipiens are considered as common to abundant ichnospecies within the three recorded oyster zones. In contrast, and with some exceptions, E. ovula, E. retiformis and Rogerella pattei are considered as frequent to common ichnospecies within the identified oyster zones. Other ichnospecies, including Caulostrepsis cretacea, G. orbicularis, Trypanites solitarius, E. geometrica and C. taeniola, are mostly recorded in rare to frequent occurrences. Careful investigation of these host shells and the preserved encrusters and/or bioerosion sculptures provided data concerning: 1) the substrate characteristics, 2) time of encrustation and bioerosion, 3) rate of sedimentation, 4) the planktonic productivity level, and 5) the general bathymetry and the rate of transgression across the substrate.

Keywords: oysters, Cenomanian, taphonomy, palaeoecology, Sinai, Egypt

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Authors: Tamar Khardziani, Violeta Berikashvili, Amrosi Chkuaseli, Vladimir Elisashvili

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The main objectives of this proposal are to develop low-cost, innovative, and competitive technologies for the production of mycelial biomass of medicinal mushrooms as a natural food supplement for poultry. To fulfill this task, industrial strains of Lentinus edodes, Ganoderma lucidum, and Pleurotus ostreatus were used in this study. The solid-state fermentation (SSF) of wheat grains, wheat bran, and soy flour was performed in flasks and bags. Among nine mushroom strains, P. ostreatus 2191 appeared to be the most productive in protein biomass accumulation in the SSF of wheat bran. All mushrooms produced exopolysaccharide with the highest yield of 5-8 mg/mL depending on fungal strain and growth substrate. Supplementation of medium with 1% glycerol and 2-4% peptone favored mushroom growth and protein accumulation. Among inorganic nitrogen sources, KNO₃ also provided high biomass and protein production. The SSF of all growth substrates was accompanied by the secretion of cellulase and xylanase activities. The highest CMCase activity (12-13 U/g) was revealed in the cultivation of P. ostreatus 2191 using wheat bran as a growth substrate and ammonium sulfate or yeast extract as a nitrogen source, whereas the highest xylanase activity was detected in the fermentation of soy flour supplemented with peptone. Acknowledgments: This work was supported by the Shota Rustaveli National Science Foundation of Georgia (Grant number STEM-22-2077).

Keywords: mushrooms, plant raw materials, fermentation, biomass protein, cellulase

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Authors: Rolivhuwa Bishop Ramagoma1*, Lynn Cairncross1, , Saartjie Roux1

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According to the world health organisation, colon cancer is among the most common cancers diagnosed in both men and women. Specifically, it is the second leading cause of cancer related deaths accounting for over 860 000 deaths worldwide in 2018. Currently, chemotherapy has become an essential component of most cancer treatments. Despite progress in cancer drug development over the previous years, traditional chemotherapeutic drugs still have low selectivity for targeting tumour tissues and are frequently constrained by dose-limiting toxicity. The creation of nanoscale delivery vehicles capable of directly directing treatment into cancer cells has recently caught the interest of researchers. Herein, the development of peptide-functionalized polyethylene glycol gold nanoparticles (Peptide-PEG-AuNPs) as a cellular probe and delivery agent is described, with the higher aim to develop a specific diagnostic prototype and assess their specificity not only against cell lines but primary human cells as well. Gold nanoparticles (AuNPs) were synthesized and stabilized through chemical conjugation. The synthesized AuNPs were characterized, stability in physiological solutions was assessed, their cytotoxicity against colon carcinoma and non-carcinoma skin fibroblasts was also studied. Furthermore, genetic effect through real-time polymerase chain reaction (RT-PCR), localization and uptake, peptide specificity were also determined. In this study, different peptide-AuNPs were found to have preferential toxicity at higher concentrations, as revealed by cell viability assays, however, all AuNPs presented immaculate stability for over 3 months following the method of synthesis. The final obtained peptide-PEG-AuNP conjugates showed good biocompatibility in the presence of high ionic solutions and biological media and good cellular uptake. Formulation of colon cancer specific targeting peptide was successful, additionally, the genes/pathways affected by the treatments were determined through RT-PCR. Primary cells study is still on going with promising results thus far.

Keywords: nanotechnology, cancer, diagnosis, therapeutics, gold nanoparticles.

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Authors: Suharni Mohamad Suharni Mohamad, Nurul Izzati Hamzan Nurul Izzati Hamzan, Norhayu Abdul Rahman Norhayu Abdul Rahman, Siti Suraiya Md Noor Siti Suraiya Md Noor

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Human papillomavirus (HPV) is an important risk factor for development of oral cancer. HPV16 is the most common type found in HPV-positive squamous cell carcinoma. In the present study, we established a real-time loop-mediated isothermal amplification (real-time LAMP) for detection of HPV16. A set of six primers was specially designed to recognize eight distinct sequences of HPV16-E6. Detection and quantification was achieved by real-time monitoring using a real-time turbidimeter based on threshold time required for turbidity in the LAMP reaction. LAMP reagents (MgSO4, dNTPs, Bst polymerase concentrations) and various incubation times and temperatures were optimized. The sensitivity was determined using 10-fold serial dilutions of HPV16 standard strain. The specificity of was evaluated using other HPV genotypes. The optimized method was established with specifically designed primers by real-time detection in approximately 30 min at 65°C. The limit of detection of HPV16 using the LAMP assay was 10 pg/ml that could be detected in 30 min. The LAMP assay was 10 times more sensitive than the conventional PCR in detecting HPV16. No cross-reactivity with other HPV genotypes was observed. This quantitative real-time LAMP assay may improve diagnostic potential for the detection and quantification of HPV16 in clinical samples and epidemiological studies due to its rapidity, simplicity, high sensitivity and specificity. This assay will be further evaluated with HPV DNAs of saliva from patients with oral squamous cell carcinoma. Acknowledgement: This study was financially supported by the ScienceFund Grant, Ministry of Science, Technology and Innovation (305/PPSG/6113219).

