Search results for: stem cell therapy

Authors: Harish K. Chandrawanshi, Mithun S. Rajput, Neelima Choure, Purnima Dey Sarkar, Shailesh Jain

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The present research study aimed to fabricate and evaluate biodegradable nanoparticles of aromatase inhibitor letrozole, intended for breast cancer therapy. Letrozole loaded poly(D,L-lactide-co-glycolide acid) nanoparticles were prepared by solvent evaporation method using dichlorometane as solvent (oil phase) and polyvinyl alcohol (PVA) as aqueous phase. Prepared nanoparticles were characterized by particle size, infrared spectra, drug loading efficiency, drug entrapment efficiency and in vitro release and also evaluated for in vivo anticancer activity. The high speed homogenizer was used to produce stable nanoparticles of mean size range 198.35 ± 0.04 nm with high entrapment efficiency (69.86 ± 2.78%). Percentage of drug and homogenization speed significantly influenced the particle size, entrapment efficiency and release (p<0.05). The nanoparticles show significant in vivo anticancer activity against Ehrlich ascites carcinoma in mice. The significant system sustained the release of letrozole drug effectively and further investigation could exhibit its potential usefulness in breast cancer therapy.

Keywords: breast cancer/therapy, letrozole, nanoparticles, PLGA

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Authors: Shaikah Alsubayae, Gianluigi Casse, Carlos Chavez, Jon Taylor, Alan Taylor, Mohammad Alsulimane

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Achieving precise radiotherapy through carbon therapy necessitates the accurate monitoring of radiation dose distribution within the patient's body. This process is pivotal for targeted tumor treatment, minimizing harm to healthy tissues, and enhancing overall treatment effectiveness while reducing the risk of side effects. In our investigation, we adopted a methodological approach to monitor secondary proton doses in carbon therapy using Monte Carlo (MC) simulations. Initially, Geant4 simulations were employed to extract the initial positions of secondary particles generated during interactions between carbon ions and water, including protons, gamma rays, alpha particles, neutrons, and tritons. Subsequently, we explored the relationship between the carbon ion beam and these secondary particles. Interaction vertex imaging (IVI) proves valuable for monitoring dose distribution during carbon therapy, providing information about secondary particle locations and abundances, particularly protons. The IVI method relies on charged particles produced during ion fragmentation to gather range information by reconstructing particle trajectories back to their point of origin, known as the vertex. In the context of carbon ion therapy, our simulation results indicated a strong correlation between some secondary particles and the range of carbon ions. However, challenges arose due to the unique elongated geometry of the target, hindering the straightforward transmission of forward-generated protons. Consequently, the limited protons that did emerge predominantly originated from points close to the target entrance. Fragment (protons) trajectories were approximated as straight lines, and a beam back-projection algorithm, utilizing interaction positions recorded in Si detectors, was developed to reconstruct vertices. The analysis revealed a correlation between the reconstructed and actual positions.

Keywords: radiotherapy, carbon therapy, monitor secondary proton doses, interaction vertex imaging

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Authors: Constant Gillot, Pauline Michaux, Julien Favresse, Jean-Michel Dogné, Jonathan Douxfils

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Introduction: NETosis has emerged as a crucial yet paradoxical factor in severe COVID-19 cases. While neutrophil extracellular traps (NETs) help contain and eliminate viral particles, excessive NET formation can lead to hyperinflammation, exacerbating tissue damage and acute respiratory distress syndrome (ARDS). Aims: This study evaluates the relationship between COVID-19-infected sera and NETosis using an ex-vivo model. Methods: Sera from 8 post-admission COVID-19 patients, after receiving corticoid therapy, were used to induce NETosis in neutrophils from a healthy donor. NET formation was tracked using fluorescent markers for DNA and neutrophil elastase (NE) every 2 minutes for 8 hours. The results were expressed as a percentage of DNA/NE released over time. Key metrics, including T50 (time to 50% release) and AUC (area under the curve), representing total NETosis potential), were calculated. A 27-cytokine screening kit was used to assess the cytokine composition of the sera. Results: COVID-19 sera induced NETosis based on their cytokine profile. The AUC of NE and DNA release decreased with time following corticoid therapy, showing a significant reduction in 6 of the 8 patients (p<0.05). T50 also decreased in parallel with AUC for both markers. Cytokines concentration decrease with time after therapy administration. There is correlation between 14 cytokines concentration and NE release. Conclusion: This ex-vivo model successfully demonstrated the induction of NETosis by COVID-19 sera using two markers. A clear decrease in NETosis potential was observed over time with glucocorticoid therapy. This model can be a valuable tool for monitoring NETosis and investigating potential NETosis inducers and inhibitors.

Keywords: NETosis, COVID-19, cytokine storm, biomarkers

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Authors: Kamesh Gupta, Nitin Jain, Anurag Rohatgi

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Objective: To study Vitamin B12 levels and presence of peripheral neuropathy among diabetes mellitus patients on metformin therapy. Method: The observational study was conducted from November 2014 to March 2015. Patients were selected from the Lady Hardinge Medical College, Delhi, India. Exhaustive history regarding dietary habits and metformin usage was taken. Lab tests including HbA1c levels and Vit B12 assays were done, on the basis of which patients were classified into subgroups. Peripheral neuropathy was detected by both clinical scoring and electrophysiological studies. Appropriate Statistical analysis for observational studies was done to evaluate the data. Results: The average duration of metformin usage was higher in patients with definite B12 deficiency (9.4y) than patients with normal B12 levels (5.6 y). Patients in the definite B12 deficiency group had much higher incidence of neuropathy (89%) than patients with no deficiency (27%). The incidence of neuropathy was higher in cases with longer metformin usage (100% with 18-22y of use and 83% with 14-17y of use) than shorter periods (29% with 2-5y of use and 75% with 6-9y of use). Conclusion: Thus patients on long-term metformin therapy are at a high risk for Vitamin B12 deficiency. Definite and possible Vitamin B12 deficiency on metformin had an earlier onset of neuropathy than the subgroup with normal Vitamin B12 levels.

