Search results for: stem cell
2973 Poly-ε-Caprolactone Nanofibers with Synthetic Growth Factor Enriched Liposomes as Controlled Drug Delivery System
Authors: Vera Sovkova, Andrea Mickova, Matej Buzgo, Karolina Vocetkova, Eva Filova, Evzen Amler
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PCL (poly-ε-caprolactone) nanofibrous scaffolds with adhered liposomes were prepared and tested as a possible drug delivery system for various synthetic growth factors. TGFβ, bFGF, and IGF-I have been shown to increase hMSC (human mesenchymal stem cells) proliferation and to induce hMSC differentiation. Functionalized PCL nanofibers were prepared with synthetic growth factors encapsulated in liposomes adhered to them in three different concentrations. Other samples contained PCL nanofibers with adhered, free synthetic growth factors. The synthetic growth factors free medium served as a control. The interaction of liposomes with the PCL nanofibers was visualized by SEM, and the release kinetics were determined by ELISA testing. The potential of liposomes, immobilized on the biodegradable scaffolds, as a delivery system for synthetic growth factors, and as a suitable system for MSCs adhesion, proliferation and differentiation in vitro was evaluated by MTS assay, dsDNA amount determination, confocal microscopy, flow cytometry and real-time PCR. The results showed that the growth factors adhered to the PCL nanofibers stimulated cell proliferation mainly up to day 11 and that subsequently their effect was lower. By contrast, the release of the lowest concentration of growth factors from liposomes resulted in gradual proliferation of MSCs throughout the experiment. Moreover, liposomes, as well as free growth factors, stimulated type II collagen production, which was confirmed by immunohistochemical staining using monoclonal antibody against type II collagen. The results of this study indicate that growth factors enriched liposomes adhered to surface of PCL nanofibers could be useful as a drug delivery instrument for application in short timescales, be combined with nanofiber scaffolds to promote local and persistent delivery while mimicking the local microenvironment. This work was supported by project LO1508 from the Ministry of Education, Youth and Sports of the Czech RepublicKeywords: drug delivery, growth factors, hMSC, liposomes, nanofibres
Procedia PDF Downloads 2872972 An Investigation of Tetraspanin Proteins’ Role in UPEC Infection
Authors: Fawzyah Albaldi
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Urinary tract infections (UTIs) are the most prevalent of infectious diseases and > 80% are caused by uropathogenic E. coli (UPEC). Infection occurs following adhesion to urothelial plaques on bladder epithelial cells, whose major protein constituent are the uroplakins (UPs). Two of the four uroplakins (UPIa and UPIb) are members of the tetraspanin superfamily. The UPEC adhesin FimH is known to interact directly with UPIa. Tetraspanins are a diverse family of transmembrane proteins that generally act as “molecular organizers” by binding different proteins and lipids to form tetraspanin enriched microdomains (TEMs). Previous work by our group has shown that TEMs are involved in the adhesion of many pathogenic bacteria to human cells. Adhesion can be blocked by tetraspanin-derived synthetic peptides, suggesting that tetraspanins may be valuable drug targets. In this study, we investigate the role of tetraspanins in UPEC adherence to bladder epithelial cells. Human bladder cancer cell lines (T24, 5637, RT4), commonly used as in-vitro models to investigate UPEC infection, along with primary human bladder cells, were used in this project. The aim was to establish a model for UPEC adhesion/infection with the objective of evaluating the impact of tetraspanin-derived reagents on this process. Such reagents could reduce the progression of UTI, particularly in patients with indwelling catheters. Tetraspanin expression on the bladder cells was investigated by q-PCR and flow cytometry, with CD9 and CD81 generally highly expressed. Interestingly, despite these cell lines being used by other groups to investigate FimH antagonists, uroplakin proteins (UPIa, UPIb and UPIII) were poorly expressed at the cell surface, although some were present intracellularly. Attempts were made to differentiate the cell lines, to induce cell surface expression of these UPs, but these were largely unsuccessful. Pre-treatment of bladder epithelial cells with anti-CD9 monoclonal antibody significantly decreased UPEC infection, whilst anti-CD81 had no effects. A short (15aa) synthetic peptide corresponding to the large extracellular region (EC2) of CD9 also significantly reduced UPEC adherence. Furthermore, we demonstrated specific binding of that fluorescently tagged peptide to the cells. CD9 is known to associate with a number of heparan sulphate proteoglycans (HSPGs) that have also been implicated in bacterial adhesion. Here, we demonstrated that unfractionated heparin (UFH)and heparin analogs significantly inhibited UPEC adhesion to RT4 cells, as did pre-treatment of the cells with heparinases. Pre-treatment with chondroitin sulphate (CS) and chondroitinase also significantly decreased UPEC adherence to RT4 cells. This study may shed light on a common pathogenicity mechanism involving the organisation of HSPGs by tetraspanins. In summary, although we determined that the bladder cell lines were not suitable to investigate the role of uroplakins in UPEC adhesion, we demonstrated roles for CD9 and cell surface proteoglycans in this interaction. Agents that target these may be useful in treating/preventing UTIs.Keywords: UTIs, tspan, uroplakins, CD9
Procedia PDF Downloads 1022971 Evaluation of Role of Surgery in Management of Pediatric Germ Cell Tumors According to Risk Adapted Therapy Protocols
Authors: Ahmed Abdallatif
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Background: Patients with malignant germ cell tumors have age distribution in two peaks, with the first one during infancy and the second after the onset of puberty. Gonadal germ cell tumors are the most common malignant ovarian tumor in females aged below twenty years. Sacrococcygeal and retroperitoneal abdominal tumors usually presents in a large size before the onset of symptoms. Methods: Patients with pediatric germ cell tumors presenting to Children’s Cancer Hospital Egypt and National Cancer Institute Egypt from January 2008 to June 2011 Patients underwent stratification according to risk into low, intermediate and high risk groups according to children oncology group classification. Objectives: Assessment of the clinicopathologic features of all cases of pediatric germ cell tumors and classification of malignant cases according to their stage, and the primary site to low, intermediate and high risk patients. Evaluation of surgical management in each group of patients focusing on surgical approach, the extent of surgical resection according to each site, ability to achieve complete surgical resection and perioperative complications. Finally, determination of the three years overall and disease-free survival in different groups and the relation to different prognostic factors including the extent of surgical resection. Results: Out of 131 cases surgically explored only 26 cases had re exploration with 8 cases explored for residual disease 9 cases for remote recurrence or metastatic disease and the other 9 cases for other complications. Patients with low risk kept under follow up after surgery, out of those of low risk group (48 patients) only 8 patients (16.5%) shifted to intermediate risk. There were 20 patients (14.6%) diagnosed as intermediate risk received 3 cycles of compressed (Cisplatin, Etoposide and Bleomycin) and all high risk group patients 69patients (50.4%) received chemotherapy. Stage of disease was strongly and significantly related to overall survival with a poorer survival in late stages (stage IV) as compared to earlier stages. Conclusion: Overall survival rate at 3 three years was (76.7% ± 5.4, 3) years EFS was (77.8 % ±4.0), however 3 years DFS was much better (89.8 ± 3.4) in whole study group with ovarian tumors had significantly higher Overall survival (90% ± 5.1). Event Free Survival analysis showed that Male gender was 3 times likely to have bad events than females. Patients who underwent incomplete resection were 4 times more than patients with complete resection to have bad events. Disease free survival analysis showed that Patients who underwent incomplete surgery were 18.8 times liable for recurrence compared to those who underwent complete surgery, and patients who were exposed to re-excision were 21 times more prone to recurrence compared to other patients.Keywords: extragonadal, germ cell tumors, gonadal, pediatric