Keywords: Oral Squamous Cell Carcinoma (OSCC), Human Papillomavirus 16 (HPV16), Loop-Mediated Isothermal Amplification (LAMP), rapid detection

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Authors: Claudio Lamilla, Andrés Santos, Vicente Llanquinao, Jocelyn Hermosilla, Leticia Barrientos

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The industry in general is very interested in improving and optimizing industrial processes in order to reduce the costs involved in obtaining raw materials and production. Thus, an interesting and cost-effective alternative is the incorporation of bioactive metabolites in such processes, being an example of this enzymes which catalyze efficiently a large number of enzymatic reactions of industrial and biotechnological interest. In the search for new sources of these active metabolites, Antarctica is one of the least explored places on our planet where the most drastic cold conditions, salinity, UVA-UVB and liquid water available are present, features that have shaped all life in this very harsh environment, especially bacteria that live in different Antarctic ecosystems, which have had to develop different strategies to adapt to these conditions, producing unique biochemical strategies. In this work the production of cellulolytic enzymes of seven bacterial strains isolated from marine sediments at different sites in the Antarctic was evaluated. Isolation of the strains was performed using serial dilutions in the culture medium at M115°C. The identification of the strains was performed using universal primers (27F and 1492R). The enzyme activity assays were performed on R2A medium, carboxy methyl cellulose (CMC)was added as substrate. Degradation of the substrate was revealed by adding Lugol. The results show that four of the tested strains produce enzymes which degrade CMC substrate. The molecular identifications, showed that these bacteria belong to the genus Streptomyces and Pseudoalteromonas, being Streptomyces strain who showed the highest activity. Only some bacteria in marine sediments have the ability to produce these enzymes, perhaps due to their greater adaptability to degrade at temperatures bordering zero degrees Celsius, some algae that are abundant in this environment and have cellulose as the main structure. The discovery of new enzymes adapted to cold is of great industrial interest, especially for paper, textiles, detergents, biofuels, food and agriculture. These enzymes represent 8% of industrial demand worldwide and is expected to increase their demand in the coming years. Mainly in the paper and food industry are required in extraction processes starch, protein and juices, as well as the animal feed industry where treating vegetables and grains helps improve the nutritional value of the food, all this clearly puts Antarctic microorganisms and their enzymes specifically as a potential contribution to industry and the novel biotechnological applications.

Keywords: antarctic, bacteria, biotechnological, cellulolytic enzymes

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Authors: Justyna Chrzanowska, Jacek Hoffman, Dariusz Garbiec, Łukasz Kurpaska, Piotr Denis, Tomasz Moscicki, Zygmunt Szymanski

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Tungsten and boron compounds belong to the group of superhard materials and its hardness could exceed 40 GPa. In this study, the properties of the tungsten boride (WB) layers deposited in magnetron sputtering process are investigated. The sputtering process occurred from specially prepared targets that were composed of boron and tungsten mixed in molar ratio of 2.5 or 4.5 and sintered in spark plasma sintering process. WB layers were deposited on silicon (100) and stainless steel 304 substrates at room temperature (RT) or in 570 °C. Layers deposited in RT and in elevated temperature varied considerably. Layers deposited in RT are amorphous and have low adhesion. In contrast, the layers deposited in 570 °C are crystalline and have good adhesion. All deposited layers have a hardness about 40 GPa. Moreover, the friction coefficient of crystalline layers is 0.22 and wear rate is about 0.67•10-6 mm3N-1m-1. After material characterization the WB layers were annealed in argon atmosphere in 1000 °C for 1 hour. On the basis of X-Ray Diffraction analysis, it has been noted that the crystalline layers are thermally stable and do not change their phase composition, whereas the amorphous layers change their phase composition. Moreover, after annealing, on the surface of WB layers some cracks were observed. It is probably connected with the differences of the thermal expansion between the layer and the substrate. Despite of the presence of cracks, the wear resistance of annealed layers is still higher than the wear resistance of uncoated substrate. The analysis of the structure and properties of tungsten boride layers lead to the discussion about the application area of this material.

Keywords: hard coatings, hard materials, magnetron sputtering, mechanical properties, tungsten boride

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Authors: Sonu Khanal, Mandira Shrestha

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Snow and Glaciers are the largest dependable reserved sources of water for the river system originating from the Himalayas so an accurate estimate of the volume of water contained in the snowpack and the rate of release of water from snow and glaciers are, therefore, needed for efficient management of the water resources. This research assess the fusion of energy exchanges between the snowpack, air above and soil below according to mass and energy balance which makes it apposite than the models using simple temperature index for the snow and glacier melt computation. UEBGrid a Distributed energy based model is used to calculate the melt which is then routed by Geo-SFM. The model robustness is maintained by incorporating the albedo generated from the Landsat-7 ETM images on a seasonal basis for the year 2002-2003 and substrate map derived from TM. The Substrate file includes predominantly the 4 major thematic layers viz Snow, clean ice, Glaciers and Barren land. This approach makes use of CPC RFE-2 and MERRA gridded data sets as the source of precipitation and climatic variables. The subsequent model run for the year between 2002-2008 shows a total annual melt of 17.15 meter is generate from the Marshyangdi Basin of which 71% is contributed by the glaciers , 18% by the rain and rest being from the snow melt. The albedo file is decisive in governing the melt dynamics as 30% increase in the generated surface albedo results in the 10% decrease in the simulated discharge. The melt routed with the land cover and soil variables using Geo-SFM shows Nash-Sutcliffe Efficiency of 0.60 with observed discharge for the study period.

Keywords: Glacier, Glacier melt, Snowmelt, Energy balance

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Authors: T. N. Tran Thi, J. Morse, C. Detlefs, P. K. Cook, C. Yıldırım, A. C. Jakobsen, T. Zhou, J. Hartwig, V. Zurbig, D. Caliste, B. Fernandez, D. Eon, O. Loto, M. L. Hicks, A. Pakpour-Tabrizi, J. Baruchel

