Search results for: cell concentration

Authors: Jamal Abd Awadallak, Thiago Olinek Reinehr, Eduardo Raizer, Deise Molinari, Edson Antonio, Camila da Silva da Silva

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The hydrolysis of soy oil catalyzed by 1,3-specific enzyme (Lecitase Ultra) in a well-stirred bioreactor was studied. Two forms of applications of the ultrasound were evaluated aiming to increase reaction rates, wherein the use of probe ultrasound associated with the use of surfactant to pre-emulsify the substrate showed the best results. Two different reaction periods were found: the first where the ultrasound has great influence on reaction rates, and the second where ultrasound influence is minimal. Studies on the time of pre-emulsification, surfactant concentration and enzyme concentration showed that the initial rate of hydrolysis depends on the interfacial area between the oil phase and the aqueous phase containing the enzyme.

Keywords: specific enzyme, free fatty acids, Hydrolysis, lecitase ultra, ultrasound

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Authors: Mokhtar Kouki Inès Rekik

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This paper focuses on the assessment of the air pollution and morbidity relationship in Tunisia. Air pollution is measured by ozone air concentration and the morbidity is measured by the number of respiratory-related restricted activity days during the 2-week period prior to the interview. Socioeconomic data are also collected in order to adjust for any confounding covariates. Our sample is composed by 407 Tunisian respondents; 44.7% are women, the average age is 35.2, near 69% are living in a house built after the 1980, and 27.8% have reported at least one day of respiratory-related restricted activity. The model consists on the regression of the number of respiratory-related restricted activity days on the air quality measure and the socioeconomic covariates. In order to correct for zero-inflation and heterogeneity, we estimate several models (Poisson, Negative binomial, Zero inflated Poisson, Poisson hurdle, Negative binomial hurdle and finite mixture Poisson models). Bootstrapping and post-stratification techniques are used in order to correct for any sample bias. According to the Akaike information criteria, the hurdle negative binomial model has the greatest goodness of fit. The main result indicates that, after adjusting for socioeconomic data, the ozone concentration increases the probability of positive number of restricted activity days.

Keywords: bootstrapping, hurdle negbin model, overdispersion, ozone concentration, respiratory-related restricted activity days

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Authors: Syed Farooq Adil, Merajuddinkhan, Mujeeb Khan, Hamad Z. Alkhathlan

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Phytochemicals from plant extracts belong to an important source of natural products which have demonstrated excellent cytotoxic activities. However, plants of different origins exhibit diverse chemical compositions and bioactivities. Therefore, the discovery of plants based new anticancer agents from different parts of the world is always challenging. In this study, methanolic extracts of different parts of 11 plants from Saudi Arabia have been tested in vitro for their anticancer potential on human liver cancer cell line (HepG2). Particularly, for this study, plants from Asteraceae, Resedaceae, and Polygonaceae families were chosen on the basis of locally available ethnobotanical data and their medicinal properties. Among 12 tested extract samples, three samples obtained from Artemisia monosperma stem, Ochradenus baccatus aerial parts, and Pulicaria glutinosa stem have demonstrated interesting cytotoxic activities with a cell viability of 29.3%, 28.4% and 24.2%, respectively. Whereas, four plant extracts including Calendula arvensis aerial parts, Scorzonera musilii whole plant, A. monosperma leaves show moderate anticancer properties bearing a cell viability ranging from 11.9 to 16.7%. The remaining extracts have shown poor cytotoxic activities. Subsequently, GC-MS analysis of methanolic extracts of the four most active plants extracts such as C. comosum, O. baccatus, P. glutinosa and A. monosperma detected the presence of 41 phytomolecules. Among which 3-(4-hydroxyphenyl) propionitrile (1), 8,11-octadecadiynoic acid methyl ester (2), 6,7-dimethoxycoumarin (3), and 1-(2-hydroxyphenyl) ethenone (4) were found to be the lead compounds of C. comosum, O. baccatus P. glutinosa and A. monosperma, respectively.

