Search results for: molecular diagnostics
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2369

Search results for: molecular diagnostics

599 Acupoint Injection of High Concentration of Glucose Attenuates Mice Chronic Pain and Depression Comorbidity

Authors: Chanya Inprasit, Yi-Wen Lin

Abstract:

Inflammation causes changes of peripheral and central nervous system properties, affecting both neuronal and non-neuronal cells, resulting in inflammatory pain. Acupoint injection (AI) was developed in the 1950s and has been widely used for relieving pain. It is an acupoint-stimulating technique that utilizes anatomically based meridians derived from Chinese medicine theory. AI has been accepted as an effective treatment and is thought to display superior results when compared to traditional acupuncture methods. However, the mechanism of AI needs to be ratified by more scientific evidence in order to support the theory and its therapeutic development. In this study, we explored the effect of AI on the comorbidity of chronic pain and depression. Mice hindpaw was injected by complete Freund’s adjuvant (CFA) to induce the condition of chronic pain. Measurements of mechanical and thermal hyperalgesia and depression-like behavior were analyzed. The results indicated a positive tendency to AI treatment. The comorbidity of chronic pain and depression was investigated with relation to transient receptor potential V1 (TRPV1) mechanism through the use of TRPV1 gene deletion. The expression of nociceptors such as voltage-gated sodium channels (Navs) or TRPV1, was significantly down-regulated by AI. The expression of inflammation-activated molecules: astrocytic marker glial fibrillary acidic protein (GFAP), the microglial marker Iba-1, S100B, and related kinases, were reversed by AI in both the peripheral and central nervous system. Taken together, these data provided a detailed molecular mechanism of AI-induced analgesia and anti-inflammatory properties. This finding may be utilized for clinical practice to treat chronic pain and depression comorbidity.

Keywords: inflammatory pain, acupoint injection, TRPV1, GFAP, S100B

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598 Sol-Gel Coated Fabric for Controlled Release of Mosquito Repellent

Authors: Bhaskar M. Murai, Neeraj Banchor, Ishveen Chabbra, Madhusudhan Nadgir, S. Vidhya

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Sol-gel technology combined with electronics and biochemistry helps to overcome the problems caused by mosquitoes by developing a portable, low-cost device which enables controlled release of trapped compound inside it. It is a wet-chemical technique which is used primarily for fabrication of silicate gel which is usually allowed to dry as per requirement. The outcome is solid rock hard material which is porous and has lots of applications in different fields. Taking porosity as a key factor, allethrin a naturally occurring synthetic compound with molecular mass 302.40 was entrapped inside the sol-gel matrix as a dopant. Allethrin is commonly used as an insecticide and is a key ingredient in commercially available mosquitoes repellent in Asian and subtropical countries. It has low toxicity for humans and birds, and are used in many household insecticides such as RAID as well as mosquito coils. They are however highly toxic to fish and bees. Insects subject to its exposure become paralyzed (nervous system effect) before dying. They are also used as an ultra-low volume spray for outdoor mosquito control. Therefore, there is a need for controlled release of allethrin in the environment. For controlled release of allethrin from sol-gel matrix, its (allethrin) we utilized temperature based controlled evaporation through porous sol-gel. Different types of fabric like cotton, Terri-cotton, polyester, surgical cap, knee-cap etc are studied and the best with maximum absorption capacity is selected to hold the sol-gel matrix with maximum quantity. For sol-gel coating 2 x 2cm cloth pieces are dipped in sol-gel solution for 10 minutes and by calculating the weight difference we concluded that Terri cotton is best suitable for our project. An electronic circuit with heating plate is developed in to test the controlled release of compound. An oscillatory circuit is used to produce the required heat.

Keywords: sol-gel, allethrin, TEOS, biochemistry

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597 Synthesis of Fullerene Nanorods for Detection of Ethylparaben an Endocrine Disruptor in Cosmetics

Authors: Jahangir Ahmad Rather, Emad A. Khudaish, Ahsanulhaq Qurashi, Palanisamy Kannan

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Chemical modification and assembling of fullerenes are fundamentally important for the application of fullerenes as functional molecules and in molecular devices and organic electronic devices. We have synthesized fullerene nanorods C60NRs conjugate via liquid-liquid interface and the synthesized C60NRs was characterized by FTIR spectroscopy, field emission electron microscopy (FESEM) and X-ray diffraction techniques. The C60NRs were immobilized on glassy carbon electrode via surface bound diazonium salts as an impact strategy. This method involves electrografting of p–nitrophenyl to give GCE–Ph–NO2 and then the terminal nitro-group was chemically reduced to GCE–Ph–NH2 in a presence of sodium borohydride/gold–polyaniline nanocomposite (NaBH4/Au–PANI). The Au–PANI composite was synthesized and characterized by FTIR, UV-vis, SEM and EDX techniques. The C60NRs were immobilized on GCE–Ph–NH2 via amination reaction which involves N-H addition across a π-bond on [60] fullerene. The immobilized C60NRs/GCE was subjected to electrochemical reduction in 1.0 M KOH to yield ERC60NRs/GCE sensor. The developed sensor shows high electrocatalytic activity for the detection of ethylparaben (EP) over a concentration range from 0.01 to 0.52 µM with a detection limit (LOD) 3.8 nM. The amount of EP present in the nourishing repair cream (OlAY®) was determined by standard addition method at the developed ERC60NRs/GCE sensor. The total concentration of EP was found to be 0.011 µM (0.1%) and is within the permissible limit of 0.19 % EP in cosmetics according to the European scientific committee (SCCS) on consumer safety on 22 March 2011 (SCCS/1348/11).

Keywords: diazonium salt reduction, ethylparaben (EP), endocrine disruptor, fullerene nanorods (C60NRs), gold–polyaniline nanocomposite (Au–PANI)

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596 Physical and Microbiological Evaluation of Chitosan Films: Effect of Essential Oils and Storage

Authors: N. Valderrama, W. Albarracín, N. Algecira

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It was studied the effect of the inclusion of thyme and rosemary essential oils into chitosan films, as well as the microbiological and physical properties when storing chitosan film with and without the mentioned inclusion. The film forming solution was prepared by dissolving chitosan (2%, w/v), polysorbate 80 (4% w/w CH) and glycerol (16% w/w CH) in aqueous lactic acid solutions (control). The thyme (TEO) and rosemary (REO) essential oils (EOs) were included 1:1 w/w (EOs:CH) on their combination 50/50 (TEO:REO). The films were stored at temperatures of 5, 20, 33°C and a relative humidity of 75% during four weeks. The films with essential oil inclusion did not show an antimicrobial activity against strains. This behavior could be explained because the chitosan only inhibits the growth of microorganisms in direct contact with the active sites. However, the inhibition capacity of TEO was higher than the REO and a synergic effect between TEO:REO was found for S. enteritidis strains in the chitosan solution. Some physical properties were modified by the inclusion of essential oils. The addition of essential oils does not affect the mechanical properties (tensile strength, elongation at break, puncture deformation), the water solubility, the swelling index nor the DSC behavior. However, the essential oil inclusion can significantly decrease the thickness, the moisture content, and the L* value of films whereas the b* value increased due to molecular interactions between the polymeric matrix, the loosing of the structure, and the chemical modifications. On the other hand, the temperature and time of storage changed some physical properties on the chitosan films. This could have occurred because of chemical changes, such as swelling in the presence of high humidity air and the reacetylation of amino groups. In the majority of cases, properties such as moisture content, tensile strength, elongation at break, puncture deformation, a*, b*, chrome, ΔE increased whereas water resistance, swelling index, L*, and hue angle decreased.