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The ability to grow boron-doped diamond epilayers of high crystalline quality is a prerequisite for the fabrication of diamond power electronic devices, in particular high voltage diodes and metal-oxide-semiconductor (MOS) transistors. Boron and intrinsic diamond layers are homoepitaxially overgrown by microwave assisted chemical vapour deposition (MWCVD) on single crystal high pressure, high temperature (HPHT) grown bulk diamond substrates. Various epilayer thicknesses were grown, with dopant concentrations ranging from 1021 atom/cm³ at nanometer thickness in the case of 'delta doping', up 1016 atom/cm³ and 50µm thickness or high electric field drift regions. The crystalline quality of these overgrown layers as regards defects, strain, distortion… is critical for the device performance through its relation to the final electrical properties (Hall mobility, breakdown voltage...). In addition to the optimization of the epilayer growth conditions in the MWCVD reactor, other important questions related to the crystalline quality of the overgrown layer(s) are: 1) what is the dependence on the bulk quality and surface preparation methods of the HPHT diamond substrate? 2) how do defects already present in the substrate crystal propagate into the overgrown layer; 3) what types of new defects are created during overgrowth, what are their growth mechanisms, and how can these defects be avoided? 4) how can we relate in a quantitative manner parameters related to the measured crystalline quality of the boron doped layer to the electronic properties of final processed devices? We describe synchrotron-based techniques developed to address these questions. These techniques allow the visualization of local defects and crystal distortion which complements the data obtained by other well-established analysis methods such as AFM, SIMS, Hall conductivity…. We have used Grazing Incidence X-ray Diffraction (GIXRD) at the ID01 beamline of the ESRF to study lattice parameters and damage (strain, tilt and mosaic spread) both in diamond substrate near surface layers and in thick (10–50 µm) overgrown boron doped diamond epi-layers. Micro- and nano-section topography have been carried out at both the BM05 and ID06-ESRF) beamlines using rocking curve imaging techniques to study defects which have propagated from the substrate into the overgrown layer(s) and their influence on final electronic device performance. These studies were performed using various commercially sourced HPHT grown diamond substrates, with the MWCVD overgrowth carried out at the Fraunhofer IAF-Germany. The synchrotron results are in good agreement with low-temperature (5°K) cathodoluminescence spectroscopy carried out on the grown samples using an Inspect F5O FESEM fitted with an IHR spectrometer.

Keywords: synchrotron X-ray diffaction, crystalline quality, defects, diamond overgrowth, rocking curve imaging

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Authors: Sanja Jovanovic, Aleksandar Simic, Ognjan Skrobic, Dragan Masulovic, Aleksandra Djuric-Stefanovic

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Introduction: Achalasia is an idiopathic primary esophageal motility disorder characterized by esophageal peristalsis and impaired swallow-induced relaxation of the lower esophageal sphincter (LES). It is a rare disease that affects both genders with an incidence of 1/100.000 and a prevalence rate of 10/100,000 per year. Objective: Laparoscopic Heller myotomy (LHM) represents a therapy of choice for patients with achalasia, providing excellent outcomes. The aim of this study was to evaluate the significance of computed tomography (CT) in analyzing achalasia subtypes and timed barium esophagogram (TBE) in evaluation of LHM success, as a part of standardized diagnostic protocol. Method: Fifty-one patients with achalasia, confirmed by manometric studies, in addition to standardized diagnostic methods, underwent CT and TBE. CT was done with multiplanar reconstruction, measuring the wall thickness above the esophago-gastric junction in the axial plane. TBE was performed preoperatively and two days postoperatively swallowing low-density barium sulfate, and plane upright frontal films were performed 1, 2 and 5 minutes after the ingestion. In all patients, LHM was done, and pre and postoperative height and weight of the barium column were compared. Results: According to CT findings we divided patients into 3 subtypes of achalasia according to wall thickness: < 4mm as subtype one, between 4 - 9mm as II, and > 10 mm as subtype 3. Correlation of manometric results, as a reference values, and CT findings indicated CT sensitivity of 90% and specificity of 70 % in establishing subtypes of achalasia. The preoperative values of TBE at 1, 2 and 5 minutes were: median barium column height 17.4 ± 7.4, 15.9 ± 6.2 and 13.9 ± 6.2 cm; median column width 5 ± 1.5, 4.7 ± 1.6 and 4.5 ± 1.8 cm respectively. LHM significantly reduced these values (height 7 ± 4.6, 5.8 ± 4.2, 3.7 ± 3.4 cm; width 2.9 ± 1.3, 2.6 ± 1.3 and 2.4 ± 1.4 cm), indicating the quantitative estimates of emptying as excellent (p value < 0.01). Conclusion: CT has high sensitivity and specificity in evaluation of achalasia subtypes, and can be introduced as an additional method for standardized evaluation of these patients. The quantitative assessment of TBE based on measurements of the barium column is an accurate and beneficial method, which adequately estimates esophageal emptying success of LHM.

Keywords: achalasia, computed tomography, esophagography, myotomy

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Authors: A. Sopharak, B. Uyyanonvara, S. Barman

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Diabetic retinopathy is characterized by the development of retinal microaneurysms. The damage can be prevented if disease is treated in its early stages. In this paper, we are comparing Support Vector Machine (SVM) and Naïve Bayes (NB) classifiers for automatic microaneurysm detection in images acquired through non-dilated pupils. The Nearest Neighbor classifier is used as a baseline for comparison. Detected microaneurysms are validated with expert ophthalmologists’ hand-drawn ground-truths. The sensitivity, specificity, precision and accuracy of each method are also compared.

Keywords: diabetic retinopathy, microaneurysm, naive Bayes classifier, SVM classifier

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Authors: Naridchaya Aberdour, Joanna Kao, Anne Miller, Timothy Shore, Richard Maher, Zhixin Liu

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Background: COVID-19 has and continues to be a strain on healthcare globally, with the number of patients requiring hospitalization exceeding the level of medical support available in many countries. As chest x-rays are the primary respiratory radiological investigation, the Radiological Assessment of Lung Edema (RALE) score was used to quantify the extent of pulmonary infection on baseline imaging. Assessment of RALE score's reproducibility and associations with clinical outcome parameters were then evaluated to determine implications for patient management and prognosis. Methods: A retrospective study was performed with the inclusion of patients testing positive for COVID-19 on nasopharyngeal swab within a single Local Health District in Sydney, Australia and baseline x-ray imaging acquired between January to June 2020. Two independent Radiologists viewed the studies and calculated the RALE scores. Clinical outcome parameters were collected and statistical analysis was performed to assess RALE score reproducibility and possible associations with clinical outcomes. Results: A total of 78 patients met inclusion criteria with the age range of 4 to 91 years old. RALE score concordance between the two independent Radiologists was excellent (interclass correlation coefficient = 0.93, 95% CI = 0.88-0.95, p<0.005). Binomial logistics regression identified a positive correlation with hospital admission (1.87 OR, 95% CI= 1.3-2.6, p<0.005), oxygen requirement (1.48 OR, 95% CI= 1.2-1.8, p<0.005) and invasive ventilation (1.2 OR, 95% CI= 1.0-1.3, p<0.005) for each 1-point increase in RALE score. For each one year increased in age, there was a negative correlation with recovery (0.05 OR, 95% CI= 0.92-1.0, p<0.01). RALE scores above three were positively associated with hospitalization (Youden Index 0.61, sensitivity 0.73, specificity 0.89) and above six were positively associated with ICU admission (Youden Index 0.67, sensitivity 0.91, specificity 0.78). Conclusion: The RALE score can be used as a surrogate to quantify the extent of COVID-19 infection and has an excellent inter-observer agreement. The RALE score could be used to prognosticate and identify patients at high risk of deterioration. Threshold values may also be applied to predict the likelihood of hospital and ICU admission.