Keywords: medicinal plants, asteraceae, polygonaceae, hepg2

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Authors: Yihenew Simegniew Birhan, Hsieh Chih Tsai

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The recent advancements in synthetic chemistry and nanotechnology fostered the development of different nanocarriers for enhanced intracellular delivery of pharmaceutical agents to tumor cells. Polymeric micelles (PMs), characterized by small size, appreciable drug loading capacity (DLC), better accumulation in tumor tissue via enhanced permeability and retention (EPR) effect, and the ability to avoid detection and subsequent clearance by the mononuclear phagocyte (MNP) system, are convenient to improve the poor solubility, slow absorption and non-selective biodistribution of payloads embedded in their hydrophobic cores and hence, enhance the therapeutic efficacy of chemotherapeutic agents. Recently, redox-responsive polymeric micelles have gained significant attention for the delivery and controlled release of anticancer drugs in tumor cells. In this study, we synthesized redox-responsive diselenide bond containing amphiphilic polymer, Bi(mPEG-PLGA)-Se₂ from mPEG-PLGA, and 3,3'-diselanediyldipropanoic acid (DSeDPA) using DCC/DMAP as coupling agents. The successful synthesis of the copolymers was verified by different spectroscopic techniques. Above the critical micelle concentration, the amphiphilic copolymer, Bi(mPEG-PLGA)-Se₂, self-assembled into stable micelles. The DLS data indicated that the hydrodynamic diameter of the micelles (123.9 ± 0.85 nm) was suitable for extravasation into the tumor cells through the EPR effect. The drug loading content (DLC) and encapsulation efficiency (EE) of DOX-loaded micelles were found to be 6.61 wt% and 54.9%, respectively. The DOX-loaded micelles showed initial burst release accompanied by sustained release trend where 73.94% and 69.54% of encapsulated DOX was released upon treatment with 6mM GSH and 0.1% H₂O₂, respectively. The biocompatible nature of Bi(mPEG-PLGA)-Se₂ copolymer was confirmed by the cell viability study. In addition, the DOX-loaded micelles exhibited significant inhibition against HeLa cells (44.46%), at a maximum dose of 7.5 µg/mL. The fluorescent microscope images of HeLa cells treated with 3 µg/mL (equivalent DOX concentration) revealed efficient internalization and accumulation of DOX-loaded Bi(mPEG-PLGA)-Se₂ micelles in the cytosol of cancer cells. In conclusion, the intelligent, biocompatible, and the redox stimuli-responsive behavior of Bi(mPEG-PLGA)-Se₂ copolymer marked the potential applications of diselenide-linked mPEG-PLGA micelles for the delivery and on-demand release of chemotherapeutic agents in cancer cells.

Keywords: anticancer drug delivery, diselenide bond, polymeric micelles, redox-responsive

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Authors: Naila Sher, Mushtaq Ahmed, Nadia Mushtaq, Rahmat Ali Khan

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In cancer therapy, nanoparticles (NPs) shall be applied possibly by inoculation in the veins of humans. This action will connect them with white (WBCs) and red blood cells (RBCs) in the bloodstream before they reach their main targeted cancer cells. However, possible effects of silver, gold, and silver/gold bimetallic NPs (Ag, Au, and Ag/Au BNPs) on baby hamster kidney-21 (BHK-21) cells were studied by MTT assay. Here, Ag, Au, and their Ag/Au BNPs (bimetallic nanoparticles) were synthesized by using Hippeastrum hybridum (HH) extract. These NPs were characterized by UV-visible spectroscopy, FT-IR, XRD, and EDX, and SEM analysis. XRD analysis conferring the crystal structure with an average size of 13.3, 10.72, and 8.34nm of Ag, Au, and Ag/Au BNPs, respectively. SEM showed that Ag, Au, and Ag/Au BNPs had irregular morphologies, with nano measures calculated sizes of 40, 30, and 20 nm respectively. EDX spectrometers confirmed the presence of elemental Ag signal of the AgNPs with 22.75%, Au signal of the AuNPs with 48.08%, Ag signal with 12%, and Au signal with 38.26% of the Ag/Au BNPs. The BHK-21cells were incubated in the existence of doxorubicin, plant extract, Ag, Au, and Ag/Au BNPs. The cytotoxic effects could be observed in a dose-dependent mode; doxorubicin and Ag/Au BNPs were more toxic than plant extract, Ag, and Au NPs. It is demonstrated that NPs interact with BHK-21cells and significantly reduce cell viability in a concentration-dependent manner. However, to reduce the potential threats of NPs further studies are recommended.

