Search results for: terrestrial microbial fuel cell

Authors: B. Nageshwar Rao

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Urinary tract infection (UTI) represents the most commonly acquired bacterial infection worldwide, with substantial morbidity, mortality, and economic burden. The objective of the study is to characterize the microbial profiles of uropathogenic in the obstetric population by RTPCR. Study design: An observational cross-sectional study was performed at a single tertiary health care hospital among 50 pregnant women with UTIs, including asymptomatic and symptomatic patients attending the outpatient department and inpatient department of Obstetrics and Gynaecology.Methods: Serotyping and genes detection of various uropathogens were studied using RTPCR. Pulse filed gel electrophoresis methods were used to determine the various genetic profiles. Results: The present study shows that CsgD protein, involved in biofilm formation in Escherichia coli, VIM1, IMP1 genes for Klebsiella were identified by using the RTPCR method. Our results showed that the prevalence of VIM1 and IMP1 genes and CsgD protein in E.coli showed a significant relationship between strong biofilm formation, and this may be due to the prevalence of specific genes. Finally, the genetic identification of RTPCR results for both bacteria was correlated with each other and concluded that the above uropathogens were common isolates in producing Biofilm in the pregnant woman suffering from urinary tract infection in our hospital observational study.

Keywords: biofilms, Klebsiella, E.coli, urinary tract infection

Procedia PDF Downloads 126

Authors: Anuj Tyagi, Balwinder Singh, Naveen Kumar B. T., Niraj K. Singh

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Characterization of diverse microbial communities in specific environment plays a crucial role in the better understanding of their functional relationship with the ecosystem. It is now well established that gut microbiome of fish is not the simple replication of microbiota of surrounding local habitat, and extensive species, dietary, physiological and metabolic variations in fishes may have a significant impact on its composition. Moreover, overuse of antibiotics in human, veterinary and aquaculture medicine has led to rapid emergence and propagation of antibiotic resistance genes (ARGs) in the aquatic environment. Microbial communities harboring specific ARGs not only get a preferential edge during selective antibiotic exposure but also possess the significant risk of ARGs transfer to other non-resistance bacteria within the confined environments. This phenomenon may lead to the emergence of habitat-specific microbial resistomes and subsequent emergence of virulent antibiotic-resistant pathogens with severe fish and consumer health consequences. In this study, gut microbiota of freshwater carp (Labeo rohita) was investigated by shotgun metagenomics to understand its taxonomic composition and functional capabilities. Metagenomic DNA, extracted from the fish gut, was subjected to sequencing on Illumina NextSeq to generate paired-end (PE) 2 x 150 bp sequencing reads. After the QC of raw sequencing data by Trimmomatic, taxonomic analysis by Kraken2 taxonomic sequence classification system revealed the presence of 36 phyla, 326 families and 985 genera in the fish gut microbiome. At phylum level, Proteobacteria accounted for more than three-fourths of total bacterial populations followed by Actinobacteria (14%) and Cyanobacteria (3%). Commonly used probiotic bacteria (Bacillus, Lactobacillus, Streptococcus, and Lactococcus) were found to be very less prevalent in fish gut. After sequencing data assembly by MEGAHIT v1.1.2 assembler and PROKKA automated analysis pipeline, pathway analysis revealed the presence of 1,608 Metacyc pathways in the fish gut microbiome. Biosynthesis pathways were found to be the most dominant (51%) followed by degradation (39%), energy-metabolism (4%) and fermentation (2%). Almost one-third (33%) of biosynthesis pathways were involved in the synthesis of secondary metabolites. Metabolic pathways for the biosynthesis of 35 antibiotic types were also present, and these accounted for 5% of overall metabolic pathways in the fish gut microbiome. Fifty-one different types of antibiotic resistance genes (ARGs) belonging to 15 antimicrobial resistance (AMR) gene families and conferring resistance against 24 antibiotic types were detected in fish gut. More than 90% ARGs in fish gut microbiome were against beta-lactams (penicillins, cephalosporins, penems, and monobactams). Resistance against tetracycline, macrolides, fluoroquinolones, and phenicols ranged from 0.7% to 1.3%. Some of the ARGs for multi-drug resistance were also found to be located on sequences of plasmid origin. The presence of pathogenic bacteria and ARGs on plasmid sequences suggested the potential risk due to horizontal gene transfer in the confined gut environment.

Keywords: antibiotic resistance, fish gut, metabolic pathways, microbial diversity

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Authors: S. Rashmi, Devashish Das, N. Spoorthi, H. V. Manasa

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Energy is essential and plays an important role for overall development of a nation. The global economy literally runs on energy. The use of fossil fuels as energy is now widely accepted as unsustainable due to depleting resources and also due to the accumulation of greenhouse gases in the environment, renewable and carbon neutral biodiesel are necessary for environment and economic sustainability. Unfortunately biodiesel produced from oil crop, waste cooking oil and animal fats are not able to replace fossil fuel. Fossil fuels remain the dominant source of primary energy, accounting for 84% of the overall increase in demand. Today biodiesel has come to mean a very specific chemical modification of natural oils. Objectives: To produce biodiesel from yellow oleander seed oil, to test the yield of biodiesel using different types of catalyst (KOH & NaOH). Methodology: Oil is extracted from dried yellow oleander seeds using Soxhlet extractor and oil expeller (bulk). The FFA content of the oil is checked and depending on the FFA value either two steps or single step process is followed to produce biodiesel. Two step processes includes esterfication and transesterification, single step includes only transesterification. The properties of biodiesel are checked. Engine test is done for biodiesel produced. Result: It is concluded that biodiesel quality parameters such as yield(85% & 90%), flash point(1710C & 1760C),fire point(1950C & 1980C), viscosity(4.9991 and 5.21 mm2/s) for the biodiesel from seed oil of Thevetiaperuviana produced by using KOH & NaOH respectively. Thus the seed oil of Thevetiaperuviana is a viable feedstock for good quality fuel.The outcomes of our project are a substitute for conventional fuel, to reduce petro diesel requirement,improved performance in terms of emissions. Future prospects: Optimization of biodiesel production using response surface method.

