Search results for: protein domains

Authors: Nicholas P. Fenlon, Aisling Lavelle, David Mclean, Margaret O'riordan

Abstract:

Background: The case for continuing professional development has been well made, and was formalized in Ireland in recent years through the enactment of the Medical Practitioner’s Act, which requires registered medical practitioners to complete a minimum of 50 hours CPD each year. The ICGP, who have been providing CPD opportunities to its members for many years, have responded to this need by developing a series of evidence-based, high-quality, multimedia modules across a range of clinical and non-clinical areas. (More traditional education opportunities are still being provided by the college also). Overview of Programme: The first module was released in September 2011, since when the eLearning program has grown steadily, and there are currently almost 20 modules available, with a further 5 in production. Each module contains three to six 10-minute video lessons, which use a combination of graphics, images, text, voice-over and clinical clips. These are supported by supplementary videos of expert pieces-to-camera, Q&As with content experts, clinical scenarios, external links and relevant documentation and other resources. Successful completion of MCQs will result in a Certificate of Completion, which can be printed or stored in Professional Competence portfolio. The Medical Practitioner’s Act requires doctors to gather CPD credits across 8 domains of practice, and various eLearning modules have been developed to address each. For instance, modules with a strong clinical content would include Management of Hypertension, Management of COPD, and Management of Asthma. Other modules focus on health promotion such as Promoting Smoking Cessation, Promoting Physical Activity, and Addressing Childhood Obesity. Modules where communication skills are keys include modules on Suicide Prevention and Management of Depression. Other modules, currently in development include non-clinical topics around risk management, including Confidentiality, Consent etc. Each module is developed by a core group, which includes where possible, a GP with a special interest in the area, and a content expert(s). The college works closely with a medical education consultant and a production company in developing and producing the modules. Modules can be accessed (with password) through the ICGP website and are available free to all ICGP members. Summary of Evaluation: There are over 1700 registered users to date (over 55% of College membership). The program was evaluated using an online survey in 2013 (N = 144/950 – 12%) and results were very positive overall but provided material for the further improvement of the program also. Future Plans: While knowledge can be imparted well through eLearning, skills and attitudes are more difficult to influence through an online environment. The college is now developing a series of linked workshops, which will lead to ICGP Professional Competence Awards. The first pilot workshop is scheduled for February 2015 and is Cardiology-themed. Participants will be required to complete the following 4 modules in advance of attending – Management of Hypertension, Management of Heart Failure, Promoting Smoking Cessation, and Promoting Physical Activity. The workshop will be case-based and interactive, addressing ECG Interpretation in General Practice. Conclusions: The ICGP have responded to members needs for high-quality evidence-based education delivered in a way that suits GPs.

Keywords: CPD opportunities, evidence-based, high quality, multimedia modules across a range of clinical and non-clinical areas, medical practitioner’s act

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Authors: Saule Musabekova

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Achievements of modern science, especially genetics, led to a sharp intensification of the process of proof. Footprints, subjected to destruction-related cause-effect relationships, are sources of evidentiary information on the circumstances it was committed and the persons committed it. Currently, with the overall growth in the number of crimes against sexual inviolability or sexual freedom, and increased the proportion of the crimes where to destroy the traces of the crime perpetrators different detergents are used. A characteristic feature of modern synthetic detergents is the presence of biological additives - enzymes that break down and gradually destroy stains of protein origin. To study the nature of the influence of modern washing powders semen stains were put kinds of fabrics and prepared in advance stained sperm of men of different groups according to ABO system. For research washing machines of known manufacturers of household appliances have been used with different production characteristics, in which the test was performed and the washing of various kinds of fabrics with semen stains. After washing the tissue with spots were tested for the presence of semen stains visually preserved, establishing in them surviving sperm or their elements, we studied the possibilities of the group diagnostics on the system ABO or molecular-genetic identification. The subsequent study of these spots by morphological method showed that 100% detection of morphological sperm cells - sperm is not possible. As a result, in 30% of further studies of these traces gave weakly positive results are obtained with an immunoassay test PSA SEMIQUANT. It is noted that the percentage of positive results obtained in the study of semen traces disposed on natural fiber fabrics is higher than sperm traces disposed on synthetic fabrics. Study traces of semen, confirmed by PSA - test 3% possible to establish a genetic profile of the person and obtain any positive findings of the molecular genetic examination. In other cases, it was not a sufficient amount of material for DNA identification. Results of research and the practical expert study found, in most cases, the conclusions of the identification of sperm traces do not seem possible. This a consequence of exposure to semen traces on the material evidence of biological additives contained in modern detergents and further the influence of other effective methods. Resulting in DNA has undergone irreversible changes (degradation) under the influence of external human factors. Using molecular genetic methods can partially solve the problems arising in the study of unlaundered physical evidence for the disclosure and investigation of crimes.

Keywords: study of sperm, modern detergents, washing powders, forensic medicine

Procedia PDF Downloads 298

Authors: Supanan Chansap, Werachon Cheukamud, Pornanan Kueakhai, Narin Changklungmoa

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Fasciola species (Fasciola spp.) is caused fasciolosis in ruminants such as cattle, sheep, and buffalo. Fasciola gigantica (F.gigantica) commonly infects tropical regions. Fasciola hepatica (F.hepatica) in temperate regions. Liver fluke infection affects livestock economically, for example, reduced milk and meat production, weight loss, sterile animals. Currently, Triclabendazole is used to treat liver flukes. However, liver flukes have also been found to be resistant to drugs in countries. Therefore, vaccination is an attractive alternative to prevent liver fluke infection. Peptide vaccines are new vaccine technologies that mimic epitope antigens that trigger an immune response. An interesting antigen used in vaccine production is catepsin L, a family of proteins that play an important role in the life of the parasite in the host. This study aims to identify immunogenic regions of protein and construct a multi-epidetope vaccine using an immunoinformatic tool. Fasciola gigantica Cathepsin L1 (FgCatL1), Fasciola gigantica Cathepsin L1G (FgCatL1G), and Fasciola gigantica Cathepsin L1H (FgCatL1H) were predicted B-cell and Helper T lymphocytes (HTL) by Immune Epitope and Analysis Resource (IEDB) servers. Both B-cell and HTL epitopes aligned with cathepsin L of the host and Fasciola hepatica (F. hepatica). Epitope groups were selected from non-conserved regions and overlapping sequences with F. hepatica. All overlapping epitopes were linked with the GPGPG and KK linker. GPGPG linker was linked between B-cell epitope. KK linker was linked between HTL epitope and B-cell and HTL epitope. The antigenic scores of multi-epitope peptide vaccine was 0.7824. multi-epitope peptide vaccine was non-allergen, non-toxic, and good soluble. Multi-epitope peptide vaccine was predicted tertiary structure and refinement model by I-Tasser and GalaxyRefine server, respectively. The result of refine structure model was good quality that was generated by Ramachandran plot analysis. Discontinuous and linear B-cell epitopes were predicted by ElliPro server. Multi-epitope peptide vaccine model was two and seven of discontinuous and linear B-cell epitopes, respectively. Furthermore, multi-epitope peptide vaccine was docked with Toll-like receptor 2 (TLR-2). The lowest energy ranged from -901.3 kJ/mol. In summary, multi-epitope peptide vaccine was antigenicity and probably immune response. Therefore, multi-epitope peptide vaccine could be used to prevent F. gigantica infections in the future.

Keywords: fasciola gigantica, Immunoinformatic tools, multi-epitope, Vaccine

Procedia PDF Downloads 79

Authors: Elena Maria Scalisi

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The increasing production and the use of TiO₂ nanoparticles (NPs) have inevitably led to their release into the environment, thereby posing a threat to organisms and also for human. Human exposure to TiO₂-NPs may occur during both manufacturing and use. TiO₂-NPs are common in consumer products for dermal application, toothpaste, food colorants, and nutritional supplements, then oral exposure may occur during use of such products. Into the body, TiO₂-NPs thanks to their small size (<100 nm), can, through testicular blood barrier inducing effect on testis and then on male reproductive health. The nanoscale size of TiO₂ increase the surface-to-volume ratio making them more reactive in a cell, then TiO₂ NPs increase their ability to produce reactive oxygen species (ROS). In male germ cells, ROS may have important implications in maintaining the normal functions of mature spermatozoa at physiological levels, moreover, in spermatozoa they are important signaling molecules for their hyperactivation and acrosome reaction. Nevertheless, an excess of ROS by external inputs such as NPs can increased the oxidative stress (OS), which results in damage DNA and apoptosis. The aim of our study has been investigate the impact of TiO₂ NPs on human spermatozoa, evaluating DNA damage and the expression of proteins involved in cell stress. According WHO guidelines 2021, we have exposed human spermatozoa in vitro to TiO₂ NP at concentrations 50 ppm, 100 ppm, 250 ppm, and 500 ppm for 1 hour (at 37°C and CO₂ at 5%). DNA damage was evaluated by Sperm Chromatin Dispersion Test (SCD) and TUNEL assay; moreover, we have evaluated the expression of biomarkers of oxidative stress like Heat Shock Protein 70 (HSP70) and Metallothioneins (MTs). Also, sperm parameters as motility viability have been evaluated. Our results not report a significant reduction in motility of spermatozoa at the end of the exposure. On the contrary, the progressive motility was increased at the highest concentration (500 ppm) and was statistically significant compared to control (p <0.05). Also, viability was not changed by exposure to TiO₂-NPs (p <0.05). However, increased DNA damage was observed at all concentrations, and the TUNEL assay highlighted the presence of single strand breaks in the DNA. The spermatozoa responded to the presence of TiO₂-NPs with the expression of Hsp70, which have a protective function because they allow the maintenance of cellular homeostasis in stressful/ lethal conditions. A positivity for MTs was observed mainly for the concentration of 4 mg/L. Although the biological and physiological function of the metallothionein (MTs) in the male genital organs is unclear, our results highlighted that the MTs expressed by spermatozoa maintain their biological role of detoxification from metals. Our results can give additional information to the data in the literature on the toxicity of TiO₂-NPs and reproduction.