Procedia PDF Downloads 2172970 Successful Cesarean Delivery with Veno-Venous Extracorporeal Membrane Oxygenation Support in a Pregnant Woman with Severe Acute Respiratory Distress Syndrome and Heart Failure Complicated by a Rare Condition of Pre-B Cell Acute Lymphoblastic Leukemia in P
Authors: Kristel Dame Bañez Sumagaysay, Marie Victoria Cruz-javier
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The current subject is a case of a 21 year-old woman at 29 1/7 weeks of gestation with Pre-B cell Acute Lymphoblastic Leukemia who was admitted to the coronary care unit (CCU) of the St. Luke’s Medical Center-Global City for Severe Acute Respiratory Distress Syndrome (ARDS) secondary to hospital-acquired pneumonia secondary to pneumocystis jiroveci; central line-associated bloodstream infection (E. aerogenes). She presented with chronic hypoxemia caused by Pulmonary edema, probably secondary to heart failure secondary to cardiomyopathy chemotherapy-induced. Due to worsening feto-maternal status, extracorporeal membrane oxygenation (ECMO) for respiratory support was instituted, and an elective cesarean section was done due to multiple maternal factors and deteriorating health status under total intravenous anesthesia assisted by veno-venous extracorporeal membrane oxygenation. She delivered a live preterm newborn male, APGAR Score: 1, 0, 0, birth weight 985 grams, birth length: 40.5cm, small for gestational age.Keywords: extracorporeal membrane oxygenation, pre-b cell acute lymphoblastic leukemia, severe acute respiratory distress syndrome, ethical dilemmas
Procedia PDF Downloads 732969 Cell Adhesion, Morphology and Cytokine Expression of Synoviocytes Can Be Altered on Different Nano-Topographic Oxidized Silicon Nanosponges
Authors: Hung-Chih Hsu, Pey-Jium Chang, Ching-Hsein Chen, Jer-Liang Andrew Yeh
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Osteoarthritis (OA) is a common disorder in rehabilitation clinic. The main characteristics include joint pain, localized tenderness and enlargement, joint effusion, cartilage destruction, loss of adhesion of perichondrium, synovium hyperplasia. Synoviocytes inflammation might be a cause of local tenderness and effusion. Inflammation cytokines might also play an important role in joint pain, cartilage destruction, decrease adhesion of perichondrium to the bone. Treatments of osteoarthritis include non-steroid anti-inflammation drugs (NSAID), glucosamine supplementation, hyaluronic acid, arthroscopic debridement, and total joint replacement. Total joint replacement is commonly used in patients with severe OA who failed respond to pharmacological treatment. However, some patients received surgery had serious adverse events, including instability of the implants due to insufficient adhesion to the adjacent bony tissue or synovial inflammation. We tried to develop ideal nano-topographic oxidized silicon nanosponges by using with various chemicals to produce thickness difference in nanometers in order to study more about the cell-environment interactions in vitro like the alterations of cell adhesion, morphology, extracellular matrix secretions in the pathogenesis of osteoarthritis. Cytokines studies like growth factor, reactive oxygen species, reactive inflammatory materials (Like nitrous oxide and prostaglandin E2), extracellular matrix (ECM) degradation enzymes, and synthesis of collagen will also be observed and discussed. Extracellular and intracellular expression transforming growth factor beta (TGF-β) will be studied by reverse transcription-polymerase chain reaction (RT-PCR). The degradation of ECM will be observed by the bioactivity ratio of matrix metalloproteinase (MMP) and tissue inhibitors of metalloproteinase by ELISA (Enzyme-linked immunosorbent assay). When rabbit synoviocytes were cultured on these nano-topographic structures, they demonstrate better cell adhesion rate, decreased expression of MMP-2,9 and PGE2, and increased expression of TGF-β when cultured in nano-topographic oxidized silicon nanosponges than in the planar oxidized silicon ones. These results show cell behavior, cytokine production can be influenced by physical characteristics from different nano-topographic structures. Our study demonstrates the possibility of manipulating cell behavior in these nano-topographic biomaterials.Keywords: osteoarthritis, synoviocyte, oxidized silicon surfaces, reactive oxygen species
Procedia PDF Downloads 3852968 Comprehensive Multi-Omics Study Highlights Osteopontin/SPP1 in Ovarian Aging Control
Authors: Chia-Jung Li, Li-Te Lin, Kuan-Hao Tsui
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The study identifies SPP1 as a potential gene associated with ovarian aging, revealing a significant decline in its expression in aged ovaries. SPP1, also known as osteopontin (OPN), is a multifunctional glycoprotein involved with regulatory proteins and pro-inflammatory immune chemokines. However, its genetic links to ovarian aging have not been extensively explored. Spatial transcriptomic analyses were conducted on ovaries from young and aged female mice, along with a sample from a 73-year-old individual. Additionally, single-cell RNA sequencing analysis was performed to identify associations between SPP1 and key genes. The study focused on crucial genes, including ITGAV, ITGB1, CD44, MMP3, and FN1, with a particular emphasis on the correlation between SPP1 and ITGB1. The findings indicate a significant decline in SPP1 expression in aged ovaries, which was consistent in the 73-year-old sample. Single-cell RNA sequencing unveiled associations between SPP1 and key genes, emphasizing a strong co-expression correlation between SPP1 and ITGB1. While the study provides valuable insights, further research is necessary to understand the broader implications and potential applications of SPP1 in ovarian aging. Translating these findings to clinical settings requires careful consideration. The identification of SPP1 as a gene implicated in ovarian aging opens new avenues for advancing precision medicine and refining treatment strategies for conditions related to ovarian aging.Keywords: SPP1, ovarian aging, spatial transcriptomic, single-cell RNA sequencing
Procedia PDF Downloads 342967 Development of Partial Sulphonated Poly(Vinylidene Fluoride - Hexafluoro Propylene)–Montmorillonite Nano-Composites as Proton Exchange Membranes
Authors: K. Selvakumar, J. Kalaiselvimary, B. Jansirani, M. Ramesh Prabhu
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Proton conducting sulphonated poly (vinylidene fluoride- hexafluoro propylene) PVdF-HFP membranes were modified with nano – sized montmorillonite (MMT) through homogeneous dispersive mixing and solution casting technique for fuel cell applications. The prepared composite membranes were characterized using Fourier Transform Infrared Spectroscopy and 1HNMR technique. The suitability of the composite membranes for fuel cell application was evaluated in terms of water uptake, swelling behavior, and proton conductivity. These composites showed good conductivities and durability and expected to be used in the development of proton exchange membrane for fuel cells.Keywords: composite, proton conduction, sulphonation, water uptake
Procedia PDF Downloads 2452966 Improving the Biocontrol of the Argentine Stem Weevil; Using the Parasitic Wasp Microctonus hyperodae
Authors: John G. Skelly, Peter K. Dearden, Thomas W. R. Harrop, Sarah N. Inwood, Joseph Guhlin