Keywords: hippeastrum hybridum, nanoparticle, BHK-21cells

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Authors: ِAbdallah A. A. Shaheen

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Gaza Strip faces economic and political issues such as conflict, siege and urbanization; all these have led to an increase in the air pollution over Gaza Strip. In this study, Particulate matter 10 (PM10) concentration over Gaza Strip has been estimated by Landsat Thematic Mapper (TM) and Landsat Enhanced Thematic Mapper Plus (ETM+) data, based on a multispectral algorithm. Simultaneously, in-situ measurements for the corresponding particulate are acquired for selected time period. Landsat and ground data for eleven years are used to develop the algorithm while four years data (2002, 2006, 2010 and 2014) have been used to validate the results of algorithm. The developed algorithm gives highest regression, R coefficient value i.e. 0.86; RMSE value as 9.71 µg/m³; P values as 0. Average validation of algorithm show that calculated PM10 strongly correlates with measured PM10, indicating high efficiency of algorithm for the mapping of PM10 concentration during the years 2000 to 2014. Overall results show increase in minimum, maximum and average yearly PM10 concentrations, also presents similar trend over urban area. The rate of urbanization has been evaluated by supervised classification of the Landsat image. Urban sprawl from year 2000 to 2014 results in a high concentration of PM10 in the study area.

Keywords: PM10, landsat, atmospheric reflectance, Gaza strip, urbanization

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Authors: Moletsane Makgotso, Mogakabe Elijah, Marrengane Zinhle

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South Africa’s water resources quality is becoming more and more weakened by eutrophication, which deteriorates its usability. Thirty five percent of fresh water resources are eutrophic to hypertrophic, including grossly-enriched reservoirs that go beyond the globally-accepted definition of hypertrophy. Failing infrastructure adds to the problem of contaminated urban runoff which encompasses an important fraction of flows to inland reservoirs, particularly in the non-coastal, economic heartland of the country. Eutrophication threatens the provision of potable and irrigation water in the country because of the dependence on fresh water resources. Eutrophicated water reservoirs increase water treatment costs, leads to unsuitability for recreational purposes and health risks to human and animal livelihood due to algal proliferation. Eutrophication is caused by high concentrations of phosphorus and nitrogen in water bodies. In South Africa, Microsystis and Anabaena are widely distributed cyanobacteria, with Microcystis being the most dominant bloom-forming cyanobacterial species associated with toxin production. Two impoundments were selected, namely the Klipvoor and Leeukraal dams as they are mainly used for fishing, recreational, agricultural and to some extent, potable water purposes. The total oxidized nitrogen and total phosphorus concentration were determined as causative nutrients for eutrophication. Chlorophyll a and total microcystins, as well as the identification of cyanobacteria was conducted as indicators of cyanobacterial infestation. The orthophosphate concentration was determined by subjecting the samples to digestion and filtration followed by spectrophotometric analysis of total phosphates and dissolved phosphates using Aquakem kits. The total oxidized nitrates analysis was conducted by initially conducting filtration followed by spectrophotometric analysis. Chlorophyll a was quantified spectrophotometrically by measuring the absorbance of before and after acidification. Microcystins were detected using the Quantiplate Microcystin Kit, as well as microscopic identification of cyanobacterial species. The Klipvoor dam was found to be hypertrophic throughout the study period as the mean Chlorophyll a concentration was 269.4µg/l which exceeds the mean value for the hypertrophic state. The mean Total Phosphorus concentration was >0.130mg/l, and the total microcystin concentration was > 2.5µg/l throughout the study. The most predominant algal species were found to be the Microcystis. The Leeukraal dam was found to be mesotrophic with the potential of it becoming eutrophic as the mean concentration for chlorophyll a was 18.49 µg/l with the mean Total Phosphorus > 0.130mg/l and the Total Microcystin concentration < 0.16µg/l. The cyanobacterial species identified in Leeukraal have been classified as those that do not pose a potential risk to any impoundment. Microcystis was present throughout the sampling period and dominant during the warmer seasons. The high nutrient concentrations led to the dominance of Microcystis that resulted in high levels of microcystins rendering the impoundments, particularly Klipvoor undesirable for utilisation.