Keywords: human spermatozoa, DNA damage, TiO₂-NPs, biomarkers

Procedia PDF Downloads 144

Authors: Bisemiire Anthony,

Abstract:

PESH Poultry is a business project proposed specifically to solve unemployment and income-related problems affecting the youths in the Busia district. The project is intended to transform the life of the youth in terms of economic, social and behavioral, as well as the domestic well-being of the community at large. PESH Poultry is a start-up poultry farm that will be engaged in the keeping of poultry birds, broilers and layers for the production of quality and affordable poultry meat and eggs respectively and other poultry derivatives targeting consumers in eastern Uganda, for example, hotels, restaurants, households and bakeries. We intend to use a semi-intensive system of farming, where water and some food are provided in a separate nighttime shelter for the birds; our location will be in Lumino, Busia district. The poultry project will be established and owned by Bisemiire Anthony, Nandera Patience, Naula Justine, Bwire Benjamin and other investors. The farm will be managed and directed by Nandera Patience, who has five years of work experience and business administration knowledge. We will sell poultry products, including poultry eggs, chicken meat, feathers and poultry manure. We also offer consultancy services for poultry farming. Our eggs and chicken meat are hygienic, rich in protein and high quality. We produce processes and packages to meet the standard organization of Uganda and international standards. The business project shall comprise five (5) workers on the key management team who will share various roles and responsibilities in the identified business functions such as marketing, finance and other related poultry farming activities. PESH Poultry seeks 30 million Ugandan shillings in long-term financing to cover start-up costs, equipment, building expenses and working capital. Funding for the launch of the business will be provided primarily by equity from the investors. The business will reach positive cash flow in its first year of operation, allowing for the expected repayment of its loan obligations. Revenue will top UGX 11,750,000, and net income will reach about UGX115 950,000 in the 1st year of operation. The payback period for our project is 2 years and 3 months. The farm plans on starting with 1000 layer birds and 1000 broiler birds, 20 workers in the first year of operation.

Keywords: chicken, pullets, turkey, ducks

Procedia PDF Downloads 95

Authors: Gizaw Mamo Gebeyehu, Marianna Pap, Geza Makkai, Tibor Z. Janosi, Shima Rashidian, Tibor A. Rauch

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Sepsis poses a significant global health threat, necessitating extensive exploration of indicators tied to its pathological mechanisms and multi-organ dysfunction. While murine studies have shed light on sepsis, the intricate cellular and molecular landscape in human sepsis remains enigmatic. Exploring the influence of activated monocyte-derived exosomes in sepsis sheds light on a promising pathway for understanding the intricate cellular and molecular mechanisms involved in this condition in humans. In sepsis, exosome-borne mRNA and miRNA orchestrate immune response gene expression in recipient cells. Yet, the specifics of exosome-mediated cell-to-cell communication, especially how mRNA cargoes modulate gene expression in recipient cells, remain poorly understood. This study aims to elucidate the precise molecular pathways through which exosomal mRNA cargo, particularly MAFB, contributes to the developing sepsis-induced molecular aberrations in liver tissues, employing rigorously defined cell culture conditions. THP-1 cells were treated with LPS to induce changes in exosomal RNA profiles. Exosomes were isolated and characterized using microscopy and mass spectrometry. RNA was extracted from exosomes and sequenced. The most abundant exosomal mRNAs were subjected to GO analysis for functional annotation analysis and KEGG database analysis to identify the involved enriched pathways. PCR (Polymerase Chain Reaction), RNA sequencing, and Western blotting were involved to analyze changes in gene expression, protein levels, and signaling pathways within the liver cells( HepG2) after exposure to exosomal MAFB. This study pinpoints exosomal MAFB as a potential key regulator linked to liver cell damage during sepsis, along with associated genes (miR155HG, H3F3A, and possibly JARD2) forming a crucial molecular pathway contributing to liver cell injury, Together, these elements indicate a vital molecular pathway that plays a significant role in the emergence of liver cell injury during sepsis.. These findings suggest the importance of further research on these components for potential therapeutic interventions in managing acute liver damage in sepsis.

Keywords: sepsis, exososome, exosomal MAFB, LPS-induced THP-1 cells, RNA profiles, sepsis-triggered liver injury

Procedia PDF Downloads 64

Authors: Patience Manjengwa-Hungwe, Carmen White

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Background: Odyssean malaria refers to malaria acquired by infected mosquito bites from malaria endemic to non-endemic regions by mechanical modes of transport, such as airplanes, water vessels, trains and vehicles. Odyssean Malaria is rare and is characterised by absence of travel history to malaria endemic areas. As not anticipated in non-endemic areas, late diagnosis and treatment lead to a high case fatality rate. On 26 September 2014, the Outbreak Response Unit at the National Institute of Communicable Diseases was notified of a suspected death from Odyssean Malaria in Johannesburg, Gauteng Province, a non-endemic area. The main objective of this investigation was to identify the etiological agent's mode and source of transmission. Methods: Epidemiological surveys were conducted with the deceased’s family and clinical details were obtained from doctors who treated the victim in Southrand, Johannesburg. Blood samples were collected prior to death and sent to the National Health Laboratory Services, Johannesburg laboratory for a full blood count, urea electrolytes, creatinine, and C-reactive protein. Environmental assessments and entomological investigations, including collection of mosquito and larvae, were conducted at the deceased’s home and surrounding areas and sent to the laboratory for analysis. Results: Epidemiological surveys revealed no travel history, no mechanical transmission through blood transfusion and no previous possible exposure of the victim to malaria mosquitoes. Laboratory findings indicated that the platelet count was low. A further smear revealed that the malaria parasite was present and malaria antigen for P. falciparum was positive. Entomological findings revealed that none of the six adult or larval mosquitoes collected on site were malaria vectors. Dumping sites found at the back of the house were identified as possible sites where mosquitoes from endemic places could possibly breed. Conclusion: Given that there was no travel history or the possibility of mechanical transmission (blood transfusion or needle), the research team concluded that it is highly probable that the infection was acquired through an infective Anopheles mosquito inadvertently translocated from a Malaria endemic area by mechanical modes of transport. We recommend that clinicians in non-endemic malaria areas be aware of this type of malaria and test for malaria in patients showing malaria-like symptoms.

Keywords: Odyssean Malaria, vector Bourne, malaria, epidemiological surveys

Procedia PDF Downloads 339

Authors: Kyung Jin Kim, Jiwon Kwak, Jae-Hoon Lee, Soo Suk Lee

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MicroRNAs (miRNA) are a class of endogenous, single-stranded, small, and non-protein coding RNA molecules typically 20-25 nucleotides long. They are thought to regulate the expression of other genes in a broad range by binding to 3’- untranslated regions (3’-UTRs) of specific mRNAs. The detection of miRNAs is very important for understanding of the function of these molecules and in the diagnosis of variety of human diseases. However, detection of miRNAs is very challenging because of their short length and high sequence similarities within miRNA families. So, a simple-to-use, low-cost, and highly sensitive method for the detection of miRNAs is desirable. In this study, we demonstrate a novel bi-directional extension (BDE) assay. In the first step, a specific linear RT primer is hybridized to 6-10 base pairs from the 3’-end of a target miRNA molecule and then reverse transcribed to generate a cDNA strand. After reverse transcription, the cDNA was hybridized to the 3’-end which is BDE sequence; it played role as the PCR template. The PCR template was amplified in an SYBR green-based quantitative real-time PCR. To prove the concept, we used human brain total RNA. It could be detected quantitatively in the range of seven orders of magnitude with excellent linearity and reproducibility. To evaluate the performance of BDE assay, we contrasted sensitivity and specificity of the BDE assay against a commercially available poly (A) tailing method using miRNAs for let-7e extracted from A549 human epithelial lung cancer cells. The BDE assay displayed good performance compared with a poly (A) tailing method in terms of specificity and sensitivity; the CT values differed by 2.5 and the melting curve showed a sharper than poly (A) tailing methods. We have demonstrated an innovative, cost-effective BDE assay that allows improved sensitivity and specificity in detection of miRNAs. Dynamic range of the SYBR green-based RT-qPCR for miR-145 could be represented quantitatively over a range of 7 orders of magnitude from 0.1 pg to 1.0 μg of human brain total RNA. Finally, the BDE assay for detection of miRNA species such as let-7e shows good performance compared with a poly (A) tailing method in terms of specificity and sensitivity. Thus BDE proves a simple, low cost, and highly sensitive assay for various miRNAs and should provide significant contributions in research on miRNA biology and application of disease diagnostics with miRNAs as targets.