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The Argentine stem weevil (ASW; L. bonariensis) is an economically important pasture pest in New Zealand, which causes about $200 million of damage per annum. Microctonus hyperodae (Mh), a parasite of the ASW in its natural range in South America, was introduced into New Zealand to curb the pasture damage caused by the ASW. Mh is an endoparasitic wasp that lays its eggs in the ASW halting its reproduction. Mh was initially successful at preventing ASW proliferation and reducing pasture damage. The effectiveness of Mh has since declined due to decreased parasitism rates and has resulted in increased pasture damage. Although the mechanism through which ASW has developed resistance to Mh has not been discovered, it has been proposed to be due to the different reproductive modes used by Mh and the ASW in New Zealand. The ASW reproduces sexually, whereas Mh reproduces asexually, which has been hypothesised to have allowed the ASW to ‘out evolve’ Mh. Other species within the Microctonus genus reproduce both sexually and asexually. Strains of Microctonus aethiopoides (Ma), a species closely related to Mh, reproduce either by sexual or asexual reproduction. Comparing the genomes of sexual and asexual Microctonus may allow for the identification of the mechanism of asexual reproduction and other characteristics that may improve Mh as a biocontrol agent. The genomes of Mh and three strains of Ma, two of which reproduce sexually and one reproduces asexually, have been sequenced and annotated. The French (MaFR) and Moroccan (MaMO) reproduce sexually, whereas the Irish strain (MaIR) reproduces asexually. Like Mh, The Ma strains are also used as biocontrol agents, but for different weevil species. The genomes of Mh and MaIR were subsequently upgraded using Hi-C, resulting in a set of high quality, highly contiguous genomes. A subset of the genes involved in mitosis and meiosis, which have been identified though the use of Hidden Markov Models generated from genes involved in these processes in other Hymenoptera, have been catalogued in Mh and the strains of Ma. Meiosis and mitosis genes were broadly conserved in both sexual and asexual Microctonus species. This implies that either the asexual species have retained a subset of the molecular components required for sexual reproduction or that the molecular mechanisms of mitosis and meiosis are different or differently regulated in Microctonus to other insect species in which these mechanisms are more broadly characterised. Bioinformatic analysis of the chemoreceptor compliment in Microctonus has revealed some variation in the number of olfactory receptors, which may be related to host preference. Phylogenetic analysis of olfactory receptors highlights variation, which may be able to explain different host range preferences in the Microctonus. Hi-C clustering implies that Mh has 12 chromosomes, and MaIR has 8. Hence there may be variation in gene regulation between species. Genome alignment of Mh and MaIR implies that there may be large scale genome structural variation. Greater insight into the genetics of these agriculturally important group of parasitic wasps may be beneficial in restoring or maintaining their biocontrol efficacy.Keywords: argentine stem weevil, asexual, genomics, Microctonus hyperodae
Procedia PDF Downloads 1542965 Thermosensitive Hydrogel Development for Its Possible Application in Cardiac Cell Therapy
Authors: Lina Paola Orozco Marin, Yuliet Montoya Osorio, John Bustamante Osorno
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Ischemic events can culminate in acute myocardial infarction by irreversible cardiac lesions that cannot be restored due to the limited regenerative capacity of the heart. Cell therapy seeks to replace these injured or necrotic cells by transplanting healthy and functional cells. The therapeutic alternatives proposed by tissue engineering and cardiovascular regenerative medicine are the use of biomaterials to mimic the native extracellular medium, which is full of proteins, proteoglycans, and glycoproteins. The selected biomaterials must provide structural support to the encapsulated cells to avoid their migration and death in the host tissue. In this context, the present research work focused on developing a natural thermosensitive hydrogel, its physical and chemical characterization, and the determination of its biocompatibility in vitro. The hydrogel was developed by mixing hydrolyzed bovine and porcine collagen at 2% w/v, chitosan at 2.5% w/v, and beta-glycerolphosphate at 8.5% w/w and 10.5% w/w in magnetic stirring at 4°C. Once obtained, the thermosensitivity and gelation time were determined, incubating the samples at 37°C and evaluating them through the inverted tube method. The morphological characterization of the hydrogels was carried out through scanning electron microscopy. Chemical characterization was carried out employing infrared spectroscopy. The biocompatibility was determined using the MTT cytotoxicity test according to the ISO 10993-5 standard for the hydrogel’s precursors using the fetal human ventricular cardiomyocytes cell line RL-14. The RL-14 cells were also seeded on the top of the hydrogels, and the supernatants were subculture at different periods to their observation under a bright field microscope. Four types of thermosensitive hydrogels were obtained, which differ in their composition and concentration, called A1 (chitosan/bovine collagen/beta-glycerolphosphate 8.5%w/w), A2 (chitosan/porcine collagen/beta-glycerolphosphate 8.5%), B1 (chitosan/bovine collagen/beta-glycerolphosphate 10.5%) and B2 (chitosan/porcine collagen/beta-glycerolphosphate 10.5%). A1 and A2 had a gelation time of 40 minutes, and B1 and B2 had a gelation time of 30 minutes at 37°C. Electron micrographs revealed a three-dimensional internal structure with interconnected pores for the four types of hydrogels. This facilitates the exchange of nutrients, oxygen, and the exit of metabolites, allowing to preserve a microenvironment suitable for cell proliferation. In the infrared spectra, it was possible to observe the interaction that occurs between the amides of polymeric compounds with the phosphate groups of beta-glycerolphosphate. Finally, the biocompatibility tests indicated that cells in contact with the hydrogel or with each of its precursors are not affected in their proliferation capacity for a period of 16 days. These results show the potential of the hydrogel to increase the cell survival rate in the cardiac cell therapies under investigation. Moreover, the results lay the foundations for its characterization and biological evaluation in both in vitro and in vivo models.Keywords: cardiac cell therapy, cardiac ischemia, natural polymers, thermosensitive hydrogel
Procedia PDF Downloads 1892964 Mannosylated Oral Amphotericin B Nanocrystals for Macrophage Targeting: In vitro and Cell Uptake Studies
Authors: Rudra Vaghela, P. K. Kulkarni
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The aim of the present research was to develop oral Amphotericin B (AmB) nanocrystals (Nc) grafted with suitable ligand in order to enhance drug transport across the intestinal epithelial barrier and subsequently, active uptake by macrophages. AmB Nc were prepared by liquid anti-solvent precipitation technique (LAS). Poloxamer 188 was used to stabilize the prepared AmB Nc and grafted with mannose for actively targeting M cells in Peyer’s patches. To prevent shedding of the stabilizer and ligand, N,N’-Dicyclohexylcarbodiimide (DCC) was used as a cross-linker. The prepared AmB Nc were characterized for particle size, PDI, zeta potential, X-ray diffraction (XRD) and surface morphology using scanning electron microscope (SEM) and evaluated for drug content, in vitro drug release and cell uptake studies using caco-2 cells. The particle size of stabilized AmB Nc grafted with WGA was in the range of 287-417 nm with negative zeta potential between -18 to -25 mV. XRD studies revealed crystalline nature of AmB Nc. SEM studies revealed that ungrafted AmB Nc were irregular in shape with rough surface whereas, grafted AmB Nc were found to be rod-shaped with smooth surface. In vitro drug release of AmB Nc was found to be 86% at the end of one hour. Cellular studies revealed higher invasion and uptake of AmB Nc towards caco-2 cell membrane when compared to ungrafted AmB Nc. Our findings emphasize scope on developing oral delivery system for passively targeting M cells in Peyer’s patches.Keywords: leishmaniasis, amphotericin b nanocrystals, macrophage targeting, LAS technique
Procedia PDF Downloads 3002963 The Effect of Acute Muscular Exercise and Training Status on Haematological Indices in Adult Males
Authors: Ibrahim Musa, Mohammed Abdul-Aziz Mabrouk, Yusuf Tanko