Keywords: nitrogen, phosphorus, cyanobacteria, microcystins

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Authors: Joseph Clement Akan, Lami Jafiya, Zaynab Muhammad Chellube, Zakari Mohammed, Fanna Inna Abdulrahman

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Five vegetables (spinach, lettuce, cabbage, tomato, and onion) were freshly harvested from the Alau Dam and Gongulong agricultural areas for the determination of some organochlorine pesticide residues (o, p-DDE, p,p’-DDD, o,p’-DDD, p,p’-DDT, α-BHC, γ-BHC, metoxichlor, lindane, endosulfan dieldrin, and aldrin.) Soil samples were also collected at different depths for the determination of the above pesticides. Samples collection and preparation were conducted using standard procedures. The concentrations of all the pesticides in the soil and vegetable samples were determined using GC/MS SHIMADZU (GC-17A) equipped with electron capture detector (ECD). The highest concentration was that of p,p’-DDD (132.4±13.45µg/g) which was observed in the leaf of cabbage, while the lowest concentration was that of p,p’-DDT (2.34µg/g) was observed in the root of spinach. Similar trends were observed at the Gongulong agricultural area, with p,p’-DDD having the highest concentration of 153.23µg/g in the leaf of cabbage, while the lowest concentration was that of p,p’-DDT (12.45µg/g) which was observed in the root of spinach. α-BHC, γ-BHC, Methoxychlor, and lindane were detected in all the vegetable samples studied. The concentrations of all the pesticides in the soil samples were observed to be higher at a depth of 21-30cm, while the lowest concentrations were observed at a depth of 0-10cm. The concentrations of all the pesticides in the vegetables and soil samples from the two agricultural sites were observed to be at alarming levels, much higher than the maximum residue limits (MRLs) and acceptable daily intake values (ADIs) .The levels of the pesticides observed in the vegetables and soil samples investigated, are of such a magnitude that calls for special attention and laws to regulate the use and circulation of such chemicals. Routine monitoring of pesticide residues in these study areas is necessary for the prevention, control and reduction of environmental pollution, so as to minimize health risks.

Keywords: Alau Dam, gongulong, organochlorine, pesticide residues, soil, vegetables

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Authors: Emmanuel Jaluchimike Iloputaife, Kelechi Nkechinyere Mbah-Omeje

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This study was designed to determine the antimicrobial activities and minimum inhibitory concentration of three different batches (Fresh, Oven dried and Sundried) of Ass Hay extracted with water, ethanol and methanolagainst selected human pathogenic microorganisms (Escherichia coli, Klebsiella Pneumonia, Staphylococcus aureus, Aspergillus niger and Candidaalbicans). All extracts were reconstituted with peptone water and tested for antimicrobial activity. The antimicrobial activity, the Minimum Inhibitory Concentration and Minimum Bactericidal/Fungicidal concentrations were determined by agar well diffusion methodagainst test organismsin which aseptic conditions were observed. The antimicrobial activities of the different batches of Ass Hay on the test organisms varied considerably. The highest inhibition zone diameter at 200 mg/ml for the different batches of Ass Hay was recorded by sundried methanol extract against Escherichia coli at 36.4 ± 0.2 mm while fresh methanol extract inhibited Klebsiela pneumonia with the least inhibition zone diameter at 20.1 ± 0.1mm. At 100 mg/ml the highest inhibition zone diameter was recorded by oven dried water extract against Escherichia coli at 30.3 ± 0.3 mm while sun dried water extract inhibited Staphylococcus aureus with the least inhibition zone diameter at 15.1 ± 0.1 mm. At 50mg/ml, the highest inhibition zone diameter was recorded by fresh water extract against Escherichia coli at 25.9 ± 0.1 mm while oven dried water extract inhibited Klebsiela pneumonia with least inhibition zone diameter at 12.1 ± 0.2 mm. At 25mg/ml, the highest inhibition zone diameter was recorded by fresh water extract against Escherichia coli at 18.3 ± 0.2 mm while sun dried ethanol extract inhibited Escherichia coli with least inhibition zone diameter at 10.1 ± 0.1 mm. The MIC and MBC result of ethanol extract of fresh Ass Hay showed a uniform value of 6.25 mg/ml and 12.5 mg/ml respectively for all test bacterial isolates. The Minimum Inhibitory concentration and Minimum bactericidal concentration results of Oven dried ethanol Ass Hay extract showed a uniform value of 3.125 mg/ml and 6.25 mg/ml respectively for all test bacterial isolates and Minimum fungicidal concentration value of 12.5 mg/ml for Aspergillus niger. Statistical analysis showed there is significant difference in mean zone inhibition diameter of the products at p < 0.05, p = 0.019. This study has shown there is antimicrobial potential in Ass Hay and at such there is need to further exploit Donkey Ass Hay in order to maximize the potential.