Keywords: diabetic neuroptahy, cobalamine deficiency, metformin, nerve conduction studies

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Authors: A. Bouloufa, F. Khaled, K. Djessas

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This paper presents a new structure of solar cell based on p-type microcrystalline silicon as an absorber and n-type aluminum doped zinc oxide (ZnO:Al) transparent conductive oxide as an optical window. The ZnO:Al layer deposited by rf-magnetron sputtering at room temperature yields a low resistivity about 7,64.10-2Ω.cm and more than 85% mean optical transmittance in the VIS–NIR range, with an optical band gap of 3.3 eV. These excellent optical properties of this layer in combination with an optimal contact at the front surface result in a superior light trapping yielding to efficiencies about 20%. In order to improve efficiency, we have used a p+-µc-Si thin layer highly doped as a back surface field which minimizes significantly the impact of rear surface recombination velocity on voltage and current leading to a high efficiency of 24%. Optoelectronic parameters were determined using the current density-voltage (J-V) curve by means of a numerical simulation with Analysis of Microelectronic and Photonic Structures (AMPS-1D) device simulator.

Keywords: optical window, thin film, solar cell, efficiency

Procedia PDF Downloads 286

Authors: Tianjiao Chu, Carla Rios Luci, Christy Maksoudian, Ara Sargsian, Bella B. Manshian, Stefaan J. Soenen

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Nanomaterials have gained high interest in their use as potent anticancer agents. Apart from delivering chemotherapeutic agents in order to reduce off-target effects, molecular agents have also been widely explored. The advances in our understanding of cell biology and cell death mechanisms1 has generated a broad library of potential therapeutic targets by siRNA, mRNA, or pDNA complexes. In the present study, we explore the ability of pDNA-polyplexes to induce tumor-specific necroptosis. This results in a cascade of effects, where immunogenic cell death potentiates anti-tumor immune responses and results in an influx of dendritic cells and cytotoxic T cells, rendering the tumor more amenable to immune checkpoint inhibition. This study aims to explore whether the induction of necroptosis in a subpopulation of tumor cells can be used to potentiate immune checkpoint inhibition studies.

Keywords: nanoparticle, MLKL, necroptosis, immunotherapy

Procedia PDF Downloads 136

Authors: Sonal Gupta, Rakhi Bansal, Javed Ali, Reema Gabrani, Shweta Dang

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Nanoemulsions (NEs) and microemulsions (MEs) have been an attractive tool for encapsulation of both hydrophilic and lipophillic actives. Both these systems are composed of oil phase, surfactant, co-surfactant and aqueous phase. Depending upon the application and intended use, both oil-in-water and water-in-oil emulsions can be designed. NEs are fabricated using high energy methods employing less percentage of surfactant as compared to MEs which are self assembled drug delivery systems. Owing to the nanometric size of the droplets these systems have been widely used to enhance solubility and bioavailability of natural as well as synthetic molecules. The aim of the present study is to assess the effect of % age of surfactants on cell viability of Vero cells (African Green Monkeys’ Kidney epithelial cells) via MTT assay. Green tea catechin (Polyphenon 60) loaded ME employing low energy vortexing and NE employing high energy ultrasonication were prepared using same excipients (labrasol as oil, cremophor EL as surfactant and glycerol as co-surfactant) however, the % age of oil and surfactant needed to prepare the ME was higher as compared to NE. These formulations along with their excipients (oilME=13.3%, SmixME=26.67%; oilNE=10%, SmixNE=13.52%) were added to Vero cells for 24 hrs. The tetrazolium dye, 3-(4,5-dimethylthia/ol-2-yl)-2,5-diphi-iiyltclrazolium bromide (MTT), is reduced by live cells and this reaction is used as the end point to evaluate the cytoxicity level of a test formulation. Results of MTT assay indicated that oil at different percentages exhibited almost equal cell viability (oilME ≅ oilNE) while surfactant mixture had a significant difference in the cell viability values (SmixME < SmixNE). Polyphenon 60 loaded ME and its PlaceboME showed higher toxicity as compared to Polyphenon 60 loaded NE and its PlaceboNE that can be attributed to the higher concentration of surfactants present in MEs. Another probable reason for high % cell viability of Polyphenon 60 loaded NE might be due to the effective release of Polyphenon 60 from NE formulation that helps in the sustenance of Vero cells.

Keywords: cell viability, microemulsion, MTT, nanoemulsion, surfactants, ultrasonication

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Authors: Shu-Hui Wen, Wei-Chuan Chang, Hsien-Chang Wu

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Background: Little is known about the benefits and risks of use of Chinese herbal medicine (CHM) in conditions related to hormone therapy (HT) use on the risk of ischemic stroke (IS). The aim of this study is to explore the risk of IS in menopausal women treated with HT and CHM. Materials and methods: A total of 32,441 menopausal women without surgical menopause aged 40- 65 years were selected from 2003 to 2010 using the 2-million random samples of the National Health Insurance Research Database in Taiwan. According to the medication usage of HT and CHM, we divided the current and recent users into two groups: an HT use-only group (n = 4,989) and an HT/CHM group (n = 9,265). Propensity-score matching samples (4,079 pairs) were further created to deal with confounding by indication. The adjusted hazard ratios (HR) of IS during HT or CHM treatment were estimated by the robust Cox proportional hazards model. Results: The incidence rate of IS in the HT/CHM group was significantly lower than in the HT group (4.5 vs. 12.8 per 1000 person-year, p < 0.001). Multivariate analysis results indicated that additional CHM use was significant with a lower risk of IS (HR = 0.3; 95% confidence interval, 0.21-0.43). Further subgroup analyses and sensitivity analyses had similar findings. Conclusion: We found that combined use of HT and CHM was associated with a lower risk for IS than HT use only. Further study is needed to examine possible mechanism underlying this association.

Keywords: Chinese herbal medicine, hormone therapy, ischemic stroke, menopause

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Authors: Travis J. Meyer, Jasodra Ramlall, Phyo Thu, Nidhi Gadura

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For centuries humans have used the antimicrobial properties of copper to their advantage. Yet, after all these years the underlying mechanisms of copper mediated cell death in various microbes remain unclear. We had explored the hypothesis that copper mediated increased levels of lipid peroxidation in the membrane fatty acids is responsible for increased killing inEscherichia coli. In this study we show that in both gram positive (Staphylococcus aureus) and gram negative (Pseudomonas aeruginosa) bacteria there is a strong correlation between copper mediated cell death and increased levels of lipid peroxidation. Interestingly, the non-spore forming gram positive bacteria as well as gram negative bacteria show similar patterns of cell death, increased levels of lipid peroxidation, as well as genomic DNA degradation, however there is some difference inloss in membrane integrity upon exposure to copper alloy surface.