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Introduction: Long term physical training affect the performance of athletes especially the females. Soccer which is a team sport, played in an outdoor field, require adequate oxygen transport system for the maximal aerobic power during exercise in order to complete 90 minutes of competitive play. Suboptimal haematological status has often been recorded in athletes with intensive physical activity. It may be due to the iron depletion caused by hemolysis or haemodilution results from plasma volume expansion. There is lack of data regarding the dynamics of red blood cell variables, in male football players. We hypothesized that, a long competitive season involving frequent matches and intense training could influence red blood cell variables, as a consequence of applying repeated physical loads when compared with sedentary. Methods: This cross sectional study was carried on 40 adult males (20 athletes and 20 non athletes) between 18-25 years of age. The 20 apparently healthy male non athletes were taken as sedentary and 20 male footballers comprise the study group. The university institutional review board (ABUTH/HREC/TRG/36) gave approval for all procedures in accordance with the Declaration of Helsinki. Red blood cell (RBC) concentration, packed cell volume (PCV), and plasma volume were measured in fasting state and immediately after exercise. Statistical analysis was done by using SPSS/ win.20.0 for comparison within and between the groups, using student’s paired and unpaired “t” test respectively. Results: The finding from our study shows that, immediately after termination of exercise, the mean RBC counts and PCV significantly (p<0.005) decreased with significant increased (p<0.005) in plasma volume when compared with pre-exercised values in both group. In addition the post exercise RBC was significantly higher in untrained (261.10±8.5) when compared with trained (255.20±4.5). However, there was no significant differences in the post exercise hematocrit and plasma volume parameters between the sedentary and the footballers. Moreover, beside changes in pre-exercise values among the sedentary and the football players, the resting red blood cell counts and Plasma volume (PV %) was significantly (p < 0.05) higher in the sedentary group (306.30±10.05 x 104 /mm3; 58.40±0.54%) when compared with football players (293.70±4.65 x 104 /mm3; 55.60±1.18%). On the other hand, the sedentary group exhibited significant (p < 0.05) decrease in PCV (41.60±0.54%) when compared with the football players (44.40±1.18%). Conclusions: It is therefore proposed that the acute football exercise induced reduction in RBC and PCV is entirely due to plasma volume expansion, and not of red blood cell hemolysis. In addition, the training status also influenced haematological indices of male football players differently from the sedentary at rest due to adaptive response. This is novel.Keywords: Haematological Indices, Performance Status, Sedentary, Male Football Players
Procedia PDF Downloads 2542962 Functional Profiling of a Circular RNA from the Huntingtin (HTT) Gene
Authors: Laura Gantley, Vanessa M. Conn, Stuart Webb, Kirsty Kirk, Marta Gabryelska, Duncan Holds, Brett W. Stringer, Simon J. Conn
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Trinucleotide repeat disorders comprise ~20 severe, inherited human neuromuscular and neurodegenerative disorders, which are a result of an abnormal expansion of repetitive sequences in the DNA. The most common of these, Huntington’s disease, results from the expansion of the CAG repeat region in exon 1 of the HTT gene via an unknown mechanism. Non-coding RNAs have been implicated in the initiation and progression of many diseases; thus, we focus on one circular RNA (circRNA) molecule arising from non-canonical splicing (back splicing) of HTT pre-mRNA. This circRNA and its mouse orthologue were transgenically overexpressed in human cells (SHSY-5Y and HEK293T) and mouse cells (Mb1), respectively. High-content imaging and flow cytometry demonstrated the overexpression of this circRNA reduces cell proliferation, reduces nuclear size independent of cellular size, and alters cell cycle progression. Analysis of protein by western blot and immunofluorescence demonstrated no change to HTT protein levels but altered nuclear-cytoplasmic distribution without impacting the expansion of the HTT repeat region. As these phenotypic and genotypic changes are found in Huntington’s disease patients, these results may suggest that this circRNA may play a functional role in the progression of Huntington’s disease.Keywords: cell biology, circular RNAs, Huntington’s disease, molecular biology, neurodegenerative disorders
Procedia PDF Downloads 982961 Antioxidant Activity of Friedelin, Eudesmic Acid and Methyl-3,4,5-Trimethoxybenzoate from Tapinanthus bangwensis (Engl., and K. Krause) [Loranthaceae] Grown in Nigeria
Authors: Odunayo Christy Atewolara-Odule, Olapeju O. Aiyelaagbe
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The search for new natural anti-oxidants has grown tremendously over the years because reactive oxygen species (ROS) production and oxidative stress have been linked to a large number of human degenerative diseases, such as cancer, cardiovascular diseases, inflammation, and diabetes. Tapinanthus bangwensis, a parasitic plant commonly known as mistletoe belonging to the Loranthaceae family, is mostly employed traditionally to treat inflammation, cancer, diabetes, and hypertension to mention a few. In this study, air-dried pulverized leaves and stem of Tapinanthus bangwensis were successively extracted with n-hexane, ethyl acetate, and methanol to give the corresponding crude extracts. The extracts were purified by column chromatography and high-performance liquid chromatography to give the isolated compounds. Structural elucidation was done using mass spectrometry, Fourier transform infra-red, 1D and 2D NMR spectroscopy. The antioxidant activity of the compounds was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ascorbic acid as standard. Three compounds; Friedelin, Eudesmic acid (3,4,5-trimethoxybenzoic) and Methyl-3,4,5-trimethoxybenzoate were isolated from the extracts of Tapinanthus bangwensis. Friedelin was isolated from the ethyl acetate extract of the stem while the two other compounds were isolated from the methanol extract of the leaves. The percentages of free radical scavenging activities of the compounds are as follows: Friedelin, 73.69%, methyl-3,4,5-trimethoxybenzoate, 79.33% and eudesmic, 87.68% anti-oxidant activity which were quite comparable to 93.96% given by ascorbic acid. We are reporting, to our best knowledge, for the first time the occurrence of friedelin and eudesmic acid in Tapinanthus bangwensis. The high anti-oxidant activity of these compounds supports the use of this plant in the management of diabetes and hypertension as they will be useful in combating complications arising from the disease.Keywords: column chromatography, eudesmic acid, friedelin, Tapinanthus bangwensis
Procedia PDF Downloads 2482960 Assessment of Cell-Rebuilding Efficacy of Selected Food Plants in the Lungs of Wild Rats Living in a Polluted Environment
Authors: Yahaya Tajudeen, Joy Okpuzor, Tolu Ajayi
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The cell-rebuilding efficacy of four food plants eating as vegetables and spices in Nigeria was assessed in the lungs of wild rats (Rattus rattus) living in a polluted environment. The plants are roselle (Hibiscus sabdarrifa), moringa (Moringa oleifera), ginger (Zingiber officinale) and ugwu (Telfairia occidentalis). Sixty rats were caught from the vicinity of a cement factory in Sagamu, Southwestern-Nigeria and grouped into 6. The control group was administered distilled water, while the test groups were given ethanolic extracts of roselle, moringa, ginger, ugwu and the mixture of the extracts for 180 days. The histopathology of the rats was conducted before and at the end of 180 days extracts administration. Before administering the extracts, the lungs of the rats showed vascular congestion, severe fibrosis and congested alveolus; all which were also observed in the lungs of control rats at the end of the treatment. However, the lungs of rats that were treated with the extracts of the plants showed moderate, mild or no histological damage compared to the control rats. The extract of the mixture of the plants performed best, followed by ginger, ugwu and roselle, respectively. These findings suggest that the food plants contain phytonutrients and phytochemicals, which repaired damaged cells and tissues in the exposed rats. Consequently, the plants could play a role in ameliorating health effects of environmental pollution.Keywords: food plants, wild rats, lung, histopathology, fibrosis, cell-rebuilding