Keywords: bi-directional extension (BDE), microRNA (miRNA), poly (A) tailing assay, reverse transcription, RT-qPCR

Procedia PDF Downloads 166

Authors: S. K. Sharma, P. Kumar, Pratibha Singh

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Consumption of SNACK is growing its popularity every day in India and a broad range of these items are available in the market. The end user interest in ready-to-eat snack foods is constantly growing mainly due to their ease, ample accessibility, appearance, taste and texture. Food extrusion has been practiced for over fifty years. Its role was initially limited to mixing and forming cereal products. Although thermoplastic extrusion has been successful for starch products, extrusion of proteins has achieved only limited success. In this study, value-added extruded prawn product was prepared with prawn flavor powder and corn flour using a twin-screw extruder. Prawn flavor concentrates prepared from fresh prawn head (Solenocera indica). To prepare flavor concentrate prawn head washed with potable water and blended with 200ml 3% salt solution per 250gm head weight to make the slurry, which was further put in muslin cloth and boiled with salt and starch solution for 10 minutes, cooled to room temperature and filtered, starch added to the filtrate and made into powder in an electrically drier at 43-450c. The mixture was passed through the twin-screw extruder (co-rotating twin screw extruder - basic technology Pvt. Ltd., Kolkata) which was operated at a particular speed of rotation, die diameter, temperature, moisture, and fish powder concentration. Many trial runs were conducted to set up the process variables. The different extrudes produced after each trail were examined for the quality and characteristics. The effect of temperature, moisture, screw speed, protein, fat, ash and thiobarbituric acid (TBA) number and expansion ratio were studied. In all the four trials, moisture, temperature, speed and die diameter used was 20%, 100°C, 350 rpm and 4 mm, respectively. The ratio of prawn powder and cornstarch used in different trials ranged between 2:98 and 10:90. The storage characteristics of the final product were studied using three different types of packaging under nitrogen flushing, i.e. a- 12-pm polyester, 12-pm metalized polyester, 60-11m polyethylene (metalized polyester a), b- 12-11m metalized polyester, 37.5-11m polyethylene (metalized polyester b), c- 12-11m polyethylene, 9-11m aluminium foil, 37.5-11m polyethylene (aluminium foil). The organoleptic analysis was carried out on a 9-point hedonic scale. The study revealed that the fried product packed in aluminum foil under nitrogen flushing would remain acceptable for more than three months.

Keywords: extruded product, prawn flavor, twin-screw extruder, storage characteristics

Procedia PDF Downloads 140

Authors: Virendra Nath, Vipin Kumar

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Background: TypeII Diabetes mellitus is a foremost health problem worldwide, predisposing to increased mortality and morbidity. Undesirable effects of the current medications have prompted the researcher to develop more potential drug(s) against the disease. The peroxisome proliferator-activated receptors (PPARs) are members of the nuclear receptors family and take part in a vital role in the regulation of metabolic equilibrium. They can induce or repress genes associated with adipogenesis, lipid, and glucose metabolism. Aims: Investigation of PPARα/γ agonistic hits were screened by hierarchical virtual screening followed by molecular dynamics simulation and knowledge-based structure-activity relation (SAR) analysis using approved PPAR α/γ dual agonist. Methods: The PPARα/γ agonistic activity of compounds was searched by using Maestro through structure-based virtual screening and molecular dynamics (MD) simulation application. Virtual screening of nuclear-receptor ligands was done, and the binding modes with protein-ligand interactions of newer entity(s) were investigated. Further, binding energy prediction, Stability studies using molecular dynamics (MD) simulation of PPARα and γ complex was performed with the most promising hit along with the structural comparative analysis of approved PPARα/γ agonists with screened hit was done for knowledge-based SAR. Results and Discussion: The silicone chip-based approach recognized the most capable nine hits and had better predictive binding energy as compared to the reference drug compound (Tesaglitazar). In this study, the key amino acid residues of binding pockets of both targets PPARα/γ were acknowledged as essential and were found to be associated in the key interactions with the most potential dual hit (ChemDiv-3269-0443). Stability studies using molecular dynamics (MD) simulation of PPARα and γ complex was performed with the most promising hit and found root mean square deviation (RMSD) stabile around 2Å and 2.1Å, respectively. Frequency distribution data also revealed that the key residues of both proteins showed maximum contacts with a potent hit during the MD simulation of 20 nanoseconds (ns). The knowledge-based SAR studies of PPARα/γ agonists were studied using 2D structures of approved drugs like aleglitazar, tesaglitazar, etc. for successful designing and synthesis of compounds PPARγ agonistic candidates with anti-hyperlipidimic potential.

Keywords: computational, diabetes, PPAR, simulation

Procedia PDF Downloads 103

Authors: Santi Maneewatchararangsri, Wanpen Chaicumpa, Patcharin Saengjaruk, Urai Chaisri

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Leptospirosis is a globally neglected disease that continues to be a significant public health and veterinary burden, with millions of cases reported each year. Early and accurate differential diagnosis of leptospirosis from other febrile illnesses and the development of a broad spectrum of leptospirosis vaccines are needed. The LipL32 outer membrane lipoprotein is a member of Leptospira adhesive matrices and has been found to exert hemolytic activity to erythrocytes in vitro. Therefore, LipL32 is regarded as a potential target for diagnosis, broad-spectrum leptospirosis vaccines, and for passive immunotherapy. In this study, we established LipL32-specific mouse monoclonal antibodies, mAbLPF1 and mAbLPF2, and their respective mouse- and humanized-engineered single chain variable fragment (ScFv). Their antibodies’ neutralizing activities against Leptospira-mediated hemolysis in vitro, and the therapeutic efficacy of mAbs against heterologous Leptospira infected hamsters were demonstrated. The epitope peptide of mAb LPF1 was mapped to a non-contiguous carboxy-terminal β-turn and amphipathic α-helix of LipL32 structure contributing to phospholipid/host cell adhesion and membrane insertion. We found that the mAbLPF2 epitope was located on the interacting loop of peptide binding groove of the LipL32 molecule responsible for interactions with host constituents. Epitope sequences are highly conserved among Leptospira spp. and are absent from the LipL32 superfamily of other microorganisms. Both epitopes are surface-exposed, readily accessible by mAbs, and immunogenic. However, they are less dominant when revealed by LipL32-specific immunoglobulins from leptospirosis-patient sera and rabbit hyperimmune serum raised by whole Leptospira. Our study also demonstrated an adhesion inhibitory activity of LipL32 protein to host membrane components and cells mediated by mAbs as well as an anti-hemolytic activity of the respective antibodies. The therapeutic antibodies, particularly the humanized-ScFv, have a potential for further development as non-drug therapeutic agent for human leptospirosis, especially in subjects allergic to antibiotics. The epitope peptides recognized by two therapeutic mAbs have potential use as tools for structure-function studies. Finally, protective peptides may be used as a target for epitope-based vaccines for control of leptospirosis.

Keywords: leptospira lipl32-specific antibodies, therapeutic epitopes, epitopes characterization, immunotherapy

Procedia PDF Downloads 297

Authors: Achiron Anat, Mathilda Mandel, Mayust Sue, Achiron Reuven, Gurevich Michael

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Introduction: As coronavirus disease 2019 (COVID-19) vaccination is currently spreading around the world, it is of importance to assess the ability of multiple sclerosis (MS) patients to mount an appropriate immune response to the vaccine in the context of disease-modifying treatments (DMT’s). Objectives: Evaluate immunity generated following COVID-19 vaccination in MS patients, and assess factors contributing to protective humoral and cellular immune responses in MS patients vaccinated against severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) virus infection. Methods: Review our recent data related to (1) the safety of PfizerBNT162b2 COVID-19 mRNA vaccine in adult MS patients; (2) the humoral post-vaccination SARS-CoV2 IgG response in MS vaccinees using anti-spike protein-based serology; and (3) the cellular immune response of memory B-cells specific for SARS-CoV-2 receptor-binding domain (RBD) and memory T-cells secreting IFN-g and/or IL-2 in response to SARS-CoV2 peptides using ELISpot/Fluorospot assays in MS patients either untreated or under treatment with fingolimod, cladribine, or ocrelizumab; (4) covariate parameters related to mounting protective immune responses. Results: COVID-19 vaccine proved safe in MS patients, and the adverse event profile was mainly characterised by pain at the injection site, fatigue, and headache. Not any increased risk of relapse activity was noted and the rate of patients with acute relapse was comparable to the relapse rate in non-vaccinated patients during the corresponding follow-up period. A mild increase in the rate of adverse events was noted in younger MS patients, among patients with lower disability, and in patients treated with DMTs. Following COVID-19 vaccination protective humoral immune response was significantly decreased in fingolimod- and ocrelizumab- treated MS patients. SARS-CoV2 specific B-cell and T-cell cellular responses were respectively decreased. Untreated MS patients and patients treated with cladribine demonstrated protective humoral and cellular immune responses, similar to healthy vaccinated subjects. Conclusions: COVID-19 BNT162b2 vaccine proved as safe for MS patients. No increased risk of relapse activity was noted post-vaccination. Although COVID-19 vaccination is new, accumulated data demonstrate differences in immune responses under various DMT’s. This knowledge can help to construct appropriate COVID-19 vaccine guidelines to ensure proper immune responses for MS patients.