Keywords: yellow oleander seeds, biodiesel, quality parameters, renewable sources

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Authors: Nivedita Sharma

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The global demand for fossil fuels is very high, but their use is not sustainable since its reserves are declining. Additionally, fossil fuels are responsible for the accumulation of greenhouse gases. The emission of greenhouse gases from the transport sector can be reduced by substituting fossil fuels by biofuels. Thus, renewable fuels capable of sequestering carbon dioxide are in high demand. Second‐generation biofuels, which require lignocellulosic biomass as a substrate and ultimately producing ethanol, fall largely in this category. Bioethanol is a favorable and near carbon-neutral renewable biofuel leading to reduction in tailpipe pollutant emission and improving the ambient air quality. Lignocellulose consists of three main components: cellulose, hemicellulose and lignin which can be converted to ethanol with the help of microbial enzymes. Enzymatic hydrolysis of lignocellulosic biomass in 1st step is considered as the most efficient and least polluting methods for generating fermentable hexose and pentose sugars which subsequently are fermented to power alcohol by yeasts in 2nd step of the process. In the present technology, a complete bioconversion process i.e. potential hydrolytic enzymes i.e. cellulase and xylanase producing microorganisms have been isolated from different niches, screened for enzyme production, identified using phenotyping and genotyping, enzyme production, purification and application of enzymes for saccharification of different lignocellulosic biomass followed by fermentation of hydrolysate to ethanol with high yield is to be presented in detail.

Keywords: cellulase, xylanase, lignocellulose, bioethanol, microbial enzymes

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Authors: Jyh-Cheng Chen, Tai-Jing Wang, Yun-Wei Lin

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Astaxanthin has been demonstrated to exhibit a wide range of beneficial effects including anti-inflammatory and anti-cancer properties. However, the molecular mechanism of astaxanthin-induced cytotoxicity in non-small cell lung cancer (NSCLC) cells has not been identified. Rad51 plays a central role in homologous recombination and high levels of Rad51 expression are observed in chemo- or radioresistant carcinomas. In this study, astaxanthin treatment inhibited cell viability and proliferation of two NSCLC cells, A549 and H1703. Treatment with astaxanthin decreased Rad51 expression and phospho-AKT protein level in a time and dose-dependent manner. Furthermore, expression of constitutively active AKT (AKT-CA) vector significantly rescued the decreased Rad51 protein and mRNA levels in astaxanthin-treated NSCLC cells. Combined treatment with PI3K inhibitors (LY294002 or wortmannin) and astaxanthin further decreased the Rad51 expression in NSCLC cells. Knockdown of Rad51 enhanced astaxanthin-induced cytotoxicity and growth inhibition in NSCLC cells. These findings may have implications for the rational design of future drug regimens incorporating astaxanthin for the treatment of NSCLC.

Keywords: astaxanthin, cytotoxicity, AKT, non-small cell lung cancer, PI3K

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Authors: Chris George, Adam Hamlin, Lily Pereg, Richard Charlesworth, Gal Winter

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Background: According to the ‘hygiene hypothesis’ the current rise in allergies and autoimmune diseases stems mainly from reduced microbial exposure due, amongst other factors, to urbanisation and distance from soil. However, this hypothesis is based on epidemiological and not biological data. Useful insights into the underlying mechanisms of this hypothesis can be gained by studying our interaction with soil. Soil microbiota may be directly ingested or inhaled by humans, enter the body through skin-soil contact or using plants as vectors. This study aims to examine the ability of soil microbiota to colonise the gut, study the interaction of soil microbes with the immune system and their potential protective activity. Method: The nutrition of the rats was supplemented daily with fresh or autoclaved soil for 21 days followed by 14 days of no supplementations. Faecal samples were collected throughout and analysed using 16S sequencing. At the end of the experiment rats were sacrificed and tissues and digesta were collected. Results/Conclusion: Results showed significantly higher richness and diversity following soil supplementation even after recovery. Specific soil microbial groups identified as able to colonise the gut. Of particular interest was the mucosal layer which emerged as a receptive host for soil microorganisms. Histological examination revealed innate and adaptive immune activation. Findings of this study reinforce the ‘hygiene hypothesis’ by demonstrating the ability of soil microbes to colonise the gut and activate the immune system. This paves the way for further studies aimed to examine the interaction of soil microorganisms with the immune system.

Keywords: gut microbiota, hygiene hypothesis, microbiome, soil

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Authors: Fuad Ameen, Ali A. Al-Homaidan

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Biochar can be used in numerous biotechnological applications due to its properties to adsorb beneficial nutrients and harmful pollutants. Objectives: We aimed to treat heavy metal polluted organic wastes using vermicomposting process and produce a fertilizer that can be used in agriculture. We improved the process by adding biochar as well as microbial inoculum and biomass into household waste or sewage sludge before vermicomposting. The earthworm Eisenia fetida used in vermicomposting was included to accumulate heavy metals, biochar to adsorb heavy metals, and the microalga Navicula sp. or the mangrove fungus Acrophialophora sp. to promote plant growth in the final product used as a seedbed for Solanaceae vegetables. We carried out vermicomposting treatments to see the effect of different amendments. Final compost quality was analyzed for maturity. The earthworms were studied for their vitality, heavy metal accumulation, and metallothionein protein content to verify their role in the process. The compost was used as a seedbed for vegetables that were inoculated with a phytopathogen Pythium sp. known to cause root rot and destroy seeds. Compost as seedbed promoted plant growth and reduced disease symptoms in leaves. In the treatment where E. fetida, 6% biochar, and Navicula sp. had been added, 90% of the seeds germinated, while less than 20% germinated in the control treatment. The experimental plants had acquired resistance against Pythium sp. The metagenomic profile of microbial communities will be reported.

Keywords: organic wastes, vermicomposting process, biochar, mangrove fungus

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Authors: Mahadehe Hassan

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Transport system and industries which are the main basis of industrial and socio-economic development of any country. It is mainly dependent on fossil fuels. Bangladesh has fossil fuel reserves of 9.51 TCF as of July 2023, and if no new gas fields are discovered in the next 7-9 years and if the existing gas consumption rate continues, the fossil fuel reserves will be exhausted. The demand for petroleum products in Bangladesh is increasing steadily, with 63% imported by BPC and 37% imported by private companies. 61.61% of BPC imported products are used in the transport sector and 5.49% in the industrial sector, which is expensive and harmful to the environment. Liquefied Petroleum Gas (LPG) should be considered as an alternative energy for Bangladesh based on Sustainable Development Goals (SDGs) criteria for sustainable, clean and affordable energy. This will not only lead to the much desired mitigation of energy famine in the country but also contribute favorably to the macroeconomic indicators. Considering the environmental and economic issues, the government has referred to CNG (compressed natural gas) as the fuel carrier since 2000, but currently due to the decline mode of gas reserves, the government of Bangladesh is thinking of new energy sources for transport and industrial sectors which will be sustainable, environmentally friendly and economically viable. Liquefied Petroleum Gas (LPG) is the best choice for fueling transport and industrial sectors in Bangladesh. At present, a total of 1.54 million metric tons of liquefied petroleum gas (LPG) is marketed in Bangladesh by the public and private sectors. 83% of it is used by households, 12% by industry and commerce and 5% by transportation. Industrial and transport sector consumption is negligible compared to household consumption. So the purpose of the research is to find out the challenges of LPG market development in transport and industrial sectors in Bangladesh and make recommendations to reduce the challenges. Secure supply chain, inadequate infrastructure, insufficient investment, lack of government monitoring and consumer awareness in the transport sector and industrial sector are major challenges for LPG market development in Bangladesh. Bangladesh government as well as private owners should come forward in the development of liquefied petroleum gas (LPG) industry to reduce the challenges of secure energy sector for sustainable development. Furthermore, ensuring adequate Liquefied Petroleum Gas (LPG) supply in Bangladesh requires government regulations, infrastructure improvements in port areas, awareness raising and most importantly proper pricing of Liquefied Petroleum Gas (LPG) to address the energy crisis in Bangladesh.