Procedia PDF Downloads 3662959 Bioactivities and Phytochemical Studies of Acrocarpus fraxinifolius Bark Wight and Arn
Authors: H. M. El-Rafie, A. H. Abou Zeid, R. S. Mohammed, A. A. Sleem
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Acrocarpus is a genus of flowering plants in the legume family Fabaceae which considered as a large and economically important family. This study aimed to investigate the phytoconstituents of the petroleum ether extract (PEE) of Acrocarpus fraxinofolius bark by Gas chromatography coupled with mass spectrometry (GC/MS) analysis of its fractions (fatty acid and unsaponifiable matter). Concerning this, identification of 52 compounds constituting 97.03 % of the total composition of the unsaponifiable matter fraction. Cycloeucalenol was found to be the major compound representing 32.52% followed by 4a, 14a-dimethyl-A8~24(28)-ergostadien (26.50%) and ß-sitosterol(13.74%), furthermore Gas liquid chromatography (GLC) analysis of the sterol fraction revealed the identification of cholesterol (7.22 %), campesterol (13.30 %), stigmasterol (10.00 %) and β - sitosterol (69.48 %). Meanwhile, the identification of 33 fatty acids representing 90.71% of the total fatty acid constituents. Methyl-9,12-octadecadienoate (40.39%) followed by methyl hexadecanoate (23.64%) were found to be the major compounds. On the other hand, column chromatography and Thin layer chromatography (TLC) fractionation of PEE separate the triterpenoid: 21β-hydroxylup-20(29)-en-3-one and β- amyrin which were structurally identified by spectroscopic analysis (NMR, MS and IR). PEE has been biologically evaluated for 1: management of diabetes in alloxan induced diabetic rats 2: cytotoxic activity against four human tumor cell lines (Cervix carcinoma cell line[HELA], Breast carcinoma cell line [MCF7], Liver carcinoma cell line[HEPG2] and Colon carcinoma cell line[HCT-116] 3: hepatoprotective activity against CCl4-induced hepatotoxicity in rats and the activity was studied by assaying the serum marker enzymes like AST, ALT, and ALP. Concerning this, the anti-diabetic activity exhibited by 100mg of PEE extract was 74.38% relative to metformin (100% potency). It also showed a significant anti-proliferative activity against MCF-7 (IC50= 2.35µg), Hela(IC50=3.85µg) and HEPG-2 (IC50= 9.54µg) compared with Doxorubicin as reference drug. The hepatoprotective activity was evidenced by significant decrease in liver function enzymes, i.e. AST, ALT and ALP by (29.18%, 28.26%, and 34.11%, respectively using silymarin as the reference drug, compared to their concentration levels in an untreated group with liver damage induced by CCl₄. This study was performed for the first time on the bark of this species.Keywords: Acrocarpus fraxinofolius, antidiabetic, cytotoxic, hepatoprotective
Procedia PDF Downloads 1952958 The Effect of High-Pressure Processing on the Inactivation of Saccharomyces cerevisiae in Different Concentration of Manuka Honey and Its Relation with ° Brix
Authors: Noor Akhmazillah Fauzi, Mohammed Mehdi Farid, Filipa V. Silva
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The aim of this paper is to investigate if different concentration of Manuka honey (as a model food) has a major influence on the inactivation of Saccharomyces cerevisiae (as the testing microorganism) after subjecting it to HPP. Honey samples with different sugar concentrations (20, 30, 40, 50, 60 and 70 °Brix) were prepared aseptically using sterilized distilled water. No dilution of honey was made for the 80 °Brix sample. For the 0 °Brix sample (control), sterilized distilled water was used. Thermal treatment at 55 °C for 10 min (conventionally applied in honey pasteurisation in industry) was carried out for comparison purpose. S. cerevisiae cell numbers in honey samples were established before and after each HPP and thermal treatment. The number of surviving cells was determined after a proper dilution of the untreated and treated samples by the viable plate count method. S. cerevisiae cells, in different honey concentrations (0 to 80 °Brix), subjected to 600 MPa (at ambient temperature) showed an increasing resistance to inactivation with °Brix. A significant correlation (p < 0.05) between cell reduction and °Brix was found. Cell reduction in high pressure-treated samples varied linearly with °Brix (R2 > 0.9), confirming that the baroprotective effect of the food is due to sugar content. This study has practical implications in establishing efficient process design for commercial manufacturing of high sugar food products and on the potential use of HPP for such products.Keywords: high pressure processing, honey, Saccharomyces cerevisiae, °Brix
Procedia PDF Downloads 3522957 Wound Healing Process Studied on DC Non-Homogeneous Electric Fields
Authors: Marisa Rio, Sharanya Bola, Richard H. W. Funk, Gerald Gerlach
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Cell migration, wound healing and regeneration are some of the physiological phenomena in which electric fields (EFs) have proven to have an important function. Physiologically, cells experience electrical signals in the form of transmembrane potentials, ion fluxes through protein channels as well as electric fields at their surface. As soon as a wound is created, the disruption of the epithelial layers generates an electric field of ca. 40-200 mV/mm, directing cell migration towards the wound site, starting the healing process. In vitro electrotaxis, experiments have shown cells respond to DC EFs polarizing and migrating towards one of the poles (cathode or anode). A standard electrotaxis experiment consists of an electrotaxis chamber where cells are cultured, a DC power source and agar salt bridges that help delaying toxic products from the electrodes to attain the cell surface. The electric field strengths used in such an experiment are uniform and homogeneous. In contrast, the endogenous electric field strength around a wound tend to be multi-field and non-homogeneous. In this study, we present a custom device that enables electrotaxis experiments in non-homogeneous DC electric fields. Its main feature involves the replacement of conventional metallic electrodes, separated from the electrotaxis channel by agarose gel bridges, through electrolyte-filled microchannels. The connection to the DC source is made by Ag/AgCl electrodes, incased in agarose gel and placed at the end of each microfluidic channel. An SU-8 membrane closes the fluidic channels and simultaneously serves as the single connection from each of them to the central electrotaxis chamber. The electric field distribution and current density were numerically simulated with the steady-state electric conduction module from ANSYS 16.0. Simulation data confirms the application of nonhomogeneous EF of physiological strength. To validate the biocompatibility of the device cellular viability of the photoreceptor-derived 661W cell line was accessed. The cells have not shown any signs of apoptosis, damage or detachment during stimulation. Furthermore, immunofluorescence staining, namely by vinculin and actin labelling, allowed the assessment of adhesion efficiency and orientation of the cytoskeleton, respectively. Cellular motility in the presence and absence of applied DC EFs was verified. The movement of individual cells was tracked for the duration of the experiments, confirming the EF-induced, cathodal-directed motility of the studied cell line. The in vitro monolayer wound assay, or “scratch assay” is a standard protocol to quantitatively access cell migration in vitro. It encompasses the growth of a confluent cell monolayer followed by the mechanic creation of a scratch, representing a wound. Hence, wound dynamics was monitored over time and compared for control and applied the electric field to quantify cellular population motility.Keywords: DC non-homogeneous electric fields, electrotaxis, microfluidic biochip, wound healing
Procedia PDF Downloads 2692956 Energy Absorption Capacity of Aluminium Foam Manufactured by Kelvin Model Loaded Under Different Biaxial Combined Compression-Torsion Conditions
Authors: H. Solomon, A. Abdul-Latif, R. Baleh, I. Deiab, K. Khanafer