Keywords: microorganisms, Ass Hay, antimicrobial activity, extracts

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Authors: Amokrane Boufares, Elise Provost, Veronique Osswald, Pascal Clain, Anthony Delahaye, Laurence Fournaison, Didier Dalmazzone

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Gas hydrates have long been considered as problematic for flow assurance in natural gas and oil transportation. On the other hand, they are now seen as future promising materials for various applications (i.e. desalination of seawater, natural gas and hydrogen storage, gas sequestration, gas combustion separation and cold storage and transport). Nonetheless, a better understanding of the crystallization mechanism of gas hydrate and of their formation kinetics is still needed for a better comprehension and control of the process. To that purpose, measuring the real-time evolution of the dissolved gas concentration in the aqueous phase during hydrate formation is required. In this work, CO₂ hydrates were formed in a stirred reactor equipped with an Attenuated Total Reflection (ATR) probe coupled to a Fourier Transform InfraRed (FTIR) spectroscopy analyzer. A method was first developed to continuously measure in-situ the CO₂ concentration in the liquid phase during solubilization, supersaturation, hydrate crystallization and dissociation steps. Thereafter, the measured concentration data were compared with those of equilibrium concentrations. It was observed that the equilibrium is instantly reached in the liquid phase due to the fast consumption of dissolved gas by the hydrate crystallization. Consequently, it was shown that hydrate crystallization kinetics is limited by the gas transfer at the gas-liquid interface. Finally, we noticed that the liquid-hydrate equilibrium during the hydrate crystallization is governed by the temperature of the experiment under the tested conditions.

Keywords: gas hydrate, dissolved gas, crystallization, infrared spectroscopy

Procedia PDF Downloads 283

Authors: Mohamad F. Jamiluddin, Emeline Sarry, Ana Bejanariu, Cécile Bauche

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Introduction: Over 360 million people are chronically infected with hepatitis B virus (HBV), of whom 1 million die each year from HBV-associated liver cirrhosis or hepatocellular carcinoma. Current treatment options for chronic hepatitis B depend on interferon-α (IFNα) or nucleos(t)ide analogs, which control virus replication but rarely eliminate the virus. Treatment with PEG-IFNα leads to a sustained antiviral response in only one third of patients. After withdrawal of the drugs, the rebound of viremia is observed in the majority of patients. Furthermore, the long-term treatment is subsequently associated with the appearance of drug resistant HBV strains that is often the cause of the therapy failure. Among the new therapeutic avenues being developed, therapeutic vaccine aimed at inducing immune responses similar to those found in resolvers is of growing interest. The high prevalence of chronic hepatitis B necessitates the design of better vaccination strategies capable of eliciting broad-spectrum of cell-mediated immunity(CMI) and humoral immune response that can control chronic hepatitis B. Induction of HBV-specific T cells and B cells by therapeutic vaccination may be an innovative strategy to overcome virus persistence. Lentiviral vectors developed and optimized by THERAVECTYS, due to their ability to transduce non-dividing cells, including dendritic cells, and induce CMI response, have demonstrated their effectiveness as vaccination tools. Method: To develop a HBV therapeutic vaccine that can induce a broad but specific immune response, we generated recombinant lentiviral vector carrying IRES(Internal Ribosome Entry Site)-containing bicistronic constructs which allow the coexpression of two vaccine products, namely HBV T- cell epitope vaccine and HBV virus like particle (VLP) vaccine. HBV T-cell epitope vaccine consists of immunodominant cluster of CD4 and CD8 epitopes with spacer in between them and epitopes are derived from HBV surface protein, HBV core, HBV X and polymerase. While HBV VLP vaccine is a HBV core protein based chimeric VLP with surface protein B-cell epitopes displayed. In order to evaluate the immunogenicity, mice were immunized with lentiviral constructs by intramuscular injection. The T cell and antibody immune responses of the two vaccine products were analyzed using IFN-γ ELISpot assay and ELISA respectively to quantify the adaptive response to HBV antigens. Results: Following a single administration in mice, lentiviral construct elicited robust antigen-specific IFN-γ responses to the encoded antigens. The HBV T- cell epitope vaccine demonstrated significantly higher T cell immunogenicity than HBV VLP vaccine. Importantly, we demonstrated by ELISA that antibodies are induced against both HBV surface protein and HBV core protein when mice injected with vaccine construct (p < 0.05). Conclusion: Our results highlight that THERAVECTYS lentiviral vectors may represent a powerful platform for immunization strategy against chronic hepatitis B. Our data suggests the likely importance of Lentiviral vector based novel bicistronic construct for further study, in combination with drugs or as standalone antigens, as a therapeutic lentiviral based HBV vaccines. THERAVECTYS bicistronic HBV vaccine will be further evaluated in animal efficacy studies.