Keywords: transportand industries fuel, LPG consumption, challenges, economical sustainability

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Authors: Eun-Soo Lim, Young-Min Kang

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M-type Sr-hexaferrite (SrFe₁₂O₁₉) have been studied during the past decades because it is the most utilized materials in permanent magnets due to their low price, outstanding chemical stability, and appropriate hard magnetic properties. Many attempts have been made to improve the intrinsic magnetic properties of M-type Sr-hexaferrites (SrM), such as by improving the saturation magnetization (MS) and crystalline anisotropy by cation substitution. It is well proved that the Ca-La-Co substitutions are one of the most successful approaches, which lead to a significant enhancement in the crystalline anisotropy without reducing MS, and thus the Ca-La-Co-doped SrM have been commercialized in high-grade magnet products. In this research, the effect of respective doping of Ca and La into the SrM lattices were studied with assumptions that these elements could substitute both of Fe and Sr sites. The hexaferrite samples of stoichiometric SrFe₁₂O₁₉ (SrM) and the Ca substituted SrM with formulae of Sr₁₋ₓCaₓFe₁₂Oₐ (x = 0.1, 0.2, 0.3, 0.4) and SrFe₁₂₋ₓCaₓOₐ (x = 0.1, 0.2, 0.3, 0.4), and also La substituted SrM of Sr₁₋ₓLaₓFe₁₂Oₐ (x = 0.1, 0.2, 0.3, 0.4) and SrFe₁₂₋ₓLaₓOₐ (x = 0.1, 0.2, 0.3, 0.4) were prepared by conventional solid state reaction processes. X-ray diffraction (XRD) with a Cu Kα radiation source (λ=0.154056 nm) was used for phase analysis. Microstructural observation was conducted with a field emission scanning electron microscopy (FE-SEM). M-H measurements were performed using a vibrating sample magnetometer (VSM) at 300 K. Almost pure M-type phase could be obtained in the all series of hexaferrites calcined at > 1250 ºC. Small amount of Fe₂O₃ phases were detected in the XRD patterns of Sr₁₋ₓCaₓFe₁₂Oₐ (x = 0.2, 0.3, 0.4) and Sr₁₋ₓLaₓFe₁₂Oₐ (x = 0.1, 0.2, 0.3, 0.4) samples. Also, small amount of unidentified secondary phases without the Fe₂O₃ phase were found in the samples of SrFe₁₂₋ₓCaₓOₐ (x = 0.4) and SrFe₁₂₋ₓLaₓOₐ (x = 0.3, 0.4). Although the Ca substitution (x) into SrM structure did not exhibit a clear tendency in the cell parameter change in both series of samples, Sr₁₋ₓCaₓFe₁₂Oₐ and SrFe₁₂₋ₓCaₓOₐ , the cell volume slightly decreased with doping of Ca in the Sr₁₋ₓCaₓFe₁₂Oₐ samples and increased in the SrFe₁₂₋ₓCaₓOₐ samples. Considering relative ion sizes between Sr²⁺ (0.113 nm), Ca²⁺ (0.099 nm), Fe³⁺ (0.064 nm), these results imply that the Ca substitutes both of Sr and Fe in the SrM. A clear tendency of cell parameter change was observed in case of La substitution into Sr site of SrM ( Sr₁₋ₓLaₓFe₁₂Oₐ); the cell volume decreased with increase of x. It is owing to the similar but smaller ion size of La³⁺ (0.106 nm) than that of Sr²⁺. In case of SrFe₁₂₋ₓLaₓOₐ, the cell volume first decreased at x = 0.1 and then remained almost constant with increase of x from 0.2 to 0.4. These results mean that La only substitutes Sr site in the SrM structure. Besides, the microstructure and magnetic properties of these samples, and correlation between them will be revealed.

Keywords: M-type hexaferrite, substitution, cell parameter, magnetic properties

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Authors: Gerhardt Boukes, Maryna Van De Venter, Sharlene Govender

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Macrofungi have been used for the past two thousand years in Asian countries, and more recently in Western countries, for their medicinal properties. Biological activities include antimicrobial, antioxidant, anti-inflammatory, antidiabetic, anticancer and immunomodulatory to name a few. Several biologically active compounds have been identified and isolated. Macrofungal research in Africa is poorly documented and to the best of our knowledge non-existent. South Africa has a rich macrofungal biodiversity, which includes endemic and exotic macrofungal species. Ethanolic extracts of 13 macrofungal species, including mushrooms, bracket fungi and puffballs, were prepared and screened for cytotoxicity against a panel of seven cell lines, including A549 (human lung adenocarcinoma), HeLa (human cervical adenocarcinoma), HT-29 (human colorectal adenocarcinoma), MCF7 (human breast adenocarcinoma), MIA PaCa-2 (human pancreatic ductal adenocarcinoma), PC-3 (human prostate adenocarcinoma) and Vero (African green monkey kidney epithelial) cells using MTT. Cell lines were chosen according to the most prevalent cancer types affecting males and females in South Africa and globally, and the mutations they contain. Preliminary results have shown that three of the macrofungal genera, i.e. Fomitopsis, Gymnopilus and Pycnoporus, have shown cytotoxic activity, ranging between IC50 ~20 and 200 µg/mL. The molecular mechanism of action contributing to cell death investigated and being investigated include apoptosis (i.e. DNA cell cycle arrest, caspase-3 activation and mitochondrial membrane potential), autophagy (i.e. acridine orange and LC3B staining) and ER stress (i.e. thioflavin T staining and caspase-12) in the presence of melphalan, chloroquine and thapsigargin/tuncamycin as positive controls, respectively. The genus, Pycnoporus, has shown the best cytotoxicity of the three macrofungal genera. Future work will focus on the identification and isolation of novel active compounds and elucidating the mechanism/s of action.