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Aluminum foams were developed and tested due to their high energy absorption abilities for multifunctional applications. The aim of this research work was to investigate experimentally the effect of quasi-static biaxial loading complexity (combined compression-torsion) on the energy absorption capacity of highly uniform architecture open-cell aluminum foam manufactured by kelvin cell model. The two generated aluminum foams have 80% and 85% porosities, spherical-shaped pores having 11mm in diameter. These foams were tested by means of several square-section specimens. A patented rig called ACTP (Absorption par Compression-Torsion Plastique), was used to investigate the foam response under quasi-static complex loading paths having different torsional components (i.e., 0°, 37° and 53°). The main mechanical responses of the aluminum foams were studied under simple, intermediate and severe loading conditions. In fact, the key responses to be examined were stress plateau and energy absorption capacity of the two foams with respect to loading complexity. It was concluded that the higher the loading complexity and the higher the relative density, the greater the energy absorption capacity of the foam. The highest energy absorption was thus recorded under the most complicated loading path (i.e., biaxial-53°) for the denser foam (i.e., 80% porosity).Keywords: open-cell aluminum foams, biaxial loading complexity, foams porosity, energy absorption capacity, characterization
Procedia PDF Downloads 1302955 Identification of the Antimicrobial Effect of Liquorice Extracts on Gram-Positive Bacteria: Determination of Minimum Inhibitory Concentration and Mechanism of Action Using a luxABCDE Reporter Strain
Authors: Madiha El Awamie, Catherine Rees
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Natural preservatives have been used as alternatives to traditional chemical preservatives; however, a limited number have been commercially developed and many remain to be investigated as sources of safer and effective antimicrobials. In this study, we have been investigating the antimicrobial activity of an extract of Glycyrrhiza glabra (liquorice) that was provided as a waste material from the production of liquorice flavourings for the food industry, and to investigate if this retained the expected antimicrobial activity so it could be used as a natural preservative. Antibacterial activity of liquorice extract was screened for evidence of growth inhibition against eight species of Gram-negative and Gram-positive bacteria, including Listeria monocytogenes, Listeria innocua, Staphylococcus aureus, Enterococcus faecalis and Bacillus subtilis. The Gram-negative bacteria tested include Pseudomonas aeruginosa, Escherichia coli and Salmonella typhimurium but none of these were affected by the extract. In contrast, for all of the Gram-positive bacteria tested, growth was inhibited as monitored using optical density. However parallel studies using viable count indicated that the cells were not killed meaning that the extract was bacteriostatic rather than bacteriocidal. The Minimum Inhibitory Concentration [MIC] and Minimum Bactericidal Concentration [MBC] of the extract was also determined and a concentration of 50 µg ml-1 was found to have a strong bacteriostatic effect on Gram-positive bacteria. Microscopic analysis indicated that there were changes in cell shape suggesting the cell wall was affected. In addition, the use of a reporter strain of Listeria transformed with the bioluminescence genes luxABCDE indicated that cell energy levels were reduced when treated with either 12.5 or 50 µg ml-1 of the extract, with the reduction in light output being proportional to the concentration of the extract used. Together these results suggest that the extract is inhibiting the growth of Gram-positive bacteria only by damaging the cell wall and/or membrane.Keywords: antibacterial activity, bioluminescence, Glycyrrhiza glabra, natural preservative
Procedia PDF Downloads 3392954 Design and in Slico Study of the Truncated Spike-M-N SARS-CoV-2 as a Novel Effective Vaccine Candidate
Authors: Aghasadeghi MR., Bahramali G., Sadat SM., Sadeghi SA., Yousefi M., Khodaei K., Ghorbani M., Sadat Larijani M.
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Background:The emerging COVID-19 pandemic is a serious concernfor the public health worldwide. Despite the many mutations in the virus genome, it is important to find an effective vaccine against viral mutations. Therefore, in current study, we aimed at immunoinformatic evaluation of the virus proteins immunogenicity to design a preventive vaccine candidate, which could elicit humoral and cellular immune responses as well. Methods:Three antigenic regions are included;Spike, Membrane, and Nucleocapsid amino acid sequences were obtained, and possible fusion proteins were assessed andcompared by immunogenicity, structural features, and population coverage. The best fusion protein was also evaluated for MHC-I and MHC-II T-cell epitopes and the linear and conformational B-cell epitopes. Results: Among the four predicted models, the truncated Spike protein in fusion with M and N proteins is composed of 24 highly immunogenic human MHC class I and 29 MHC class II, along with 14 B-cell linear and 61 discontinues epitopes. Also, the selected protein has high antigenicity and acceptable population coverage of 82.95% in Iran and 92.51% in Europe. Conclusion: The data indicate that the truncated Spike-M-N SARS-CoV-2form which could be potential targets of neutralizing antibodies. The protein also has the ability to stimulate humoral and cellular immunity. The in silico study provided the fusion protein as a potential preventive vaccine candidate for further in vivo evaluation.Keywords: SARS-CoV-2, immunoinformatic, protein, vaccine
Procedia PDF Downloads 2222953 Using Atomic Force Microscope to Investigate the Influence of UVA Radiation and HA on Cell Behaviour and Elasticity of Dermal Fibroblasts
Authors: Pei-Hsiu Chiang, Ling Hong Huang, Hsin-I Chang
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In this research, we used UVA irradiation, which can penetrate into dermis and fibroblasts, the most abundant cells in dermis, to investigate the effect of UV light on dermis, such as inflammation, ECM degradation and elasticity loss. Moreover, this research is focused on the influence of hyaluronic acid (HA) on UVA treated dermal fibroblasts. We aim to establish whether HA can effectively relief ECM degradation, and restore the elasticity of UVA-damaged fibroblasts. Prolonged exposure to UVA radiation can damage fibroblasts and led variation in cell morphology and reduction in cell viability. Besides, UVA radiation can induce IL-1β expression on fibroblasts and then promote MMP-1 and MMP-3 expression, which can accelerate ECM degradation. On the other hand, prolonged exposure to UVA radiation reduced collagen and elastin synthesis on fibroblasts. Due to the acceleration of ECM degradation and the reduction of ECM synthesis, Atomic force microscope (AFM) was used to analyze the elasticity reduction on UVA-damaged fibroblasts. UVA irradiation causes photoaging on fibroblasts. UVA damaged fibroblasts with HA treatment can down-regulate the gene expression of MMP-1, MMP-3, and then slow down ECM degradation. On the other hand, HA may restore elastin and collagen synthesis in UV-damaged fibroblasts. Based on the slowdown of ECM degradation, UVA-damaged fibroblast elasticity can be effectively restored by HA treatment. In summary, HA can relief the photoaging conditions on fibroblasts, but may not be able to return fibroblasts to normal, healthy state. Although HA cannot fully recover UVA-damaged fibroblasts, HA is still potential for repairing photoaging skin.Keywords: atomic force microscope, hyaluronic acid, UVA radiation, dermal fibroblasts
Procedia PDF Downloads 3902952 Coordinated Interference Canceling Algorithm for Uplink Massive Multiple Input Multiple Output Systems
Authors: Messaoud Eljamai, Sami Hidouri
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Massive multiple-input multiple-output (MIMO) is an emerging technology for new cellular networks such as 5G systems. Its principle is to use many antennas per cell in order to maximize the network's spectral efficiency. Inter-cellular interference remains a fundamental problem. The use of massive MIMO will not derogate from the rule. It improves performances only when the number of antennas is significantly greater than the number of users. This, considerably, limits the networks spectral efficiency. In this paper, a coordinated detector for an uplink massive MIMO system is proposed in order to mitigate the inter-cellular interference. The proposed scheme combines the coordinated multipoint technique with an interference-cancelling algorithm. It requires the serving cell to send their received symbols, after processing, decision and error detection, to the interfered cells via a backhaul link. Each interfered cell is capable of eliminating intercellular interferences by generating and subtracting the user’s contribution from the received signal. The resulting signal is more reliable than the original received signal. This allows the uplink massive MIMO system to improve their performances dramatically. Simulation results show that the proposed detector improves system spectral efficiency compared to classical linear detectors.Keywords: massive MIMO, COMP, interference canceling algorithm, spectral efficiency