Keywords: chronic hepatitis B, lentiviral vectors, therapeutic vaccine, virus-like particle

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Authors: Nattawit Thiapairat

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Multiple diseases have been linked to excessive levels of free radicals, which cause tissue or cell damage as a result of oxidative stress. Many plants are sources of high antioxidant activity. Derris scandens has a high amount of phenolic and flavonoid contents which demonstrated good biological activities. This study focused on the antioxidant activity of polyphenols extracted from D. scandens. This study performs total flavonoids content and various antioxidant assays, which were 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging capacity assays. The total flavonoid content of D. scandens extract was determined and expressed as quercetin equivalents (QE)/g measured by the aluminum chloride colorimetric method. The antioxidant activity of D. scandens extract was also determined by DPPH and ABTS assays. In the DPPH assay, vitamin C was used as a positive control, whereas Trolox was used as a positive control in the ABTS assay. The half-maximal inhibitory concentration (IC50) values for D. scandens extract from DPPH and ABTS assays were 41.79 μg/mL ± 0.783 and 29.42 μg/mL ± 0.890, respectively, in the DPPH assay. To conclude, D. scandens extract consists of a high amount of total phenolic content, which exhibits a significant antioxidant activity. However, further investigation regarding antioxidant activity such as SOD, ROS, and RNS scavenging assays and in vivo experiments should be performed.

Keywords: ABTS assay, antioxidant activity, Derris scandens, DPPH assays, total flavonoid content

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Authors: Ann D. Christy, Beenish Saba

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The Microbial fuel Cell (MFC) is a successful integrated technology for power production and wastewater treatment. MFCs are recognized for their dual function, but research in this field is still ongoing to increase efficiency and power output. One such effort is successful integration of phototrophic and autotrophic microorganisms to create bacterio-algal MFCs for sustainable electricity production along with wastewater treatment and algal biomass production. An MFC is typically configured with an anaerobic anodic chamber containing exoelectrogenic microorganisms separated by a cation exchange membrane from an adjacent aerobic cathodic chamber. The two electrodes are connected by an external circuit. This conventional MFC can be converted into a phototrophic MFC by introducing photosynthetic microorganisms into the cathode chamber. This study examines adding a third desalination chamber to a two-chamber bacterio-algal MFC. Successful results have been observed from these three-chamber MFCs demonstrating wastewater treatment in the anodic chamber, phototrophic algal growth in the cathodic chamber, and desalination in the middle chamber. The present article will summarize successful results of the bacterio-algal fuel cells and offer insights about the mechanisms involved. Tables summarizing the input substrate along with optimized operational conditions and output performance in terms of power production and efficiencies of water and wastewater treatment will be presented. The negative impacts and challenges will be discussed, along with possible future research directions. Results suggest that the three chamber bacterio-algal desalination cell has potential as a feasible technology for power production, wastewater treatment and desalination, but it needs further investigation under optimized conditions.

Keywords: bacterio-algal MFC, three chamber, microbial fuel cell, wastewater treatment and desalination

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Authors: Iannello G., Coppa F., Pennisi S., Giuffrida G., Lo Faro R., Cartelli S., Ferruggia G., Brundo M. V.

