Search results for: adipose tissue derived stromal/stem cell
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 7464

Search results for: adipose tissue derived stromal/stem cell

6504 Multi-Channel Charge-Coupled Device Sensors Real-Time Cell Growth Monitor System

Authors: Han-Wei Shih, Yao-Nan Wang, Ko-Tung Chang, Lung-Ming Fu

Abstract:

A multi-channel cell growth real-time monitor and evaluation system using charge-coupled device (CCD) sensors with 40X lens integrating a NI LabVIEW image processing program is proposed and demonstrated. The LED light source control of monitor system is utilizing 8051 microprocessor integrated with NI LabVIEW software. In this study, the same concentration RAW264.7 cells growth rate and morphology in four different culture conditions (DMEM, LPS, G1, G2) were demonstrated. The real-time cells growth image was captured and analyzed by NI Vision Assistant every 10 minutes in the incubator. The image binarization technique was applied for calculating cell doubling time and cell division index. The cells doubling time and cells division index of four group with DMEM, LPS, LPS+G1, LPS+G2 are 12.3 hr,10.8 hr,14.0 hr,15.2 hr and 74.20%, 78.63%, 69.53%, 66.49%. The image magnification of multi-channel CCDs cell real-time monitoring system is about 100X~200X which compares with the traditional microscope.

Keywords: charge-coupled device (CCD), RAW264.7, doubling time, division index

Procedia PDF Downloads 353
6503 On Virtual Coordination Protocol towards 5G Interference Mitigation: Modelling and Performance Analysis

Authors: Bohli Afef

Abstract:

The fifth-generation (5G) wireless systems is featured by extreme densities of cell stations to overcome the higher future demand. Hence, interference management is a crucial challenge in 5G ultra-dense cellular networks. In contrast to the classical inter-cell interference coordination approach, which is no longer fit for the high density of cell-tiers, this paper proposes a novel virtual coordination based on the dynamic common cognitive monitor channel protocol to deal with the inter-cell interference issue. A tractable and flexible model for the coverage probability of a typical user is developed through the use of the stochastic geometry model. The analyses of the performance of the suggested protocol are illustrated both analytically and numerically in terms of coverage probability.

Keywords: ultra dense heterogeneous networks, dynamic common channel protocol, cognitive radio, stochastic geometry, coverage probability

Procedia PDF Downloads 319
6502 Providing Energy Management of a Fuel Cell-Battery Hybrid Electric Vehicle

Authors: Fatma Keskin Arabul, Ibrahim Senol, Ahmet Yigit Arabul, Ali Rifat Boynuegri

Abstract:

On account of the concern of the fossil fuel is depleting and its negative effects on the environment, interest in alternative energy sources is increasing day by day. However, considering the importance of transportation in human life, instead of oil and its derivatives fueled vehicles with internal combustion engines, electric vehicles which are sensitive to the environment and working with electrical energy has begun to develop. In this study, simulation was carried out for providing energy management and recovering regenerative braking in fuel cell-battery hybrid electric vehicle. The main power supply of the vehicle is fuel cell on the other hand not only instantaneous power is supplied by the battery but also the energy generated due to regenerative breaking is stored in the battery. Obtained results of the simulation is analyzed and discussed.

Keywords: electric vehicles, fuel cell, battery, regenerative braking, energy management

Procedia PDF Downloads 705
6501 Expanding Access and Deepening Engagement: Building an Open Source Digital Platform for Restoration-Based Stem Education in the Largest Public-School System in the United States

Authors: Lauren B. Birney

Abstract:

This project focuses upon the expansion of the existing "Curriculum and Community Enterprise for the Restoration of New York Harbor in New York City Public Schools" NSF EHR DRL 1440869, NSF EHR DRL 1839656 and NSF EHR DRL 1759006. This project is recognized locally as “Curriculum and Community Enterprise for Restoration Science,” or CCERS. CCERS is a comprehensive model of ecological restoration-based STEM education for urban public-school students. Following an accelerated rollout, CCERS is now being implemented in 120+ Title 1 funded NYC Department of Education middle schools, led by two cohorts of 250 teachers, serving more than 11,000 students in total. Initial results and baseline data suggest that the CCERS model, with the Billion Oyster Project (BOP) as its local restoration ecology-based STEM curriculum, is having profound impacts on students, teachers, school leaders, and the broader community of CCERS participants and stakeholders. Students and teachers report being receptive to the CCERS model and deeply engaged in the initial phase of curriculum development, citizen science data collection, and student-centered, problem-based STEM learning. The BOP CCERS Digital Platform will serve as the central technology hub for all research, data, data analysis, resources, materials and student data to promote global interactions between communities, Research conducted included qualitative and quantitative data analysis. We continue to work internally on making edits and changes to accommodate a dynamic society. The STEM Collaboratory NYC® at Pace University New York City continues to act as the prime institution for the BOP CCERS project since the project’s inception in 2014. The project continues to strive to provide opportunities in STEM for underrepresented and underserved populations in New York City. The replicable model serves as an opportunity for other entities to create this type of collaboration within their own communities and ignite a community to come together and address the notable issue. Providing opportunities for young students to engage in community initiatives allows for a more cohesive set of stakeholders, ability for young people to network and provide additional resources for those students in need of additional support, resources and structure. The project has planted more than 47 million oysters across 12 acres and 15 reef sites, with the help of more than 8,000 students and 10,000 volunteers. Additional enhancements and features on the BOP CCERS Digital Platform will continue over the next three years through funding provided by the National Science Foundation, NSF DRL EHR 1759006/1839656 Principal Investigator Dr. Lauren Birney, Professor Pace University. Early results from the data indicate that the new version of the Platform is creating traction both nationally and internationally among community stakeholders and constituents. This project continues to focus on new collaborative partners that will support underrepresented students in STEM Education. The advanced Digital Platform will allow for us connect with other countries and networks on a larger Global scale.

Keywords: STEM education, environmental restoration science, technology, citizen science

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6500 Ethiopian Women in Science, Technology, Engineering, and Mathematics Higher Education: Insights Gained Through an Onsite Culturally Embedded Workshop

Authors: Araceli Martinez Ortiz, Gillian U Bayne, Solomon Abraham

Abstract:

This paper describes research led by faculty from three American universities and four Ethiopian universities on the delivery of professional leadership development for early-career female Ethiopian university instructors in the Science, Technology, Engineering, and Mathematics (STEM) fields. The objective was to carry out a case study focused on the impact of an innovative intervention program designed to assist in the empowerment and leadership development related to teaching effectiveness, scholarly activity participation, and professional service participation by female instructors. This research was conducted utilizing a case study methodology for the weeklong intervention and a survey to capture the voices of the leadership program participants. The data regarding insights into the challenges and opportunities for women in these fields is presented. The research effort project expands upon existing linkages between universities to support professional development and research effort in this region of the world. Findings indicate the positive reception of this kind of professional development by the participating women. Survey data also reflects the educational technology and cultural challenges professional women in STEM education face in Ethiopia as well as the global challenges of balancing family expectations with career development.