Keywords: cancer, cytotoxicity, macrofungi, mechanism/s of action

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Authors: Ketsada Sutthiumporn, Sittichot Thongkaw, Malee Santikunaporn

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In Thailand, biodiesel has been utilized as an attractive substitute of petroleum diesel and the government imposes a mandatory biodiesel blending requirement in transport sector to improve energy security, support agricultural sector and reduce emissions. Though biodiesel blend has many advantages over diesel fuel such as improved lubricity, low sulfur content and higher flash point, there are still some technical problems such as oxidative stability, poor cold- flow properties and impurity. Such problems were related to the fatty acid composition in feedstock. Moreover, Thailand has announced the use of low sulfur diesel as a base diesel and will be continually upgrading to EURO 5 in 2023. With ultra low sulfur content, it may affect the diesel fuel properties especially lubricity as well. Therefore, in this study, the physical and chemical properties of palm oil-based biodiesel in low sulfur diesel blends from different producers will be investigated by standard methods per ASTM and EN. Also, its economic benefits based on diesel price structure in Thailand will be highlighted. The appropriate biodiesel blend ratio can affect the physico-chemical properties and reasonable price in the country. Properties of biodiesel, including specific gravity, kinematic viscosity, FAME composition, flash point, sulfur, water, oxidation stability and lubricity were measured by standard methods of ASTM and EN. The results show that the FAME composition of biodiesel has the fatty acid of C12:0 to C20:1, mostly in C16:0, C18:0, C18:1, and C18:2, which were main characteristic compositions of palm biodiesel. The physical and chemical properties of biodiesel blended diesel was found to be increases with an increasing amount of biodiesel such as specific gravity, flash point and kinematic viscosity while sulfur value was decreased. Moreover, in this study, the various properties of each biodiesel blends were plotted to determine the appropriate proportional range of biodiesel-blended diesel with an optimum fuel price.It can be seen that the amount of B100 can be filled from 1% up to 7% in which the quality was in accordance with Notification of the department of Energy business.The understanding of relation between physico-chemical properties of palm oil-based biodiesel and pricing is beneficial to guide the better development of desired feedstock in Thailand and to implement biodiesel blends with comparative price and diesel engine performance.

Keywords: fatty acid methyl ester, biodiesel, fuel price structure, palm oil in Thailand

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Authors: V. Yurina, H. Sujuti, E. Rahmani, A. R. Nopitasari

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Breast cancer has the highest prevalence cancer in women. Moringa leaves (M. oleifera) contain quercetin, kaempferol, and benzyl isothiocyanate which can enhance induction of apoptosis. This research aimed to study the role of the leaf extract of Moringa to increase apoptosis in breast cancer cell line, MCF-7 cells. This research used in vitro experimental, post-test only, control group design on breast cancer cells MCF-7 in vitro. Moringa leaves were extracted by maceration method with ethanol 70%. Cells were treated with drumstick leaves extract on 1100, 2200, and 4400 μg/ml for Hsp27 and caspase-9 expression (immunocytochemistry) and apoptosis (TUNEL assay) test. The results of this study found that the IC50 2200 µg/ml. Moringa leaves extract can significantly increase the expression of caspase-9 (p<0.05) and decreased Hsp 27 expression (p<0.05). Moreover it can increase apoptosis (p<0.05) significantly in MCF-7 cells. The conclusion of this study is Moringa leaves extract is able to increase the expression of caspase-9, decrease Hsp27 expression and increase apoptosis in breast cancer cell-line MCF-7.

Keywords: apoptosis, breast cancer, caspase-9, Hsp27, Moringa oleifera

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Authors: Amir Raeisdana, Davood Sohrabi, Mojtaba Nohekhan, Ameneh Kargarian, Maryam Ghapanvari, Alireza Aslezaeem

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Improvement of zirconium properties by chromium coating and nitrogen implantation is ideal to protect the nuclear fuel rods against corrosion and secondary hydrogenation. Metallic chromium (Cr) has attracted attention as a potential coating material on zirconium alloys, to limit external cladding corrosion. In this research, high energy plasma focus device was used to coat the chromium and implant the nitrogen ions in the zirconium substrate. This device emits high-energy nitrogen ions of 10 keV-1 MeV and with a flux of 10^16 ions/cm^2 in each shot toward the target so it is attractive for implantation on the substrate materials at the room temperature. Six zirconium samples in 2cm×2cm dimensions with 1mm thickness were located at a distance of 20cm from the place where the pinch is formed. The experiments are carried out in 0.5 mbar of the nitrogen gas pressure and 15 kV of the charging voltage. Pure Cr disc was installed on the anode head for sputtering of the chromium and deposition on zirconium substrate. When the pinch plasma column decays due to various instabilities, intense and high-energy N2 ions are accelerated towards the zirconium substrate also sputtered Cr is deposited on the zirconium substrate. XRD and XRF analysis were used to study the structural properties of the samples. XRF analysis indicates 77.1% of Zr and 11.1% of Cr in the surface of the sample. XRD spectra shows the formation of ZrN, CrN and CrZr composites after nitrogen implantation and chromium coating. XRD spectra shows the chromium peak height equal to 152.80 a.u. for the major sample (θ=0֯) and 92.99 a.u. for the minor sample (θ=6֯), so implantation and coating along the main axis of the device is significantly more than other directions.

Keywords: ZrN and CrN and CrZr composites, angular distribution for Cr deposition rate, zirconium corrosion resistance, nuclear fuel rods, plasma focus device

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Authors: Isil Gazioglu, Abdulselam Ertas

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Aflatoxins (AFs) are secondary toxic metabolites of Aspergillus flavus and A. parasiticus. AFs can be absorbed through the skin. Potent carcinogens like AFs should be completely absent from cosmetics, this can be achieved by careful quality control of the raw plant materials. Regulatory limits for aflatoxins have been established in many countries, and reliable testing methodology is needed to implement and enforce the regulatory limits. In this study, ten medicinal plant samples (Bundelia tournefortti, Capsella bursa-pastoris, Carduus tenuiflorus, Cardaria draba, Malva neglecta, Malvella sharardiana, Melissa officinalis, Sideritis libanotica, Stakys thirkei, Thymus nummularius) were investigated for aflatoxin (AF) contaminations by employing an HPLC assay for the determination of AFB1, B2, G1 and G2. The samples were extracted with 70% (v/v) methanol in water before further cleaned up with an immunoaffinity column and followed by the detection of AFs by using an electrochemically post-column derivatization with Kobra-Cell and fluorescence detector. The extraction procedure was optimized in order to obtain the best recovery. The method was successfully carried out with all medicinal plant samples. The results revealed that five (50%) of samples were contaminated with AFs. The association between particular samples and the AF contaminated could not be determined due to the low frequency of positive samples.