Keywords: antimicrobial, copper, gram positive, gram negative

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Authors: Mohammadreza Delfieh, Seyed Ali Mohammad Modarres Sanavy, Rouzbeh Farhoudi

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Fennel is of most important medicinal plants which is widely used in food and pharmaceutical industries. In order to investigate the effect of different nitrogen nutritional systems including chemical, organic and biologic ones at different plant densities on yield, yield components and seed essential oil content and yield of this valuable medicinal plant, a field experiment was carried out in 2013-2014 agricultural season at Islamic Azad University of Shoushtar agricultural college in split plot design with 18 treatments and based on completely randomized blocks design. Different nitrogen system treatments consisting of: 1. N1 or control (Uniformly spreading urea fertilizer in the plot, 50% at planting time and 50% at stem elongation), 2. N2 (Uniformly spreading 50% of urea fertilizer in the plot at planting time and spraying the other 50% of urea fertilizer at stem elongation on fennel foliage), 3. N3 or cow manure, 4. N4 or biofertilizer (Inoculation of fennel seeds with Azotobacter and Azospirillum), 5. N5 or Integrated-1 (Cow manure + uniformly spreading urea fertilizer in the plot at stem elongation), 6. N6 or Integrated-2 (Cow manure + Inoculation of fennel seeds with Azotobacter and Azospirillum) were applied to the main plots. Three fennel densities consisting of: 1. FD1 (60 plant/m2), 2. FD2 (80 plant/m2) and 3. FD3 (100 plant/m2) were applied to subplots. Results showed that all of the traits were significantly affected by applied treatments (P 0.01). The interaction between treatments also were significant at 5 percent level for shoot dry weight and at 1 percent level for other traits. Based on the results, using the Integrated-1 treatment at 100 plant per m2 produced 94.575 g/m2 seed yield containing 3.375 percent of essential oil. Utilization of such combination not only could lead to a desirable fennel quantity and quality, but also is more consistent with environment.

Keywords: fennel (foeniculum vulgare mill.), nutritional system, nitrogen, biofertilizer, organic fertilizer, chemical fertilizer, density

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Authors: Bin Liu

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MicroRNAs (miRNAs) are small non-coding RNA molecules, functioning in transcriptional and post-transcriptional regulation of gene expression. The discrimination of the real pre-miRNAs from the false ones (such as hairpin sequences with similar stem-loops) is necessary for the understanding of miRNAs’ role in the control of cell life and death. Since both their small size and sequence specificity, it cannot be based on sequence information alone but requires structure information about the miRNA precursor to get satisfactory performance. Kmers are convenient and widely used features for modeling the properties of miRNAs and other biological sequences. However, Kmers suffer from the inherent limitation that if the parameter K is increased to incorporate long range effects, some certain Kmer will appear rarely or even not appear, as a consequence, most Kmers absent and a few present once. Thus, the statistical learning approaches using Kmers as features become susceptible to noisy data once K becomes large. In this study, we proposed a Gapped k-mer approach to overcome the disadvantages of Kmers, and applied this method to the field of miRNA prediction. Combined with the structure status composition, a classifier called imiRNA-GSSC was proposed. We show that compared to the original imiRNA-kmer and alternative approaches. Trained on human miRNA precursors, this predictor can achieve an accuracy of 82.34 for predicting 4022 pre-miRNA precursors from eleven species.

Keywords: gapped k-mer, imiRNA-GSSC, microRNA precursor, support vector machine

Procedia PDF Downloads 159

Authors: Bounar Nedjemeddine

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Materials with the pyrochlore lattice structure have attracted much recent attention due to their wide applications in ceramic thermal barrier coatings, high-permittivity dielectrics, and potential solid electrolytes in solid-oxide fuel cells. The work described in this paper is devoted to the synthesis and characterization of a pyrochlore structure based on lanthanum (La₂O₃) and tin (SnO₂) oxides of general formula La₂Sn₂O₇, substituted by Sr at the site La. Their structures were determined from X-ray powder diffraction using CELFER analysis. All the compositions present the space group Fd-3m. The substitution of La by Sr in the La₂Sn₂O₇ compound causes a variation of the cell parameters. The difference in charge between La³⁺ and Sr²⁺ and the difference in size cause the cell parameters to decrease from a=10.7165 A° to a=10.6848 A° for the substitution rates (x = 0.05, 0.1, 0.15 ...), which leads to a decrease in the volume of the mesh. For a substitution rate x = 0.25, there is an increase in the cell parameters (a=10.7035A°), which can be explained by a competitiveness of the size effect and the presence of a gap in the structure which go in the opposite direction.

Keywords: solid-oxide fuel cells, structure, pyrochlore, X-ray diffraction

Procedia PDF Downloads 125

Authors: M. H. Rajikin, S. M. M. Syairah, A. R. Sharaniza

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Effects of nicotine on pre-partum body weight and preimplantation embryonic development has been reported previously. Present study was conducted to determine the effects of annatto (Bixa orellana)-derived delta-tocotrienol (TCT) (with presence of 10% gamma-TCT isomer) on the nicotine-induced reduction in body weight and 8-cell embryonic growth in mice. Twenty four 6-8 weeks old (23-25g) female balb/c mice were randomly divided into four groups (G1-G4; n=6). Those groups were subjected to the following treatments for 7 consecutive days: G1 (control) were gavaged with 0.1 ml tocopherol stripped corn oil, G2 was subcutaneously (s.c.) injected with 3 mg/kg/day of nicotine, G3 received concurrent treatment of nicotine (3 mg/kg/day) and 60 mg/kg/day of δ-TCT mixture (contains 90% delta & 10% gamma isomers) and G4 was given 60 mg/kg/day of δ-TCT mixture alone. Body weights were recorded daily during the treatment. On Day 8, females were superovulated with 5 IU Pregnant Mare’s Serum Gonadotropin (PMSG) for 48 hours followed with 5 IU human Chorionic Gonadotropin (hCG) before mated with males at the ratio of 1:1. Females were sacrificed by cervical dislocation for embryo collection 48 hours post-coitum. Collected embryos were cultured in vitro. Results showed that throughout Day 1 to Day 7, the body weight of nicotine treated group (G2) was significantly lower (p<0.05) than that of G1, G3 and G4. Intervention with δ-TCT mixture (G3) managed to increase the body weight close to the control group. This is also observed in the group treated with δ-TCT mixture alone (G4). The development of 8-cell embryos following in vitro culture (IVC) was totally inhibited in G2. Intervention with δ-TCT mixture (G3) resulted in the production of 8-cell embryos, although it was not up to that of the control group. Treatment with δ-TCT mixture alone (G4) caused significant increase in the average number of produced 8-cell embryo compared to G1. Present data indicated that δ-TCT mixture was able to reverse the body weight loss in nicotine treated mice and the development of 8-cell embryos was also improved.