Keywords: women, STEM education, higher education, Ethiopia

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6499 Electromagnetic-Mechanical Stimulation on PC12 for Enhancement of Nerve Axonal Extension

Authors: E. Nakamachi, K. Matsumoto, K. Yamamoto, Y. Morita, H. Sakamoto

Abstract:

In recently, electromagnetic and mechanical stimulations have been recognized as the effective extracellular environment stimulation technique to enhance the defected peripheral nerve tissue regeneration. In this study, we developed a new hybrid bioreactor by adopting 50 Hz uniform alternative current (AC) magnetic stimulation and 4% strain mechanical stimulation. The guide tube for nerve regeneration is mesh structured tube made of biodegradable polymer, such as polylatic acid (PLA). However, when neural damage is large, there is a possibility that peripheral nerve undergoes necrosis. So it is quite important to accelerate the nerve tissue regeneration by achieving enhancement of nerve axonal extension rate. Therefore, we try to design and fabricate the system that can simultaneously load the uniform AC magnetic field stimulation and the stretch stimulation to cells for enhancement of nerve axonal extension. Next, we evaluated systems performance and the effectiveness of each stimulation for rat adrenal pheochromocytoma cells (PC12). First, we designed and fabricated the uniform AC magnetic field system and the stretch stimulation system. For the AC magnetic stimulation system, we focused on the use of pole piece structure to carry out in-situ microscopic observation. We designed an optimum pole piece structure using the magnetic field finite element analyses and the response surface methodology. We fabricated the uniform AC magnetic field stimulation system as a bio-reactor by adopting analytically determined design specifications. We measured magnetic flux density that is generated by the uniform AC magnetic field stimulation system. We confirmed that measurement values show good agreement with analytical results, where the uniform magnetic field was observed. Second, we fabricated the cyclic stretch stimulation device under the conditions of particular strains, where the chamber was made of polyoxymethylene (POM). We measured strains in the PC12 cell culture region to confirm the uniform strain. We found slightly different values from the target strain. Finally, we concluded that these differences were allowable in this mechanical stimulation system. We evaluated the effectiveness of each stimulation to enhance the nerve axonal extension using PC12. We confirmed that the average axonal extension length of PC12 under the uniform AC magnetic stimulation was increased by 16 % at 96 h in our bio-reactor. We could not confirm that the axonal extension enhancement under the stretch stimulation condition, where we found the exfoliating of cells. Further, the hybrid stimulation enhanced the axonal extension. Because the magnetic stimulation inhibits the exfoliating of cells. Finally, we concluded that the enhancement of PC12 axonal extension is due to the magnetic stimulation rather than the mechanical stimulation. Finally, we confirmed that the effectiveness of the uniform AC magnetic field stimulation for the nerve axonal extension using PC12 cells.

Keywords: nerve cell PC12, axonal extension, nerve regeneration, electromagnetic-mechanical stimulation, bioreactor

Procedia PDF Downloads 261
6498 Analysis of Osmotin as Transcription Factor/Cell Signaling Modulator Using Bioinformatic Tools

Authors: Usha Kiran, M. Z. Abdin

Abstract:

Osmotin is an abundant cationic multifunctional protein discovered in cells of tobacco (Nicotiana tabacum L. var Wisconsin 38) adapted to an environment of low osmotic potential. It provides plants protection from pathogens, hence placed in the PRP family of proteins. The osmotin induced proline accumulation has been reported in plants including transgenic tomato and strawberry conferring tolerance against both biotic and abiotic stresses. The exact mechanism of induction of proline by osmotin is however, not known till date. These observations have led us to hypothesize that osmotin induced proline accumulation could be due to its involvement as transcription factor and/or cell signal pathway modulator in proline biosynthesis. The present investigation was therefore, undertaken to analyze the osmotin protein as transcription factor /cell signalling modulator using bioinformatics tools. The results of available online DNA binding motif search programs revealed that osmotin does not contain DNA-binding motifs. The alignment results of osmotin protein with the protein sequence from DATF showed the homology in the range of 0-20%, suggesting that it might not contain a DNA binding motif. Further to find unique DNA-binding domain, the superimposition of osmotin 3D structure on modeled Arabidopsis transcription factors using Chimera also suggested absence of the same. We, however, found evidence implicating osmotin in cell signaling. With these results, we concluded that osmotin is not a transcription factor but regulating proline biosynthesis and accumulation through cell signaling during abiotic stresses.

Keywords: osmotin, cell signaling modulator, bioinformatic tools, protein

Procedia PDF Downloads 262
6497 Wound Healing Process Studied on DC Non-Homogeneous Electric Fields

Authors: Marisa Rio, Sharanya Bola, Richard H. W. Funk, Gerald Gerlach

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Cell migration, wound healing and regeneration are some of the physiological phenomena in which electric fields (EFs) have proven to have an important function. Physiologically, cells experience electrical signals in the form of transmembrane potentials, ion fluxes through protein channels as well as electric fields at their surface. As soon as a wound is created, the disruption of the epithelial layers generates an electric field of ca. 40-200 mV/mm, directing cell migration towards the wound site, starting the healing process. In vitro electrotaxis, experiments have shown cells respond to DC EFs polarizing and migrating towards one of the poles (cathode or anode). A standard electrotaxis experiment consists of an electrotaxis chamber where cells are cultured, a DC power source and agar salt bridges that help delaying toxic products from the electrodes to attain the cell surface. The electric field strengths used in such an experiment are uniform and homogeneous. In contrast, the endogenous electric field strength around a wound tend to be multi-field and non-homogeneous. In this study, we present a custom device that enables electrotaxis experiments in non-homogeneous DC electric fields. Its main feature involves the replacement of conventional metallic electrodes, separated from the electrotaxis channel by agarose gel bridges, through electrolyte-filled microchannels. The connection to the DC source is made by Ag/AgCl electrodes, incased in agarose gel and placed at the end of each microfluidic channel. An SU-8 membrane closes the fluidic channels and simultaneously serves as the single connection from each of them to the central electrotaxis chamber. The electric field distribution and current density were numerically simulated with the steady-state electric conduction module from ANSYS 16.0. Simulation data confirms the application of nonhomogeneous EF of physiological strength. To validate the biocompatibility of the device cellular viability of the photoreceptor-derived 661W cell line was accessed. The cells have not shown any signs of apoptosis, damage or detachment during stimulation. Furthermore, immunofluorescence staining, namely by vinculin and actin labelling, allowed the assessment of adhesion efficiency and orientation of the cytoskeleton, respectively. Cellular motility in the presence and absence of applied DC EFs was verified. The movement of individual cells was tracked for the duration of the experiments, confirming the EF-induced, cathodal-directed motility of the studied cell line. The in vitro monolayer wound assay, or “scratch assay” is a standard protocol to quantitatively access cell migration in vitro. It encompasses the growth of a confluent cell monolayer followed by the mechanic creation of a scratch, representing a wound. Hence, wound dynamics was monitored over time and compared for control and applied the electric field to quantify cellular population motility.