Keywords: covid-19, vaccination, multiple sclerosis, IgG

Procedia PDF Downloads 139

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

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

Keywords: antarctic, bacteria, biotechnological, cellulolytic enzymes

Procedia PDF Downloads 297

Authors: Kusmartono, Siti Chuzaemi, Hartutik dan Mashudi

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Two studies were conducted involving an in vitro (Expt 1) and in vivo (Expt 2) measurements. Expt 1. aimed to evaluate effects of adding CT extracts on gas production and efficiency of microbial protein synthesis (EMPS), Expt 2 aimed to evaluate effects of supplementing shrub/tree leaves as CT source on feed consumption, digestibility, N retention, body weight gain and dressing percentage of growing sheep fed on elephant grass (EG) as a basal diet.Ten shrub and tree leaves used as CT sources were wild sunflower (Tithonia diversifolia), mulberry (Morus macroura), cassava (Manihot utilissima), avicienna (Avicennia marina), calliandra (Calliandra calothyrsus), sesbania (Sesbania grandiflora), acacia (acacia vilosa), glyricidia (Glyricidia sepium), jackfruit (Artocarpus heterophyllus), moringa (Moringa oleifera). The treatments applied in Expt 1 were: T1=Elephant grass (60%)+concentrate (40%); T2 = T1 + CT (3% DM); T3= T2 + PEG; T4 = T1 + CT (3.5% DM); T5 = T4 + PEG; T6 = T1 + CT (4% DM) and T7 = T6 + PEG. Data obtained were analysed using Randomized Block Design. Statistical analyses showed that treatments significanty affected (P<0.05) total gas production and EMPS. The lowest values of total gas production (45.9 ml/500 mg DM) and highest value of EMPS (64.6 g/kg BOTR) were observed in the treatment T4 (3.5% CT from cassava leave extract). Based on this result it was concluded that this treatment was the best and was chosen for further investigation using in vivo method. The treatmets applied for in vivo trial were: T1 = EG (60%) + concentrate (40%); T2 = T1 + dried cassava leave (equivalent to 3.5% CT); T3 = T2 + PEG. 18 growing sheep aging of 8-9 months and weighing of 23.67kg ± 1.23 were used in Expt 2. Results of in vivo study showed that treatments significanty affected (P<0.05) nutrients intake and digestibility (DM, OM and CP). N retention for sheep receiving treatment T2 were significantly higher (P<0.05; 15.6 g/d) than T1 (9.1 g/d) and T3 (8.53 g/d). Similar results were obtained for daily weight gain where T2 were the highest (62.79 g/d), followed by T1 (51.9 g/d) and T3 (52.85 g/d). Dressing percentage of T2 was the highest (51.54%) followed by T1 (49.61%) and T3 (49.32%). It can be concluded that adding adding dried cassava leaves did not reduce palatability due to CT, but rather increased OM digestibility and hence feed consumption was improved. N retention was increased due to the action of CT in the cassava leaves and this may have explained a higher input of N into duodenum which was further led to higer daily weight gain and dressing percentage.

Keywords: in vitro gas production, sheep, shrub and tree leaves, condensed tannin

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Authors: Dhekra Khazri, Hakim Gabtni

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Because of Sidi Bouzid water table overexploitation, this study aims at integrating geophysical methods to determinate aquifers geometry assessing their geological situation and geophysical characteristics. However in highly tectonic zones controlled by Atlassic structural features with NE-SW major directions (central Tunisia), Bouguer gravimetric responses of some areas can be as much dominated by the regional structural tendency, as being non-identified or either defectively interpreted such as the case of Sidi Bouzid basin. This issue required a residual gravity anomaly elaboration isolating the Sidi Bouzid basin gravity response ranging between -8 and -14 mGal and crucial for its aquifers geometry characterization. Several gravity techniques helped constructing the Sidi Bouzid basin's residual gravity anomaly, such as Upwards continuation compared to polynomial regression trends and power spectrum analysis detecting deep basement sources at (3km), intermediate (2km) and shallow sources (1km). A 3D Euler Deconvolution was also performed detecting deepest accidents trending NE-SW, N-S and E-W with depth values reaching 5500 m and delineating the main outcropping structures of the study area. Further gravity treatments highlighted the subsurface geometry and structural features of Sidi Bouzid basin over Horizontal and vertical gradient, and also filters based on them such as Tilt angle and Source Edge detector locating rooted edges or peaks from potential field data detecting a new E-W lineament compartmentalizing the Sidi Bouzid gutter into two unequally residual anomaly and subsiding domains. This subsurface morphology is also detected by the used 2D seismic reflection sections defining the Sidi Bouzid basin as a deep gutter within a tectonic set of negative flower structures, and collapsed and tilted blocks. Furthermore, these structural features were confirmed by forward gravity modeling process over several modeled residual gravity profiles crossing the main area. Sidi Bouzid basin (central Tunisia) is also of a big interest cause of the unknown total thickness and the undefined substratum of its siliciclastic Tertiary package, and its aquifers unbounded structural subsurface features and deep accidents. The Combination of geological, hydrogeological and geophysical methods is then of an ultimate need. Therefore, a geophysical methods integration based on gravity survey supporting available seismic data through forward gravity modeling, enhanced lateral and vertical extent definition of the basin's complex sedimentary fill via 3D gravity models, improved depth estimation by a depth to basement modeling approach, and provided 3D isochronous seismic mapping visualization of the basin's Tertiary complex refining its geostructural schema. A subsurface basin geomorphology mapping, over an ultimate matching between the basin's residual gravity map and the calculated theoretical signature map, was also displayed over the modeled residual gravity profiles. An ultimate multidisciplinary geophysical study of the Sidi Bouzid basin aquifers can be accomplished via an aeromagnetic survey and a 4D Microgravity reservoir monitoring offering temporal tracking of the target aquifer's subsurface fluid dynamics enhancing and rationalizing future groundwater exploitation in this arid area of central Tunisia.

Keywords: aquifer geometry, geophysics, 3D gravity modeling, improved depths, source edge detector

Procedia PDF Downloads 284

Authors: Ainur Sharip, Jose E. Perez, Nouf Alsharif, Aldo I. M. Bandeas, Enzo D. Fabrizio, Timothy Ravasi, Jasmeen S. Merzaban, Jürgen Kosel

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Bone marrow-derived human mesenchymal stem cells (MSCs) are attractive candidates for tissue engineering and regenerative medicine, due to their ability to differentiate into osteoblasts, chondrocytes or adipocytes. Differentiation is influenced by biochemical and biophysical stimuli provided by the microenvironment of the cell. Thus, altering the mechanical characteristics of a cell culture scaffold can directly influence a cell’s microenvironment and lead to stem cell differentiation. Mesenchymal stem cells were cultured on densely packed, vertically aligned magnetic iron nanowires (NWs) and the effect of NWs on the cell cytoskeleton rearrangement and differentiation were studied. An electrochemical deposition method was employed to fabricate NWs into nanoporous alumina templates, followed by a partial release to reveal the NW array. This created a cell growth substrate with free-standing NWs. The Fe NWs possessed a length of 2-3 µm, with each NW having a diameter of 33 nm on average. Mechanical stimuli generated by the physical movement of these iron NWs, in response to a magnetic field, can stimulate osteogenic differentiation. Induction of osteogenesis was estimated using an osteogenic marker, osteopontin, and a reduction of stem cell markers, CD73 and CD105. MSCs were grown on the NWs, and fluorescent microscopy was employed to monitor the expression of markers. A magnetic field with an intensity of 250 mT and a frequency of 0.1 Hz was applied for 12 hours/day over a period of one week and two weeks. The magnetically activated substrate enhanced the osteogenic differentiation of the MSCs compared to the culture conditions without magnetic field. Quantification of the osteopontin signal revealed approximately a seven-fold increase in the expression of this protein after two weeks of culture. Immunostaining staining against CD73 and CD105 revealed the expression of antibodies at the earlier time point (two days) and a considerable reduction after one-week exposure to a magnetic field. Overall, these results demonstrate the application of a magnetic NW substrate in stimulating the osteogenic differentiation of MSCs. This method significantly decreases the time needed to induce osteogenic differentiation compared to commercial biochemical methods, such as osteogenic differentiation kits, that usually require more than two weeks. Contact-free stimulation of MSC differentiation using a magnetic field has potential uses in tissue engineering, regenerative medicine, and bone formation therapies.