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Skin is the largest multifunctional human organ and possesses a complex, multilayered structure with the ability to regenerate and renew. The key role in skin regeneration is played by fibroblasts, which also occupy an important role in the wound healing process. Different methods, including dynamic light scattering, scanning electron microscopy, ELISA, and MTT assay, were employed to evaluate on fibroblasts the in vitro effects of plant-derived nanovesicles and cord blood stem cells‐derived exosomes. We compared the results with those of cells exposed to a technology called AMPLEX PLUS, containing a mixture of 20 different biologically active factors (GF20) and exosomes isolated and purified from bovine colostrum. AMPLEX PLUS was able to significantly enhance the cell proliferation status of cells at both 24 and 48 hours compared to untreated cells (control). The obtained results suggest how AMPLEX PLUS could be potentially effective in treating skin rejuvenation.

Keywords: AMPLEX PLUS, cell vitality, colostrum, nanovesicles

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Authors: Pinar Erturk, Sevde Altuntas, Fatih Buyukserin

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In order to obtain an effective integration between an implant and a bone, implant surfaces should have similar properties to bone tissue surfaces. Especially mimicry of the chemical, mechanical and topographic properties of the implant to the bone is crucial for fast and effective osseointegration. Titanium-based biomaterials are more preferred in clinical use, and there are studies of coating these implants with oxide layers that have chemical/nanotopographic properties stimulating cell interactions for enhanced osseointegration. There are low success rates of current implantations, especially in craniofacial implant applications, which are large and vital zones, and the oxide layer coating increases bone-implant integration providing long-lasting implants without requiring revision surgery. Our aim in this study is to examine bone-cell behavior on titanium implants with an aluminum oxide layer (AAO) on effective osseointegration potential in the deformation of large zones with difficult spontaneous healing. In our study, aluminum layer coated titanium surfaces were anodized in sulfuric, phosphoric, and oxalic acid, which are the most common used AAO anodization electrolytes. After morphologic, chemical, and mechanical tests on AAO coated Ti substrates, viability, adhesion, and mineralization of adult bone cells on these substrates were analyzed. Besides with atomic layer deposition (ALD) as a sensitive and conformal technique, these surfaces were coated with pure alumina (5 nm); thus, cell studies were performed on ALD-coated nanoporous oxide layers with suppressed ionic content too. Lastly, in order to investigate the effect of the topography on the cell behavior, flat non-porous alumina layers on silicon wafers formed by ALD were compared with the porous ones. Cell viability ratio was similar between anodized surfaces, but pure alumina coated titanium and anodized surfaces showed a higher viability ratio compared to bare titanium and bare anodized ones. Alumina coated titanium surfaces, which anodized in phosphoric acid, showed significantly different mineralization ratios after 21 days over other bare titanium and titanium surfaces which anodized in other electrolytes. Bare titanium was the second surface that had the highest mineralization ratio. Otherwise, titanium, which is anodized in oxalic acid electrolyte, demonstrated the lowest mineralization. No significant difference was shown between bare titanium and anodized surfaces except AAO titanium surface anodized in phosphoric acid. Currently, osteogenic activities of these cells on the genetic level are investigated by quantitative real-time polymerase chain reaction (qRT-PCR) analysis results of RUNX-2, VEGF, OPG, and osteopontin genes. Also, as a result of the activities of the genes mentioned before, Western Blot will be used for protein detection. Acknowledgment: The project is supported by The Scientific and Technological Research Council of Turkey.

Keywords: alumina, craniofacial implant, MG-63 cell line, osseointegration, oxalic acid, phosphoric acid, sulphuric acid, titanium

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Authors: Gizem Buldum, Alexander Bismarck, Athanasios Mantalaris