Keywords: DC non-homogeneous electric fields, electrotaxis, microfluidic biochip, wound healing

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6496 Surface Display of Lipase on Yarrowia lipolytica Cells

Authors: Evgeniya Y. Yuzbasheva, Tigran V. Yuzbashev, Natalia I. Perkovskaya, Elizaveta B. Mostova

Abstract:

Cell-surface display of lipase is of great interest as it has many applications in the field of biotechnology owing to its unique advantages: simplified product purification, and cost-effective downstream processing. One promising area of application for whole-cell biocatalysts with surface displayed lipase is biodiesel synthesis. Biodiesel is biodegradable, renewable, and nontoxic alternative fuel for diesel engines. Although the alkaline catalysis method has been widely used for biodiesel production, it has a number of limitations, such as rigorous feedstock specifications, complicated downstream processes, including removal of inorganic salts from the product, recovery of the salt-containing by-product glycerol, and treatment of alkaline wastewater. Enzymatic synthesis of biodiesel can overcome these drawbacks. In this study, Lip2p lipase was displayed on Yarrowia lipolytica cells via C- and N-terminal fusion variant. The active site of lipase is located near the C-terminus, therefore to prevent the activity loosing the insertion of glycine-serine linker between Lip2p and C-domains was performed. The hydrolytic activity of the displayed lipase reached 12,000–18,000 U/g of dry weight. However, leakage of enzyme from the cell wall was observed. In case of C-terminal fusion variant, the leakage was occurred due to the proteolytic cleavage within the linker peptide. In case of N-terminal fusion variant, the leaking enzyme was presented as three proteins, one of which corresponded to the whole hybrid protein. The calculated number of recombinant enzyme displayed on the cell surface is approximately 6–9 × 105 molecules per cell, which is close to the theoretical maximum (2 × 106 molecules/cell). Thus, we attribute the enzyme leakage to the limited space available on the cell surface. Nevertheless, cell-bound lipase exhibited greater stability to short-term and long-term temperature treatment than the native enzyme. It retained 74% of original activity at 60°C for 5 min of incubation, and 83% of original activity after incubation at 50°C during 5 h. Cell-bound lipase had also higher stability in organic solvents and detergents. The developed whole-cell biocatalyst was used for recycling biodiesel synthesis. Two repeated cycles of methanolysis yielded 84.1–% and 71.0–% methyl esters after 33–h and 45–h reactions, respectively.

Keywords: biodiesel, cell-surface display, lipase, whole-cell biocatalyst

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6495 Effect of Ultrasound and Enzyme on the Extraction of Eurycoma longifolia (Tongkat Ali)

Authors: He Yuhai, Ahmad Ziad Bin Sulaiman

Abstract:

Tongkat Ali, or Eurycoma longifolia, is a traditional Malay and Orang Asli herb used as aphrodisiac, general tonic, anti-Malaria, and anti-Pyretic. It has been recognized as a cashcrop by Malaysia due to its high value for the pharmaceutical use. In Tongkat Ali, eurycomanone, a quassinoid is usually chosen as a marker phytochemical as it is the most abundant phytochemical. In this research, ultrasound and enzyme were used to enhance the extraction of Eurycomanone from Tongkat Ali. Ultrasonic assisted extraction (USE) enhances extraction by facilitating the swelling and hydration of the plant material, enlarging the plant pores, breaking the plant cell, reducing the plant particle size and creating cavitation bubbles that enhance mass transfer in both the washing and diffusion phase of extraction. Enzyme hydrolyses the cell wall of the plant, loosening the structure of the cell wall, releasing more phytochemicals from the plant cell, enhancing the productivity of the extraction. Possible effects of ultrasound on the activity of the enzyme during the hydrolysis of the cell wall is under the investigation by this research. The extracts was analysed by high performance liquid chromatography for the yields of Eurycomanone. In this whole process, the conventional water extraction was used as a control of comparing the performance of the ultrasound and enzyme assisted extraction.

Keywords: ultrasound, enzymatic, extraction, Eurycoma longifolia

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6494 Case Report on ‘Primary Adenocarcinoma of Aberrant HER2+ Anogenital Mammary-like Glands in a Male'

Authors: Shivani Kuttuva, James Sampson, Timothy Simmons, Vinayak Thattaruparambil, Holly Burton, Peter Coyne

Abstract:

Anogenital mammary-like glands were established to be embryological remnants of breast tissue due to failed resolution of the ectodermal mammary ridge. However, recent studies are now considering this to represent normal constituents of the anogenital area with histological resemblance to the orthotopic breast tissue with multiple benign and malignant lesions arising from it. The incidence of the above has been predominant in females in the vulval region. Due to the paucity of cases reported in men, this poses a diagnostic and therapeutic challenge resulting in a delay in treatment and, thereby, poor outcomes. Our patient presented to the dermatology clinic with an itchy, purplish lesion in the peri-anal region which, on punch biopsy, was diagnosed to be Extra-mammary Paget’s disease and taken up for Wide local excision. Immunochemically, staining was positive for HER2, ER and Cytokeratin 7, keeping with the presence of actual breast tissue with no primary breast carcinoma. Due to the invasive nature of the disease, he required Abdominoperineal resection with flap reconstruction. Despite complete surgical clearance and adjuvant radiotherapy, the disease progressed to adjacent inguinal and obturator lymph nodes with origin resembling anogenital type mammary glands but histology negative for hormonal receptors of the breast.

Keywords: anogenital mammary-like glands, abdominoperineal resection, ectopic breast tissue, ectopic male breast carcinoma, peri-anal skin lesion

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6493 Comparisons between Student Leaning Achievements and Their Problem Solving Skills on Stoichiometry Issue with the Think-Pair-Share Model and Stem Education Method

Authors: P. Thachitasing, N. Jansawang, W. Rakrai, T. Santiboon

Abstract:

The aim of this study is to investigate of the comparing the instructional design models between the Think-Pair-Share and Conventional Learning (5E Inquiry Model) Processes to enhance students’ learning achievements and their problem solving skills on stoichiometry issue for concerning the 2-instructional method with a sample consisted of 80 students in 2 classes at the 11th grade level in Chaturaphak Phiman Ratchadaphisek School. Students’ different learning outcomes in chemistry classes with the cluster random sampling technique were used. Instructional Methods designed with the 40-experimenl student group by Think-Pair-Share process and the 40-controlling student group by the conventional learning (5E Inquiry Model) method. These learning different groups were obtained using the 5 instruments; the 5-lesson instructional plans of Think-Pair-Share and STEM Education Method, students’ learning achievements and their problem solving skills were assessed with the pretest and posttest techniques, students’ outcomes of their instructional the Think-Pair-Share (TPSM) and the STEM Education Methods were compared. Statistically significant was differences with the paired t-test and F-test between posttest and pretest technique of the whole students in chemistry classes were found, significantly. Associations between student learning outcomes in chemistry and two methods of their learning to students’ learning achievements and their problem solving skills also were found. The use of two methods for this study is revealed that the students perceive their learning achievements to their problem solving skills to be differently learning achievements in different groups are guiding practical improvements in chemistry classrooms to assist teacher in implementing effective approaches for improving instructional methods. Students’ learning achievements of mean average scores to their controlling group with the Think-Pair-Share Model (TPSM) are lower than experimental student group for the STEM education method, evidence significantly. The E1/E2 process were revealed evidence of 82.56/80.44, and 83.02/81.65 which results based on criteria are higher than of 80/80 standard level with the IOC, consequently. The predictive efficiency (R2) values indicate that 61% and 67% and indicate that 63% and 67% of the variances in chemistry classes to their learning achievements on posttest in chemistry classes of the variances in students’ problem solving skills to their learning achievements to their chemistry classrooms on Stoichiometry issue with the posttest were attributable to their different learning outcomes for the TPSM and STEMe instructional methods.