Keywords: chest radiography, coronavirus, COVID-19, RALE score

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Authors: Yingwei Hou, Tao Liu, Yong Pang

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The paper explored the three-dimensional deformation of mussel plaques anchor to wet polydimethylsiloxane (PDMS) substrates under tension stress with different angles. Mussel plaques exhibiting natural adhesive structures, have attracted significant attention for their remarkable adhesion properties. Understanding their behavior under mechanical stress, particularly in a three-dimensional context, holds immense relevance for biomimetic material design and bio-inspired adhesive development. This study employed a novel approach to investigate the 3D deformation of the PDMS substrates anchored by mussel plaques subjected to controlled tension. Utilizing our customized stereo digital image correlation technique and mechanical mechanics analyses, we found the distributions of the displacement and resultant force on the substrate became concentrated under the plaque. Adhesion and sucking mechanisms were analyzed for the mussel plaque-substrate system under tension until detachment. The experimental findings were compared with a developed model using finite element analysis and the results provide new insights into mussels’ attachment mechanism. This research not only contributes to the fundamental understanding of biological adhesion but also holds promising implications for the design of innovative adhesive materials with applications in fields such as medical adhesives, underwater technologies, and industrial bonding. The comprehensive exploration of mussel plaque behavior in three dimensions is important for advancements in biomimicry and materials science, fostering the development of adhesives that emulate nature's efficiency.

Keywords: adhesion mechanism, mytilus edulis, mussel plaque, stereo digital image correlation

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Authors: Jyosthna Khanna Goli, Shaik Naseeruddin, Hameeda Bee

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Employing lignocellulosic biomass in fermentative production of xylitol and bioethanol is gaining interest as it is renewable, cheap, and abundantly available. Xylitol is a polyol, gaining its importance in the food and pharmacological industry due to its low calorific value and anti-cariogenic nature. Bioethanol from lignocellulosic biomass is widely accepted as an alternative fuel for transportation with reduced CO₂ emissions, thus reducing the greenhouse effect. Typha latifolia, an aquatic weed, was found to be promising lignocellulosic substrate as it posses a high amount of sugars and does not compete with arable lands and interfere with food and feed competition. In the present study, xylose from hemicellulosic fraction of typha is converted to xylitol by isolate Jfh5 (Candida. tropicalis) and cellulose part to ethanol using Saccharomyces cerevisiaeVS3. Initially, alkali pretreatment of typha using sodium hydroxide, potassium hydroxide, ammonium hydroxide, calcium hydroxide, sodium bisulphate and sodium dithionate for overnight (18h) at room temperature (28 ± 2°C), resulted in maximum delignification of 75% with 2% (v/v) sodium bisulphate. Later, pretreated biomass was subjected to acid hydrolysis with 1%, 1.5%, 2%, and 3% H₂SO₄ at 110 °C and 121°C for 30 and 60 min, respectively. 2% H₂SO₄ at 121°C for 60 min was found to release 13.5 g /l sugars, which on detoxification and fermentation produced 8.1g/l xylitol with yield and productivity of 0.65g/g and 0.112g/l/h respectively. Further enzymatic hydrolysis of the residual substrate obtained after acid hydrolysis released 11g/l sugar, which on fermentation with VS3 produced 4.9g/l ethanol with yield and productivity of 0.22g/g and 0.136g/l/h respectively.

Keywords: delignification, xylitol, bioethanol, acid hydrolysis, enzyme hydrolysis

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Authors: Qiao Pei Wen, Ng Seng Lee, Sae Tae Veera, Chiu Ah Fong, Loke Weng Onn

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Thin film thickness uniformity is crucial to get consistent film etch rate and device yield across the wafer. In the capacitive-coupled parallel plate PECVD system; the film thickness uniformity can be affected by many factors such as the heater temperature uniformity, the spacing between top and bottom electrode, RF power, pressure, gas flows and etc. In this paper, we studied how the PECVD SiN film thickness uniformity is affected by the substrate electrical conductivity and the RF power coupling efficiency. PECVD SiN film was deposited on 150-mm sapphire wafers in 200-mm Lam Sequel tool, fork marks were observed on the wafers. On the fork marks area SiN film thickness is thinner than that on the non-fork area. The forks are the wafer handler inside the process chamber to move the wafers from one station to another. The sapphire wafers and the ceramic forks both are insulator. The high resistivity of the sapphire wafers and the forks inhibits the RF power coupling efficiency during PECVD deposition, thereby reducing the deposition rate. Comparing between the high frequency and low frequency RF power (HFRF and LFRF respectively), the LFRF power coupling effect on the sapphire wafers is more dominant than the HFRF power on the film thickness. This paper demonstrated that the SiN thickness uniformity on sapphire wafers can be improved by depositing a thin TiW layer on the wafer before the SiN deposition. The TiW layer can be on the wafer surface, bottom or any layer before SiN deposition.