Keywords: chitosan, food additives, modified films, polymers

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595 Combining Transcriptomics, Bioinformatics, Biosynthesis Networks and Chromatographic Analyses for Cotton Gossypium hirsutum L. Defense Volatiles Study

Authors: Ronald Villamar-Torres, Michael Staudt, Christopher Viot

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Cotton Gossypium hirsutum L. is one of the most important industrial crops, producing the world leading natural textile fiber, but is very prone to arthropod attacks that reduce crop yield and quality. Cotton cultivation, therefore, makes an outstanding use of chemical pesticides. In reaction to herbivorous arthropods, cotton plants nevertheless show natural defense reactions, in particular through volatile organic compounds (VOCs) emissions. These natural defense mechanisms are nowadays underutilized but have a very high potential for cotton cultivation, and elucidating their genetic bases will help to improve their use. Simulating herbivory attacks by mechanical wounding of cotton plants in greenhouse, we studied by qPCR the changes in gene expression for genes of the terpenoids biosynthesis pathway. Differentially expressed genes corresponded to higher levels of the terpenoids biosynthesis pathway and not to enzymes synthesizing particular terpenoids. The genes were mapped on the G. hirsutum L. reference genome; their global relationships inside the general metabolic pathways and the biosynthesis of secondary metabolites were visualized with iPath2. The chromatographic profiles of VOCs emissions indicated first monoterpenes and sesquiterpenes emissions, dominantly four molecules known to be involved in plant reactions to arthropod attacks. As a result, the study permitted to identify potential key genes for the emission of volatile terpenoids by cotton plants in reaction to an arthropod attack, opening possibilities for molecular-assisted cotton breeding in benefit of smallholder cotton growers.

Keywords: biosynthesis pathways, cotton, mechanisms of plant defense, terpenoids, volatile organic compounds

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594 Assessing Diagnostic and Evaluation Tools for Use in Urban Immunisation Programming: A Critical Narrative Review and Proposed Framework

Authors: Tim Crocker-Buque, Sandra Mounier-Jack, Natasha Howard

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Background: Due to both the increasing scale and speed of urbanisation, urban areas in low and middle-income countries (LMICs) host increasingly large populations of under-immunized children, with the additional associated risks of rapid disease transmission in high-density living environments. Multiple interdependent factors are associated with these coverage disparities in urban areas and most evidence comes from relatively few countries, e.g., predominantly India, Kenya, Nigeria, and some from Pakistan, Iran, and Brazil. This study aimed to identify, describe, and assess the main tools used to measure or improve coverage of immunisation services in poor urban areas. Methods: Authors used a qualitative review design, including academic and non-academic literature, to identify tools used to improve coverage of public health interventions in urban areas. Authors selected and extracted sources that provided good examples of specific tools, or categories of tools, used in a context relevant to urban immunization. Diagnostic (e.g., for data collection, analysis, and insight generation) and programme tools (e.g., for investigating or improving ongoing programmes) and interventions (e.g., multi-component or stand-alone with evidence) were selected for inclusion to provide a range of type and availability of relevant tools. These were then prioritised using a decision-analysis framework and a tool selection guide for programme managers developed. Results: Authors reviewed tools used in urban immunisation contexts and tools designed for (i) non-immunization and/or non-health interventions in urban areas, and (ii) immunisation in rural contexts that had relevance for urban areas (e.g., Reaching every District/Child/ Zone). Many approaches combined several tools and methods, which authors categorised as diagnostic, programme, and intervention. The most common diagnostic tools were cross-sectional surveys, key informant interviews, focus group discussions, secondary analysis of routine data, and geographical mapping of outcomes, resources, and services. Programme tools involved multiple stages of data collection, analysis, insight generation, and intervention planning and included guidance documents from WHO (World Health Organisation), UNICEF (United Nations Children's Fund), USAID (United States Agency for International Development), and governments, and articles reporting on diagnostics, interventions, and/or evaluations to improve urban immunisation. Interventions involved service improvement, education, reminder/recall, incentives, outreach, mass-media, or were multi-component. The main gaps in existing tools were an assessment of macro/policy-level factors, exploration of effective immunization communication channels, and measuring in/out-migration. The proposed framework uses a problem tree approach to suggest tools to address five common challenges (i.e. identifying populations, understanding communities, issues with service access and use, improving services, improving coverage) based on context and available data. Conclusion: This study identified many tools relevant to evaluating urban LMIC immunisation programmes, including significant crossover between tools. This was encouraging in terms of supporting the identification of common areas, but problematic as data volumes, instructions, and activities could overwhelm managers and tools are not always suitably applied to suitable contexts. Further research is needed on how best to combine tools and methods to suit local contexts. Authors’ initial framework can be tested and developed further.

Keywords: health equity, immunisation, low and middle-income countries, poverty, urban health

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593 Antifungal Susceptibility of Saprolegnia parasitica Isolated from Rainbow Trout and Its Host Pathogen Interaction in Zebrafish Disease Model

Authors: Sangyeop Shin, D. C. M. Kulatunga, S. H. S. Dananjaya, Chamilani Nikapitiya, Jehee Lee, Mahanama De Zoysa

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Saprolegniasis is one of the most devastating fungal diseases in freshwater fish which is caused by species in the genus Saprolegnia including Saprolegnia parasitica. In this study, we isolated the strain of S. parasitica from diseased rainbow trout in Korea. Morphological and molecular based identification confirmed that isolated fungi belong to the member of S. parasitica, supported by its typical fungal features including cotton-like whitish mycelium, zoospores (primary and secondary) and phylogenetic analysis with internal transcribed spacer (ITS) region. Pathogenicity of isolated S. parasitica was developed in embryo, larvae, juvenile and adult zebrafish as a disease model. Up regulation of host genes encoding ZfTnf-α, Zfc-Rel, ZfIl-12, ZfLyz-c, Zfβ-def, and ZfHsp-70 was identified in zebrafish larvae after experimental challenge of S. parasitica showing the host immune responses against the S. parasitica. Survival of the juveniles upon fungal infection might be due to the increased immune protection in the host. Investigation of antifungal susceptibility of S. parasitica with natural lawsone (2-hydroxy-1,4-naphthoquinone) revealed the minimum inhibitory concentration (MIC) and percentage inhibition of radial growth (PIRG %) as 200 µg/mL and 31.8%, respectively. Lawsone was able to change the membrane permeability, and cause irreversible damage and disintegration to the cellular membranes of S. parasitica which might have effect on fungi growth inhibition. Moreover, the mycelium exposed to lawsone (MIC level) changed the transcriptional responses of S. parasitica genes. Overall results indicate that lawsone could be a potential and novel anti-S. parasitica agent for controlling S. parasitica infection.