Keywords: comparisons, students’ learning achievements, think-pare-share model (TPSM), stem education, problem solving skills, chemistry classes, stoichiometry issue

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6492 Analyzing the Effects of a Psychological Intervention on Black Students’ Sense of Belonging in Physics and Math: Exploring Differential Impacts for Historically Black Colleges and Universities and Predominantly White Institutions

Authors: Terrell Strayhorn

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The lack of diversity in science, technology, engineering, and mathematics (STEM) fields is a persistent and concerning issue. One contributing factor to the underrepresentation of minority groups in STEM fields is a lack of sense of belonging, which can lead to lower levels of academic engagement, motivation, and achievement. In particular, Black students have been shown to experience lower levels of sense of belonging in STEM compared to their white peers. This study aimed to explore the effects of a psychological intervention on Black students' sense of belonging in physics and math courses at historically Black colleges and universities (HBCUs) and predominantly white institutions (PWIs). The study used a randomized controlled trial design and included 305 Black undergraduate students enrolled in physics or math courses at HBCUs and PWIs in the United States. Participants were randomly assigned to either an intervention group or a control group. The intervention consisted of a brief psychological, video-based intervention designed to enhance sense of belonging, which was delivered in a single session. The control group received no intervention. The primary outcome measure was sense of belonging in physics and math courses, as assessed by a validated self-report measure. Other outcomes included academic engagement, motivation, and achievement as measured by physics and math (course) grades. Preliminary results show that the intervention has a significant positive effect on Black students' sense of belonging in physics and math courses, with a moderate effect size. The intervention also had a significant positive effect on academic engagement and motivation, but not on academic achievement. Importantly, the effects of the intervention were larger for Black students enrolled at PWIs compared to those enrolled at HBCUs. Findings, at present, suggest that a brief psychological web-based intervention can enhance Black students' sense of belonging in physics and math courses, and that the effects may be particularly strong for Black students enrolled at PWIs, although they are not negligible for Black students at HBCUs. This is an important finding given the persistent underrepresentation of Black students in STEM fields, the growing number of Black students at PWIs, and the potential for enhancing sense of belonging to improve academic outcomes and increase diversity in these fields. The study has several limitations, including a relatively small sample size and a lack of long-term follow-up. Future research could explore the generalizability of these findings to other minority groups and other STEM fields, as well as the potential for longer-term interventions to sustain and enhance the effects observed in this study. Overall, this study highlights the potential for psychological interventions to enhance sense of belonging and improve academic outcomes for Black students in STEM courses, and underscores the importance of addressing sense of belonging as a key factor in promoting diversity and equity in STEM fields.

Keywords: sense of belonging, achievement, racial equity, postsecondary education, intervention

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6491 Investigation of Alumina Membrane Coated Titanium Implants on Osseointegration

Authors: Pinar Erturk, Sevde Altuntas, Fatih Buyukserin

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In order to obtain an effective integration between an implant and a bone, implant surfaces should have similar properties to bone tissue surfaces. Especially mimicry of the chemical, mechanical and topographic properties of the implant to the bone is crucial for fast and effective osseointegration. Titanium-based biomaterials are more preferred in clinical use, and there are studies of coating these implants with oxide layers that have chemical/nanotopographic properties stimulating cell interactions for enhanced osseointegration. There are low success rates of current implantations, especially in craniofacial implant applications, which are large and vital zones, and the oxide layer coating increases bone-implant integration providing long-lasting implants without requiring revision surgery. Our aim in this study is to examine bone-cell behavior on titanium implants with an aluminum oxide layer (AAO) on effective osseointegration potential in the deformation of large zones with difficult spontaneous healing. In our study, aluminum layer coated titanium surfaces were anodized in sulfuric, phosphoric, and oxalic acid, which are the most common used AAO anodization electrolytes. After morphologic, chemical, and mechanical tests on AAO coated Ti substrates, viability, adhesion, and mineralization of adult bone cells on these substrates were analyzed. Besides with atomic layer deposition (ALD) as a sensitive and conformal technique, these surfaces were coated with pure alumina (5 nm); thus, cell studies were performed on ALD-coated nanoporous oxide layers with suppressed ionic content too. Lastly, in order to investigate the effect of the topography on the cell behavior, flat non-porous alumina layers on silicon wafers formed by ALD were compared with the porous ones. Cell viability ratio was similar between anodized surfaces, but pure alumina coated titanium and anodized surfaces showed a higher viability ratio compared to bare titanium and bare anodized ones. Alumina coated titanium surfaces, which anodized in phosphoric acid, showed significantly different mineralization ratios after 21 days over other bare titanium and titanium surfaces which anodized in other electrolytes. Bare titanium was the second surface that had the highest mineralization ratio. Otherwise, titanium, which is anodized in oxalic acid electrolyte, demonstrated the lowest mineralization. No significant difference was shown between bare titanium and anodized surfaces except AAO titanium surface anodized in phosphoric acid. Currently, osteogenic activities of these cells on the genetic level are investigated by quantitative real-time polymerase chain reaction (qRT-PCR) analysis results of RUNX-2, VEGF, OPG, and osteopontin genes. Also, as a result of the activities of the genes mentioned before, Western Blot will be used for protein detection. Acknowledgment: The project is supported by The Scientific and Technological Research Council of Turkey.

Keywords: alumina, craniofacial implant, MG-63 cell line, osseointegration, oxalic acid, phosphoric acid, sulphuric acid, titanium

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6490 Cytotoxic Activity of Extracts from Hibiscus sabdariffa Leaves against Women’s Cancer Cell Lines

Authors: Patsorn Worawattananutai, Srisopa Ruangnoo, Arunporn Itharat

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Hibiscus sabdariffa (HS) leaves are vegetables which are extensively used as blood tonic and laxatives in Thai traditional medicine. They are popularly used as healthy sour soup for prevention of chronic diseases such as cancer. Therefore, the cytotoxic activity of different extracts of fresh and dried Hibiscus sabdariffa leaves were investigated via the sulforhodamine B (SRB) assay against three types of women’s cancer cell lines, namely the human cervical adenocarcinoma cell line (HeLa), the human ovarian adenocarcinoma cell line (SKOV-3), and the human breast adenocarcinoma cell line (MCF-7). Extraction methods were squeezing, boiling with water and maceration with 95% or 50% ethanol. The 95% ethanolic extracts of Hibiscus sabdariffa dry leaves (HSDE95) showed the highest cytotoxicity against all types of women’s cancer cell lines with the IC50 values in range 7.51±0.33 to 12.13±1.85 µg/ml. Its IC50 values against SKOV-3, HeLa and MCF-7 were 7.51±0.33, 9.44±1.41 and 12.13±1.85 µg/ml, respectively. In these results, this extract can be classified as “active” according to the NCI guideline which indicated that IC50 values of the active cytotoxic plant extracts have to be beneath 20 µg/ml. Thus, HSDE95 was concluded to be a potent cytotoxic drug for all women’s cancer cells. This extract should be further investigated to isolate active compounds against women’s cancer cells.