Keywords: PECVD SiN deposition, sapphire wafer, substrate electrical conductivity, RF power coupling, high frequency RF power, low frequency RF power, film deposition rate, thickness uniformity

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Authors: Mirosław Krzemieniewski, Marcin Zieliński, Marcin Dębowski

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Electrokinetic disintegration is one of the high-voltage electric methods. The design of systems is exceptionally simple. Biomass flows through a system of pipes with alongside mounted electrodes that generate an electric field. Discharges in the electric field deform cell walls and lead to their successive perforation, thereby making their contents easily available to bacteria. The spark-over occurs between electrode surface and pipe jacket which is the second pole and closes the circuit. The value of voltage ranges from 10 to 100kV. Electrodes are supplied by normal “power grid” monophase electric current (230V, 50Hz). Next, the electric current changes into direct current of 24V in modules serving for particular electrodes, and this current directly feeds the electrodes. The installation is completely safe because the value of generated current does not exceed 250mA and because conductors are grounded. Therefore, there is no risk of electric shock posed to the personnel, even in the case of failure or incorrect connection. Low values of the electric current mean small energy consumption by the electrode which is extremely low – only 35W per electrode – compared to other methods of disintegration. Pipes with electrodes with diameter of DN150 are made of acid-proof steel and connected from both sides with 90º elbows ended with flanges. The available S and U types of pipes enable very convenient fitting with system construction in the existing installations and rooms or facilitate space management in new applications. The system of pipes for electrokinetic disintegration may be installed horizontally, vertically, askew, on special stands or also directly on the wall of a room. The number of pipes and electrodes is determined by operating conditions as well as the quantity of substrate, type of biomass, content of dry matter, method of disintegration (single or circulatory), mounting site etc. The most effective method involves pre-treatment of substrate that may be pumped through the disintegration system on the way to the fermentation tank or recirculated in a buffered intermediate tank (substrate mixing tank). Biomass structure destruction in the process of electrokinetic disintegration causes shortening of substrate retention time in the tank and acceleration of biogas production. A significant intensification of the fermentation process was observed in the systems operating in the technical scale, with the greatest increase in biogas production reaching 18%. The secondary, but highly significant for the energetic balance, effect is a tangible decrease of energy input by agitators in tanks. It is due to reduced viscosity of the biomass after disintegration, and may result in energy savings reaching even 20-30% of the earlier noted consumption. Other observed phenomena include reduction in the layer of surface scum, reduced sewage capability for foaming and successive decrease in the quantity of bottom sludge banks. Considering the above, the system for electrokinetic disintegration seems a very interesting and valuable solutions meeting the offer of specialist equipment for the processing of plant biomass, including Virginia fanpetals, before the process of methane fermentation.

Keywords: electrokinetic disintegration, biomass, biogas production, fermentation, Virginia fanpetals

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Authors: L. Mentar, O. Baka, M. R. Khelladi, A. Azizi

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Zinc oxide (ZnO), as a transparent semiconductor with a wide band gap of 3.4 eV and a large exciton binding energy of 60 meV at room temperature, is one of the most promising materials for a wide range of modern applications. With the development of film growth technologies and intense recent interest in nanotechnology, several varieties of ZnO nanostructured materials have been synthesized almost exclusively by thermal evaporation methods, particularly chemical vapor deposition (CVD), which generally require a high growth temperature above 550 °C. In contrast, wet chemistry techniques such as hydrothermal synthesis and electro-deposition are promising alternatives to synthesize ZnO nanostructures, especially at a significantly lower temperature (below 200°C). In this study, the electro-deposition method was used to produce zinc oxide (ZnO) nanostructures on fluorine-doped tin oxide (FTO)-coated conducting glass substrate from chloride bath. We present the influence of KCl concentrations on the electro-deposition process, morphological, structural and optical properties of ZnO nanostructures. The potentials of electro-deposition of ZnO were determined using the cyclic voltammetry. From the Mott-Schottky measurements, the flat-band potential and the donor density for the ZnO nanostructure are determined. Field emission scanning electron microscopy (FESEM) images showed different sizes and morphologies of the nanostructures which depends on the concentrations of Cl-. Very netted hexagonal grains are observed for the nanostructures deposited at 0.1M of KCl. X-ray diffraction (XRD) study confirms the Wurtzite phase of the ZnO nanostructures with a preferred oriented along (002) plane normal to the substrate surface. UV-Visible spectra showed a significant optical transmission (~80%), which decreased with low Cl-1 concentrations. The energy band gap values have been estimated to be between 3.52 and 3.80 eV.

Keywords: Cl-, electro-deposition, FESEM, Mott-Schottky, XRD, ZnO

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Authors: Fayad Ghawbar

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The significance of the Multiple Input Multiple Output (MIMO) system in the 5G wireless communication system is essential to enhance channel capacity and provide a high data rate resulting in a need for dual-polarization in vertical and horizontal. Furthermore, size reduction is critical in a MIMO system to deploy more antenna elements requiring a compact, low-profile design. A compact dual-band 4-MIMO antenna system has been presented in this paper with pattern and polarization diversity. The proposed single antenna structure has been designed using two antenna layers with a C shape in the front layer and a partial slot with a U-shaped cut in the ground to enhance isolation. The single antenna is printed on an FR4 dielectric substrate with an overall size of 18 mm×18 mm×1.6 mm. The 4-MIMO antenna elements were printed orthogonally on an FR4 substrate with a size dimension of 36 × 36 × 1.6 mm3 with zero edge-to-edge separation distance. The proposed compact 4-MIMO antenna elements resonate at 3.4-3.6 GHz and 4.8-5 GHz. The s-parameters measurement and simulation results agree, especially in the lower band with a slight frequency shift of the measurement results at the upper band due to fabrication imperfection. The proposed design shows isolation above -15 dB and -22 dB across the 4-MIMO elements. The MIMO diversity performance has been evaluated in terms of efficiency, ECC, DG, TARC, and CCL. The total and radiation efficiency were above 50 % across all parameters in both frequency bands. The ECC values were lower than 0.10, and the DG results were about 9.95 dB in all antenna elements. TARC results exhibited values lower than 0 dB with values lower than -25 dB in all MIMO elements at the dual-bands. Moreover, the channel capacity losses in the MIMO system were depicted using CCL with values lower than 0.4 Bits/s/Hz.