Keywords: δ-tocotrienol, body weight, nicotine, preimplantation embryonic development

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Authors: Laura Gleason, Sahithi Talasila, Lauren Banner, Ladan Afifi, Neda Nikbakht

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Primary cutaneous anaplastic large cell lymphoma (pcALCL) accounts for 9% of all cutaneous T-cell lymphomas. pcALCL is classically characterized as a solitary papulonodule that often enlarges, ulcerates, and can be locally destructive, but overall exhibits an indolent course with overall 5-year survival estimated to be 90%. Distinguishing pcALCL from systemic ALCL (sALCL) is essential as sALCL confers a poorer prognosis with average 5-year survival being 40-50%. Although extremely rare, there have been several cases of ALK-positive ALCL diagnosed on skin biopsy without evidence of systemic involvement, which poses several challenges in the classification, prognostication, treatment, and follow-up of these patients. Objectives: We present a case of cutaneous ALK-positive ALCL without evidence of systemic involvement, and a narrative review of the literature to further characterize that ALK-positive ALCL limited to the skin is a distinct variant with a unique presentation, history, and prognosis. A 30-year-old woman presented for evaluation of an erythematous-violaceous papule present on her right chest for two months. With the development of multifocal disease and persistent lymphadenopathy, a bone marrow biopsy and lymph node excisional biopsy were performed to assess for systemic disease. Both biopsies were unrevealing. The patient was counseled on pursuing systemic therapy consisting of Brentuximab, Cyclophosphamide, Doxorubicin, and Prednisone given the concern for sALCL. Apropos of the patient we searched for clinically evident, cutaneous ALK-positive ALCL cases, with and without systemic involvement, in the English literature. Risk factors, such as tumor location, number, size, ALK localization, ALK translocations, and recurrence, were evaluated in cases of cutaneous ALK-positive ALCL. The majority of patients with cutaneous ALK-positive ALCL did not progress to systemic disease. The majority of cases that progressed to systemic disease in adults had recurring skin lesions and cytoplasmic localization of ALK. ALK translocations did not influence disease progression. Mean time to disease progression was 16.7 months, and significant mortality (50%) was observed in those cases that progressed to systemic disease. Pediatric cases did not exhibit a trend similar to adult cases. In both the adult and pediatric cases, a subset of cutaneous-limited ALK-positive ALCL were treated with chemotherapy. All cases treated with chemotherapy did not progress to systemic disease. Apropos of an ALK-positive ALCL patient with clinical cutaneous limited disease in the histologic presence of systemic markers, we discussed the literature data, highlighting the crucial issues related to developing a clinical strategy to approach this rare subtype of ALCL. Physicians need to be aware of the overall spectrum of ALCL, including cutaneous limited disease, systemic disease, disease with NPM-ALK translocation, disease with ALK and EMA positivity, and disease with skin recurrence.

Keywords: anaplastic large cell lymphoma, systemic, cutaneous, anaplastic lymphoma kinase, ALK, ALCL, sALCL, pcALCL, cALCL

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Authors: Sanjay Shukla, A. Nayak, R. K. Sharma, A. P. Singh, S. P. Tiwari

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Excessive use of antibiotics is a major problem in the treatment of wounds and other chronic infections, and antibiotic treatment is frequently non-curative, thus alternative treatment is necessary. Phage therapy is considered one of the most promising approaches to treat multi-drug resistant bacterial pathogens. Infections caused by Staphylococcus aureus are very efficiently controlled with phage cocktails, containing a different individual phages lysate infecting a majority of known pathogenic S. aureus strains. The aim of the present study was to evaluate the efficacy of a purified phage cocktail for prophylactic as well as therapeutic application in mouse model and in large animals with chronic septic infection of wounds. A total of 150 sewage samples were collected from various livestock farms. These samples were subjected for the isolation of bacteriophage by the double agar layer method. A total of 27 sewage samples showed plaque formation by producing lytic activity against S. aureus in the double agar overlay method out of 150 sewage samples. In TEM, recovered isolates of bacteriophages showed hexagonal structure with tail fiber. In the bacteriophage (ØVS) had an icosahedral symmetry with the head size 52.20 nm in diameter and long tail of 109 nm. Head and tail were held together by connector and can be classified as a member of the Myoviridae family under the order of Caudovirale. Recovered bacteriophage had shown the antibacterial activity against the S. aureus in vitro. Cocktail (ØVS1, ØVS5, ØVS9, and ØVS 27) of phage lysate were tested to know in vivo antibacterial activity as well as the safety profile. Result of mice experiment indicated that the bacteriophage lysate were very safe, did not show any appearance of abscess formation, which indicates its safety in living system. The mice were also prophylactically protected against S. aureus when administered with cocktail of bacteriophage lysate just before the administration of S. aureuswhich indicates that they are good prophylactic agent. The S. aureusinoculated mice were completely recovered by bacteriophage administration with 100% recovery, which was very good as compere to conventional therapy. In the present study, ten chronic cases of the wound were treated with phage lysate, and follow up of these cases was done regularly up to ten days (at 0, 5, and 10 d). The result indicated that the six cases out of ten showed complete recovery of wounds within 10 d. The efficacy of bacteriophage therapy was found to be 60% which was very good as compared to the conventional antibiotic therapy in chronic septic wounds infections. Thus, the application of lytic phage in single dose proved to be innovative and effective therapy for the treatment of septic chronic wounds.