Keywords: cell substrate, magnetic nanowire, mesenchymal stem cell, stem cell differentiation

Procedia PDF Downloads 197

Authors: Gail Louw, Helena Boshoff, Taeksun Song, Clifton Barry

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Drug resistance in Mycobacterium tuberculosis is primarily attributed to mutations in target genes. These mutations incur a fitness cost and result in bacterial generations that are less fit, which subsequently acquire compensatory mutations to restore fitness. We hypothesize that mutations in specific drug target genes influence bacterial metabolism and cellular function, which affects its ability to develop subsequent resistance to additional agents. We aim to determine whether the sequential acquisition of drug resistance and specific mutations in a well-defined clinical M. tuberculosis strain promotes or limits the development of additional resistance. In vitro mutants resistant to pretomanid, linezolid, moxifloxacin, rifampicin and kanamycin were generated from a pan-susceptible clinical strain from the Beijing lineage. The resistant phenotypes to the anti-TB agents were confirmed by the broth microdilution assay and genetic mutations were identified by targeted gene sequencing. Growth of mono-resistant mutants was done in enriched medium for 14 days to assess in vitro fitness. Double resistant mutants were generated against anti-TB drug combinations at concentrations 5x and 10x the minimum inhibitory concentration. Subsequently, mutation frequencies for these anti-TB drugs in the different mono-resistant backgrounds were determined. The initial level of resistance and the mutation frequencies observed for the mono-resistant mutants were comparable to those previously reported. Targeted gene sequencing revealed the presence of known and clinically relevant mutations in the mutants resistant to linezolid, rifampicin, kanamycin and moxifloxacin. Significant growth defects were observed for mutants grown under in vitro conditions compared to the sensitive progenitor. Mutation frequencies determination in the mono-resistant mutants revealed a significant increase in mutation frequency against rifampicin and kanamycin, but a significant decrease in mutation frequency against linezolid and sutezolid. This suggests that these mono-resistant mutants are more prone to develop resistance to rifampicin and kanamycin, but less prone to develop resistance against linezolid and sutezolid. Even though kanamycin and linezolid both inhibit protein synthesis, these compounds target different subunits of the ribosome, thereby leading to different outcomes in terms of fitness in the mutants with impaired cellular function. These observations showed that oxazolidinone treatment is instrumental in limiting the development of multi-drug resistance in M. tuberculosis in vitro.

Keywords: oxazolidinones, mutations, resistance, tuberculosis

Procedia PDF Downloads 163

Authors: Boutheina Ben Abdelmoumen Mardassi, Salim Chibani, Safa Boujemaa, Amaury Vaysse, Julien Guglielmini, Elhem Yacoub

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Background and aim: Mycoplasma hominis (MH) is a pathogenic bacterium belonging to the Mollicutes class. It causes a wide range of gynecological infections and infertility among adults. Recently, we have explored for the first time the phylodistribution of Tunisian M. hominis clinical strains using an expanded MLST. We have demonstrated their distinction into two pure lineages, which each corresponding to a specific pathotype: genital infections and infertility. The aim of this project is to gain further insight into the evolutionary dynamics and the specific genetic factors that distinguish MH pathotypes Methods: Whole genome sequencing of Mycoplasma hominis clinical strains was performed using illumina Miseq. Denovo assembly was performed using a publicly available in-house pipeline. We used prokka to annotate the genomes, panaroo to generate the gene presence matrix and Jolytree to establish the phylogenetic tree. We used treeWAS to identify genetic loci associated with the pathothype of interest from the presence matrix and phylogenetic tree. Results: Our results revealed a clear categorization of the 62 MH clinical strains into two distinct genetic lineages, with each corresponding to a specific pathotype.; gynecological infections and infertility[AV1] . Genome annotation showed that GC content is ranging between 26 and 27%, which is a known characteristic of Mycoplasma genome. Housekeeping genes belonging to the core genome are highly conserved among our strains. TreeWas identified 4 virulence genes associated with the pathotype gynecological infection. encoding for asparagine--tRNA ligase, restriction endonuclease subunit S, Eco47II restriction endonuclease, and transcription regulator XRE (involved in tolerance to oxidative stress). Five genes have been identified that have a statistical association with infertility, tow lipoprotein, one hypothetical protein, a glycosyl transferase involved in capsule synthesis, and pyruvate kinase involved in biofilm formation. All strains harbored an efflux pomp that belongs to the family of multidrug resistance ABC transporter, which confers resistance to a wide range of antibiotics. Indeed many adhesion factors and lipoproteins (p120, p120', p60, p80, Vaa) have been checked and confirmed in our strains with a relatively 99 % to 96 % conserved domain and hypervariable domain that represent 1 to 4 % of the reference sequence extracted from gene bank. Conclusion: In summary, this study led to the identification of specific genetic loci associated with distinct pathotypes in M hominis.

Keywords: mycoplasma hominis, infertility, gynecological infections, virulence genes, antibiotic resistance

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Authors: Haji Mohammad Zulfan Farhi Bin Haji Sulaini, Mohammad Azeezudde’en Bin Mohd Ismaon

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The demands placed on military athletes extend beyond physical prowess to encompass cognitive resilience in high-stress environments. This study investigates the effects of resilience training on the cognitive performance of military athletes, shedding light on the potential benefits and implications for optimizing their overall readiness. In a rapidly evolving global landscape, armed forces worldwide are recognizing the importance of cognitive resilience alongside physical fitness. The study employs a mixed-methods approach, incorporating quantitative cognitive assessments and qualitative data from military athletes undergoing resilience training programs. Cognitive performance is evaluated through a battery of tests, including measures of memory, attention, decision-making, and reaction time. The participants, drawn from various branches of the military, are divided into experimental and control groups. The experimental group undergoes a comprehensive resilience training program, while the control group receives traditional physical training without a specific focus on resilience. The initial findings indicate a substantial improvement in cognitive performance among military athletes who have undergone resilience training. These improvements are particularly evident in domains such as attention and decision-making. The experimental group demonstrated enhanced situational awareness, quicker problem-solving abilities, and increased adaptability in high-stress scenarios. These results suggest that resilience training not only bolsters mental toughness but also positively impacts cognitive skills critical to military operations. In addition to quantitative assessments, qualitative data is collected through interviews and surveys to gain insights into the subjective experiences of military athletes. Preliminary analysis of these narratives reveals that participants in the resilience training program report higher levels of self-confidence, emotional regulation, and an improved ability to manage stress. These psychological attributes contribute to their enhanced cognitive performance and overall readiness. Moreover, this study explores the potential long-term benefits of resilience training. By tracking participants over an extended period, we aim to assess the durability of cognitive improvements and their effects on overall mission success. Early results suggest that resilience training may serve as a protective factor against the detrimental effects of prolonged exposure to stressors, potentially reducing the risk of burnout and psychological trauma among military athletes. This research has significant implications for military organizations seeking to optimize the performance and well-being of their personnel. The findings suggest that integrating resilience training into the training regimen of military athletes can lead to a more resilient and cognitively capable force. This, in turn, may enhance mission success, reduce the risk of injuries, and improve the overall effectiveness of military operations. In conclusion, this study provides compelling evidence that resilience training positively impacts the cognitive performance of military athletes. The preliminary results indicate improvements in attention, decision-making, and adaptability, as well as increased psychological resilience. As the study progresses and incorporates long-term follow-ups, it is expected to provide valuable insights into the enduring effects of resilience training on the cognitive readiness of military athletes, contributing to the ongoing efforts to optimize military personnel's physical and mental capabilities in the face of ever-evolving challenges.

Keywords: military athletes, cognitive performance, resilience training, cognitive enhancement program

Procedia PDF Downloads 80

Authors: Ali R. Alshawawreh, Francisco Liébana-Cabanillas, Francisco J. Blanco-Encomienda

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Collaboration between marketing and business intelligence (BI) is crucial in today's ever-evolving business landscape. These two domains play pivotal roles in molding customers' experiential knowledge. Marketing insights offer valuable information regarding customer needs, preferences, and behaviors. Conversely, BI facilitates data-driven decision-making, leading to heightened operational efficiency, product quality, and customer satisfaction. Customer experiential knowledge (CEK) encompasses customers' implicit comprehension of consumption experiences influenced by diverse factors, including social and cultural influences. This study primarily focuses on telecommunications companies in Jordan, scrutinizing how experiential customer knowledge mediates the relationship between marketing intelligence and business intelligence. Drawing on theoretical frameworks such as the resource-based view (RBV) and service-dominant logic (SDL), the research aims to comprehend how organizations utilize their resources, particularly knowledge, to foster Evolution. Employing a quantitative research approach, the study collected and analyzed primary data to explore hypotheses. Structural equation modeling (SEM) facilitated by Smart PLS software evaluated the relationships between the constructs, followed by mediation analysis to assess the indirect associations in the model. The study findings offer insights into the intricate dynamics of organizational Creation, uncovering the interconnected relationships between business intelligence, customer experiential knowledge-based innovation (CEK-DI), marketing intelligence (MI), and product and service innovation (PSI), underscoring the pivotal role of advanced intelligence capabilities in developing innovative practices rooted in a profound understanding of customer experiences. Furthermore, the positive impact of BI on PSI reaffirms the significance of data-driven decision-making in shaping the innovation landscape. The significant impact of CEK-DI on PSI highlights the critical role of customer experiences in driving an organization. Companies that actively integrate customer insights into their opportunity creation processes are more likely to create offerings that match customer expectations, which drives higher levels of product and service sophistication. Additionally, the positive and significant impact of MI on CEK-DI underscores the critical role of market insights in shaping evolutionary strategies. While the relationship between MI and PSI is positive, the slightly weaker significance level indicates a subtle association, suggesting that while MI contributes to the development of ideas, In conclusion, the study emphasizes the fundamental role of intelligence capabilities, especially artificial intelligence, emphasizing the need for organizations to leverage market and customer intelligence to achieve effective and competitive innovation practices. Collaborative efforts between marketing and business intelligence serve as pivotal drivers of development, influencing customer experiential knowledge and shaping organizational strategies and practices. Future research could adopt longitudinal designs and gather data from various sectors to offer broader insights. Additionally, the study focuses on the effects of marketing intelligence, business intelligence, customer experiential knowledge, and innovation, but other unexamined variables may also influence innovation processes. Future studies could investigate additional factors, mediators, or moderators, including the role of emerging technologies like AI and machine learning in driving innovation.