Keywords: compact antennas, MIMO antenna system, 5G communication, dual band, ECC, DG, TARC

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Authors: Chitralekha Ngangbam, Aniruddha Mondal, Naorem Khelchand Singh

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Aluminium (Al) doped In2O3 (Indium Oxide) nanocolumn array was synthesized by glancing angle deposition (GLAD) technique on Si (n-type) substrate for photodetector application. The sample was characterized by scanning electron microscopy (SEM). The average diameter of the nanocolumn was calculated from the top view of the SEM image and found to be ∼80 nm. The length of the nanocolumn (~500 nm) was calculated from cross sectional SEM image and it shows that the nanocolumns are perpendicular to the substrate. The EDX analysis confirmed the presence of Al (Aluminium), In (Indium), O (Oxygen) elements in the samples. The XRD patterns of the Al-doped In2O3 nanocolumn show the presence of different phases of the Al doped In2O3 nanocolumn i.e. (222) and (622). Three different peaks were observed from the PL analysis of Al doped In2O3 nanocolumn at 365 nm, 415 nm and 435 nm respectively. The peak at PL emission at 365 nm can be attributed to the near band gap transition of In2O3 whereas the peaks at 415 nm and 435 nm can be attributed to the trap state emissions due to oxygen vacancies and oxygen–indium vacancy centre in Al doped In2O3 nanocolumn. The current-voltage (I–V) characteristics of the Al doped In2O3 nanocolumn based detector was measured through the Au Schottky contact. The devices were then examined under the halogen light (20 W) illumination for photocurrent measurement. The Al-doped In2O3 nanocolumn based optical detector showed high conductivity and low turn on voltage at 0.69 V under white light illumination. A maximum photoresponsivity of 82 A/W at 380 nm was observed for the device. The device shows a high internal gain of ~267 at UV region (380 nm) and ∼127 at visible region (760 nm). Also the rise time and fall time for the device at 650 nm is 0.15 and 0.16 sec respectively which makes it suitable for fast response detector.

Keywords: glancing angle deposition, nanocolumn, semiconductor, photodetector, indium oxide

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Authors: Wen-Jay Lee, Kuo-Ning Chiang

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The graphene binding with silicon substrate has apparently Schottky barriers property, which can be used in the application of solar cell and light source. Because graphene has only one atom layer, the atomistic structure of graphene binding with the silicon surface plays an important role to affect the properties of graphene. In this work, temperature effect on the morphology of graphene sheet attached on different crystal planes of silicon substrates are investigated by Molecular dynamics (MD) (LAMMPS, developed by Sandia National Laboratories). The results show that the covered graphene sheet would cause the structural deformation of the surface Si atoms of stubtrate. To achieve a stable state in the binding process, the surface Si atoms would adjust their position and fit the honeycomb structure of graphene after the graphene attaches to the Si surface. The height contour of graphene on different plane of silicon surfaces presents different pattern, leading the local residual stress at the interface. Due to the high density of dangling bond on the Si (111)7x7 surface, the surface of Si(111)7x7 is not matching with the graphene so well in contrast with Si(100)2x1and Si(111)2x1. Si(111)7x7 is found that only partial silicon adatoms are rearranged on surface after the attachment when the temperature is lower than 200K, As the temperature gradually increases, the deformation of surface structure becomes significant, as well as the residue stress. With increasing temperature till the 815K, the graphene sheet begins to destroy and mixes with the silicon atoms. For the Si(100)2x1 and Si(111)2x1, the silicon surface structure keep its structural arrangement with a higher temperature. With increasing temperature, the residual stress gradually decrease till a critical temperatures. When the temperature is higher than the critical temperature, the residual stress gradually increases and the structural deformation is found on the surface of the Si substrates.

Keywords: molecular dynamics, graphene, silicon, Schottky barriers, interface

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Authors: Y. Saylan, F. Yılmaz, A. Denizli

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Rheumatoid arthritis (RA) which is the most common autoimmune disorder of the body's own immune system attacking healthy cells. RA has both articular and systemic effects.Until now romatiod factor (RF) assay is used the most commonly diagnosed RA but it is not specific. Anti-cyclic citrullinated peptide (anti-CCP) antibodies are IgG autoantibodies which recognize citrullinated peptides and offer improved specificity in early diagnosis of RA compared to RF. Anti-CCP antibodies have specificity for the diagnosis of RA from 91 to 98% and the sensitivity rate of 41-68%. Molecularly imprinted polymers (MIP) are materials that are easy to prepare, less expensive, stable have a talent for molecular recognition and also can be manufactured in large quantities with good reproducibility. Molecular recognition-based adsorption techniques have received much attention in several fields because of their high selectivity for target molecules. Quartz crystal microbalance (QCM) is an effective, simple, inexpensive approach mass changes that can be converted into an electrical signal. The applications for specific determination of chemical substances or biomolecules, crystal electrodes, cover by the thin films for bind or adsorption of molecules. In this study, we have focused our attention on combining of molecular imprinting into nanofilms and QCM nanosensor approaches and producing QCM nanosensor for anti-CCP, chosen as a model protein, using anti-CCP imprinted nanofilms. For this aim, anti-CCP imprinted QCM nanosensor was characterized by Fourier transform infrared spectroscopy, atomic force microscopy, contact angle measurements and ellipsometry. The non-imprinted nanosensor was also prepared to evaluate the selectivity of the imprinted nanosensor. Anti-CCP imprinted QCM nanosensor was tested for real-time detection of anti-CCP from aqueous solution. The kinetic and affinity studies were determined by using anti-CCP solutions with different concentrations. The responses related with mass shifts (Δm) and frequency shifts (Δf) were used to evaluate adsorption properties and to calculate binding (Ka) and dissociation (Kd) constants. To show the selectivity of the anti-CCP imprinted QCM nanosensor, competitive adsorption of anti-CCP and IgM was investigated.The results indicate that anti-CCP imprinted QCM nanosensor has a higher adsorption capabilities for anti-CCP than for IgM, due to selective cavities in the polymer structure.

Keywords: anti-CCP, molecular imprinting, nanosensor, rheumatoid arthritis, QCM

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Authors: A.Boularouk, F. Chouikh, M. Lamri, H. Moualkia, Y. Bouznit

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The principal aim of this study is to considerably reduce the resistivity of the SnO2 thin layers. In this order, we have doped tin oxide with aluminum and antimony incorporation with different atomic percentages (0 and 4%). All the pure and doped SnO2 films were grown by simple, flexible and cost-effective Automatic Spray Pyrolysis Method (ASPM) on glass substrates at a temperature of 350 °C. The microstructural, optical, morphological and electrical properties of the films have been studied. The XRD results demonstrate that all films have polycrystalline nature with a tetragonal rutile structure and exhibit the (200) preferential orientation. It has been observed that all the dopants are soluble in the SnO2 matrix without forming secondary phases. However, dopant introduction does not modify the film growth orientation. The crystallite size of the pure SnO2 film is about 36 nm. The films are highly transparent in the visible region with an average transmittance reaching up to 80% and it slightly reduces with increasing doping concentration (Al and Sb). The optical band gap value was evaluated between 3.60 eV and 3.75 eV as a function of doping. The SEM image reveals that all films are nanostructured, densely continuous, with good adhesion to the substrate. We note again that the surface morphology change with the type and concentration dopant. The minimum resistivity is 0.689*10-4, which is observed for SnO2 film doped 4% Al. This film shows better properties and is considered the best among all films. Finally, we concluded that the physical properties of the pure and doped SnO2 films grown on a glass substrate by ASPM strongly depend on the type and concentration dopant (Al and Sb) and have highly desirable optical and electrical properties and are promising materials for several applications.