Keywords: aflatoxin B1, HPLC-FLD, KOBRA-Cell, mycotoxin

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Authors: P. Akbartehrani, M. Khaledi Pour, M. Amini, M. Khani, M. Mohajeri Tehrani, E. Ghasemi, P. Charipoor, B. Shokri

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A common problem between diabetic patients is foot ulcers which are chronic and require specialized treatment. Previous studies illustrate that Cold atmospheric plasma (CAP) has beneficial effects on wound healing and infection. Nevertheless, the comparison of different cap exposure protocols in diabetic ulcer wound healing remained to be studied. This study aims to determine the effect of two different exposure protocols on wound healing in diabetic ulcers. A prospective, randomized clinical trial was conducted at two clinics. Diabetic patients with G1 and G2 wanger classification diabetic foot ulcers were divided into two groups of study. One group was treated by the first protocol, which was treating wounds by argon-generated cold atmospheric plasma jet once a week for five weeks in a row. The other group was treated by the second protocol, which was treating wounds every three days for five weeks in a row. The wounds were treated for 40 seconds/cubic centimeter, while the nozzle tip was moved nonlocalized 1 cm above the wounds. A patient with one or more wounds could participate in different groups as wounds were separately randomized, which allow a participant to be treated several times during the study. The study's significant findings were two different reductions rate in wound size, microbial load, and two different healing speeds. This study concludes that CAP therapy by the second protocol yields more effective healing speeds, reduction in wound sizes, and microbial loads of foot ulcers in diabetic patients.

Keywords: wound healing, diabetic ulcers, cold atmospheric plasma, cold argon jet

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Authors: Colin S. Chen, Chien-Jung Tien, Hsin-Jan Huang

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For weathering studies, the change of chemical constituents by biodegradation effect in diesel-contaminated soils are important factors to be considered, especially when there is a prolonged period of weathering processes. The objective was to evaluate biodegradation effects onto hydrocarbon fingerprinting and distribution patterns of diesel fuels, fuel source screening and differentiation, source-specific marker compounds, and diagnostic ratios of diesel fuel constituents by laboratory and field studies. Biodegradation processes of diesel contaminated soils were evaluated by experiments lasting for 15 and 12 months, respectively. The degradation of diesel fuel in top soils was affected by organic carbon content and biomass of microorganisms in soils. Higher depletion of total petroleum hydrocarbon (TPH), n-alkanes, and polynuclear aromatic hydrocarbons (PAHs) and their alkyl homologues was observed in soils containing higher organic carbon content and biomass. Decreased ratio of selected isoprenoids (i.e., pristane (Pr) and phytane (Ph)) including n-C17/pristane and n-C18/phytane was observed. The ratio of pristane/phytane was remained consistent for a longer period of time. At the end of the experimental period, a decrease of pristane/phytane was observed. Biomarker compounds of bicyclic sesquiterpanes (BS) were less susceptible to the effects of biodegradation. The ratios of characteristic factors such as C15 sesquiterpane/ 8β(H)-drimane (BS3/BS5), C15 sesquiterpane/ 8β(H)-drimane (BS4/BS5), 8β(H)-drimane/8β(H)-homodrimane (BS5/BS10), and C15 sesquiterpane/8β(H)-homodrimane (BS3/BS10) could be adopted for source identification of diesel fuels in top soil. However, for biodegradation processes lasted for six months but shorter than nine months, only BS3/BS5 and BS3/BS10 could be distinguished in two diesel fuels. In subsoil experiments (contaminated soil located 50 cm below), the ratios of characteristic factors including BS3/BS5, BS4/BS5, and BS5/BS10 were valid for source identification of two diesel fuels for nine month biodegradation. At the early stage of contamination, biomass of soil decreased significantly. However, 6 and 7 dominant species were found in soils in top soil experiments, respectively. With less oxygen and nutrients in subsoil, less biomass of microorganisms was observed in subsoils. Only 2 and 4 diesel-degrading species of microorganisms were identified in two soils, respectively. Parameters of double ratio such as fluorene/C1-fluorene: C2-phenanthrene/C3-phenanthrene (C0F/C1F:C2P/C3P) in both top and subsoil, C2-naphthalene/C2-phenanthrene: C1-phenanthrene/C3-phenanthrene (C2N/C2P:C1P/C3P), and C1-phenanthrene/C1-fluorene: C3-naphthalene/C3-phenanthrene (C1P/C1F:C3N/C3P) in subsoil could serve as forensic indicators in diesel contaminated sites. BS3/BS10:BS4/BS5 could be used in 6 to 9 months of biodegradation processes. Results of principal component analysis (PCA) indicated that source identification of diesel fuels in top soil could only be perofrmed for weathering process less than 6 months. For subsoil, identification can be conducted for weathering process less than 9 months. Ratio of isoprenoids (pristane and phytane) and PAHs might be affected by biodegradation in spilled sites. The ratios of bicyclic sesquiterpanes could serve as forensic indicators in diesel-contaminated soils. Finally, source identification was attemped for samples collected from different fuel contaminated sites by using the unique pattern of sesquiterpanes. It was anticipated that the information generated from this study would be adopted by decision makers to evaluate the liability of cleanup in diesel contaminated sites.

Keywords: biodegradation, diagnostic ratio, diesel fuel, environmental forensics

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Authors: D. C. Asebiah, D. Saranin, S. Karazhanov, A. R. Tameev, M. Kah

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In this paper, the mesoscopic NiO was used as a hole transport layer in the inverted planar organometallic hybrid perovskite solar cell to study the effect of hysteresis. The devices we fabricated have the structures Fluorine Tin Oxide (FTO)/mesoscopic NiO/perovskite/[6,6]-phenyl C₆₁-butyric acid methyl ester (PC₆₁BM) photovoltaic device. The perovskite solar cell was done by toluene air (TLA) method and horn sonication for the dispersion of the NiO nanoparticles in deionized water. The power conversion efficiency was 12.07% under 1.5 AM illumination. We report hysteresis in the in current-voltage dependence of the solar cells with mesoscopic NiO as a hole transport layer.

Keywords: perovskite, mesoscopic, hysteresis, toluene air

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Authors: Nishanthi N. S., Srikanth Vedantam

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Intracellular delivery of materials has always proved to be a challenge in research and therapeutic applications. Usually, vector-based methods, such as liposomes and polymeric materials, and physical methods, such as electroporation and sonoporation have been used for introducing nucleic acids or proteins. Reliance on exogenous materials, toxicity, off-target effects was the short-comings of these methods. Microinjection was an alternative process which addressed the above drawbacks. However, its low throughput had hindered its adoption widely. Mechanical deformation of cells by squeezing them through constriction channel can cause the temporary development of pores that would facilitate non-targeted diffusion of materials. Advantages of this method include high efficiency in intracellular delivery, a wide choice of materials, improved viability and high throughput. This cell squeezing process can be studied deeper by employing simple models and efficient computational procedures. In our current work, we present a finite sized dissipative particle dynamics (FDPD) model to simulate the dynamics of the cell flowing through a constricted channel. The cell is modeled as a capsule with FDPD particles connected through a spring network to represent the membrane. The total energy of the capsule is associated with linear and radial springs in addition to constraint of the fixed area. By performing detailed simulations, we studied the strain on the membrane of the capsule for channels with varying constriction heights. The strain on the capsule membrane was found to be similar though the constriction heights vary. When strain on the membrane was correlated to the development of pores, we found higher porosity in capsule flowing in wider channel. This is due to localization of strain to a smaller region in the narrow constriction channel. But the residence time of the capsule increased as the channel constriction narrowed indicating that strain for an increased time will cause less cell viability.