Keywords: host-pathogen interactions, lawsone, rainbow trout, Saprolegnia parasitica, Saprolegniasis, zebrafish

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592 Developing a Cloud Intelligence-Based Energy Management Architecture Facilitated with Embedded Edge Analytics for Energy Conservation in Demand-Side Management

Authors: Yu-Hsiu Lin, Wen-Chun Lin, Yen-Chang Cheng, Chia-Ju Yeh, Yu-Chuan Chen, Tai-You Li

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Demand-Side Management (DSM) has the potential to reduce electricity costs and carbon emission, which are associated with electricity used in the modern society. A home Energy Management System (EMS) commonly used by residential consumers in a down-stream sector of a smart grid to monitor, control, and optimize energy efficiency to domestic appliances is a system of computer-aided functionalities as an energy audit for residential DSM. Implementing fault detection and classification to domestic appliances monitored, controlled, and optimized is one of the most important steps to realize preventive maintenance, such as residential air conditioning and heating preventative maintenance in residential/industrial DSM. In this study, a cloud intelligence-based green EMS that comes up with an Internet of Things (IoT) technology stack for residential DSM is developed. In the EMS, Arduino MEGA Ethernet communication-based smart sockets that module a Real Time Clock chip to keep track of current time as timestamps via Network Time Protocol are designed and implemented for readings of load phenomena reflecting on voltage and current signals sensed. Also, a Network-Attached Storage providing data access to a heterogeneous group of IoT clients via Hypertext Transfer Protocol (HTTP) methods is configured to data stores of parsed sensor readings. Lastly, a desktop computer with a WAMP software bundle (the Microsoft® Windows operating system, Apache HTTP Server, MySQL relational database management system, and PHP programming language) serves as a data science analytics engine for dynamic Web APP/REpresentational State Transfer-ful web service of the residential DSM having globally-Advanced Internet of Artificial Intelligence (AI)/Computational Intelligence. Where, an abstract computing machine, Java Virtual Machine, enables the desktop computer to run Java programs, and a mash-up of Java, R language, and Python is well-suited and -configured for AI in this study. Having the ability of sending real-time push notifications to IoT clients, the desktop computer implements Google-maintained Firebase Cloud Messaging to engage IoT clients across Android/iOS devices and provide mobile notification service to residential/industrial DSM. In this study, in order to realize edge intelligence that edge devices avoiding network latency and much-needed connectivity of Internet connections for Internet of Services can support secure access to data stores and provide immediate analytical and real-time actionable insights at the edge of the network, we upgrade the designed and implemented smart sockets to be embedded AI Arduino ones (called embedded AIduino). With the realization of edge analytics by the proposed embedded AIduino for data analytics, an Arduino Ethernet shield WizNet W5100 having a micro SD card connector is conducted and used. The SD library is included for reading parsed data from and writing parsed data to an SD card. And, an Artificial Neural Network library, ArduinoANN, for Arduino MEGA is imported and used for locally-embedded AI implementation. The embedded AIduino in this study can be developed for further applications in manufacturing industry energy management and sustainable energy management, wherein in sustainable energy management rotating machinery diagnostics works to identify energy loss from gross misalignment and unbalance of rotating machines in power plants as an example.

Keywords: demand-side management, edge intelligence, energy management system, fault detection and classification

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591 Spectrum and Prevalence of Candida Infection in Diabetic Foot Ulcers

Authors: Seyed Reza Aghili, Tahereh Shokohi, Lotfollah Davoodi, Zahra Kashi, Azam Moslemi, Mahdi Abastabar, Iman Haghani, Sabah Mayahi, Asoudeh A.

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Introduction: In diabetic foot ulcers, if fungal agents such as Candida species penetrate into the cutaneous or depth of ulcer, can increase the degree of the wound and cause Candia infection and make it more difficult to heal. Material & Methods: A cross-sectional study was performed on 100 diabetic foot ulcer patients in 2020 in Sari, Iran. patient's data and wound grade were recorded in a questionnaire. Candida infection was diagnosed with direct microscopic examination and culture of samples. Colony-PCR molecular method was used for ITS region of DNA and then PCR-RFLP with Msp1 enzyme and using HWP1 specific gene to determine species of Candida agent. Results: Of 100 patients, the mean age 62.1 ± 10.8 years, 95% type 2 diabetes, 83%>10 years duration diabetes, 59% male, 66%> poor education level, 99% married, 52% rural, 95% neuropathic symptoms, 88% using antibiotics, 69%HbA1C >9%, and mean ulcer degree 2.6±1.05 were. Candida infection was seen in 13% of the deep tissue of the wound and 7% cutaneous around the wound. The predominant Candida isolated was C. parapsilosis (71.5%), C .albicans (14.3%). Fungal infections caused by mold fungi were not detected. There was a statistically significant relationship between yeast infection and gender, rural, HbA1C and ulcer degree. Conclusion: Mycological evaluations often are ignored. Candida parapsilosis is the most common infectious agent in these patients and may require specific treatment. Therefore, more attention of physicians to Candida infections particularly, early diagnosis and effective treatment can help faster recovery and prevent amputation.

Keywords: diabetic foot ulcer, candida infection, risk factors, c. parapsilosis

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590 Prognostic Value of Tumor Markers in Younger Patients with Breast Cancer

Authors: Lola T. Alimkhodjaeva, Lola T. Zakirova, Soniya S. Ziyavidenova

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Background: Breast cancer occupies the first place among the cancer in women in the world. It is urgent today to study the role of molecular markers which are capable of predicting the dynamics and outcome of the disease. The aim of this study is to define the prognostic value of the content of estrogen receptor (ER), progesterone receptor (PgR), and amplification of HER-2 / neu oncoprotein by studying 3 and 5-year overall and relapse-free survival in 470 patients with primary operable and 280 patients with locally–advanced breast cancer. Materials and methods: Study results of 3 and 5-year overall and relapse-free survival, depending on the content of RE, PgR in primary operable patients showed that ER positive (+) and PgR (+) survival was 100 (96.2%) and 97.3 (94.6%), for ER negative (-) and PgR (-) - 69.2 (60.3%) and 65.4 (57.7%), for ER positive (+) and negative PgR (-) 87.4 (80.1%) and 81.5 (79.3%), for ER negative (-) and positive PgR (+) - 97.4 (93.4%) and 90.4 (88.5%), respectively. Survival results depended also on the level of HER-2 / neu expression. In patients with HER-2 / neu negative the survival rates were as follows: 98.6 (94.7%) and 96.2 (92.3%). In group of patients with the level of HER-2 / neu (2+) expression these figures were: 45.3 (44.3%) and 45.1 (40.2%), and in group of patients with the level of HER-2 / neu (3+) expression - 41.2 (33.1%) and 34.3 (29.4%). The combination of ER negative (-), PgR (-), HER-2 / neu (-) they were 27.2 (25.4%) and 19.5 (15.3%), respectively. In patients with locally-advanced breast cancer the results of 3 and 5-year OS and RFS for ER (+) and PgR (+) were 76.3 (69.3%) and 62.2 (61.4%), for ER (-) and RP (-) 29.1 (23.7%) and 18.3 (12.6%), for ER (+) and PgR (-) 61.2 (47.2%) and 39.4 (25.6%), for ER (-) and PgR (+) 54.3 (43.1%) and 41.3 (18.3%), respectively. The level of HER-2 / neu expression also affected the survival results. Therefore, in HER-2/ neu negative patients the survival rate was 74.1 (67.6%) and 65.1 (57.3%), with the level of expression (2+) 20.4 (14.2%) and 8.6 (6.4%), with the level of expression (3+) 6.2 (3.1%) and 1.2 (1.5%), respectively. The combination for ER, PgR, HER-2 / neu negative was 22.1 (14.3%) and 8.4 (1.2%). Conclusion: Thus, the presence of steroid hormone receptors in breast tumor tissues at primary operable and locally- advanced process as the lack of HER-2/neu oncoprotein correlates with the highest rates of 3- and 5-year overall and relapse-free survival. The absence of steroid hormone receptors as well as of HER-2/neu overexpression in malignant breast tissues significantly degrades the 3- and 5-year overall and relapse-free survival. Tumors with ER, PgR and HER-2/neu negative have the most unfavorable prognostics.