Keywords: breast adenocarcinoma, cervical adenocarcinoma, cytotoxic activity, Hibiscus sabdariffa, ovarian adenocarcinoma

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6489 Fabrication and Characterization of PPy/rGO|PPy/ZnO Composite with Varying Zno Concentration as Anode for Fuel Cell Applications

Authors: Bryan D. Llenarizas, Maria Carla F. Manzano

Abstract:

The rapid growth of electricity demand has led to a pursuit of alternative energy sources with high power output and not harmful to the environment. The fuel cell is a device that generates electricity via chemical reactions between the fuel and oxidant. Fuel cells have been known for decades, but the development of high-power output and durability was still one of the drawbacks of this energy source. This study investigates the potential of layer-by-layer composite for fuel cell applications. A two-electrode electrochemical cell was used for the galvanostatic electrochemical deposition method to fabricate a Polypyrrole/rGO|Polypyrrole/ZnO layer-by-layer composite material for fuel cell applications. In the synthesis, the first layer comprised 0.1M pyrrole monomer and 1mg of rGO, while the second layer had 0.1M pyrrole monomer and variations of ZnO concentration ranging from 0.08M up to 0.12M. A constant current density of 8mA/cm² was applied for 1 hour in fabricating each layer. Scanning electron microscopy (SEM) for the fabricated LBL material shows a globular surface with white spots. These white spots are the ZnO particles confirmed by energy-dispersive X-ray spectroscopy, indicating a successful deposition of the second layer onto the first layer. The observed surface morphology was consistent for each variation of ZnO concentrations. AC measurements were conducted to obtain the AC resistance of the fabricated film. Results show a decrease in AC resistance as the concentration of ZnO increases.

Keywords: anode, composite material, electropolymerization, fuel cell, galvanostatic, polypyrrole

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6488 The Type II Immune Response in Acute and Chronic Pancreatitis Mediated by STAT6 in Murine

Authors: Hager Elsheikh

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Context: Pancreatitis is a condition characterized by inflammation in the pancreas, which can lead to serious complications if untreated. Both acute and chronic pancreatitis are associated with immune reactions and fibrosis, which further damage the pancreas. The type 2 immune response, primarily driven by alternative activated macrophages (AAMs), plays a significant role in the development of fibrosis. The IL-4/STAT6 pathway is a crucial signaling pathway for the activation of M2 macrophages. Pancreatic fibrosis is induced by dysregulated inflammatory responses and can result in the autodigestion and necrosis of pancreatic acinar cells. Research Aim: The aim of this study is to investigate the impact of STAT6, a crucial molecule in the IL-4/STAT6 pathway, on the severity and development of fibrosis during acute and chronic pancreatitis. The research also aims to understand the influence of the JAK/STAT6 signaling pathway on the balance between fibrosis and regeneration in the presence of different macrophage populations. Methodology: The research utilizes murine models of acute and chronic pancreatitis induced by cerulean injection. Animal models will be employed to study the effect of STAT6 knockout on disease severity and fibrosis. Isolation of acinar cells and cell culture techniques will be used to assess the impact of different macrophage populations on wound healing and regeneration. Various techniques such as PCR, histology, immunofluorescence, and transcriptomics will be employed to analyze the tissues and cells. Findings: The research aims to provide insights into the mechanisms underlying tissue fibrosis and wound healing during acute and chronic pancreatitis. By investigating the influence of the JAK/STAT6 signaling pathway and different macrophage populations, the study aims to understand their impact on tissue fibrosis, disease severity, and pancreatic regeneration. Theoretical Importance: This research contributes to our understanding of the role of specific signaling pathways, macrophage polarization, and the type 2 immune response in pancreatitis. It provides insights into the molecular mechanisms underlying tissue fibrosis and the potential for targeted therapies. Data Collection and Analysis Procedures: Data will be collected through the use of murine models, isolation and culture of acinar cells, and various experimental techniques such as PCR, histology, immunofluorescence, and transcriptomics. Data will be analyzed using appropriate statistical methods and techniques, and the findings will be interpreted in the context of the research objectives. Conclusion: By investigating the mechanisms of tissue fibrosis and wound healing during acute and chronic pancreatitis, this research aims to enhance our understanding of the disease progression and potential therapeutic targets. The findings have theoretical importance in expanding our knowledge of pancreatic fibrosis and the role of macrophage polarization in the context of the type 2 immune response.

Keywords: immunity in chronic diseases, pancreatitis, macrophages, immune response

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6487 [Keynote Speech]: Curiosity, Innovation and Technological Advancements Shaping the Future of Science, Technology, Engineering and Mathematics Education

Authors: Ana Hol

Abstract:

We live in a constantly changing environment where technology has become an integral component of our day to day life. We rely heavily on mobile devices, we search for data via web, we utilise smart home sensors to create the most suited ambiences and we utilise applications to shop, research, communicate and share data. Heavy reliance on technology therefore is creating new connections between STEM (Science, Technology, Engineering and Mathematics) fields which in turn rises a question of what the STEM education of the future should be like? This study was based on the reviews of the six Australian Information Systems students who undertook an international study tour to India where they were given an opportunity to network, communicate and meet local students, staff and business representatives and from them learn about the local business implementations, local customs and regulations. Research identifies that if we are to continue to implement and utilise electronic devices on the global scale, such as for example implement smart cars that can smoothly cross borders, we will need the workforce that will have the knowledge about the cars themselves, their parts, roads and transport networks, road rules, road sensors, road monitoring technologies, graphical user interfaces, movement detection systems as well as day to day operations, legal rules and regulations of each region and country, insurance policies, policing and processes so that the wide array of sensors can be controlled across country’s borders. In conclusion, it can be noted that allowing students to learn about the local conditions, roads, operations, business processes, customs and values in different countries is giving students a cutting edge advantage as such knowledge cannot be transferred via electronic sources alone. However once understanding of each problem or project is established, multidisciplinary innovative STEM projects can be smoothly conducted.

Keywords: STEM, curiosity, innovation, advancements

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6486 Expression of miRNA 335 in Gall Bladder Cancer: A Correlative Study

Authors: Naseem Fatima, A. N. Srivastava, Tasleem Raza, Vijay Kumar

Abstract:

Introduction: Carcinoma gallbladder is third most common gastrointestinal lethal disease with the highest incidence and mortality rate among women in Northern India. Scientists have found several risk factors that make a person more likely to develop gallbladder cancer; among these risk factors, deregulation of miRNAs has been demonstrated to be one of the most crucial factors. The changes in the expression of specific miRNA genes result in the control of inflammation, cell cycle regulation, stress response, proliferation, differentiation, apoptosis and invasion thus mediate the process in tumorgenesis. The aim of this study was to investigate the role of MiRNA-335 and may as a molecular marker in early detection of gallbladder cancer in suspected cases. Material and Methods: A total of 20 consecutive patients with gallbladder cancer aged between 30-75 years were registered for the study. Total RNA was extracted from tissue by using the mirVANA MiRNA isolation Kit according to the manufacturer’s protocol. The MiRNA- 335 and U6 snRNA-specific cDNA were reverse-transcribed from total RNA using Taqman microRNA reverse-transcription kit according to the manufacturer’s protocol. TaqMan MiRNA probes hsa-miR-335 and Taqman Master Mix without AmpEase UNG, Individual real-time PCR assays were performed in a 20 μL reaction volume on a Real-Time PCR system (Applied Biosystems StepOnePlus™) to detect MiRNA-335 expression in tissue. Relative quantification of target MiRNA expression was evaluated using the comparative cycle threshold (CT) method. The correlation was done in between cycle threshold (CT Value) of target MiRNA in gallbladder cancer with respect to non-cancerous Cholelithiasis gallbladder. Each sample was examined in triplicate. The Newman-Keuls Multiple Comparison Test was used to determine the expression of miR-335. Results: MiRNA335 was found to be significantly downregulated in the gallbladder cancer tissue (P<0.001), when compared with non-cancerous Cholelithiasis gallbladder cases. Out of 20 cases, 75% showed reduced expression of MiRNA335, were at last stage of disease with low overall survival rate and remaining 25% were showed up-regulated expression of MiRNA335 with high survival rate. Conclusion: The present study showed that reduced expression of MiRNA335 is associated with the advancement of the disease, and its deregulation may provide important clues to understanding it as a prognostic marker and opportunities for future research.