Keywords: capsule, cell squeezing, dissipative particle dynamics, intracellular delivery, microfluidics, numerical simulations

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Authors: Elena Petersen, Inna Kornienko, Svetlana Guryeva, Sergey Dobdin, Anatoly Skripal, Andrey Usanov, Dmitry Usanov

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One of the most important unsolved problems in modern medicine is the increase of chronic diseases that lead to organ dysfunction or even complete loss of function. Current methods of treatment do not result in decreased mortality and disability statistics. Currently, the best treatment for many patients is still transplantation of organs and/or tissues. Therefore, finding a way of correct artificial matrix biofabrication in case of limited number of natural organs for transplantation is a critical task. One important problem that needs to be solved is development of a nondestructive and noninvasive method to analyze dynamic changes of mechanical characteristics of a matrix with minimal side effects on the growing cells. This research was focused on investigating the properties of matrix as a marker of graft condition. In this study, the collagen gel with human primary dermal fibroblasts in suspension (60, 120, 240*103 cells/mL) and collagen gel with cell spheroids were used as model objects. The stiffness and elasticity characteristics were evaluated by a semiconductor laser autodyne. The time and cell concentration dependency of the stiffness and elasticity were investigated. It was shown that these properties changed in a non-linear manner with respect to cell concentration. The maximum matrix stiffness was observed in the collagen gel with the cell concentration of 120*103 cells/mL. This study proved the opportunity to use the mechanical properties of matrix as a marker of graft condition, which can be measured by noninvasive semiconductor laser autodyne technique.

Keywords: graft, matrix, noninvasive method, regenerative medicine, semiconductor laser autodyne

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Authors: E. T. El Shenawy, O. N. A. Esmail, Adel A. Elbaset, Hesham F. A. Hamed

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Generation of the electrical energy based on photovoltaic (PV) technology has been increased over the world due to either the continuous reduction in the traditional energy sources in addition to the pollution problems related to their usage, or the clean nature and safe usage of the PV technology. Also, PV systems can generate clean electricity in the site of use without any transmission, which can be considered cost effective than other generation systems. The performance of the PV system is highly affected by the amount of solar radiation incident on it. Completely or partially shaded PV systems can affect its output. The PV system can be shaded by trees, buildings, dust, incorrect system configuration, or other obstacles. The present paper studies the effect of the partial shading on the performance of a thin film PV module under climatic conditions of Cairo, Egypt. This effect was measured and evaluated according to practical measurement of the characteristic curves such as current-voltage and power-voltage for two identical PV modules (with and without shading) placed at the same time on one mechanical structure for comparison. The measurements have been carried out for the following shading patterns; half cell (bottom, middle, and top of the PV module); complete cell; and two adjacent cells. The results showed that partially shading the PV module changes the shapes of the I-V and P-V curves and produces more than one maximum power point, that can disturb the traditional maximum power point trackers. Also, the output power from the module decreased according to the incomplete solar radiation reaching the PV module due to shadow patterns. The power loss due shading was 7%, 22%, and 41% for shading of half-cell, one cell, and two adjacent cells of the PV module, respectively.

Keywords: I-V measurements, PV module characteristics, PV module power loss, PV module shading

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Authors: Samia Dardouri, Ridha Bouallegue

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Scheduling is the process of dynamically allocating physical resources to User Equipment (UE) based on scheduling algorithms implemented at the LTE base station. Various algorithms have been proposed by network researchers as the implementation of scheduling algorithm which represents an open issue in Long Term Evolution (LTE) standard. This paper makes an attempt to study and compare the performance of PF, MLWDF and EXP/PF scheduling algorithms. The evaluation is considered for a single cell with interference scenario for different flows such as Best effort, Video and VoIP in a pedestrian and vehicular environment using the LTE-Sim network simulator. The comparative study is conducted in terms of system throughput, fairness index, delay, packet loss ratio (PLR) and total cell spectral efficiency.

Keywords: LTE, multimedia flows, scheduling algorithms, mobile computing

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Authors: Majid Vafaeipour, Mohamed El Baghdadi, Florian Verbelen, Peter Sergeant, Joeri Van Mierlo, Kurt Stockman, Omar Hegazy

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The Electrical Variable Transmission (EVT), an electromechanical device, can be considered as an alternative solution to the conventional transmission system utilized in Hybrid Electric Vehicles (HEVs). This study present comparisons in terms of fuel consumption, power split, and state of charge (SoC) of an HEV containing an EVT to a conventional parallel topology and a series topology. To this end, corresponding simulations of these topologies are all performed in presence of control strategies enabling battery charge-sustaining and efficient power split. The power flow through the components of the vehicle are attained, and fuel consumption results of the considered cases are compared. The investigation of the results indicates utilizing EVT can provide significant added values in HEV configurations. The outcome of the current research paves its path for implementation of design optimization approaches on such systems in further research directions.

Keywords: Electrical Variable Transmission (EVT), Hybrid Electric Vehicle (HEV), parallel, series, modeling

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Authors: Oshma Chakoory, Sophie Comtet-Marre, Pierre Peyret

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Metagenomic classifiers are widely used for the taxonomic profiling of metagenomic data and estimation of taxa relative abundance. Small subunit rRNA genes are nowadays a gold standard for the phylogenetic resolution of complex microbial communities, although the power of this marker comes down to its use as full-length. We benchmarked the performance and accuracy of rRNA-specialized versus general-purpose read mappers, reference-targeted assemblers and taxonomic classifiers. We then built a pipeline called RiboTaxa to generate a highly sensitive and specific metataxonomic approach. Using metagenomics data, RiboTaxa gave the best results compared to other tools (Kraken2, Centrifuge (1), METAXA2 (2), PhyloFlash (3)) with precise taxonomic identification and relative abundance description, giving no false positive detection. Using real datasets from various environments (ocean, soil, human gut) and from different approaches (metagenomics and gene capture by hybridization), RiboTaxa revealed microbial novelties not seen by current bioinformatics analysis opening new biological perspectives in human and environmental health. In a study focused on corals’ health involving 20 metagenomic samples (4), an affiliation of prokaryotes was limited to the family level with Endozoicomonadaceae characterising healthy octocoral tissue. RiboTaxa highlighted 2 species of uncultured Endozoicomonas which were dominant in the healthy tissue. Both species belonged to a genus not yet described, opening new research perspectives on corals’ health. Applied to metagenomics data from a study on human gut and extreme longevity (5), RiboTaxa detected the presence of an uncultured archaeon in semi-supercentenarians (aged 105 to 109 years) highlighting an archaeal genus, not yet described, and 3 uncultured species belonging to the Enorma genus that could be species of interest participating in the longevity process. RiboTaxa is user-friendly, rapid, allowing microbiota structure description from any environment and the results can be easily interpreted. This software is freely available at https://github.com/oschakoory/RiboTaxa under the GNU Affero General Public License 3.0.