Keywords: phage therapy, S aureus, antimicrobial resistance, lytic phage, and bacteriophage

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Authors: Soultana Konstantinidou, Tiziana Schmidt, Elena Landi, Alessandro De Carli, Giovanni Maltinti, Darius Witt, Alicja Dziadosz, Agnieszka Lindstaedt, Michele Lai, Mauro Pistello, Valentina Cappello, Luciana Dente, Chiara Gabellini, Piotr Barski, Vittoria Raffa

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CRISPR/Cas9 technology has gained the interest of researchers in the field of biotechnology for genome editing. Since its discovery as a microbial adaptive immune defense, this system has been widely adopted and is acknowledged for having a variety of applications. However, critical barriers related to safety and delivery are persisting. Here, we propose a new concept of genome engineering, which is based on a nano-formulation of Cas9. The Cas9 enzyme was conjugated to a gold nanoparticle (AuNP-Cas9). The AuNP-Cas9 maintained its cleavage efficiency in vitro, to the same extent as the ribonucleoprotein, including non-conjugated Cas9 enzyme, and showed high gene editing efficiency in vivo in zebrafish embryos. Since CRISPR/Cas9 technology is extensively used in cancer research, melanoma was selected as a validation target. Cell studies were performed in A375 human melanoma cells. Particles per se had no impact on cell metabolism and proliferation. Intriguingly, the AuNP-Cas9 internalized spontaneously in cells and localized as a single particle in the cytoplasm and organelles. More importantly, the AuNP-Cas9 showed a high nuclear localization signal. The AuNP-Cas9, overcoming the delivery difficulties of Cas9, could be used in cellular biology and localization studies. Taking advantage of the plasmonic properties of gold nanoparticles, this technology could potentially be a bio-tool for combining gene editing and photothermal therapy in cancer cells. Further work will be focused on intracellular interactions of the nano-formulation and characterization of the optical properties.

Keywords: CRISPR/Cas9, gene editing, gold nanoparticles, nanotechnology

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Authors: Xing Lu, Yi-ming Wu, Yan-hong Zhu, Zhen-feng Chen, Hong Liang, Yan Peng

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[Eu(BMQ)2(NO3)3(CH3OH)(H2O)] (1),and [Er(BMQ)2(NO3)3(CH3OH)(H2O)] (2),were synthesized. Compounds 1 and 2 exhibit a certain extent cytotoxicity against Hep G2, Hela 229, MGC80-3 and BEL-7404 cell lines invitro, with IC50 values in the14.51±1.41μM to 52.49±4.01μM range. Compound 1 exhibited significantly enhanced cytotoxicity against MGC80-3 cell line, comparing with free 3-(1H-benzimidazol-2-yl)-6-methyl-2(1H)- quinolinone. The binding abilities of 1 to DNA were stronger than that of 2. Intercalation is the most probable binding mode for both the complexes.

Keywords: quinolinone, Eu(II) complex, Er(III) complex, cytotoxicity.

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Authors: M. Karzhauov, А. Mukhambetova, M. Sarsenova, E. Raimagambetov, V. Ogay

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Regeneration of injured articular cartilage remains one of the most difficult and unsolved problems in traumatology and orthopedics. Currently, for the treatment of cartilage defects surgical techniques for stimulation of the regeneration of cartilage in damaged joints such as multiple microperforation, mosaic chondroplasty, abrasion and microfractures is used. However, as shown by clinical practice, they can not provide a full and sustainable recovery of articular hyaline cartilage. In this regard, the current high hopes in the regeneration of cartilage defects reasonably are associated with the use of tissue engineering approaches to restore the structural and functional characteristics of damaged joints using stem cells, growth factors and biopolymers or scaffolds. The purpose of the present study was to investigate the effects of chondroinductive growth factors and synovium-derived mesenchymal stem cells (SD-MSCs) on the regeneration of cartilage defects in rabbits. SD-MSCs were isolated from the synovium membrane of Flemish giant rabbits, and expanded in complete culture medium α-MEM. Rabbit SD-MSCs were characterized by CFU-assay and by their ability to differentiate into osteoblasts, chondrocytes and adipocytes. The effects of growth factors (TGF-β1, BMP-2, BMP-4 and IGF-I) on MSC chondrogenesis were examined in micromass pellet cultures using histological and biochemical analysis. Articular cartilage defect (4mm in diameter) in the intercondylar groove of the patellofemoral joint was performed with a kit for the mosaic chondroplasty. The defect was made until subchondral bone plate. Delivery of SD-MSCs and growth factors was conducted in combination with hyaloronic acid (HA). SD-MSCs, growth factors and control groups were compared macroscopically and histologically at 10, 30, 60 and 90 days aftrer intra-articular injection. Our in vitro comparative study revealed that TGF-β1 and BMP-4 are key chondroinductive factors for both the growth and chondrogenesis of SD-MSCs. The highest effect on MSC chondrogenesis was observed with the synergistic interaction of TGF-β1 and BMP-4. In addition, biochemical analysis of the chondrogenic micromass pellets also revealed that the levels of glycosaminoglycans and DNA after combined treatment with TGF-β1 and BMP-4 was significantly higher in comparison to individual application of these factors. In vivo study showed that for complete regeneration of cartilage defects with intra-articular injection of SD-MSCs with HA takes time 90 days. However, single injection of SD-MSCs in combiantion with TGF-β1, BMP-4 and HA significantly promoted regeneration rate of the cartilage defects in rabbits. In this case, complete regeneration of cartilage defects was observed in 30 days after intra-articular injection. Thus, our in vitro and in vivo study demonstrated that combined application of rabbit SD-MSC with chondroinductive growth factors and HA results in strong synergistic effect on the chondrogenesis significantly enhancing regeneration of the damaged cartilage.