Procedia PDF Downloads 1462951 Immunoinformatic Design and Evaluation of an Epitope-Based Tetravalent Vaccine against Human Hand, Foot, and Mouth Disease
Authors: Aliyu Maje Bello, Yaowaluck Maprang Roshorm
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Hand, foot, and mouth disease (HFMD) is a highly contagious viral infection affecting mostly infants and children. Although the Enterovirus A71 (EV71) is usually the major causative agent of HFMD, other enteroviruses such as coxsackievirus A16, A10, and A6 are also found in some of the recent outbreaks. The commercially available vaccines have demonstrated their effectiveness against only EV71 infection but no protection against other enteroviruses. To address the limitation of the monovalent EV71 vaccine, the present study thus designed a tetravalent vaccine against the four major enteroviruses causing HFMD and primarily evaluated the designed vaccine using an immunoinformatics approach. The immunogen was designed to contain the EV71 VP1 protein and multiple reported epitopes from all four distinct enteroviruses and thus designated a tetravalent vaccine. The 3D structure of the designed tetravalent vaccine was modeled, refined, and validated. Epitope screening showed the presence of B-cell, CTL, CD4 T cell, and IFN epitopes with vast application among the Asian population. Docking analysis confirmed the stable and strong binding interactions between the immunogen and immune receptor B-cell receptor (BCR). In silico cloning and immune simulation analyses guaranteed high efficiency and sufficient expression of the vaccine candidate in humans. Overall, the promising results obtained from the in-silico studies of the proposed tetravalent vaccine make it a potential candidate worth further experimental validation.Keywords: enteroviruses, coxsackieviruses, hand foot and mouth disease, immunoinformatics, tetravalent vaccine
Procedia PDF Downloads 702950 The Antibacterial and Anticancer Activity of Marine Actinomycete Strain HP411 Isolated in the Northern Coast of Vietnam
Authors: Huyen T. Pham, Nhue P. Nguyen, Tien Q. Phi, Phuong T. Dang, Hy G. Le
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Since the marine environmental conditions are extremely different from the other ones, so that marine actinomycetes might produce novel bioactive compounds. Therefore, actinomycete strains were screened from marine water and sediment samples collected from the coastal areas of Northern Vietnam. Ninety-nine actinomycete strains were obtained on starch-casein agar media by dilution technique, only seven strains, named HP112, HP12, HP411, HPN11, HP 11, HPT13 and HPX12, showed significant antibacterial activity against both gram-positive and gram-negative bacteria (Bacillus subtilis ATCC 6633, Staphylococcus epidemidis ATCC 12228, Escherichia coli ATCC 11105). Further studies were carried out with the most active HP411strain against Candida albicans ATCC 10231. This strain could grow rapidly on starch casein agar and other media with high salt containing 7-10% NaCl at 28-30oC. Spore-chain of HP411 showed an elongated and circular shape with 10 to 30 spores/chain. Identification of the strain was carried out by employing the taxonomical studies including the 16S rRNA sequence. Based on phylogenetic and phenotypic evidence it is proposed that HP411 to be belongs to species Streptomyces variabilis. The potent of the crude extract of fermentation broth of HP411that are effective against wide range of pathogens: both gram-positive, gram-negative and fungi. Further studies revealed that the crude extract HP411 could obtain the anticancer activity for cancer cell lines: Hep-G2 (liver cancer cell line); RD (cardiac and skeletal muscle letters cell line); FL (membrane of the uterus cancer cell line). However, the actinomycetes from marine ecosystem will be useful for the discovery of new drugs in the furture.Keywords: marine actinomycetes, antibacterial, anticancer, Streptomyces variabilis
Procedia PDF Downloads 4182949 Rule Insertion Technique for Dynamic Cell Structure Neural Network
Authors: Osama Elsarrar, Marjorie Darrah, Richard Devin
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This paper discusses the idea of capturing an expert’s knowledge in the form of human understandable rules and then inserting these rules into a dynamic cell structure (DCS) neural network. The DCS is a form of self-organizing map that can be used for many purposes, including classification and prediction. This particular neural network is considered to be a topology preserving network that starts with no pre-structure, but assumes a structure once trained. The DCS has been used in mission and safety-critical applications, including adaptive flight control and health-monitoring in aerial vehicles. The approach is to insert expert knowledge into the DCS before training. Rules are translated into a pre-structure and then training data are presented. This idea has been demonstrated using the well-known Iris data set and it has been shown that inserting the pre-structure results in better accuracy with the same training.Keywords: neural network, self-organizing map, rule extraction, rule insertion
Procedia PDF Downloads 1702948 Research Project of National Interest (PRIN-PNRR) DIVAS: Developing Methods to Assess Tree Vitality after a Wildfire through Analyses of Cambium Sugar Metabolism
Authors: Claudia Cocozza, Niccolò Frassinelli, Enrico Marchi, Cristiano Foderi, Alessandro Bizzarri, Margherita Paladini, Maria Laura Traversi, Eleftherious Touloupakis, Alessio Giovannelli
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The development of tools to quickly identify the fate of injured trees after stress is highly relevant when biodiversity restoration of damaged sites is based on nature-based solutions. In this context, an approach to assess irreversible physiological damages within trees could help to support planning management decisions of perturbed sites to restore biodiversity, for the safety of the environment and understanding functionality adjustments of the ecosystems. Tree vitality can be estimated by a series of physiological proxies like cambium activity, starch, and soluble sugars amount in C-sinks whilst the accumulation of ethanol within the cambial cells and phloem is considered an alert of cell death. However, their determination requires time-consuming laboratory protocols, which makes the approach unfeasible as a practical option in the field. The project aims to develop biosensors to assess the concentration of soluble sugars and ethanol in stem tissues. Soluble sugars and ethanol concentrations will be used to define injured trees to discriminate compromised and recovering trees in the forest directly. To reach this goal, we select study sites subjected to prescribed fires or recent wildfires as experimental set-ups. Indeed, in Mediterranean countries, forest fire is a recurrent event that must be considered as a central component of regional and global strategies in forest management and biodiversity restoration programs. A biosensor will be developed through a multistep process related to target analytes characterization, bioreceptor selection, and, finally, calibration/testing of the sensor. To validate biosensor signals, soluble sugars and ethanol will be quantified by HPLC and GC using synthetic media (in lab) and phloem sap (in field) whilst cambium vitality will be assessed by anatomical observations. On burnt trees, the stem growth will be monitored by dendrometers and/or estimated by tree ring analyses, whilst the tree response to past fire events will be assessed by isotopic discrimination. Moreover, the fire characterization and the visual assessment procedure will be used to assign burnt trees to a vitality class. At the end of the project, a well-defined procedure combining biosensor signal and visual assessment will be produced and applied to a study case. The project outcomes and the results obtained will be properly packaged to reach, engage and address the needs of the final users and widely shared with relevant stakeholders involved in the optimal use of biosensors and in the management of post-fire areas. This project was funded by National Recovery and Resilience Plan (NRRP), Mission 4, Component C2, Investment 1.1 - Call for tender No. 1409 of 14 September 2022 – ‘Progetti di Ricerca di Rilevante interesse Nazionale – PRIN’ of Italian Ministry of University and Research funded by the European Union – NextGenerationEU; Grant N° P2022Z5742, CUP B53D23023780001.Keywords: phloem, scorched crown, conifers, prescribed burning, biosensors