Keywords: marketing intelligence, business intelligence, product, customer experiential knowledge-driven innovation

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Authors: Sultan Z. Mahmud, Emily C. Graff, Adil Bashir

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Phosphoprotein enriched in astrocytes 15 kDa (PEA-15) is a multifunctional adapter protein which is associated with the regulation of apoptotic cell death. Recently it has been discovered that PEA-15 is crucial in normal neurodevelopment of domestic cats, a gyrencephalic animal model, although the exact function of PEA-15 in neurodevelopment is unknown. This study investigates how PEA-15 affects the blood-brain barrier (BBB) permeability in cat brain, which can cause abnormalities in tissue metabolite and energy supplies. Severe polymicrogyria and microcephaly have been observed in cats with a loss of function PEA-15 mutation, affecting the normal neurodevelopment of the cat. This suggests that the vital role of PEA-15 in neurodevelopment is associated with gyrification. Neurodevelopment is a highly energy demanding process. The mammalian brain depends on glucose as its main energy source. PEA-15 plays a very important role in glucose uptake and utilization by interacting with phospholipase D1 (PLD1). Mitochondria also plays a critical role in bioenergetics and essential to supply adequate energy needed for neurodevelopment. Cerebral blood flow regulates adequate metabolite supply and recent findings also showed that blood plasma contains mitochondria as well. So the BBB can play a very important role in regulating metabolite and energy supply in the brain. In this study the blood-brain permeability in cat brain was measured using MRI magnetization transfer (MT) effect on the perfusion signal. Perfusion is the tissue mass normalized supply of blood to the capillary bed. Perfusion also accommodates the supply of oxygen and other metabolites to the tissue. A fraction of the arterial blood can diffuse to the tissue, which depends on the BBB permeability. This fraction is known as water extraction fraction (EF). MT is a process of saturating the macromolecules, which has an effect on the blood that has been diffused into the tissue while having minimal effect on intravascular blood water that has not been exchanged with the tissue. Measurement of perfusion signal with and without MT enables to estimate the microvascular blood flow, EF and permeability surface area product (PS) in the brain. All the experiments were performed with Siemens 7T Magnetom with 32 channel head coil. Three control cats and three PEA-15 mutant cats were used for the study. Average EF in white and gray matter was 0.9±0.1 and 0.86±0.15 respectively, perfusion in white and gray matter was 85±15 mL/100g/min and 97±20 mL/100g/min respectively, PS in white and gray matter was 201±25 mL/100g/min and 225±35 mL/100g/min respectively for control cats. For PEA-15 mutant cats, average EF in white and gray matter was 0.81±0.15 and 0.77±0.2 respectively, perfusion in white and gray matter was 140±25 mL/100g/min and 165±18 mL/100g/min respectively, PS in white and gray matter was 240±30 mL/100g/min and 259±21 mL/100g/min respectively. This results show that BBB is compromised in PEA-15 mutant cat brain, where EF is decreased and perfusion as well as PS are increased in the mutant cats compared to the control cats. This findings might further explain the function of PEA-15 in neurodevelopment.

Keywords: BBB, cat brain, magnetization transfer, PEA-15

Procedia PDF Downloads 144

Authors: Benedicta N. Nkeh-Chungag, Constance R. Sewani-Rusike, Isaac M. Malema, Daniel T. Goon, Oladele V. Adeniyi, Idowu A. Ajayi

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Background: Childhood and adolescent obesity is an important predictor of adult cardiometabolic diseases. Current data on age- and gender-specific cardiometabolic risk factors are lacking in the peri-urban Eastern Cape Province, South Africa. However, such information is important in designing innovative strategies to promote healthy living among children and adolescents. The purpose of this pilot study was to compare and determine the extent of cardiometabolic risk factors between samples of lean and overweight/obese adolescent population in a peri-urban township of South Africa. Methods: In this case-control study, age-matched, non-pregnant and non-lactating female adolescents consisting of equal number of cases (50 overweight/obese) and control (50 lean) participated in the study. Fasting venous blood samples were obtained for total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), triglyceride (Trig), highly sensitive C-reactive protein (hsCRP) and blood sugar. Anthropometric measurements included weight, height, waist and hip circumferences. Body mass index was calculated. Blood pressure was measured; and metabolic syndrome was assessed using appropriate diagnostic criteria for children and adolescents. Results: Of the 76 participants with complete data, 12/38 of the overweight/obese and 1/38 of the lean group met the criteria for adolescent metabolic syndrome. All cardiometabolic risk factors were elevated in the overweight/obese group compared with the lean group: low HDL-C (RR = 2.21), elevated TC (RR = 1.23), elevated LDL-C (RR = 1.42), elevated Trig (RR = 1.73), and elevated hsCRP (RR = 1.9). There were significant atherosclerotic indices among the overweight/obese group compared with the lean group: TC/HDL and LDL/HDL (2.99±0.91 vs 2.63±0.48; p=0.016 and 1.73±0.61 vs 1.41±0.46; p= 0.014, respectively). Conclusion: There are multiple cardiometabolic risk factors among the overweight/obese female adolescent group compared with lean adolescent group in the study. Female adolescent who are overweight and obese have higher relative risks of developing cardiometabolic diseases compared with their lean counterparts in the peri-urban Mthatha, South Africa. School health programme focusing on promoting physical exercise, healthy eating and keeping appropriate weight are needed in the country.

Keywords: adolescents, cardiometabolic risk factors, obesity, peri-urban South Africa

Procedia PDF Downloads 474

Authors: Catherine P. Bradshaw, Elise T. Pas, Katrina J. Debnam, Jessika H. Bottiani, Michael Rosenberg

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Worldwide, ethnically and culturally diverse students are at increased risk for school failure, discipline problems, and dropout. Despite decades of concern about this issue of disparities in education and other fields (e.g., 'school to prison pipeline'), there has been limited empirical examination of models that can actually reduce these gaps in schools. Moreover, few studies have examined the effectiveness of in-service teacher interventions and supports specifically designed to reduce discipline disparities and improve student engagement. This session provides an overview of the evidence-based Double Check model which serves as a framework for teachers to use culturally-responsive strategies to engage ethnically and culturally diverse students in the classroom and reduce discipline problems. Specifically, Double Check is a school-based prevention program which includes three core components: (a) enhancements to the school-wide Positive Behavioral Interventions and Supports (PBIS) tier-1 level of support; (b) five one-hour professional development training sessions, each of which addresses five domains of cultural competence (i.e., connection to the curriculum, authentic relationships, reflective thinking, effective communication, and sensitivity to students’ culture); and (c) coaching of classroom teachers using an adapted version of the Classroom Check-Up, which intends to increase teachers’ use of effective classroom management and culturally-responsive strategies using research-based motivational interviewing and data-informed problem-solving approaches. This paper presents findings from a randomized controlled trial (RCT) testing the impact of Double Check, on office discipline referrals (disaggregated by race) and independently observed and self-reported culturally-responsive practices and classroom behavior management. The RCT included 12 elementary and middle schools; 159 classroom teachers were randomized either to receive coaching or serve as comparisons. Specifically, multilevel analyses indicated that teacher self-reported culturally responsive behavior management improved over the course of the school year for teachers who received the coaching and professional development. However, the average annual office discipline referrals issued to black students were reduced among teachers who were randomly assigned to receive coaching relative to comparison teachers. Similarly, observations conducted by trained external raters indicated significantly more teacher proactive behavior management and anticipation of student problems, higher student compliance, less student non-compliance, and less socially disruptive behaviors in classrooms led by coached teachers than classrooms led teachers randomly assigned to the non-coached condition. These findings indicated promising effects of the Double Check model on a range of teacher and student outcomes, including disproportionality in office discipline referrals among Black students. These results also suggest that the Double Check model is one of only a few systematic approaches to promoting culturally-responsive behavior management which has been rigorously tested and shown to be associated with improvements in either student or staff outcomes indicated significant reductions in discipline problems and improvements in behavior management. Implications of these findings are considered within the broader context of globalization and demographic shifts, and their impacts on schools. These issues are particularly timely, given growing concerns about immigration policies in the U.S. and abroad.