Keywords: breast cancer, estrogen receptor, oncoprotein, progesterone receptor

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589 Organic Geochemistry of the Late Cenomanian–Early Turonian Source Rock in Central and Northern Tunisia

Authors: Belhaj Mohamed, M. Saidi, I. Bouazizi, M. Soussi, M. Ben Jrad

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The Late Cenomanian-Early Turonian laminated, black, organic-rich limestones were described in Central Tunisia and attributed to the Bahloul Formation. It covers central and northern Tunisia, and the northern part of the Gulf of Gabes. The Bahloul Formation is considered as one of the main source rocks in Tunisia and is composed of outer-shelf to slop-laminated and dark-gray to black-colored limestones and marls. This formation had been deposited in a relatively deep-marine, calm, and anoxic environment. Rock-Eval analysis and vitrinite reflectance (Ro) measurements were performed on the basis of the organic carbon content. Several samples were chosen for molecular organic geochemistry. Saturate and aromatic hydrocarbons were analyzed by gas chromatography (GC) and GC–mass spectrometry. Geochemical data of the Bahloul Formation in northern and central Tunisia show this level to be a good potential source rock as indicated by the high content of type II organic matter. This formation exhibits high total organic carbon contents (as much as 14%), with an average value of 2% and a good to excellent petroleum potential, ranging between 2 and 50 kg of hydrocarbon/ton of rock. The extracts from the Bahloul Formation are characterized by Pr/Ph ratios ranging between 1.5 and 3, a moderate diasterane content, a C27 sterane approximately equal to C29 sterane, a high C28/C29 ratio, low gammacerane index, a C35/C34 homohopane ratio less than 1 and carbon isotope compositions between -24 and -26‰. The thermal maturity is relatively low, corresponding to the beginning of the oil window in the western area near the Algerian border, in the oil window in the eastern area (Sahel basin) and late mature in northern part.

Keywords: biomarkers, organic geochemistry, source rock, Tunisia

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588 Integrative Transcriptomic Profiling of NK Cells and Monocytes: Advancing Diagnostic and Therapeutic Strategies for COVID-19

Authors: Salma Loukman, Reda Benmrid, Najat Bouchmaa, Hicham Hboub, Rachid El Fatimy, Rachid Benhida

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In this study, it use integrated transcriptomic datasets from the GEO repository with the purpose of investigating immune dysregulation in COVID-19. Thus, in this context, we decided to be focused on NK cells and CD14+ monocytes gene expression, considering datasets GSE165461 and GSE198256, respectively. Other datasets with PBMCs, lung, olfactory, and sensory epithelium and lymph were used to provide robust validation for our results. This approach gave an integrated view of the immune responses in COVID-19, pointing out a set of potential biomarkers and therapeutic targets with special regard to standards of physiological conditions. IFI27, MKI67, CENPF, MBP, HBA2, TMEM158, THBD, HBA1, LHFPL2, SLA, and AC104564.3 were identified as key genes from our analysis that have critical biological processes related to inflammation, immune regulation, oxidative stress, and metabolic processes. Consequently, such processes are important in understanding the heterogeneous clinical manifestations of COVID-19—from acute to long-term effects now known as 'long COVID'. Subsequent validation with additional datasets consolidated these genes as robust biomarkers with an important role in the diagnosis of COVID-19 and the prediction of its severity. Moreover, their enrichment in key pathophysiological pathways presented them as potential targets for therapeutic intervention.The results provide insight into the molecular dynamics of COVID-19 caused by cells such as NK cells and other monocytes. Thus, this study constitutes a solid basis for targeted diagnostic and therapeutic development and makes relevant contributions to ongoing research efforts toward better management and mitigation of the pandemic.

Keywords: SARS-COV-2, RNA-seq, biomarkers, severity, long COVID-19, bio analysis

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587 Cas9-Assisted Direct Cloning and Refactoring of a Silent Biosynthetic Gene Cluster

Authors: Peng Hou

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Natural products produced from marine bacteria serve as an immense reservoir for anti-infective drugs and therapeutic agents. Nowadays, heterologous expression of gene clusters of interests has been widely adopted as an effective strategy for natural product discovery. Briefly, the heterologous expression flowchart would be: biosynthetic gene cluster identification, pathway construction and expression, and product detection. However, gene cluster capture using traditional Transformation-associated recombination (TAR) protocol is low-efficient (0.5% positive colony rate). To make things worse, most of these putative new natural products are only predicted by bioinformatics analysis such as antiSMASH, and their corresponding natural products biosynthetic pathways are either not expressed or expressed at very low levels under laboratory conditions. Those setbacks have inspired us to focus on seeking new technologies to efficiently edit and refractor of biosynthetic gene clusters. Recently, two cutting-edge techniques have attracted our attention - the CRISPR-Cas9 and Gibson Assembly. By now, we have tried to pretreat Brevibacillus laterosporus strain genomic DNA with CRISPR-Cas9 nucleases that specifically generated breaks near the gene cluster of interest. This trial resulted in an increase in the efficiency of gene cluster capture (9%). Moreover, using Gibson Assembly by adding/deleting certain operon and tailoring enzymes regardless of end compatibility, the silent construct (~80kb) has been successfully refactored into an active one, yielded a series of analogs expected. With the appearances of the novel molecular tools, we are confident to believe that development of a high throughput mature pipeline for DNA assembly, transformation, product isolation and identification would no longer be a daydream for marine natural product discovery.

Keywords: biosynthesis, CRISPR-Cas9, DNA assembly, refactor, TAR cloning

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586 Effect of a GABA/5-HTP Mixture on Behavioral Changes and Biomodulation in an Invertebrate Model

Authors: Kyungae Jo, Eun Young Kim, Byungsoo Shin, Kwang Soon Shin, Hyung Joo Suh

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Gamma-aminobutyric acid (GABA) and 5-hydroxytryptophan (5-HTP) are amino acids of digested nutrients or food ingredients and these can possibly be utilized as non-pharmacologic treatment for sleep disorder. We previously investigated the GABA/5-HTP mixture is the principal concept of sleep-promoting and activity-repressing management in nervous system of D. melanogaster. Two experiments in this study were designed to evaluate sleep-promoting effect of GABA/5-HTP mixture, to clarify the possible ratio of sleep-promoting action in the Drosophila invertebrate model system. Behavioral assays were applied to investigate distance traveled, velocity, movement, mobility, turn angle, angular velocity and meander of two amino acids and GABA/5-HTP mixture with caffeine treated flies. In addition, differentially expressed gene (DEG) analyses from next generation sequencing (NGS) were applied to investigate the signaling pathway and functional interaction network of GABA/5-HTP mixture administration. GABA/5-HTP mixture resulted in significant differences between groups related to behavior (p < 0.01) and significantly induced locomotor activity in the awake model (p < 0.05). As a result of the sequencing, the molecular function of various genes has relationship with motor activity and biological regulation. These results showed that GABA/5-HTP mixture administration significantly involved the inhibition of motor behavior. In this regard, we successfully demonstrated that using a GABA/5-HTP mixture modulates locomotor activity to a greater extent than single administration of each amino acid, and that this modulation occurs via the neuronal system, neurotransmitter release cycle and transmission across chemical synapses.