Keywords: carcinoma gallbladder, downregulation, MiRNA-335, RT-PCR assay

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6485 The Comparison of Primary B-Cell and NKT-Cell Non-Hodgkin Lymphomas in Nasopharynx, Nasal Cavity, and Paranasal Sinuses

Authors: Jiajia Peng, Jianqing Qiu, Jianjun Ren, Yu Zhao

Abstract:

Background: We aimed to compare clinical and survival differences between B-cell (B-NHL) and NKT-cell non-Hodgkin lymphomas (NKT-NHL) located in the nasal cavity, nasopharynx and paranasal sinuses, which are always categorized as one sinonasal type. Methods: Patients diagnosed with primary B-NHL and NKT-NHL in the nasal cavity, nasopharynx, and paranasal sinuses from the SEER database were included. We identified these patients based on histological types and anatomical sites and subsequently conducted univariate and multivariate Cox regression and Kaplan–Meier analyses to examine cancer-special survival (CSS) outcomes. Results: Overall, most B-NHL cases originated from the nasopharynx, while the majority of NKT-NHL cases occurred in the nasal cavity. Notably, the CSS outcomes improved significantly in all sinonasal B-NHL cases over time, whereas no such improvement trend was observed in each sinonasal NKT-NHL type. Additionally, increasing age was linked with an elevated risk of death in B-NHL, particularly in the nasal cavity (HR:3.37), rather than in NKT-NHL. Compared with B-NHL, the adverse effect of the higher stage on CSS was more evident in NKT-NHL, particularly in its nasopharynx site (HR: 5.12). Furthermore, radiotherapy was beneficial for survival in patients with sinonasal B-NHL and NKT-NHL, except in those with NKT-NHL in the nasopharynx site. However, chemotherapy has only been beneficial for CSS in patients with B-NHL in paranasal sinuses (HR: 0.42) since 2010, rather than in other types of B-NHL or NKT-NHL. Conclusions: Although B-NHL and NKT-NHL in the nasal cavity, nasopharynx and paranasal sinuses have similar anatomical locations, their clinic demographics and prognoses are largely different and should be treated and studied as distinct diseases.

Keywords: B-cell non-Hodgkin lymphomas, NKT-cell non-Hodgkin lymphomas, nasal cavity lymphomas, nasal sinuses lymphomas, nasopharynx lymphomas

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6484 Collagen Gel in Hip Cartilage Repair: in vivo Preliminary Study

Authors: A. Bajek, J. Skopinska-Wisniewska, A. Rynkiewicz, A. Jundzill, M. Bodnar, A. Marszalek, T. Drewa

Abstract:

Traumatic injury and age-related degenerative diseases associated with cartilage are major health problems worldwide. The articular cartilage is comprised of a relatively small number of cells, which have a relatively slow rate of turnover. Therefore, damaged articular cartilage has a limited capacity for self-repair. New clinical methods have been designed to achieve better repair of injured cartilage. However, there is no treatment that enables full restoration of it. The aim of this study was to evaluate how collagen gel with bone marrow mesenchymal stem cells (MSCs) and collagen gel alone will influence on the hip cartilage repair after injury. Collagen type I was isolated from rats’ tails and cross-linked with N-hydroxysuccinimide in 24-hour process. MSCs were isolated from rats’ bone marrow. The experiments were conducted according to the guidelines for animal experiments of Ethics Committee. Fifteen 8-week-old Wistar rats were used in this study. All animals received hip joint surgery with a total of 30 created cartilage defects. Then, animals were randomly divided into three groups and filled, respectively, with collagen gel (group 1), collagen gel cultured with MSCs (group II) or left untreated as a control (control group). Immunohistochemy and radiological evaluation was carried out 11 weeks post implantation. It has been proved that the surface of the matrix is non-toxic, and its porosity promotes cell adhesion and growth. However, the in vivo regeneration process was poor. We observed the low integration rate of biomaterial. Immunohistochemical evaluation of cartilage after 11 weeks of treatment showed low II and high X collagen expression in two tested groups in comparison to the control one, in which we observed the high II collagen expression. What is more, after radiological analysis, we observed the best regeneration process in control group. The biomaterial construct and mesenchymal stem cells, as well as the use of the biomaterial itself was not sufficient to regenerate the hip cartilage surfaces. These results suggest that the collagen gel based biomaterials, even with MSCs, are not satisfactory in repar of hip cartilage defect. However, additional evaluation is needed to confirm these results.

Keywords: collafen gel, MSCs, cartilage repair, hip cartilage

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6483 The Inhibitory Effect of Weissella koreensis 521 Isolated from Kimchi on 3T3-L1 Adipocyte Differentiation

Authors: Kyungbae Pi, Kibeom Lee, Yongil Kim, Eun-Jung Lee

Abstract:

Abnormal adipocyte growth, in terms of increased cell numbers and increased cell differentiation, is considered to be a major pathological feature of obesity. Thus, the inhibition of preadipocyte mitogenesis and differentiation could help prevent and suppress obesity. The aim of this study was to assess whether extracts from Weissella koreensis 521 cells isolated from kimchi could exert anti-adipogenic effects in 3T3-L1 cells (fat cells). Differentiating 3T3-L1 cells were treated with W. koreensis 521 cell extracts (W. koreensis 521_CE), and cell viability was assessed by MTT assays. At concentrations below 0.2 mg/ml, W. koreensis 521_CE did not exert any cytotoxic effect in 3T3-L1 cells. However, treatment with W. koreensis 521_CE significantly inhibited adipocyte differentiation, as assessed by morphological analysis and Oil Red O staining of fat. W. koreensis 521_CE treatment (0.2 mg/ml) also reduced lipid accumulation by 24% in fully differentiated 3T3-L1 adipocytes. These findings collectively indicate that Weissella koreensis 521 may help prevent obesity.

Keywords: Weissella koreensis 521, 3T3-L1 cells, adipocyte differentiation, obesity

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6482 Latest Generation Conducted Electrical Weapon Dart Design: Signature Marking and Removal for the Emergency Medicine Professional

Authors: J. D. Ho, D. M. Dawes, B. Driver

Abstract:

Introduction: TASER Conducted Electrical Weapons (CEWs) are the dominant CEWs in use and have been used in modern police and military operations since the late 1990s as a form of non-lethal weaponry. The 3rd generation of CEWs has been recently introduced and is known as The TASER 7. This new CEW will be replacing current CEW technology and has a new dart design that is important for emergency medical professionals to be familiar with because it requires a different method of removal and will leave a different marking pattern in human tissue than they may have been previously familiar with. features of this new dart design include: higher velocity impact, larger impact surface area, break away dart body segment, dual back-barb retention, newly designed removal process. As the TASER 7 begins to be deployed by the police and military personnel, these new features make it imperative that emergency medical professionals become familiar with the signature markings that this new dart design will make on human tissue and how to remove them. Methods: Multiple observational studies using high speed photography were used to record impact patterns of the new dart design on fresh tissue and also the newly recommended dart removal process. Both animal and human subjects were used to test this dart design prior to production release. Results: Data presented will include dart design overview, flight pattern accuracy, impact analysis, and dart removal example. Tissue photographs will be presented to demonstrate examples of signature TASER 7 dart markings that emergency medical professionals can expect to see. Conclusion: This work will provide the reader with an understanding of this newest generation CEW dart design, its key features, its signature marking pattern that can be expected and a recommendation of how to remove it from human tissue.