Procedia PDF Downloads 152947 Induction of Callus and Expression of Compounds in Capsicum Frutescens Supplemented with of 2, 4-D
Authors: Jamilah Syafawati Yaacob, Muhammad Aiman Ramli
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Cili padi or Capsicum frutescens is one of capsicum species from nightshade family, Solanaceae. It is famous in Malaysia and is widely used as a food ingredient. Capsicum frutescens also possess vast medicinal properties. The objectives of this study are to determine the most optimum 2,4-D hormone concentration for callus induction from stem explants C. frutescens and the effects of different 2,4-D concentrations on expression of compounds from C. frutescens. Seeds were cultured on MS media without hormones (MS basal media) to yield aseptic seedlings of this species, which were then used to supply explant source for subsequent tissue culture experiments. Stem explants were excised from aseptic seedlings and cultured on MS media supplemented with various concentrations (0.1, 0.3 and 0.5 mg/L) of 2,4-D to induce formation of callus. Fresh weight, dry weight and callus growth percentage in all samples were recorded. The highest mean of dry weight was observed in MS media supplemented with 0.5 mg/L 2,4-D, where 0.4499 ± 0.106 g of callus was produced. The highest percentage of callus growth (16.4%) was also observed in cultures supplemented with 0.5 mg/L 2,4-D. The callus samples were also subjected to HPLC-MS to evaluate the effect of hormone concentration on expression of bio active compounds in different samples. Results showed that caffeoylferuloylquinic acids were present in all samples, but was most abundant in callus cells supplemented with 0.3 & 0.5 mg/L 2,4-D. Interestingly, there was an unknown compound observed to be highly expressed in callus cells supplemented with 0.1 mg/L 2,4-D, but its presence was less significant in callus cells supplemented with 0.3 and 0.5 mg/L 2,4-D. Furthermore, there was also a compound identified as octadecadienoic acid, which was uniquely expressed in callus supplemented with 0.5 mg/L 2,4-D, but absent in callus cells supplemented with 0.1 and 0.3 mg/L 2,4-D. The results obtained in this study indicated that plant growth regulators played a role in expression of secondary metabolites in plants. The increase or decrease of these growth regulators may have triggered a change in the secondary metabolite biosynthesis pathways, thus causing differential expression of compounds in this plant.Keywords: callus, in vitro, secondary metabolite, 2, 4-Dichlorophenoxyacetic acid
Procedia PDF Downloads 3732946 Pain Management Strategies for Effective Coping with Sickle Cell Disease: The Perspective of Patients in Ghana
Authors: V. A. Adzika, D. Ayim-Aboagye, T. Gordh
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Background and aims: Prevalence of Sickle Cell Disease (SCD) is high in Ghana but not much is known in terms of research into non-medical strategies for managing and coping with the pain associated with SCD. This study was carried out to examine effective non-medical related strategies patients use to cope and manage their SCD condition. Methods: SCD patients (387) consisting of 180 males and 204 females between 18-65 years old years participated in the study. A cross-sectional research design was used in which participants completed questionnaires on pain, non-medical coping and management strategies, anxiety, and depression. Results of multiple regression analysis showed that socio-demographic characteristics contributed to the variance in the pain associated with SCD. Results: Over 90% of participants reported that pains associated with SCD were the main reason for seeking treatment in SCD crisis. In terms of non-medical related coping strategies, attending a place of worship and praying were the main coping strategies used in SCD crises, suggesting that patients’ beliefs, particularly in a supernatural being, served as a mitigating factor in the process of coping with the pain associated with SCD crisis. Also, avoidance and withdrawal from people and social activities were reported to be strategies used to cope effectively with the pain associated with SCD crisis. Conclusion: This indicates that it is imperative to incorporate non-medical related coping and management strategies, especially religious beliefs and psychosocial factors, to coping and management of the pain associated with SCD.Keywords: anxiety, depression, sickle cell disease, quality of life, socio-demographic characteristics, Ghana
Procedia PDF Downloads 4082945 Some Studies on Endometritis in Pure Arabian Mares
Authors: Khairi El Battawy, Monika Skalicki
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The present investigation has been done on pure Egyptian Arabian mares that reared in private horse studs. Fifty non-pregnant mares were selected and examined to classify them as either being reproductively healthy or subfertile mares including clinical endometritis, early embryonic death, granulosa cell tumor, repeat breeder (post-breeding endometritis), and anoestrus mares. The purpose of the study was to assess oxidative/antioxidant biochemical metabolites, lipogram, trace elements and reproductive hormones throughout reproductive conditions in mares during regular estrous, anestrum, early pregnancy, granulose cell tumor, ovulation failure, and endometritis. Results showed intensification of the free radical-dependent process in the blood of infertile mare, especially mares with endometritis. Ultrasonography as a diagnostic tool diagnosis of endometritis in mares was an important step as it revealed much information concerning infertility problem.Keywords: endometritis, ovulation, oxidative, mare
Procedia PDF Downloads 1762944 Clara Cell Secretory Protein 16 Serum Level Decreases in Patients with Non-Smoking-Related Chronic Obstructive Pulmonary Diseases (COPD)
Authors: Lian Wu, Mervyn Merrilees
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Chronic Obstructive Pulmonary Disease (COPD) is a worldwide problem, characterized by irreversible and progressive airflow obstruction. In New Zealand, it is currently the 4th commonest cause of death and exacerbations of COPD are a frequent cause of admission to hospital. Serum levels of Clara cell secretory protein-16 (CC-16) are believed to represent Clara cell toxicity. More recently, CC-16 has been found to be associated with smoker COPD. It is produced almost exclusively by non-ciliated Clara cells in the airways, and its primary function is to protect the lungs against oxidative stress and carcinogenesis. After acute exposure to cigarette smoke, serum levels of CC-16 become elevated. CC16 is a potent natural immune-suppressor and anti-inflammatory agent. In vitro, CC16 inhibits both monocyte and polymorphonuclear neutrophils chemotaxis and phagocytosis. CC16 also inhibits fibroblast chemotaxis. However, the role of CC-16 in non-smoking related COPD is still not clear. In this study, we investigated serum CC-16 levels in non-smoking related COPD. Methods: We compared non-smoker patients with COPD (FEV1<60% of predicted, FEV1/FVC <0.7, n=100) and individuals with normal lung function FEV1≥ 80% of predicted and FEV1/FVC≥ 0.7, n=80). All subjects had no smoking history. CC-16 was measured by ELISA. Results and conclusion: Serum CC-16 levels are reduced in individuals with non-smoking related COPD, and there is a weak correlation with disease severity in non-smoking related COPD group compared to non-smoker controls.Keywords: COPD, CC-16, ELISA, non-smoking-related COPD
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