Keywords: sleep, γ-aminobutyric acid, 5-hydroxytryptophan, Drosophila melanogaster

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585 [Keynote Talk]: Three Dimensional Finite Element Analysis of Functionally Graded Radiation Shielding Nanoengineered Sandwich Composites

Authors: Nasim Abuali Galehdari, Thomas J. Ryan, Ajit D. Kelkar

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In recent years, nanotechnology has played an important role in the design of an efficient radiation shielding polymeric composites. It is well known that, high loading of nanomaterials with radiation absorption properties can enhance the radiation attenuation efficiency of shielding structures. However, due to difficulties in dispersion of nanomaterials into polymer matrices, there has been a limitation in higher loading percentages of nanoparticles in the polymer matrix. Therefore, the objective of the present work is to provide a methodology to fabricate and then to characterize the functionally graded radiation shielding structures, which can provide an efficient radiation absorption property along with good structural integrity. Sandwich structures composed of Ultra High Molecular Weight Polyethylene (UHMWPE) fabric as face sheets and functionally graded epoxy nanocomposite as core material were fabricated. A method to fabricate a functionally graded core panel with controllable gradient dispersion of nanoparticles is discussed. In order to optimize the design of functionally graded sandwich composites and to analyze the stress distribution throughout the sandwich composite thickness, a finite element method was used. The sandwich panels were discretized using 3-Dimensional 8 nodded brick elements. Classical laminate analysis in conjunction with simplified micromechanics equations were used to obtain the properties of the face sheets. The presented finite element model would provide insight into deformation and damage mechanics of the functionally graded sandwich composites from the structural point of view.

Keywords: nanotechnology, functionally graded material, radiation shielding, sandwich composites, finite element method

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584 Field Theories in Chiral Liquid Crystals: A Theory for Helicoids and Skyrmions

Authors: G. De Matteis, L. Martina, V. Turco

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The work is focused on determining and comparing special nonlinear static configurations in cholesteric liquid crystals (CLCs), confined between two parallel plates and in the presence of an external static electric/magnetic field. The solutions are stabilised by topological and non-topological conservation laws since they are described in terms of integrable or partially integrable nonlinear boundary value problems. In cholesteric liquid crystals which are subject to geometric frustration; anchoring conditions at boundaries, i.e., homeotropic conditions, are incompatible with the cholesteric twist. This aspect turns out to be essential in the admissible classes of solutions, allowing also for disclination type singularities. Within the framework of Frank-Oseen theory, we study the static configurations for CLCs. First, we find numerical solutions for isolated axisymmetric states in confined CLCs with weak homeotropic anchoring at the boundaries. These solutions describe 3-dimensional modulations, namely spherulites or cholesteric bubbles, actually observed in these systems, of standard baby skyrmions. Relations with well-known nonlinear integrable systems are found and are used to explore the asymptotic behavior of the solutions. Then we turn our attention to extended periodic static configurations called Helicoids or cholesteric fingers, described by an elliptic sine-Gordon model with appropriate boundary conditions, showing how their period and energies are determined by both the thickness of the cell and the intensity of the external electric/magnetic field. We explicitly show that helicoids with π or 2π of rotations of the molecular director are different in many aspects and are not simply algebraically related. The behaviour of the solutions, their energy and the properties of the associated disclinations are discussed in detail, both analytically and numerically.

Keywords: cholesteric liquid crystals, geometric frustration, helicoids, skyrmions

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583 Elastic Collisions of Electrons with DNA and Water From 10 eV to 100 KeV: Scar Macro Investigation

Authors: Aouina Nabila Yasmina, Zine El Abidine Chaoui

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Recently, understanding the interactions of electrons with the DNA molecule and its components has attracted considerable interest because DNA is the main site damaged by ionizing radiation. The interactions of radiation with DNA induce a variety of molecular damage such as single-strand breaks, double-strand breaks, basic damage, cross-links between proteins and DNA, and others, or the formation of free radicals, which, by chemical reactions with DNA, can also lead to breakage of the strand. One factor that can contribute significantly to these processes is the effect of water hydration on the formation and reaction of radiation induced by these radicals in and / or around DNA. B-DNA requires about 30% by weight of water to maintain its native conformation in the crystalline state. The transformation depends on various factors such as sequence, ion composition, concentration and water activity. Partial dehydration converts it to DNA-A. The present study shows the results of theoretical calculations for positrons and electrons elastic scattering with DNA medium and water over a broad energy range from 10 eV to 100 keV. Indeed, electron elastic cross sections and elastic mean free paths are calculated using a corrected form of the independent atom method, taking into account the geometry of the biomolecule (SCAR macro). Moreover, the elastic scattering of electrons and positrons by atoms of the biomolecule was evaluated by means of relativistic (Dirac) partial wave analysis. Our calculated results are compared with theoretical data available in the literature in the absence of experimental data, in particular for positron. As a central result, our electron elastic cross sections are in good agreement with existing theoretical data in the range of 10 eV to 1 keV.

Keywords: elastic cross scrion, elastic mean free path, scar macro method, electron collision

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582 Sensitivity Enhancement of Photonic Crystal Fiber Biosensor

Authors: Mohamed Farhat O. Hameed, Yasamin K. A. Alrayk, A. A Shaalan, S. S. A. Obayya

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The surface plasmon resonance (SPR) sensors are widely used due to its high sensitivity with molecular labels free. The commercial SPR sensors depend on the conventional prism-coupled configuration. However, this type of configuration suffers from miniaturization and integration. Therefore, the search for compact, portable and highly sensitive SPR sensors becomes mandatory.In this paper, sensitivity enhancement of a novel photonic crystal fiber biosensoris introduced and studied. The suggested design has microstructure of air holes in the core region surrounded by two large semicircular metallized channels filled with the analyte. The inner surfaces of the two channels are coated by a silver layer followed by a gold layer.The simulation results are obtained using full vectorial finite element methodwith perfect matched layer (PML) boundary conditions. The proposed design depends on bimetallic configuration to enhance the biosensor sensitivity. Additionally, the suggested biosensor can be used for multi-channel/multi-analyte sensing. In this study, the sensor geometrical parameters are studied to maximize the sensitivity for the two polarized modes. The numerical results show that high refractive index sensitivity of 4750 nm/RIU (refractive index unit) and 4300 nm/RIU can be achieved for the quasi (transverse magnetic) TM and quasi (transverse electric) TE modes of the proposed biosensor, respectively. The reportedbiosensor has advantages of integration of microfluidics setup, waveguide and metallic layers into a single structure. As a result, compact biosensor with better integration compared to conventional optical fiber SPR biosensors can be obtained.