Keywords: TASER 7, conducted electrical weapon, dart mark, dart removal

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6481 Effect of Exercise Training on Body Composition and Metabolic Profile in Older Adults during Cancer Treatment

Authors: Adeline Fontvieille, Hugo Parent-Roberge, Marie-France Langlois, Tamas Fulop, Michel Pavic, Eleonor Riesco

Abstract:

Introduction: Total lean body mass is reduced during cancer treatment. This loss is called cancer cachexia and is accompanied by a progressive loss of fat mass. In older adults, these body composition changes can have a larger impact on metabolic health, physical autonomy, and cancer survival. Although currently untreatable, exercise training could reduce these effects. Hence, the objective of this pilot study is to investigate if 12 weeks of exercise training during cancer treatment can mitigate the loss of muscle mass and fat mass in older adults. Methods: A total of 40 older adults (65-80 years) with an ongoing treatment for a curable cancer are currently recruited and randomised in two groups: 1) Combined training (EX, n=20) and 2) Control group (CON, n=20). All variables are measured before and after 12 weeks of intervention: Anthropometry (weight, height, body mass index), body composition (total fat mass, visceral adipose tissue, total and appendicular muscle mass; DXA), metabolic profile (HDL-C and LDL-C, triglycerides, glucose and insulin levels). Results: Preliminary analyses revealed no impact of exercise training on appendicular muscle mass (p=0,31) and fat mass (p=0,31). Furthermore, total body weight, waist circumference, HDL-cholesterol, LDL-cholesterol, glucose and insulin levels remained unchanged (all p ≥ 0.79) after 12 weeks of training. However, statistical analyses revealed that triglyceride levels slightly increased (p=0.03), irrespective of the group. Conclusion: Preliminary analyses did not reveal any impact of aerobic and resistance exercise training on body composition in oncogeriatric patients. Furthermore, exercise training seems not efficient to prevent the cancer treatment-related triglyceride levels increase.

Keywords: muscle mass, fat mass, metabolic profile, combined training, aging, cancer

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6480 Central Vascular Function and Relaxibility in Beta-thalassemia Major Patients vs. Sickle Cell Anemia Patients by Abdominal Aorta and Aortic Root Speckle Tracking Echocardiography

Authors: Gehan Hussein, Hala Agha, Rasha Abdelraof, Marina George, Antoine Fakhri

Abstract:

Background: β-Thalassemia major (TM) and sickle cell disease (SCD) are inherited hemoglobin disorders resulting in chronic hemolytic anemia. Cardiovascular involvement is an important cause of morbidity and mortality in these groups of patients. The narrow border is between overt myocardial dysfunction and clinically silent left ventricular (LV) and / or right ventricular (RV) dysfunction in those patients. 3 D Speckle tracking echocardiography (3D STE) is a novel method for the detection of subclinical myocardial involvement. We aimed to study myocardial affection in SCD and TM using 3D STE, comparing it with conventional echocardiography, correlate it with serum ferritin level and lactate dehydrogenase (LDH). Methodology: Thirty SCD and thirty β TM patients, age range 4-18 years, were compared to 30 healthy age and sex matched control group. Cases were subjected to clinical examination, laboratory measurement of hemoglobin level, serum ferritin, and LDH. Transthoracic color Doppler echocardiography, 3D STE, tissue Doppler echocardiography, and aortic speckle tracking were performed. Results: significant reduction in global longitudinal strain (GLS), global circumferential strain (GCS), and global area strain (GAS) in SCD and TM than control (P value <0.001) there was significantly lower aortic speckle tracking in patients with TM and SCD than control (P value< 0.001). LDH was significantly higher in SCD than both TM and control and it correlated significantly positive mitral inflow E, (p value:0.022 and 0.072. r: 0.416 and -0.333 respectively) lateral E/E’ (p value.<0.001and 0.818. r. 0.618 and -0. 044.respectively) and septal E/E’ (p value 0.007 and 0.753& r value 0.485 and -0.060 respectively) in SCD but not TM and significant negative correlation between LDH and aortic root speckle tracking (value 0.681& r. -0.078.). The potential diagnostic accuracy of LDH in predicting vascular dysfunction as represented by aortic root GCS with a sensitivity 74% and aortic root GCS was predictive of LV dysfunction in SCD patients with sensitivity 100% Conclusion: 3D STE LV and RV systolic dysfunction in spite of their normal values by conventional echocardiography. SCD showed significantly lower right ventricular dysfunction and aortic root GCS than TM and control. LDH can be used to screen patients for cardiac dysfunction in SCD, not in TM

Keywords: thalassemia major, sickle cell disease, 3d speckle tracking echocardiography, LDH

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6479 Effect of Far Infrared and Endothelial Cell Growth Supplement on Human Umbilical Vascular Endothelial Cells

Authors: Ming-Tzu Tsai, Jui-Ting Hsu, Chia-Chieh Lin, Feng-Tsai Chiang, Cheng-Chin Huang

Abstract:

Far infrared (FIR), an invisible and short electromagnetic waves ranges from 6-14 μm also defines as the “growth ray.” Although the mechanism of FIR is still unknown, most data have suggested that FIR could accelerate the skin microcirculation by elevating the blood flow and nitric-oxide (NO) synthesis. In this present work, the effect of FIR irradiation and endothelial cell growth supplement (ECGS) on human umbilical vascular endothelial cells (HUVECs) was evaluated. To understand whether the cell viability and NO production of HUVECs affected by NO, cells with/without ECGS were treated in the presence or absence of L-NAME, an eNOS inhibitor. For FIR exposure, FIR-emitted ceramic powders consisted of a variety of well-mixed metal oxides were developed. The results showed that L-NAME did had a strong effect on the inhibition of NO production, especially in the ECGS-treated group. However, the cell viability of each group was rarely affected in the presence of L-NAME. Cells with the incubation of ECGS showed much higher cell viability compared to the control. Moreover, NO production of HUVECs exposed to FIR irradiation was significantly inhibited in the presence of L-NAME. It suggested that NO could play a role modulating the downstream signals of HUVECs during FIR exposure.