Keywords: metagenomics profiling, microbial diversity, SSU rRNA genes, full-length phylogenetic marker

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Authors: Thomas Louis, Irina Primac, Florent Morfoisse, Tania Durre, Silvia Blacher, Agnes Noel

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In vitro and in vivo lymphangiogenesis assays are essential for the identification of potential lymphangiogenic agents and the screening of pharmacological inhibitors. In the present study, we analyse three biological models: in vitro lymphatic endothelial cell spheroids, in vivo ear sponge assay, and in vivo lymph node colonisation by tumour cells. These assays provide suitable 3D models to test pro- and anti-lymphangiogenic factors or drugs. 3D images were acquired by confocal laser scanning and light sheet fluorescence microscopy. Virtual scan microscopy followed by 3D reconstruction by image aligning methods was also used to obtain 3D images of whole large sponge and ganglion samples. 3D reconstruction, image segmentation, skeletonisation, and other image processing algorithms are described. Fixed and time-lapse imaging techniques are used to analyse lymphatic endothelial cell spheroids behaviour. The study of cell spatial distribution in spheroid models enables to detect interactions between cells and to identify invasion hierarchy and guidance patterns. Global measurements such as volume, length, and density of lymphatic vessels are measured in both in vivo models. Branching density and tortuosity evaluation are also proposed to determine structure complexity. Those properties combined with vessel spatial distribution are evaluated in order to determine lymphangiogenesis extent. Lymphatic endothelial cell invasion and lymphangiogenesis were evaluated under various experimental conditions. The comparison of these conditions enables to identify lymphangiogenic agents and to better comprehend their roles in the lymphangiogenesis process. The proposed methodology is validated by its application on the three presented models.

Keywords: 3D image segmentation, 3D image skeletonisation, cell invasion, confocal microscopy, ear sponges, light sheet microscopy, lymph nodes, lymphangiogenesis, spheroids

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Authors: Jianli Dong, Fangling Xu, Gengming Huang

Abstract:

Human endogenous retroviral elements (HERVs) comprise approximately 8% of the human genome. They are thought to be germline-integrated genetic remnants of retroviral infections. Although HERV sequences are highly defective, some, especially the K type (HERV-K), have been shown to be expressed and may have biological activities in the pathogenesis of cancer, chronic inflammation and autoimmune diseases. We found that HERV-K GAG and ENV proteins were strongly expressed in pleomorphic melanoma cells. We also detected a critical role of HERV-K ENV in mediating intercellular fusion and colony formation of melanoma cells. Interestingly, we found that levels of HERV-K GAG and ENV expression correlated with the activation of ERK and loss of p16INK4A in melanoma cells, and inhibition of MEK or CDK4, especially in combination, reduced HERV-K expression in melanoma cells. We also performed a reverse transcription-polymerase chain reaction (RT-PCR) assay using DNase I digestion to remove “contaminating” HERV-K genomic DNA and examined HERV-K RNA expression in plasma samples from HIV-1 infected individuals. We found a covariation between HERV-K RNA expression and CD4 cell counts in HIV-1 positive samples. Although a causal link between HERV-K activation and melanoma development, and between HERV-K activation, HIV-1 infection and CD4 cell count have yet to be determined, existing data support the further research efforts in HERV-K.

Keywords: CD4 cell, HERV-K, HIV-1, melanoma

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Authors: Mona Hamdy, Iman Shaheen, Hadeel Seif Eldin, Basma Ali, Omnia Abdeldayem

Abstract:

Summary: Bone involvement of sickle cell disease (SCD) patients varies from acute clinical manifestations of painful vaso-occlusive crises or osteomyelitis to more chronic affection of bone mineral density (BMD) and debilitating osteonecrosis and osteoporosis. Secreted klotho protein is involved in calcium (Ca) reabsorption in the kidney. This study aimed to measure serum klotho levels in children with SCD to determine the possibility of using it as a marker of low BMD in children with SCD in correlation with a dual-energy radiograph absorptiometry scan. This study included 60 sickle disease patients and 30 age-matched and sex-matched control participants without SCD. A highly statistically significant difference was found between patients with normal BMD and those with low BMD, with serum Ca and klotho levels being lower in the latter group. Klotho serum level correlated positively with both serum Ca and BMD. Serum klotho level showed 94.9% sensitivity and 95.2% specificity in the detection of low BMD. Both serum Ca and klotho serum levels may be useful markers for detection of low BMD related to SCD with high sensitivity and specificity; however, klotho may be a better indicator as it is less affected by the nutritional and endocrinal status of patients or by intake of Ca supplements.

Keywords: sickle cell disease, BMD, osteoporosis, DEXA, klotho

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Authors: Mubashir Hussain, Mu Lv, Xiaohan Dong, Zhiyang Li, Bin Liu, Nongyue He

Abstract:

Detection and classification of microbes have a vast range of applications in biomedical engineering especially in detection, characterization, and quantification of bacterial contaminants. For identification of pathogens, different techniques are emerging in the field of biomedical engineering. Latest technology uses light scattering, capable of identifying different pathogens without any need for biochemical processing. Bacterial Pathogens Identification System (BPIS) which uses a laser beam, passes through the sample and light scatters off. An assembly of photodetectors surrounded by the sample at different angles to detect the scattering of light. The algorithm of the system consists of two parts: (a) Library files, and (b) Comparator. Library files contain data of known species of bacterial microbes in the form of binned plots, while comparator compares data of unknown sample with library files. Using collected data of unknown bacterial species, highest voltage values stored in the form of peaks and arranged in 3D histograms to find the frequency of occurrence. Resulting data compared with library files of known bacterial species. If sample data matching with any library file of known bacterial species, sample identified as a matched microbe. An experiment performed to identify three different bacteria particles: Enterococcus faecalis, Pseudomonas aeruginosa, and Escherichia coli. By applying algorithm using library files of given samples, results were compromising. This system is potentially applicable to several biomedical areas, especially those related to cell morphology.