Keywords: photonic crystal fibers, gold, silver, surface plasmon, biosensor

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581 Layer-by-Layer Modified Ceramic Membranes for Micropollutant Removal

Authors: Jenny Radeva, Anke-Gundula Roth, Christian Goebbert, Robert Niestroj-Pahl, Lars Daehne, Axel Wolfram, Juergen Wiese

Abstract:

Ceramic membranes for water purification combine excellent stability with long-life characteristics and high chemical resistance. Layer-by-Layer coating is a well-known technique for customization and optimization of filtration properties of membranes but is mostly used on polymeric membranes. Ceramic membranes comprising a metal oxide filtration layer of Al2O3 or TiO2 are charged and therefore highly suitable for polyelectrolyte adsorption. The high stability of the membrane support allows efficient backwash and chemical cleaning of the membrane. The presented study reports metal oxide/organic composite membrane with an increased rejection of bivalent salts like MgSO4 and the organic micropollutant Diclofenac. A self-build apparatus was used for applying the polyelectrolyte multilayers on the ceramic membrane. The device controls the flow and timing of the polyelectrolytes and washing solutions. As support for the Layer-by-Layer coat, ceramic mono-channel membranes were used with an inner capillary of 8 mm diameter, which is connected to the coating device. The inner wall of the capillary is coated subsequently with polycat- and anions. The filtration experiments were performed with a feed solution of MgSO4 and Diclofenac. The salt content of the permeate was detected conductometrically and Diclofenac was measured with UV-Adsorption. The concluded results show retention values of magnesium sulfate of 70% and diclofenac retention of 60%. Further experimental research studied various parameters of the composite membrane-like Molecular Weight Cut Off and pore size, Zeta potential and its mechanical and chemical robustness.

Keywords: water purification, polyelectrolytes, membrane modification, layer-by-layer coating, ceramic membranes

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580 Exploring the Bifunctional Organocatalysts for Asymmetric Synthesis of 3-Substituted-3-Aminooxindoles

Authors: Jasneet Kaur, Swapandeep Singh Chimni

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The unfavorable use of metal-based catalysts that are often extortionate and toxic can be overcome by using small organic molecules known as organocatalysts. A variety of small organic molecules, including Brønsted/Lewis bases and acids, based on sulfonic acids, phosphoric acids, amines, phosphines or carbenes, Cinchona alkaloids, have been used as organocatalysts. One of the key reasons for using organocatalysis is their ability to be effectively removed from the final product in comparison to the metallic counterparts, which are exceedingly difficult to remove. The present investigation seeks to explore the catalytic nature of Cinchona alkaloids as an organocatalyst for enantioselective synthesis of 3-substituted-3-aminooxindole, which is known to exhibit a variety of biological activities and pharmacological activities. In this context, an organocatalytic asymmetric route for the synthesis of 3-aminooxindoles via reaction of isatin imine with α-acetoxy-β-ketoesters has been developed. The bifunctional Cinchona derived thiourea catalyzed the reaction of α-acetoxy-β-ketoesters derivatives with isatin imine to afford 3-substituted-aminooxindole derivatives in up to 93% yield, 95% enantiomeric excess and >20:1 diastereomeric ratio. The reaction was performed at room temperature for two hours using 10 mol% of catalyst, in the presence of 4Å molecular sieves in tetrahydrofuran as a solvent at ambient temperature. After the completion of the reaction, the pure product could be easily separated by using column chromatography using hexane and ethyl acetate as solvents. In conclusion, the catalytic potential of Cinchona derived chiral thiourea-tertiary amine catalyst was explored for an organocatalytic enantioselective Mannich reaction of β-ketoester derivatives with various isatin imine derivatives under mild conditions.

Keywords: asymmetric synthesis, aminooxindoles, enantioselective, isatin imine

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579 Influence of Cyperus Rotundus Active Principles Inhibit Viral Multiplication and Stimulate Immune System in Indian White Shrimp Fenneropenaeus Indicus against White Spot Syndrome Virus Infection

Authors: Thavasimuthu Citarasu, Mariavincent Michaelbabu, Vikram Vakharia

Abstract:

The rhizome of Java grass, Cyperus rotundus was extracted different organic polar and non-polar solvents and performed the in vitro antiviral and immunostimulant activities against White Spot Syndrome Virus (WSSV) and Vibrio harveyi respectively. Based on the initial screening the ethyl acetate extract of C. rotundus was strong activities and further it was purified through silica column chromatography and the fractions were screened again for antiviral and immunostimulant activity. Among the different fractions screened against the WSSV and V. harveyi, the fractions, F-III to FV had strong activities. In order to study the in vivo influence of C. rotundus, the fractions (F-III to FV) were pooled and delivered to the F. indicus through artificial feed for 30 days. After the feeding trail the experimental and control diet fed F. indicus were challenged with virulent WSSV and studied the survival, molecular diagnosis, biochemical, haematological and immunological parameters. Surprisingly, the pooled fractions (F-III to FV) incorporated diets helped to significantly (P < 0.01) suppressed viral multiplication, showed significant (P < 0.01) differences in protein and glucose levels, improved total haemocyte count (THC), coagulase activity, significantly increased (P < =0.001) prophenol oxidase and intracellular superoxide anion production compared to the control shrimps. Based on the results, C. rotundus extracts effectively suppressed WSSV multiplication and improve the immune system in F. indicus against WSSV infection and this knowledge will helps to develop novel drugs from C. rotundus against WSSV.

Keywords: antiviral drugs, cyperus rotundus, fenneropenaeus indicus, WSSV

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578 MicroRNA-211 Regulates Oxidative Phosphorylation and Energy Metabolism in Human Vitiligoa

Authors: Anupama Sahoo, Bongyong Lee, Katia Boniface, Julien Seneschal, Sanjaya K. Sahoo, Tatsuya Seki, Chunyan Wang, Soumen Das, Xianlin Han, Michael Steppie, Sudipta Seal, Alain Taieb, Ranjan J. Perera

Abstract:

Vitiligo is a common, chronic skin disorder characterized by loss of epidermal melanocytes and progressive depigmentation. Vitiligo has a complex immune, genetic, environmental, and biochemical etiology, but the exact molecular mechanisms of vitiligo development and progression, particularly those related to metabolic control, are poorly understood. Here we characterized the human vitiligo cell line PIG3V and the normal human melanocytes, HEM-l by RNA-sequencing, targeted metabolomics, and shotgun lipidomics. Melanocyte-enriched miR-211, a known metabolic switch in non-pigmented melanoma cells, was severely downregulated in vitiligo cell line PIG3V and skin biopsies from vitiligo patients, while its novel predicted targets transcriptional co-activator PGC1-α (PPARGC1A), ribonucleotide reductase regulatory subunit M2 (RRM2), and serine-threonine protein kinase TAO1 (TAOK1) were reciprocally upregulated. miR-211 binds to PGC1-α 3’UTR locus and represses it. Although mitochondrial numbers were constant, mitochondrial complexes I, II, and IV and respiratory responses were defective in vitiligo cells. Nanoparticle-coated miR-211 partially augmented the oxygen consumption rate in PIG3V cells. The lower oxygen consumption rate, changes in lipid and metabolite profiles, and increased reactive oxygen species production observed in vitiligo cells appear to be partly due to abnormal regulation of miR-211 and its target genes. These genes represent potential biomarkers and therapeutic targets in human vitiligo.