Keywords: Mesenchymal stem cells, synovium, chondroinductive factors, TGF-β1, BMP-2, BMP-4, IGF-I

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Authors: Prasanthi Malapati, R. Reshma, Vijay Soni, Perumal Yogeeswari, Dharmarajan Sriram

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In the present study, we attempted to develop Mycobacterium tuberculosis (Mtb) inhibitors by exploring the pharmaceutically underexploited enzyme targets which are majorly involved in cell wall biosynthesis of mycobacteria. For this purpose, glutamate racemase (coded by MurI gene) was selected. This enzyme racemize L-glutamate to D-glutamate required for the construction of peptidoglycan in the bacterial cell wall synthesis process. Furthermore this enzyme is neither expressed nor its product, D-glutamate is normally found in mammals, and hence designing inhibitors against this enzyme will not affect the host system as well act as potential antitubercular drugs. A library of BITS in house compounds were screened against Mtb MurI enzyme. Based on docking score, interactions and synthetic feasibility one hit lead was identified. Further optimization of lead was attempted and its derivatives were synthesized. Forty eight derivatives of 2-phenylbenzo[d]oxazole and 2-phenylbenzo[d]thiazole were synthesized and evaluated for Mtb MurI inhibition study, in vitro activities against Mtb, cytotoxicity against RAW 264.7 cell line. Chemical derivatization of the lead resulted in compounds NR-1213 AND NR-1124 as the potent M. tuberculosis glutamate racemase inhibitors with IC50 of 4-5µM which are remarkable and were found to be non-cytotoxic. Molecular dynamics, dormant models and cardiotoxicity studies of the most active molecules are in process.

Keywords: cell wall biosynthesis, dormancy, glutamate racemase, tuberculosis

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Authors: Wei-Wen Hu, Yong-Zhi Zhong

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Depositing conductive polypyrrole (PPy) onto elastic polydimethylsiloxane (PDMS) substrate can obtain a highly stretchable conductive film, which can be used to construct a bioreactor to cyclically stretch and electrically stimulate surface cells. However, how to completely harvest these stimulated muscle tissue to repair damaged muscle is a challenge. To address this concern, N-isopropylacrylamide (NIPAAm), a monomer of temperature-sensitive polymer, was added during the polymerization of pyrrole on PDMS so that the resulting P(Py-co-NIPAAm)/PDMS should own both conductivity and thermo-sensitivity. Therefore, cells after stimulation can be completely harvested as cell sheets by reducing temperature. Mouse skeletal myoblast, C2C12 cells, were applied to examine our hypothesis. In electrical stimulation, C2C12 cells on P(Py-co-NIPAAm)/PDMS demonstrated the best myo-differentiation under the electric field of 1 V/cm. Regarding cyclic stretching, the strain equal to or higher than 9% can highly align C2C12 perpendicular to the stretching direction. The Western blotting experiments demonstrated that the cell sheets harvested by cooling reserved more extracellular matrix (ECM) than cells collected by the traditional trypsin digestion method. Immunostaining of myosin heavy chain protein (MHC) indicated that both mechanical and electrical stimuli effectively increased the number of myotubes and the differentiation ratio, and the myotubes can be aligned by cyclic stretching. Stimulated cell sheets can be harvested by cooling, and the alignment of myotubes was still maintained. These results suggested that the deposition of P(Py-co-NIPAAm) on PDMS can be applied to harvest intact cell sheets after cyclic stretching and electrical stimulation, which increased the feasibility of bioreactor for the application of tissue engineering and regenerative medicine.

Keywords: bioreactor, cell sheet, conductive polymer, cyclic stretching, electrical stimulation, muscle tissue engineering, myogenesis, thermosensitive hydrophobicity

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Authors: Rachel Jones

Abstract:

Systemic Family therapy in the Queensland Foster Care System is the implementation of Integrative Practice as a purposeful intervention implemented with complex ‘family’ systems (by expanding the traditional concept of family to include all relevant stakeholders for a child) and is shown to improve the overall wellbeing of children (with developmental delays and trauma) in Queensland out of home care contexts. The importance of purposeful integrative practice in the field of systemic family therapy has been highlighted in achieving change in complex family systems. Essentially, it is the purposeful use of multiple interventions designed to meet the myriad of competing needs apparent for a child (with developmental delays resulting from early traumatic experiences - both in utero and in their early years) and their family. In the out-of-home care context, integrative practice is particularly useful to promote positive change for the child and what is an extended concept of whom constitutes their family. Traditionally, a child’s family may have included biological and foster care family members, but when this concept is extended to include all their relevant stakeholders (including biological family, foster carers, residential care workers, child safety, school representatives, Health and Allied Health staff, police and youth justice staff), the use of integrative family therapy can produce positive change for the child in their overall wellbeing, development, risk profile, social and emotional functioning, mental health symptoms and relationships across domains. By tailoring therapeutic interventions that draw on systemic family therapies from the first and second-order schools of family therapy, neurobiology, solution focussed, trauma-informed, play and art therapy, and narrative interventions, disability/behavioural interventions, clinicians can promote change by mixing therapeutic modalities with the individual and their stakeholders. This presentation will unpack the implementation of systemic family therapy using this integrative approach to formulation and treatment for a child in out-of-home care in Queensland (experiencing developmental delays resulting from trauma). It considers the need for intervention for the individual and in the context of the environment and relationships. By reviewing a case example, this study aims to highlight the simultaneous and successful use of pharmacological interventions, psychoeducational programs for carers and school staff, parenting programs, cognitive-behavioural and trauma-informed interventions, traditional disability approaches, play therapy, mapping genograms and meaning-making, and using family and dyadic sessions for the system associated with the foster child. These elements of integrative systemic family practice have seen success in the reduction of symptoms and improved overall well-being of foster children and their stakeholders. Accordingly, a model for best practice using this integrative systemic approach is presented for this population group and preliminary findings for this approach over four years of local data have been reviewed.

Keywords: systemic family therapy, treating families of children with delays, trauma and attachment in families systems, improving practice and functioning of children and families

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Authors: Sree Vennela P., V. Samyuktha Bhardwaj

Abstract:

Hyponatremia is an electrolyte disturbance in which the sodium ion concentration in the serum is lower than normal. Sodium is the dominant extracellular cation (positive ion) and cannot freely cross from the interstitial space through the cell membrane, into the cell. Its homeostasis (stability of concentration) inside the cell is vital to the normal function of any cell. Normal serum sodium levels are between 135 and 145 mEq/L. Hyponatremia is defined as a serum level of less than 135 mEq/L and is considered severe when the serum level is below 125 mEq/L. In the vast majority of cases, Hyponatremia occurs as a result of excess body water diluting the serum sodium (salt level in the blood). Hyponatremia is often a complication of other medical illnesses in which excess water accumulates in the body at a higher rate than can be excreted (for example in congestive heart failure, syndrome of inappropriate antidiuretic hormone, SIADH, or polydipsia). Sometimes it may be a result of over-hydration (drinking too much water).Lack of sodium (salt) is very rarely the cause of Hyponatremia, although it can promote Hyponatremia indirectly. In particular, sodium loss can lead to a state of volume depletion (loss of blood volume in the body), with volume depletion serving as a signal for the release of ADH (anti-diuretic hormone). As a result of ADH-stimulated water retention (too much water in the body), blood sodium becomes diluted and Hyponatremia results.