Keywords: far-infrared irradiation (FIR), nitric oxide (NO), endothelial nitric oxide synthase (eNOS), endothelial cell growth supplement (ECGS)

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6478 Biophysical Consideration in the Interaction of Biological Cell Membranes with Virus Nanofilaments

Authors: Samaneh Farokhirad, Fatemeh Ahmadpoor

Abstract:

Biological membranes are constantly in contact with various filamentous soft nanostructures that either reside on their surface or are being transported between the cell and its environment. In particular, viral infections are determined by the interaction of viruses (such as filovirus) with cell membranes, membrane protein organization (such as cytoskeletal proteins and actin filament bundles) has been proposed to influence the mechanical properties of lipid membranes, and the adhesion of filamentous nanoparticles influence their delivery yield into target cells or tissues. The goal of this research is to integrate the rapidly increasing but still fragmented experimental observations on the adhesion and self-assembly of nanofilaments (including filoviruses, actin filaments, as well as natural and synthetic nanofilaments) on cell membranes into a general, rigorous, and unified knowledge framework. The global outbreak of the coronavirus disease in 2020, which has persisted for over three years, highlights the crucial role that nanofilamentbased delivery systems play in human health. This work will unravel the role of a unique property of all cell membranes, namely flexoelectricity, and the significance of nanofilaments’ flexibility in the adhesion and self-assembly of nanofilaments on cell membranes. This will be achieved utilizing a set of continuum mechanics, statistical mechanics, and molecular dynamics and Monte Carlo simulations. The findings will help address the societal needs to understand biophysical principles that govern the attachment of filoviruses and flexible nanofilaments onto the living cells and provide guidance on the development of nanofilament-based vaccines for a range of diseases, including infectious diseases and cancer.

Keywords: virus nanofilaments, cell mechanics, computational biophysics, statistical mechanics

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6477 PARP1 Links Transcription of a Subset of RBL2-Dependent Genes with Cell Cycle Progression

Authors: Ewelina Wisnik, Zsolt Regdon, Kinga Chmielewska, Laszlo Virag, Agnieszka Robaszkiewicz

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Apart from protecting genome, PARP1 has been documented to regulate many intracellular processes inter alia gene transcription by physically interacting with chromatin bound proteins and by their ADP-ribosylation. Our recent findings indicate that expression of PARP1 decreases during the differentiation of human CD34+ hematopoietic stem cells to monocytes as a consequence of differentiation-associated cell growth arrest and formation of E2F4-RBL2-HDAC1-SWI/SNF repressive complex at the promoter of this gene. Since the RBL2 complexes repress genes in a E2F-dependent manner and are widespread in the genome in G0 arrested cells, we asked (a) if RBL2 directly contributes to defining monocyte phenotype and function by targeting gene promoters and (b) if RBL2 controls gene transcription indirectly by repressing PARP1. For identification of genes controlled by RBL2 and/or PARP1,we used primer libraries for surface receptors and TLR signaling mediators, genes were silenced by siRNA or shRNA, analysis of gene promoter occupation by selected proteins was carried out by ChIP-qPCR, while statistical analysis in GraphPad Prism 5 and STATISTICA, ChIP-Seq data were analysed in Galaxy 2.5.0.0. On the list of 28 genes regulated by RBL2, we identified only four solely repressed by RBL2-E2F4-HDAC1-BRM complex. Surprisingly, 24 out of 28 emerged genes controlled by RBL2 were co-regulated by PARP1 in six different manners. In one mode of RBL2/PARP1 co-operation, represented by MAP2K6 and MAPK3, PARP1 was found to associate with gene promoters upon RBL2 silencing, which was previously shown to restore PARP1 expression in monocytes. PARP1 effect on gene transcription was observed only in the presence of active EP300, which acetylated gene promoters and activated transcription. Further analysis revealed that PARP1 binding to MA2K6 and MAPK3 promoters enabled recruitment of EP300 in monocytes, while in proliferating cancer cell lines, which actively transcribe PARP1, this protein maintained EP300 at the promoters of MA2K6 and MAPK3. Genome-wide analysis revealed a similar distribution of PARP1 and EP300 around transcription start sites and the co-occupancy of some gene promoters by PARP1 and EP300 in cancer cells. Here, we described a new RBL2/PARP1/EP300 axis which controls gene transcription regardless of the cell type. In this model cell, cycle-dependent transcription of PARP1 regulates expression of some genes repressed by RBL2 upon cell cycle limitation. Thus, RBL2 may indirectly regulate transcription of some genes by controlling the expression of EP300-recruiting PARP1. Acknowledgement: This work was financed by Polish National Science Centre grants nr DEC-2013/11/D/NZ2/00033 and DEC-2015/19/N/NZ2/01735. L.V. is funded by the National Research, Development and Innovation Office grants GINOP-2.3.2-15-2016-00020 TUMORDNS, GINOP-2.3.2-15-2016-00048-STAYALIVE and OTKA K112336. AR is supported by Polish Ministry of Science and Higher Education 776/STYP/11/2016.

Keywords: retinoblastoma transcriptional co-repressor like 2 (RBL2), poly(ADP-ribose) polymerase 1 (PARP1), E1A binding protein p300 (EP300), monocytes

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6476 Protective Role of Autophagy Challenging the Stresses of Type 2 Diabetes and Dyslipidemia

Authors: Tanima Chatterjee, Maitree Bhattacharyya

Abstract:

The global challenge of type 2 diabetes mellitus is a major health concern in this millennium, and researchers are continuously exploring new targets to develop a novel therapeutic strategy. Type 2 diabetes mellitus (T2DM) is often coupled with dyslipidemia increasing the risks for cardiovascular (CVD) complications. Enhanced oxidative and nitrosative stresses appear to be the major risk factors underlying insulin resistance, dyslipidemia, β-cell dysfunction, and T2DM pathogenesis. Autophagy emerges to be a promising defense mechanism against stress-mediated cell damage regulating tissue homeostasis, cellular quality control, and energy production, promoting cell survival. In this study, we have attempted to explore the pivotal role of autophagy in T2DM subjects with or without dyslipidemia in peripheral blood mononuclear cells and insulin-resistant HepG2 cells utilizing flow cytometric platform, confocal microscopy, and molecular biology techniques like western blotting, immunofluorescence, and real-time polymerase chain reaction. In the case of T2DM with dyslipidemia higher population of autophagy, positive cells were detected compared to patients with the only T2DM, which might have resulted due to higher stress. Autophagy was observed to be triggered both by oxidative and nitrosative stress revealing a novel finding of our research. LC3 puncta was observed in peripheral blood mononuclear cells and periphery of HepG2 cells in the case of the diabetic and diabetic-dyslipidemic conditions. Increased expression of ATG5, LC3B, and Beclin supports the autophagic pathway in both PBMC and insulin-resistant Hep G2 cells. Upon blocking autophagy by 3-methyl adenine (3MA), the apoptotic cell population increased significantly, as observed by caspase‐3 cleavage and reduced expression of Bcl2. Autophagy has also been evidenced to control oxidative stress-mediated up-regulation of inflammatory markers like IL-6 and TNF-α. To conclude, this study elucidates autophagy to play a protective role in the case of diabetes mellitus with dyslipidemia. In the present scenario, this study demands to have a significant impact on developing a new therapeutic strategy for diabetic dyslipidemic subjects by enhancing autophagic activity.

Keywords: autophagy, apoptosis, dyslipidemia, reactive oxygen species, reactive nitrogen species, Type 2 diabetes

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6475 Characterization of Fe Doped ZnO Synthesised by Sol-Gel and Combustion Routes

Authors: M. Ravindiran, P. Shankar

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This paper deals with the comparison of two synthesis methods, namely, sol-gel, and combustion to prepare Fe doped ZnO nano material. Characterization results for structural, optical and magnetic properties were analyzed for the sol gel and combustion synthesis derived materials. Magnetic studies of the prepared compounds reveal that the combustion synthesis derived material has good magnetization of 50 emu/gm with a better hysteresis loop curve.

Keywords: DMS, combustion, ferromagnetic, synthesis methods

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