Keywords: metabolism, microRNA, mitochondria, vitiligo

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577 Identification and Validation of Co-Dominant Markers for Selection of the CO-4 Anthracnose Disease Resistance Gene in Common Bean Cultivar G2333

Authors: Annet Namusoke, Annet Namayanja, Peter Wasswa, Shakirah Nampijja

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Common bean cultivar G2333 which offers broad resistance for anthracnose has been widely used as a source of resistance in breeding for anthracnose resistance. The cultivar is pyramided with three genes namely CO-4, CO-5 and CO-7 and of these three genes, the CO-4 gene has been found to offer the broadest resistance. The main aim of this work was to identify and validate easily assayable PCR based co-dominant molecular markers for selection of the CO-4 gene in segregating populations derived from crosses of G2333 with RWR 1946 and RWR 2075, two commercial Andean cultivars highly susceptible to anthracnose. Marker sequences for the study were obtained by blasting the sequence of the COK-4 gene in the Phaseolus gene database. Primer sequence pairs that were not provided from the Phaseolus gene database were designed by the use of Primer3 software. PCR conditions were optimized and the PCR products were run on 6% HPAGE gel. Results of the polymorphism test indicated that out of 18 identified markers, only two markers namely BM588 and BM211 behaved co-dominantly. Phenotypic evaluation for reaction to anthracnose disease was done by inoculating 21days old seedlings of three parents, F1 and F2 populations with race 7 of Colletotrichum lindemuthianum in the humid chamber. DNA testing of the BM588 marker onto the F2 segregating population of the crosses RWR 1946 x G 2333 and RWR 2075 x G2333 further revealed that the marker BM588 co-segregated with disease resistance with co-dominance of two alleles of 200bp and 400bp, fitting the expected segregation ratio of 1:2:1. The BM588 marker was significantly associated with disease resistance and gave promising results for marker assisted selection of the CO-4 gene in the breeding lines. Activities to validate the BM211 marker are also underway.

Keywords: codominant, Colletotrichum lindemuthianum, MAS, Phaseolus vulgaris

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576 Microbubbles Enhanced Synthetic Phorbol Ester Degradation by Ozonolysis

Authors: D. Kuvshinov, A. Siswanto, W. Zimmerman

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A phorbol-12-myristate-13-acetate (TPA) is a synthetic analogue of phorbol ester (PE), a natural toxic compound of Euphorbiaceae plant. The oil extracted from plants of this family is useful source for primarily biofuel. However this oil can also be used as a food stock due to its significant nutrition content. The limitations for utilizing the oil as a food stock are mainly due to a toxicity of PE. Nowadays a majority of PE detoxification processes are expensive as include multi steps alcohol extraction sequence. Ozone is considered as a strong oxidative agent. It reaction with PE it attacks the carbon double bond of PE. This modification of PE molecular structure results into nontoxic ester with high lipid content. This report presents data on development of simple and cheap PE detoxification process with water application as a buffer and ozone as reactive component. The core of this new technique is a simultaneous application of new microscale plasma unit for ozone production and patented gas oscillation technology. In combination with a reactor design the technology permits ozone injection to the water-TPA mixture in form of microbubbles. The efficacy of a heterogeneous process depends on diffusion coefficient which can be controlled by contact time and interface area. The low velocity of rising microbubbles and high surface to volume ratio allow fast mass transfer to be achieved during the process. Direct injection of ozone is the most efficient process for a highly reactive and short lived chemical. Data on the plasma unit behavior are presented and influence of the gas oscillation technology to the microbubbles production mechanism has been discussed. Data on overall process efficacy for TPA degradation is shown.

Keywords: microbubble, ozonolysis, synthetic phorbol ester, chemical engineering

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575 Record Peak Current Density in AlN/GaN Double-Barrier Resonant Tunneling Diodes on Free-Standing Gan Substrates by Modulating Barrier Thickness

Authors: Fang Liu, Jia Jia Yao, Guan Lin Wu, Ren Jie Liu, Zhuang Guo

Abstract:

Leveraging plasma-assisted molecular beam epitaxy (PA-MBE) on c-plane free-standing GaN substrates, this work demonstrates high-performance AlN/GaN double-barrier resonant tunneling diodes (RTDs) featuring stable and repeatable negative differential resistance (NDR) characteristics at room temperature. By scaling down the barrier thickness of AlN and the lateral mesa size of collector, a record peak current density of 1551 kA/cm2 is achieved, accompanied by a peak-to-valley current ratio (PVCR) of 1.24. This can be attributed to the reduced resonant tunneling time under thinner AlN barrier and the suppressed external incoherent valley current by reducing the dislocation number contained in the RTD device with the smaller size of collector. Statistical analysis of the NDR performance of RTD devices with different AlN barrier thicknesses reveals that, as the AlN barrier thickness decreases from 1.5 nm to 1.25 nm, the average peak current density increases from 145.7 kA/cm2 to 1215.1 kA/cm2, while the average PVCR decreases from 1.45 to 1.1, and the peak voltage drops from 6.89 V to 5.49 V. The peak current density obtained in this work represents the highest value reported for nitride-based RTDs to date, while maintaining a high PVCR value simultaneously. This illustrates that an ultra-scaled RTD based on a vertical quantum-well structure and lateral collector size is a valuable approach for the development of nitride-based RTDs with excellent NDR characteristics, revealing their great potential applications in high-frequency oscillation sources and high-speed switch circuits.

Keywords: GaN resonant tunneling diode, peak current density, peak-to-valley current ratio, negative differential resistance

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574 Characterization, Antibacterial and Cytotoxicity Evaluation of Silver Nanoparticles Synthesised Using Grewia lasiocarpa E. Mey. Ex Harv. Plant Extracts

Authors: Nneka Augustina Akwu, Yougasphree Naidoo

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Molecular advancement in technology has created a means whereby the atoms and molecules (solid forms) of certain materials such as plants, can now be reduced to a range of 1-100 nanometres. Green synthesis of silver nanoparticles (AgNPs) was carried out at room temperature (RT) 25 ± 2°C and 80°C, using the metabolites in the aqueous extracts of the leaves and stem bark of Grewia lasiocarpa as reductants and stabilizing agents. The biosynthesized AgNPs were characterized by UV-Vis spectrophotometry, attenuated total reflectance - Fourier transforms infrared (ATR-FTIR) spectroscopy, nanoparticle tracking analysis (NTA), Energy Dispersive X-ray fluorescence scanning electron microscope (SEM-EDXRF) and high-resolution transmission electron microscopy (HRTEM). The AgNPs were biologically evaluated for antioxidant, antibacterial and cytotoxicity activities. The phytochemical and FTIR analyses revealed the presence of metabolites that act as reducing and capping agents, while the UV-Vis spectroscopy of the biosynthesized NPs showed absorption between 380-460 nm, confirming AgNP synthesis. The Zeta potential values were between -9.1 and -20.6 mV with a hydrodynamics diameter ranging from 38.3 to 46.7 nm. SEM and HRTEM analyses revealed that AgNPs were predominately spherical with an average particle size of 2- 31 nm for the leaves and 5-27 nm for the stem bark. The cytotoxicity IC50 values of the AgNPs against HeLa, Caco-2 and MCF-7 were >1 mg/mL. The AgNPs were sensitive to all strains of bacteria used, with methicillin-resistant Staphylococcus aureus (MRSA), Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922) being more sensitive to the AgNPs. Our findings propose that antibacterial and anticancer agents could be derived from these AgNPs of G. lasiocarpa, and warrant their further investigation.