Keywords: Tolvaptan, hyponatremia, syndrome of insufficient anti diuretic hormone (SIADH), euvolemic hyponatremia

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Authors: Syeda Kiran Shahzadi, Nasir Mahmood, Muhammad Abdul Qadir

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In the current research, three recombinant human interferon alpha-2b proteins (two modified and one normal form) were produced and Pegylated with an aim to produce more effective drugs against viral infections and cancers. The modified recombinant human interferon alpha-2b proteins were produced by site-directed modifications of interferon alpha 2b gene, targeting the amino acids at positions ‘R23’ and ‘H34’. The resulting chemically modified and unmodified forms of human interferon alpha 2b were conjugated with methoxy-polyethylene glycol propanealdehyde (400 KDa) and methoxy-polyethylene glycol succinimidyl succinate (400 KDa). Pegylation of normal and modified forms of Interferon alpha-2b prolong their release time and enhance their efficacy. The conjugation of PEG with modified and unmodified human interferon alpha 2b protein drugs was also characterized with 1H-NMR, HPLC, and SDS-PAGE. Antiproliferative assays of modified and unmodified forms of drugs were performed in cell based bioassays using MDBK cell lines. The results indicated that experimentally produced recombinant human interferon alpha-2b proteins were biologically active and resulted in significant inhibition of cell growth.

Keywords: protein refolding, antiproliferative activities, biomedical applications, human interferon alpha-2b, pegylation, mPEG-propionaldehyde, site directed mutagenesis, E. coli expression

Procedia PDF Downloads 175

Authors: Hady Atef, Zinab Helmy, Heba Abdeen, Mostafa Fadel

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Background and purpose: After the success of the first trials of this experiment which were done on rabbits, a new study were conducted on dogs to ensure the past results; in a step forward to use low-level LASER therapy in the treatment of congenital septal defects in infants. The aim of this study was to investigate the effect of low-level LASER irradiation on congenital septal defects in dogs. Subjects and Methodology: six male dogs who have congenital septal defects in their hearts -with age ranged 6-10 months- enrolled in this study for one and half months. They were assigned into two groups: Group (A): The study group consisted of 3 canine hearts who received routine animal care associated with LASER irradiation. Group (B): The control group consisted of 3 canine hearts who received only routine animal care. Sizes of the septal defects were measured for both groups at the beginning and after the end of the study. Results: There was a significant decrease in the size of the diameter of the congenital septal defect with the study group (percentage of improvement was 42.19%) when compared with control group. Conclusion: It was concluded that low-level LASER therapy can be considered as a promising therapy for congenital heart defects in animals and to be examined on children with similar congenital lesions after then.

Keywords: laser, congenital septal defects, dogs, infants

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Authors: Yuvarani Naidu, Yasmeen Siddiqui, Mohd Yusof Rafii , Abu Seman Idris

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Sustainability of oil palm production in Southeast Asia, especially in Indonesia and Malaysia, is jeopardized by Ganoderma boninense, the fungus which causes basal stem rot (BSR) in oil palm. The root contact with unattended infected debris left in the plantations during replanting is known to be the primary source of inoculum. Abiding by the law, potentially effective technique of managing Ganoderma infected oil palm debris is deemed necessary because of the zero-burning policy in Malaysian oil palm plantations. White-rot hymenomycetes antagonistic to Ganoderma sp were selected to test their efficacy as log treatments in degrading Ganoderma infected oil palm logs and to minimize the survival of Ganoderma inoculum. Decay rate in terms of mass loss was significantly higher after the application of solid-state cultivation (SSC) of Trametes lactinea FBW (64% ±1.2), followed by Pycnoporus sanguineus FBR (55% ±1.7) in infected log block tissues, after 10 months of treatments. The degradation pattern was clearly distinguished between the treated and non-treated log blocks with the developed SSC formulations. The control infected log blocks showed the highest, whereas infected log blocks treated with either P. sanguineus FBR or T. lactinea FBW SSC formulations exhibited statistically lowest number of Ganoderma spp. recovery on Ganoderma Selective Medium (GSM), after 8 months of treatment. Out of that, the lowest recovery of Ganoderma spp. was reported in infected log blocks inoculated with the strain T. lactinea FBW (21% ± 0.9) followed by P. sanguineus FBR (33% ± 2.2), after 8 months, Further, no recovery of Ganoderma was noticeable, 10 months after treatment applications in log blocks treated with both of the formulations. This is the first nursery-base study to substantiate the initial colonization of white-rot hymenomycetes on oil palm log blocks previously infected with BSR pathogen, G. boninense. The present study has indicated that log blocks treatment with white-rot hymenomycetes significantly affected the mass loss of diseased and healthy log block tissues. This study provides a basis of biotechnological approaches inefficient degradation of oil palm-generated crop debris, under natural conditions with an ultimate aim of reducing the Ganoderma inoculum under heavy BSR infection pressure in eco-friendly manner.

Keywords: basal stem rot disease, ganoderma boninense, oil palm, white-rot fungi

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Authors: S. C. Santos, S. R. Dionísio, A. L. D. De Andrade, L. R. Roque, A. C. Da Costa, J. L. Ienczak

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In this work, two fermentations at different temperatures (25 and 30 ºC), with cell recycling, were accomplished to produce ethanol, using a mix of commercial substrates, xylose (70%) and glucose (30%), as organic source for Scheffersomyces stipitis. Five consecutive fermentations of 80 g L-1 (1º, 2º and 3º recycles), 96 g L-1 (4º recycle) and 120 g L-1 (5º recycle)reduced sugars led to a final maximum ethanol concentration of 17.2 and 34.5 g L-1, at 25 and 30 ºC, respectively. Glucose was the preferred substrate; moreover xylose startup degradation was initiated after a remaining glucose presence in the medium. Results showed that yeast acid treatment, performed before each cycle, provided improvements on cell viability, accompanied by ethanol productivity of 2.16 g L-1 h-1 at 30 ºC. A maximum 36% of xylose was retained in the fermentation medium and after five-cycle fermentation an ethanol yield of 0.43 g ethanol/g sugars was observed. S. stipitis fermentation capacity and tolerance showed better results at 30 ºC with 83.4% of theoretical yield referenced on initial biomass.