Keywords: ethnically and culturally diverse students, student engagement, school-based prevention, academic achievement

Procedia PDF Downloads 282

Authors: G. J. Fan, T. T. Lee, M. H. Chen, T. F. Shiao, B. Yu, C. F. Lee

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Rice straw with poor feed quality and spent mushroom substrate are both the most abundant agricultural residues in Taiwan. Edible mushrooms from white rot fungi possess lignocellulase activity. It was expected to improve the feeding value of rice straw for ruminant by solid-state fermentation pretreatment using spent mushroom substrate. Six varieties or subspecies of spent edible mushrooms (Pleurotus ostreatus (blue or white color), P. sajor-caju, P. citrinopileatus, P. eryngii and Ganoderma lucidium) substrate were evaluated in solid-state fermentation process with rice straw for 8 wks. Quality improvement of fermented rice straw was determined by its in vitro digestibility, lignocellulose degradability, and cell wall breakdown checked by scanning electron microscope. Results turned out that Pleurotus ostreatus (white color) and P. sajor-caju had the better lignocellulose degradation effect than the others and was chosen for advance in vivo study. Rice straw fermented with spent Pleurotus ostreatus or Pleurotus sajor-caju mushroom substrate 8 wks was prepared for growing and lactating feeding trials of dairy goat, respectively. Pangolagrass hay at 15% diet dry matter was the control diet. Fermented or original rice straw was added to substitute pangolagrass hay in both feeding trials. A total of 30 head of Alpine castrated ram were assigned into three groups for 11 weeks, 5 pens (2 head/pen) each group. A total of 21 head of Saanen and Alpine goats were assigned into three treatments and individually fed in two repeat lactating trials with 28-d each. In castrated ram study, results showed that fermented rice straw by spent Pleurotus ostreatus mushroom substrate attributed the higher daily dry matter intakes (DMI, 1.53 vs. 1.20 kg) and body weight gain (138 vs. 101 g) than goats fed original rice straw. DMI (2.25 vs. 1.81 kg) and milk yield (3.31 vs. 3.02 kg) of lactating goats fed control pangolagrass diet and fermented rice straw by spent Pleurotus sajor-caju mushroom substrate were also higher than those fed original rice straw diet (P < 0.05). Milk compositions, milk fat, protein, total solid and lactose, were similar among treatments. In conclusion, solid-state fermentation by spent Pleurotus ostreatus or Pleurotus sajor-caju mushroom substrate could effectively improve the feeding value of rice straw. Fermented rice straw is a good alternative fiber feed resource for growing and lactating dairy goats and 15% in diet dry matter is recommended.

Keywords: feeding value, fermented rice straw, growing and lactating dairy goat, spent Pleurotus ostreatus and Pleurotus sajor-caju mushroom substrate

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Authors: Shraddha Rane, Richard Warren, Stephen Eyre

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L-23 is a pro-inflammatory molecule that signals T cells to release cytokines such as IL-17A and IL-22. Psoriasis is driven by a dysregulated immune response, within which IL-23 is now thought to play a key role. Genome-wide association studies (GWAS) have identified a number of genetic risk loci that support the involvement of IL-23 signalling in psoriasis; in particular a robust susceptibility locus at a gene encoding a subunit of the IL-23 receptor (IL23R) (Stuart et al., 2015; Tsoi et al., 2012). The lead psoriasis-associated SNP rs9988642 is located approximately 500 bp downstream of IL23R but is in tight linkage disequilibrium (LD) with a missense SNP rs11209026 (R381Q) within IL23R (r2 = 0.85). The minor (G) allele of rs11209026 is present in approximately 7% of the population and is protective for psoriasis and several other autoimmune diseases including IBD, ankylosing spondylitis, RA and asthma. The psoriasis-associated missense SNP R381Q causes an arginine to glutamine substitution in a region of the IL23R protein between the transmembrane domain and the putative JAK2 binding site in the cytoplasmic portion. This substitution is expected to affect the receptor’s surface localisation or signalling ability, rather than IL23R expression. Recent studies have also identified a psoriatic arthritis (PsA)-specific signal at IL23R; thought to be independent from the psoriasis association (Bowes et al., 2015; Budu-Aggrey et al., 2016). The lead PsA-associated SNP rs12044149 is intronic to IL23R and is in LD with likely causal SNPs intersecting promoter and enhancer marks in memory CD8+ T cells (Budu-Aggrey et al., 2016). It is therefore likely that the PsA-specific SNPs affect IL23R function via a different mechanism compared with the psoriasis-specific SNPs. It could be hypothesised that the risk allele for PsA located within the IL23R promoter causes an increase IL23R expression, relative to the protective allele. An increased expression of IL23R might then lead to an exaggerated immune response. The independent genetic signals identified for psoriasis and PsA in this locus indicate that different mechanisms underlie these two conditions; although likely both affecting the function of IL23R. It is very important to further characterise these mechanisms in order to better understand how the IL-23 receptor and its downstream signalling is affected in both diseases. This will help to determine how psoriasis and PsA patients might differentially respond to therapies, particularly IL-23 biologics. To investigate this further we have developed an in vitro model using CD4 T cells which express either wild type IL23R and IL12Rβ1 or mutant IL23R (R381Q) and IL12Rβ1. Model expressing different isotypes of IL23R is also underway to investigate the effects on IL23R expression. We propose to further investigate the variants for Ps and PsA and characterise key intracellular processes related to the variants.

Keywords: IL23R, psoriasis, psoriatic arthritis, SNP

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Authors: Mohammad Afzal Khan, Fatimah Alanazi, Hala Abdalrahman Ahmed, Talal Shamma, Kilian Kelly, Mohammed A. Hammad, Abdullah O. Alawad, Abdullah Mohammed Assiri, Dieter Clemens Broering

Abstract:

Lung transplantation is a life-saving surgical replacement of diseased lungs in patients with end-stage respiratory malfunctions. Despite the remarkable short-term recovery, long-term lung survival continues to face several significant challenges, including chronic rejection and severe toxic side-effects due to global immunosuppression. Stem cell-based immunotherapy has been recognized as a crucial immunoregulatory regimen in various preclinical and clinical studies. Despite initial therapeutic outcomes, conventional stem cells face key limitations. The Cymerus™ manufacturing facilitates the production of a virtually limitless supply of consistent human induced pluripotent stem cell (iPSC)-derived mesenchymal stem cells, which could play a key role in selective immunosuppression and graft repair during rejection. Here, we demonstrated the impact of iPSC-derived human MSCs on the development of immune-tolerance and long-term graft survival in mouse orthotopic airway allografts. BALB/c→C57BL/6 allografts were reconstituted with iPSC-derived MSCs (2 million/transplant/ at d0), and allografts were examined for regulatory T cells (Tregs), oxygenation, microvascular blood flow, airway epithelium and collagen deposition during rejection. We demonstrated that iPSC-derived MSC treatment leads to significant increase in tissue expression of hTSG-6 protein, followed by an upregulation of mouse Tregs and IL-5, IL-10, IL-15 cytokines, which augments graft microvascular blood flow and oxygenation, and thereby maintained a healthy airway epithelium and prevented the subepithelial deposition of collagen at d90 post-transplantation. Collectively, these data confirmed that iPSC-derived MSC-mediated immunosuppression has potential to establish immune-tolerance and rescue allograft from sustained hypoxic/ischemic phase and subsequently limits long-term airway epithelial injury and collagen progression, which therapeutically warrant a study of Cymerus iPSC-derived MSCs as a potential management option for immunosuppression in transplant recipients.

Keywords: stem cell therapy, immunotolerance, regulatory T cells, hypoxia and ischemia, microvasculature

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Authors: Stefano Scippo, Damiana Luzzi, Stefano Cuomo, Maria Ranieri

Abstract:

Background: In response to the rise in natural disasters linked to climate change, numerous studies on Disaster Risk Reduction Education (DRRE) have emerged since the '90s, mainly using a didactic transmission-based approach. Effective DRRE should align with an interactive, experiential, and participatory educational model, which can be costly and risky. A potential solution is using simulations facilitated by eXtended Reality (XR). Research Question: This study aims to conduct a scoping review to explore educational methodologies that use XR to enhance knowledge among teachers, students, and citizens about environmental risks, natural disasters (including climate-related ones), and their management. Method: A search string of 66 keywords was formulated, spanning three domains: 1) education and target audience, 2) environment and natural hazards, and 3) technologies. On June 21st, 2023, the search string was used across five databases: EBSCOhost, IEEE Xplore, PubMed, Scopus, and Web of Science. After deduplication and removing papers without abstracts, 2,152 abstracts (published between 2013 and 2023) were analyzed and 2,062 papers were excluded, followed by the exclusion of 56 papers after full-text scrutiny. Excluded studies focused on unrelated technologies, non-environmental risks, and lacked educational outcomes or accessible texts. Main Results: The 34 reviewed papers were analyzed for context, risk type, research methodology, learning objectives, XR technology use, outcomes, and educational affordances of XR. Notably, since 2016, there has been a rise in scientific publications, focusing mainly on seismic events (12 studies) and floods (9), with a significant contribution from Asia (18 publications), particularly Japan (7 studies). Methodologically, the studies were categorized into empirical (26) and non-empirical (8). Empirical studies involved user or expert validation of XR tools, while non-empirical studies included systematic reviews and theoretical proposals without experimental validation. Empirical studies were further classified into quantitative, qualitative, or mixed-method approaches. Six qualitative studies involved small groups of users or experts, while 20 quantitative or mixed-method studies used seven different research designs, with most (17) employing a quasi-experimental, one-group post-test design, focusing on XR technology usability over educational effectiveness. Non-experimental studies had methodological limitations, making their results hypothetical and in need of further empirical validation. Educationally, the learning objectives centered on knowledge and skills for surviving natural disaster emergencies. All studies recommended XR technologies for simulations or serious games but did not develop comprehensive educational frameworks around these tools. XR-based tools showed potential superiority over traditional methods in teaching risk and emergency management skills. However, conclusions were more valid in studies with experimental designs; otherwise, they remained hypothetical without empirical evidence. The educational affordances of XR, mainly user engagement, were confirmed by the studies. Authors’ Conclusions: The analyzed literature lacks specific educational frameworks for XR in DRRE, focusing mainly on survival knowledge and skills. There is a need to expand educational approaches to include uncertainty education, developing competencies that encompass knowledge, skills, and attitudes like risk perception.