Keywords: antioxidant, cytotoxicity, Grewia lasiocarpa, silver nanoparticles, Zeta potentials

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573 Selection of a Potential Starter Culture for Milk Fermentation

Authors: Stephen Olusanmi Akintayo, Ilesanmi Fadahunsi

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The ability of Lactic acid bacteria (LAB) to grow and survive in milk is being exploited in industrial and biotechnological applications. Although considerable studies have been reported on the fermentation of milk, however, not so much work has been documented on the selection of LAB strains from milk of the Nigerian local cattle breeds for their starter culture potentials. A total of 110 LAB were isolated from raw milk of Sokoto gudali cattle breed. The isolates were screened for their proteolytic activities on skimmed milk media with isolates A07, F06 and A01 showing the highest zone of clearance of 18.5mm, 18.5mm, and 18.0mm respectively and were selected for the studies of their growth in different constituents of milk. A01, F06, and A07 were identified as Pediococcus acidilactici, Lactococcus raffinolactis, and Leuconostoc mesenteriodes respectively using cultural, biochemical, physiological and molecular characterization techniques. Leuconostoc mesenteriodes showed the highest growth in all the milk components that were used in this study. The three LAB species selected showed a growth range of 6.46 log cfu/ml to 10.91 log cfu/ml in lactose with Leuconostoc mesenteriodes showing the highest growth of 10.91 log cfu/ml while Pediococcus acidilactici recorded the lowest growth of 9.78 log cfu/ml. In medium containing leucine as the only amino acid, the viable counts of Pediococcus acidilactici, Lactococcus raffinolactis and Leuconostoc mesenteriodes in log cfu/ml at zero hour were 6.39, 6.36 and 6.38 respectively which increased to 9.31 log cfu/ml, 9.21 log cfu/ml, 9.92 log cfu/ml respectively after 24 hours. Similarly, in all other substrates (casein, lysine, glutamic acid, aspartic acid, stearic acid and oleic acid ) tested in this study, Leuconostoc mesenteriodes showed the highest growth. It was observed that the highest quantity of lactic acid (15.31mg/ml) was produced by Leuconostoc mesenteriodes. The same trend was also observed in the production of diacetyl and hydrogen peroxide by the three tested microorganisms. Due to its ability to grow maximally in milk components, Leuconostoc mesenteriodes shows potential as starter culture for milk fermentation.

Keywords: Leuconostoc mesenteriodes, lactic acid bacteria, Sokoto gudali, starter culture

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572 Modeling of Gas Migration in High-Pressure–High-Temperature Fields

Authors: Deane Roehl, Roberto Quevedo

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Gas migration from pressurized formations is a problem reported in the oil and gas industry. This means increased risks for drilling, production, well integrity, and hydrocarbon escape. Different processes can contribute to the development of pressurized formations, particularly in High-Pressure–High-Temperature (HPHT) gas fields. Over geological time-scales, the different formations of those fields have maintained and/or developed abnormal pressures owing to low permeability and the presence of an impermeable seal. However, if this seal is broken, large volumes of gas could migrate into other less pressurized formations. Three main mechanisms for gas migration have been identified in the literature –molecular diffusion, continuous-phase flow, and continuous-phase flow coupled with mechanical effects. In relation to the latter, gas migration can occur as a consequence of the mechanical effects triggered by reservoir depletion. The compaction of the reservoir can redistribute the in-situ stresses sufficiently to induce deformations that may increase the permeability of rocks and lead to fracture processes or reactivate nearby faults. The understanding of gas flow through discontinuities is still under development. However, some models based on porosity changes and fracture aperture have been developed in order to obtain enhanced permeabilities in numerical simulations. In this work, a simple relationship to integrate fluid flow through rock matrix and discontinuities has been implemented in a fully thermo-hydro-mechanical simulator developed in-house. Numerical simulations of hydrocarbon production in an HPHT field were carried out. Results suggest that rock permeability can be considerably affected by the deformation of the field, creating preferential flow paths for the transport of large volumes of gas.

Keywords: gas migration, pressurized formations, fractured rocks, numerical modeling

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571 Combined Effect of Gluten-Free Superfoods and by-Products from Ecuador to Evaluate the Functional and Sensory Properties of Breadmaking

Authors: Andrea Vasquez, Pedro Maldonado-Alvarado

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In general, 'gluten-free' foods like breadmaking products provide functional or nutraceutical benefits for the consumer's health and increased their demand on the market. In Ecuador, there is an overproduction of superfoods, and the food by-products are undervalued. For the first time, to the author's best knowledge, gluten-free bread mixtures from quinoa and banana flour, cassava starch, lupine flour (LF), or whey protein (WP) with hydroxypropylmethylcellulose (HPMC) and transglutaminase (TG) were evaluated on their functional and sensory properties. Free amino groups and thiols, rheology, and electrophoresis SDS PAGE were performed to analyze the crosslinking of TG at different concentrations with HC or PL proteins. Dough characterization, pasting properties were evaluated, respectively, by a MIXOLAB and a rheometer with a pasting cell. The texture, porosity, and loaf volume were characterized using a texturometer, ImageJ software, and breadmaking ability, respectively. Finally, a breadmaking aptitude and sensorial bread acceptability were performed. A significant decrease in the content of free amino groups (0.16 to 0.11 and 0.46 to 0.36 mM/mg of protein) and free thiol groups (0.37 to 0.21 and 1.79 to 1.32 mM/mg protein) was observed when 1.0% and 0.5% TG were added to LF and WP, respectively. In apparent viscosity analysis, the action of TG on HC proteins changes their viscosity, while the viscosity of LF is not modified by TG. Results of electrophoresis in PL showed bands of higher molecular weight of different fragments of proteins with 1% TG. Formulation with 59.8, 39.9, 160.8, 6.0, 1.0, and 1.5% of, respectively, QF, BF, CS, LF or WP, TG, and HPMC had the best properties in dough parameters, pasting parameters (lower pasting temperature and higher peak viscosity), best crumb structure, lower crumb hardness and higher loaf volume (2.24 and 2.28 mL/g). All the loaves of bread were acceptable in baking aptitude and general acceptability.

Keywords: breadmaking, gluten-free, superfoods, by-products, Ecuador

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570 PCR Detection, Histopathological Characterization, and Autogenous Immunization of Bovine Papillomatosis (Wart) in Cattle, in Mekelle, Northern Ethiopia

Authors: Kidane Workelul, Yohans Tekle, Guesh Negash, Haftay Abraha, Nigus Abebe Shumuye, Yisehak Tsegaye Redda

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Bovine papillomatosis (wart) is one of the economically important bovine skin diseases worldwide, caused by a group of viruses named papillomaviruses (PVs). However, it has often been misdiagnosed as other skin diseases and remained untreated. In order to determine the status of the diseases, twenty-two farms were visited, and fourteen infected cattle with cutaneous papillomatosis were identified from a total of 235. Papilloma biopsies were taken for molecular and histopathological characterization, the therapeutic trial of an autogenous vaccine was evaluated on infected animals. The overall status of bovine papillomatosis in this study was calculated as 5.96% (14/235). The disease was found to be statistically significant in the age groups less than two years (X² = 26.69, P = 0.0001). The more prominent histologically characterized lesions in the sampled tissue were identified as squamous papilloma and fibro-papilloma. The Polymerase Chain Reaction (PCR) based identification revealed that all the clinically and histo-pathologically characterized papillomatosis cases were found to be infected with Bovine Papilloma Virus1(BPV1), indicating that BPV1 was the most common and sole causative agent of the diseases in the study area. In immunizing active bovine papillomatosis, an autogenous vaccine therapeutic trial demonstrated excellent results, with practically full recovery and no recurrence of the infection. Hence, it is concluded that bovine papillomatosis is an economically important disease of young age group cattle as well as a treatable disease. So, the production of marketable autogenous vaccines against bovine papillomatosis should be started and given at an early stage.

Keywords: autogenous vaccine, bovine papillomatosis, bovine papilloma virus1 clinical-pathology, polymerase chine reaction, wart

Procedia PDF Downloads 80