Keywords: 5-carbon sugar, cell recycling fermenter, mixed sugars, xylose-fermenting yeast

Procedia PDF Downloads 416

Authors: Evgeniia Surkova, Manal Assaad, Hleb Makeyeu, Juho Makio

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The main objective of STIMEY (Science, Technology, Innovation, Mathematics, Engineering for the Young) project is the delivery of a hybrid learning environment that combines multi-level components such as social media concepts, robotic artefacts, and radio, among others. It is based on a well-researched pedagogical framework to attract European youths to STEM (science, technology, engineering, and mathematics) education and careers. To develop and integrate these various components, STIMEY is executed in iterative research cycles leading to progressive improvements. Scrum was the development methodology of choice in the project, as studies indicated its benefits as an agile methodology in global software development, especially of e-learning and integrated learning projects. This paper describes the project partners’ experience with the Scrum framework, discussing the challenges faced in its implementation and the mitigation practices employed. The authors conclude with exploring user experience tools and principles for future research, as a novel direction in supporting the Scrum development team.

Keywords: e-learning, global software development, scrum, STEM education

Procedia PDF Downloads 178

Authors: Usman Asif, Klaus Schmidt

Abstract:

In recent years, the development of alternate fuel vehicles has helped to reduce carbon emissions worldwide. As the number of vehicles will continue to increase in the future, the energy demand will also increase. Therefore, we must consider automotive technologies that are efficient and less harmful to the environment in the long run. Battery Electric Vehicles (BEVs) have gained popularity in recent years because of their lower maintenance, lower fuel costs, and lower carbon emissions. Nevertheless, BEVs show several disadvantages, such as slow charging times and lower range than traditional combustion-powered vehicles. These factors keep many people from switching to BEVs. The authors of this research believe that these limitations can be overcome by using fuel cell technology. Fuel cell technology converts chemical energy into electrical energy from hydrogen power and therefore serves as fuel to power the motor and thus replacing heavy lithium batteries that are expensive and hard to recycle. Also, in contrast to battery-powered electric vehicle technology, Fuel Cell Electric Vehicles (FCEVs) offer higher ranges and lower fuel-up times and therefore are more competitive with electric vehicles. However, FCEVs have not gained the same popularity as electric vehicles due to stringent legal frameworks, underdeveloped infrastructure, high fuel transport, and storage costs plus the expense of fuel cell technology itself. This research will focus on the legal frameworks for hydrogen-powered vehicles, and how a change in these policies may affect and improve hydrogen fueling infrastructure and lower hydrogen transport and storage costs. These policies may also facilitate reductions in fuel cell technology costs. In order to attain a better framework, a number of countries have developed conceptual roadmaps. These roadmaps have set out a series of objectives to increase the access of FCEVs to their respective markets. This research will specifically focus on policies in Japan, Europe, and the USA in their attempt to shape the automotive industry of the future. The researchers also suggest additional policies that may help to accelerate the advancement of FCEVs to mass-markets. The approach was to provide a solid literature review using resources from around the globe. After a subsequent analysis and synthesis of this review, the authors concluded that in spite of existing legal challenges that have hindered the advancement of fuel-cell technology in the automobile industry in the past, new initiatives that enhance and advance the very same technology in the future are underway.

Keywords: fuel cell electric vehicles, fuel cell technology, legal frameworks, policies and regulations

Procedia PDF Downloads 114

Authors: Liang Jiansheng, Zhu Wei

Abstract:

Light and brassinosteroids are essential external and internal cues for plant survival. Although the coordination of light with phytohormone signals is crucial for plant growth and development, the molecular connection between light and brassinosteroid signaling during root soil penetration remains elusive. Here, we reveal that light-stabilized RACK1 couples a brassinosteroid signaling cascade to drive cell division in root meristems. RACK1 family scaffold proteins positively regulate light-induced the promotion of root elongation during soil penetration. Under the light condition, RACK1A interacts with both phyB and SPA1, then reinforces the phyB-SPA1 association to accumulate its abundance in roots. In response to brassinosteroid signals, RACK1A competes with BKI1 to attenuate the BRI1-BKI1 interaction, thereby leading to activating BRI1 actions in root development. Furthermore, RACK1A binds to BES1 to repress its DNA binding activity toward the target gene CYCD3;1. This ultimately allows to release the inhibition of CYCD3;1 transcription, and promotes cell division during root growth. Our study illustrates a new mechanistic model of how plants engage scaffold proteins in transducing light information to facilitate brassinosteroid signaling for root growth in the soil.

Keywords: root growth, cell division, light signaling, brassinosteroid signaling, soil penetration, scaffold protein, RACK1

Procedia PDF Downloads 77

Authors: John K. Mensah, Mary A. Sword Sayer, Ryan L. Nadel, George Matusick, Zhaofei Fan, Lori G. Eckhardt

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Leptographium terebrantis is an opportunistic root pathogen commonly associated with loblolly pine (Pinus taeda L.) stands that are undergoing a loss of vigor in the southeastern US. In order to understand the relationship between L. terebrantis inoculum density and host physiology, an artificial inoculation study was conducted in a five-year-old naturally regenerated loblolly pine stand over a 24 week period in a completely randomized design. L. terebrantis caused sapwood occlusions that increased in severity as inoculum density increased. The occlusions significantly reduced water transport through the stem but did not interfere with fascicle-level stomatal conductance or induce moisture stress in the saplings. The resilience of stomatal conductance among pathogen-infested saplings is attributed to the growth and hydraulic function of new sapwood that developed after artificial inoculation. Results demonstrate that faster-growing families of loblolly pine may be capable of tolerating the vascular root disease when the formation of new sapwood is supported by sustained crown health.

Keywords: hydraulic conductance, inoculum density, Leptographium terebrantis, Pinus taeda, sapwood occlusion

Procedia PDF Downloads 318