Keywords: tin oxide, automatic spray, Al and Sb doped, transmittance, MEB, XRD and UV-VIS

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Authors: Melanie Macgregor-Ramiasa, Isabel Hopp, Patricia Murray, Krasimir Vasilev

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Stem cells based therapies are one of the greatest promises of new-age medicine due to their potential to help curing most dreaded conditions such as cancer, diabetes and even auto-immune disease. However, establishing suitable in vitro culture materials allowing to control the fate of stem cells remain a challenge. Amongst the factor influencing stem cell behavior, substrate chemistry and nanotopogaphy are particularly critical. In this work, we used plasma assisted surface modification methods to produce model substrates with tailored nanotopography and controlled chemistry. Three different sizes of gold nanoparticles were bound to amine rich plasma polymer layers to produce homogeneous and gradient surface nanotopographies. The outer chemistry of the substrate was kept constant for all substrates by depositing a thin layer of our patented biocompatible polyoxazoline plasma polymer on top of the nanofeatures. For the first time, protein adsorption and stem cell behaviour (mouse kidney stem cells and mesenchymal stem cells) were evaluated on nanorough plasma deposited polyoxazoline thin films. Compared to other nitrogen rich coatings, polyoxazoline plasma polymer supports the covalent binding of proteins. Moderate surface nanoroughness, in both size and density, triggers cell proliferation. In association with polyoxazoline coating, cell proliferation is further enhanced on nanorough substrates. Results are discussed in term of substrates wetting properties. These findings provide valuable insights on the mechanisms governing the interactions between stem cells and their growth support.

Keywords: nanotopography, stem cells, differentiation, plasma polymer, oxazoline, gold nanoparticles

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Authors: Steven Arcidiacono, Robert Stote, Erin Anderson, Molly Richards

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There are numerous standard test methods for antimicrobial textiles that measure activity against specific microorganisms. However, many times these results do not translate to the performance of treated textiles when worn by individuals. Standard test methods apply a single target organism grown under optimal conditions to a textile, then recover the organism to quantitate and determine activity; this does not reflect the actual performance environment that consists of polymicrobial communities in less than optimal conditions or interaction of the textile with the skin substrate. Here we propose the development of in vitro skin model method to bridge the gap between lab testing and wear studies. The model will consist of a defined polymicrobial community of 5-7 commensal microbes simulating the skin microbiome, seeded onto a solid tissue platform to represent the skin. The protocol would entail adding a non-commensal test organism of interest to the defined community and applying a textile sample to the solid substrate. Following incubation, the textile would be removed and the organisms recovered, which would then be quantitated to determine antimicrobial activity. Important parameters to consider include identification and assembly of the defined polymicrobial community, growth conditions to allow the establishment of a stable community, and choice of skin surrogate. This model could answer the following questions: 1) is the treated textile effective against the target organism? 2) How is the defined community affected? And 3) does the textile cause unwanted effects toward the skin simulant? The proposed model would determine activity under conditions comparable to the intended application and provide expanded knowledge relative to current test methods.

Keywords: antimicrobial textiles, defined polymicrobial community, in vitro skin model, skin microbiome

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Authors: Aneesh Joshi, Sagil James

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Cold Spray (CS) process is deposition of solid particles over a substrate above a certain critical impact velocity. Unlike thermal spray processes, CS process does not melt the particles thus retaining their original physical and chemical properties. These characteristics make CS process ideal for various engineering applications involving metals, polymers, ceramics and composites. The bonding mechanism involved in CS process is extremely complex considering the dynamic nature of the process. Though CS process offers great promise for several engineering applications, the realization of its full potential is limited by the lack of understanding of the complex mechanisms involved in this process and the effect of critical process parameters on the deposition efficiency. The goal of this research is to understand the complex nanoscale mechanisms involved in CS process. The study uses Molecular Dynamics (MD) simulation technique to understand the material deposition phenomenon during the CS process. Impact of a single crystalline copper nanoparticle on copper substrate is modelled under varying process conditions. The quantitative results of the impacts at different velocities, impact angle and size of the particles are evaluated using flattening ratio, von Mises stress distribution and local shear strain. The study finds that the flattening ratio and hence the quality of deposition was highest for an impact velocity of 700 m/s, particle size of 20 Å and an impact angle of 90°. The stress and strain analysis revealed regions of shear instabilities in the periphery of impact and also revealed plastic deformation of the particles after the impact. The results of this study can be used to augment our existing knowledge in the field of CS processes.

Keywords: cold spray process, molecular dynamics simulation, nanoparticles, particle impact

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Authors: Ghebremedhin Tsegay, Weldu Tesfagaber, Yuanmao Zhu, Xijun He, Wan Wang, Zhenjiang Zhang, Encheng Sun, Jinya Zhang, Yuntao Guan, Fang Li, Renqiang Liu, Zhigao Bu, Dongming Zhao*

Abstract:

African swine fever (ASF) is a highly infectious viral disease of pigs, resulting in significant economic loss worldwide. As there is no approved vaccines and treatments, the control of ASF entirely depends on early diagnosis and culling of infected pigs. Thus, highly specific and sensitive diagnostic assays are required for accurate and early diagnosis of ASF virus (ASFV). Currently, only a few recombinant proteins have been tested and validated for use as reagents in ASF diagnostic assays. The most promising ones for ASFV antibody detection were p72, p30, p54, and pp62. So far, three ELISA kits based on these recombinant proteins have been commercialized. Due to the complex nature of the virus and variety forms of the disease, robust serodiagnostic assays are still required. ASFV p22 protein, encoded by KP177R gene, is located in the inner membrane of viral particle and appeared transiently in the plasma membrane early after virus infection. The p22 protein interacts with numerous cellular proteins, involved in processes of phagocytosis and endocytosis through different cellular pathways. However, p22 does not seem to be involved in virus replication or swine pathogenicity. In this study, E.coli expressed recombinant p22 protein was used to generate a monoclonal antibody (mAb), and its potential use for the development of blocking ELISA (bELISA) was evaluated. A total of 806 pig serum samples were tested to evaluate the bELISA. Acording the ROC (Reciever operating chracteristic) analysis, 100% sensitivity and 98.10% of specificity was recorded when the PI cut-off value was set at 47%. The novel assay was able to detect the antibodies as early as 9 days post infection. Finaly, a highly sensitive, specific and rapid novel p22-mAb based bELISA assay was developed, and optimized for detection of antibodies against genotype I and II ASFVs. It is a promising candidate for an early and acurate detection of the antibodies and is highly expected to have a valuable role in the containment and prevention of ASF.

Keywords: ASFV, blocking ELISA, diagnosis, monoclonal antibodies, sensitivity, specificity

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Authors: Hanjun Yun, Gun Bae, Nabin Paul, Cheolhee Moon

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Flexible OLED is an important technology for the next generation display over various kinds of applications. However, the organic materials of OLEDs degrade rapidly under the invasion of oxygen and water moisture. The degradation causes the formation of non-emitting areas which gradually suppress the device brightness, ultimately the lifetime of the device decreasing rapidly. Until now, the most suitable sealing process of the flexible OLED devices is a thin film encapsulation (TFE). However, TFE consists of a multilayer thin-film structure with organic-inorganic materials, so the cost is expensive and the process time is long. Another problem is that the blocking characteristics from the moisture and oxygen are not perfect. Therefore, the encapsulation of the flexible OLED device is a still key technical issue for the successful market entry. In this study, we are to introduce an auxiliary sealing line between the two flexible substrates. The electrode lines were formed on the substrates which have a SiNx barrier coating layer. To induce the solid phase diffusion process between the SiNx layer and the electrode lines, the electrode materials were determined as Al-Si composition. Thermal energy was supplied for both the SiNx layer and Al-Si electrode lines within the furnace to induce the interfacial bonding through the solid phase diffusion of Si. We printed a test pattern for the edge of the flexible PET substrate of 3cm*3cm size. Experimental conditions such as heating temperature, heating time were optimized to get enough adhesion strength which was estimated through the competitive bending test. Finally, OLED devices with flexible PET substrate of 3cm*3cm size were manufactured to investigate the blocking characteristics as an encapsulation layer.

Keywords: barrier, encapsulation, OLED, solid phase diffusion

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Authors: Lay Poh Tan, Chor Yong Tay, Haiyang Yu

Abstract:

Micro-contact printing is a form of soft lithography that uses the relief patterns on a master polydimethylsiloxane (PDMS) stamp to form patterns of self-assembled monolayers (SAMs) of ink on the surface of a substrate through conformal contact technique. Here, we adopt this method to print proteins of different dimensions on our biodegradable polymer substrates. We started off with printing 20-500 μm scale lanes of fibronectin to engineer the shape of bone marrow derived human mesenchymal stem cell (hMSCs). After 8 hours of culture, the hMSCs adopted elongated shapes, and upon analysis of the gene expressions, genes commonly associated with myogenesis (GATA-4, MyoD1, cTnT and β-MHC) and neurogenesis (NeuroD, Nestin, GFAP, and MAP2) were up-regulated but gene expression associated to osteogenesis (ALPL, RUNX2, and SPARC) were either down modulated or remained at the nominal level. This is the first evidence that cellular morphology control via micropatterning could be used to modulate stem cell fate without external biochemical stimuli. We further our studies to modulate the focal adhesion (FA) instead of the macro shape of cells. Micro-contact printed islands of different smaller dimensions were investigated. We successfully regulated the FAs into dense FAs and elongated FAs by micropatterning. Additionally, the combined effects of hard (40.4 kPa), and intermediate (10.6 kPa) PA gel and FAs patterning on hMSCs differentiation were studied. Results showed that FA and matrix compliance plays an important role in hMSCs differentiation, and there is a cross-talk between different physical stimulants and the significance of these stimuli can only be realized if they are combined at the optimum level.

Keywords: micro-contact printing, polymer substrate, cell-material interaction, stem cell differentiation

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Authors: Iman Farasat, Howard M. Salis

Abstract:

The CRISPR/Cas9 system has revolutionized genome engineering by enabling site-directed and high-throughput genome editing, genome insertion, and gene knockdowns in several species, including bacteria, yeast, flies, worms, and human cell lines. This technology has the potential to enable human gene therapy to treat genetic diseases and cancer at the molecular level; however, the current CRISPR/Cas9 system suffers from seemingly sporadic off-target genome mutagenesis that prevents its use in gene therapy. A comprehensive mechanistic model that explains how the CRISPR/Cas9 functions would enable the rational design of the guide-RNAs responsible for target site selection while minimizing unexpected genome mutagenesis. Here, we present the first quantitative model of the CRISPR/Cas9 genome mutagenesis system that predicts how guide-RNA sequences (crRNAs) control target site selection and cleavage activity. We used statistical thermodynamics and law of mass action to develop a five-step biophysical model of cas9 cleavage, and examined it in vivo and in vitro. To predict a crRNA's binding specificities and cleavage rates, we then compiled a nearest neighbor (NN) energy model that accounts for all possible base pairings and mismatches between the crRNA and the possible genomic DNA sites. These calculations correctly predicted crRNA specificity across 5518 sites. Our analysis reveals that cas9 activity and specificity are anti-correlated, and, the trade-off between them is the determining factor in performing an RNA-mediated cleavage with minimal off-targets. To find an optimal solution, we first created a scheme of safe-design criteria for Cas9 target selection by systematic analysis of available high throughput measurements. We then used our biophysical model to determine the optimal Cas9 expression levels and timing that maximizes on-target cleavage and minimizes off-target activity. We successfully applied this approach in bacterial and mammalian cell lines to reduce off-target activity to near background mutagenesis level while maintaining high on-target cleavage rate.

Keywords: biophysical model, CRISPR, Cas9, genome editing

Procedia PDF Downloads 377