Keywords: disaster risk reduction education, educational technologies, scoping review, XR technologies

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Authors: Maria Ostroukhova, Zhanneta Zalutskaya, Elena Ermilova

Abstract:

The alternative oxidase (AOX) mediates cyanide-resistant respiration, which bypasses proton-pumping complexes III and IV of the cytochrome pathway to directly transfer electrons from reduced ubiquinone to molecular oxygen. In Chlamydomonas reinhardtii, AOX is a monomeric protein that is encoded by two genes of discrete subfamilies, AOX1 and AOX2. Although AOX has been proposed to play essential roles in stress tolerance of organisms, the role of subfamily AOX2 is largely unknown. In C. reinhardtii, AOX2 was initially identified as one of constitutively low expressed genes. Like other photosynthetic organisms C. reinhardtii cells frequently experience periods of hypoxia. To examine AOX2 transcriptional regulation and role of AOX2 in hypoxia adaptation, real-time PCR analysis and artificial microRNA method were employed. Two experimental approaches have been used to induce the anoxic conditions: dark-anaerobic and light-anaerobic conditions. C. reinhardtii cells exposed to the oxygen deprivation have shown increased AOX2 mRNA levels. By contrast, AOX1 was not an anoxia-responsive gene. In C. reinhardtii, a subset of genes is regulated by transcription factor CRR1 in anaerobic conditions. Notable, the AOX2 promoter region contains the potential motif for CRR1 binding. Therefore, the role of CRR1 in the control of AOX2 transcription was tested. The CRR1-underexpressing strains, that were generated and characterized in this work, exhibited low levels of AOX2 transcripts under anoxic conditions. However, the transformants still slightly induced AOX2 gene expression in the darkness. These confirmed our suggestions that darkness is a regulatory stimulus for AOX genes in C. reinhardtii. Thus, other factors must contribute to AOX2 promoter activity under dark-anoxic conditions. Moreover, knock-down of CRR1 caused a complete reduction of AOX2 expression under light-anoxic conditions. These results indicate that (1) CRR1 is required for AOX2 expression during hypoxia, and (2) AOX2 gene is regulated by CRR1 together with yet-unknown regulatory factor(s). In addition, the AOX2-underexpressing strains were generated. The analysis of amiRNA-AOX2 strains suggested a role of this alternative oxidase in hypoxia adaptation of the alga. In conclusion, the results reported here show that C. reinhardtii AOX2 gene is stress inducible. CRR1 transcriptional factor is involved in the regulation of the AOX2 gene expression in the absence of oxygen. Moreover, AOX2 but not AOX1 functions under oxygen deprivation. This work was supported by Russian Science Foundation (research grant № 16-14-10004).

Keywords: alternative oxidase 2, artificial microRNA approach, chlamydomonas reinhardtii, hypoxia

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Authors: Mustafa M. Donma, Orkide Donma

Abstract:

Obesity is a clinical state associated with low-grade inflammation. It is also a major risk factor for insulin resistance (IR). In its advanced stages, metabolic syndrome (MetS), a much more complicated disease which may lead to life-threatening problems, may develop. Obesity-mediated IR seems to correlate with the inflammation. Human studies performed particularly on pediatric population are scarce. The aim of this study is to detect possible associations between inflammation and IR in terms of some related ratios. 549 children were grouped according to their age- and sex-based body mass index (BMI) percentile tables of WHO. MetS components were determined. Informed consent and approval from the Ethics Committee for Clinical Investigations were obtained. The principles of the Declaration of Helsinki were followed. The exclusion criteria were infection, inflammation, chronic diseases and those under drug treatment. Anthropometric measurements were obtained. Complete blood cell, fasting blood glucose, insulin, and C-reactive protein (CRP) analyses were performed. Homeostasis model assessment of insulin resistance (HOMA-IR), systemic immune inflammation (SII) index, tense index, alanine aminotransferase to aspartate aminotransferase ratio (ALT/AST), neutrophils to lymphocyte (NLR), platelet to lymphocyte, and lymphocyte to monocyte ratios were calculated. Data were evaluated by statistical analyses. The degree for statistical significance was 0.05. Statistically significant differences were found among the BMI values of the groups (p < 0.001). Strong correlations were detected between the BMI and waist circumference (WC) values in all groups. Tense index values were also correlated with both BMI and WC values in all groups except overweight (OW) children. SII index values of children with normal BMI were significantly different from the values obtained in OW, obese, morbid obese and MetS groups. Among all the other lymphocyte ratios, NLR exhibited a similar profile. Both HOMA-IR and ALT/AST values displayed an increasing profile from N towards MetS3 group. BMI and WC values were correlated with HOMA-IR and ALT/AST. Both in morbid obese and MetS groups, significant correlations between CRP versus SII index as well as HOMA-IR versus ALT/AST were found. ALT/AST and HOMA-IR values were correlated with NLR in morbid obese group and with SII index in MetS group, (p < 0.05), respectively. In conclusion, these findings showed that some parameters may exhibit informative differences between the early and late stages of obesity. Important associations among HOMA-IR, ALT/AST, NLR and SII index have come to light in the morbid obese and MetS groups. This study introduced the SII index and NLR as important inflammatory markers for the discrimination of normal and obese children. Interesting links were observed between inflammation and IR in morbid obese children and those with MetS, both being late stages of obesity.

Keywords: children, inflammation, insulin resistance, metabolic syndrome, obesity

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Authors: Sulalit Bandyopadhyay, Muhammad Awais Ashfaq Alvi, Anuvansh Sharma, Wilhelm R. Glomm

Abstract:

Release of drugs from nanogels and nanogel-based systems can occur under the influence of external stimuli like temperature, pH, magnetic fields and so on. pNIPAm-AAc nanogels respond to the combined action of both temperature and pH, the former being mostly determined by hydrophilic-to-hydrophobic transitions above the volume phase transition temperature (VPTT), while the latter is controlled by the degree of protonation of the carboxylic acid groups. These nanogels based systems are promising candidates in the field of drug delivery. Combining nanogels with magneto-plasmonic nanoparticles (NPs) introduce imaging and targeting modalities along with stimuli-response in one hybrid system, thereby incorporating multifunctionality. Fe@Au core-shell NPs possess optical signature in the visible spectrum owing to localized surface plasmon resonance (LSPR) of the Au shell, and superparamagnetic properties stemming from the Fe core. Although there exist several synthesis methods to control the size and physico-chemical properties of pNIPAm-AAc nanogels, yet, there is no comprehensive study that highlights the dependence of incorporation of one or more layers of NPs to these nanogels. In addition, effective determination of volume phase transition temperature (VPTT) of the nanogels is a challenge which complicates their uses in biological applications. Here, we have modified the swelling-collapse properties of pNIPAm-AAc nanogels, by combining with Fe@Au NPs using different solution based methods. The hydrophilic-hydrophobic transition of the nanogels above the VPTT has been confirmed to be reversible. Further, an analytical method has been developed to deduce the average VPTT which is found to be 37.3°C for the nanogels and 39.3°C for nanogel coated Fe@Au NPs. An opposite swelling –collapse behaviour is observed for the latter where the Fe@Au NPs act as bridge molecules pulling together the gelling units. Thereafter, Cyt C, a model protein drug and L-Dopa, a drug used in the clinical treatment of Parkinson’s disease were loaded separately into the nanogels and nanogel coated Fe@Au NPs, using a modified breathing-in mechanism. This gave high loading and encapsulation efficiencies (L Dopa: ~9% and 70µg/mg of nanogels, Cyt C: ~30% and 10µg/mg of nanogels respectively for both the drugs. The release kinetics of L-Dopa, monitored using UV-vis spectrophotometry was observed to be rather slow (over several hours) with highest release happening under a combination of high temperature (above VPTT) and acidic conditions. However, the release of L-Dopa from nanogel coated Fe@Au NPs was the fastest, accounting for release of almost 87% of the initially loaded drug in ~30 hours. The chemical structure of the drug, drug incorporation method, location of the drug and presence of Fe@Au NPs largely alter the drug release mechanism and the kinetics of these nanogels and Fe@Au NPs coated with nanogels.

Keywords: controlled release, nanogels, volume phase transition temperature, l-dopa

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