Keywords: microbial identification, laser scattering, peak identification, binned plots classification

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Authors: Snehal D. Ganjave, Hardik Dodia, Avinash V. Sunder, Swati Madhu, Pramod P. Wangikar

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Batch cultivation of recombinant bacteria in shake flasks results in low cell density due to nutrient depletion. Previous protocols on high cell density cultivation in shake flasks have relied mainly on controlled release mechanisms and extended cultivation protocols. In the present work, we report an optimized fed-batch process for high cell density cultivation of recombinant E. coli BL21(DE3) for protein production. A cybernetic model-based, multi-objective optimization strategy was implemented to obtain the optimum operating variables to achieve maximum biomass and minimized substrate feed rate. A syringe pump was used to feed a mixture of glycerol and yeast extract into the shake flask. Preliminary experiments were conducted with online monitoring of dissolved oxygen (DO) and offline measurements of biomass and glycerol to estimate the model parameters. Multi-objective optimization was performed to obtain the pareto front surface. The selected optimized recipe was tested for a range of proteins that show different extent soluble expression in E. coli. These included eYFP and LkADH, which are largely expressed in soluble fractions, CbFDH and GcanADH , which are partially soluble, and human PDGF, which forms inclusion bodies. The biomass concentrations achieved in 24 h were in the range 19.9-21.5 g/L, while the model predicted value was 19.44 g/L. The process was successfully reproduced in a standard laboratory shake flask without online monitoring of DO and pH. The optimized fed-batch process showed significant improvement in both the biomass and protein production of the tested recombinant proteins compared to batch cultivation. The proposed process will have significant implications in the routine cultivation of E. coli for various applications.

Keywords: cybernetic model, E. coli, high cell density cultivation, multi-objective optimization

Procedia PDF Downloads 258

Authors: Ipsita Biswas, Arnab Sarkar, Ashikur Rahaman, Gopeswar Mukherjee, Subhrangsu Chatterjee, Shamee Bhattacharjee, Deba Prasad Mandal

Abstract:

One of the major reasons for the high mortality rate of lung cancer is the substantial delays in disease detection at late metastatic stages. It is of utmost importance to understand the detailed molecular signaling and detect the molecular markers that can be used for the early diagnosis of cancer. Several studies explored the emerging roles of long noncoding RNAs (lncRNAs) in various cancers as well as lung cancer. A long non-coding RNA LINC00273 was recently discovered to promote cancer cell migration and invasion, and its positive correlation with the pathological stages of metastasis may prove it to be a potential target for inhibiting cancer cell metastasis. Comparing real-time expression of LINC00273 in various human clinical cancer tissue samples with normal tissue samples revealed significantly higher expression in cancer tissues. This long intergenic noncoding RNA was found to be highly expressed in human liver tumor-initiating cells, human gastric adenocarcinoma AGS cell line, as well as human non-small cell lung cancer A549 cell line. SiRNA and shRNA-induced knockdown of LINC00273 in both in vitro and in vivo nude mice significantly subsided AGS and A549 cancer cell migration and invasion. LINC00273 knockdown also reduced TGF-β induced SNAIL, SLUG, VIMENTIN, ZEB1 expression, and metastasis in A549 cells. Plenty of reports have suggested the role of microRNAs of the miR200 family in reversing epithelial to mesenchymal transition (EMT) by inhibiting ZEB transcription factors. In this study, hsa-miR-200a-3p was predicted via IntaRNA-Freiburg RNA tools to be a potential target of LINC00273 with a negative free binding energy of −8.793 kcal/mol, and this interaction was verified as a confirmed target of LINC00273 by RNA pulldown, real-time PCR and luciferase assay. Mechanistically, LINC00273 accelerated TGF-β induced EMT by sponging hsa-miR-200a-3p which in turn liberated ZEB1 and promoted prometastatic functions in A549 cells in vitro as verified by real-time PCR and western blotting. The similar expression patterns of these EMT regulatory pathway molecules, viz. LINC00273, hsa-miR-200a-3p, ZEB1 and TGF-β, were also detected in various clinical samples like breast cancer tissues, oral cancer tissues, lung cancer tissues, etc. Overall, this LINC00273 mediated EMT regulatory signaling can serve as a potential therapeutic target for the prevention of lung cancer metastasis.

Keywords: epithelial to mesenchymal transition, long noncoding RNA, microRNA, non-small-cell lung carcinoma

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Authors: Z. Karahaliloğlu, E. Yalçın, M. Demirbilek, E.B. Denkbaş

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Critical-sized bone defects caused by trauma, bone diseases, prosthetic implant revision or tumor excision cannot be repaired by physiological regenerative processes. Current orthopedic applications for critical-sized bone defects are to use autologous bone grafts, bone allografts, or synthetic graft materials. However, these strategies are unable to solve completely the problem, and motivate the development of novel effective biological scaffolds for tissue engineering applications and regenerative medicine applications. In particular, scaffolds combined with a variety of bio-agents as fundamental tools emerge to provide the regeneration of damaged bone tissues due to their ability to promote cell growth and function. In this study, a magnetic silk fibroin (SF) hydrogel scaffold was prepared by electrogelation process of the concentrated Bombxy mori silk fibroin (8 %wt) aqueous solution. For enhancement of osteoblast-like cells (SaOS-2) growth and adhesion, basal fibroblast growth factor (bFGF) were conjugated physically to the HSA-coated magnetic nanoparticles (Fe3O4) and magnetic SF e-gel scaffolds were prepared by incorporation of Fe3O4, HSA (human serum albumin)=Fe3O4 and HSA=Fe3O4-bFGF nanoparticles. HSA=Fe3O4, HSA=Fe3O4-bFGF loaded and bare SF e-gels scaffolds were characterized using scanning electron microscopy (SEM.) For cell studies, human osteoblast-like cell line (SaOS-2) was used and an MTT assay was used to assess the cytotoxicity of magnetic silk fibroin e-gel scaffolds and cell density on these surfaces. For the evaluation osteogenic activation, ALP (alkaline phosphatase), the amount of mineralized calcium, total protein and collagen were studied. Fe3O4 nanoparticles were successfully synthesized and bFGF was conjugated to HSA=Fe3O4 nanoparticles with %97.5 of binding yield which has a particle size of 71.52±2.3 nm. Electron microscopy images of the prepared HSA and bFGF incorporated SF e-gel scaffolds showed a 3D porous morphology. In terms of water uptake results, bFGF conjugated HSA=Fe3O4 nanoparticles has the best water absorbability behavior among all groups. In the in-vitro cell culture studies realized using SaOS-2 cell line, the coating of Fe3O4 nanoparticles surface with a protein enhance the cell viability and HSA coating and bFGF conjugation, the both have an inductive effect in the cell proliferation. One of the markers of bone formation and osteoblast differentiation, according to the ALP activity and total protein results, HSA=Fe3O4-bFGF loaded SF e-gels had significantly enhanced ALP activity. Osteoblast cultured HSA=Fe3O4-bFGF loaded SF e-gels deposited more calcium compared with SF e-gel. The proposed magnetic scaffolds seem to be promising for bone tissue regeneration and used in future work for various applications.

Keywords: basic fibroblast growth factor (bFGF), e-gel, iron oxide nanoparticles, silk fibroin

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