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Cellulose is the most abundant biopolymer on earth and it is currently being utilised in a multitude of industrial applications. Over the last 30 years, attention has been paid to the bacterial cellulose (BC), since BC exhibits unique physical, chemical and mechanical properties when compared to plant-based cellulose, including high purity and biocompatibility. Although Acetobacter xylinum is the most efficient producer of BC, it’s long doubling time results in insufficient yields of the cellulose production. This limits widespread and continued use of BC. In this study, E. coli BL21 (DE3) or E. coli HMS cells are selected as host organisms for the expression of bacterial cellulose synthase operon (bcs) of A.xylinum. The expression system is created based on pET-Duet1 and pCDF plasmid vectors, which carry bcs operon. The results showed that all bcs genes were successfully transferred and expressed in E.coli strains. The expressions of bcs proteins were shown by SDS and Native page analyses. The functionality of the bcs operon was proved by congo red binding assay. The effect of culturing temperature and the inducer concentration (IPTG) on cell growth and plasmid stability were monitored. The percentage of plasmid harboring cells induced with 0.025 mM IPTG was obtained as 85% at 22˚C in the end of 10-hr culturing period. It was confirmed that the high output cellulose production machinery of A.xylinum can be transferred into other organisms.

Keywords: bacterial cellulose, biopolymer, recombinant expression system, production

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Authors: Alpana Saini, Sanghamitra Barman

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Investigations were carried out to determine whether low cost peanut hull as adsorbent hold promise in removal of thionine dyes in the biomedical industries. Pollution of water due to presence of colorants is a severe socio-environmental problem caused by the discharge of industrial wastewater. In view of their toxicity, non-biodegradability and persistent nature, their removal becomes an absolute necessity. For the removal of Thionine Dye using Peanut Hull, the 10mg/L concentration of dyes, 0.5g/l of adsorbent and 200 rpm agitation speed are found to be optimum for the adsorption studies. The Spectrophotometric technique was adopted for the measurement of concentration of dyes before and after adsorption at ʎmax 598nm. The adsorption data has been fitted well to Langmuir isotherm than to Freundlich adsorption isotherm. The adsorbent was characterized by Scanning Electron Microscopy (SEM).

Keywords: adsorption, langmuir isotherm, peanut hull, thionine

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Authors: G. Krishnamoorthy, S. Anandhakumar

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The effect of D-Lysine (D-Lys) on collagen with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide(EDC)/N-hydroxysuccinimide(NHS) initiated cross linking using experimental and modelling tools are evaluated. The results of the Coll-D-Lys-EDC/NHS scaffold also indicate an increase in the tensile strength (TS), percentage of elongation (% E), denaturation temperature (Td), and decrease the decomposition rate compared to L-Lys-EDC/NHS. Scanning electron microscopic (SEM) and atomic force microscopic (AFM) analyses revealed a well ordered with properly oriented and well-aligned structure of scaffold. The D-Lys stabilizes the scaffold against degradation by collagenase than L-Lys. The cell assay showed more than 98% fibroblast viability (NIH3T3) and improved cell adhesions, protein adsorption after 72h of culture when compared with native scaffold. Cell attachment after 74h was robust, with cytoskeletal analysis showing that the attached cells were aligned along the fibers assuming a spindle-shape appearance, despite, gene expression analyses revealed no apparent alterations in mRNA levels, although cell proliferation was not adversely affected. D-Lysine (D-Lys) plays a pivotal role in the self-assembly and conformation of collagen fibrils. The D-Lys assisted EDC/NHS initiated cross-linking induces the formation of an carboxamide by the activation of the side chain -COOH group, followed by aminolysis of the O-iso acylurea intermediates by the -NH2 groups are directly joined via an isopeptides bond. This leads to the formation of intra- and inter-helical cross links. Modeling studies indicated that D-Lys bind with collagen-like peptide (CLP) through multiple H-bonding and hydrophobic interactions. Orientational changes in collagenase on CLP-D-Lys are observed which may decrease its accessibility to degradation and stabilize CLP against the action of the former. D-Lys has lowest binding energy and improved fibrillar-assembly and staggered alignment without the undesired structural stiffness and aggregations. The proteolytic machinery is not well equipped to deal with Coll-D-Lys than Coll-L-Lys scaffold. The information derived from the present study could help in designing collagenolytically stable heterochiral collagen based scaffold for biomedical applications.

Keywords: collagen, collagenase, collagen like peptide, D-lysine, heterochiral collagen scaffold

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Authors: Akhmadjon Sultanov, Eun-Ho Lee, Hye-Jin Park, Young-Je Cho

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This study tested the anti-inflammatory effect of wild indigo (Baptisia tinctoria) root in Raw 264.7 cells. We prepared two extracts of B. tinctoria; one in water and the other in 50% ethanol. Then we evaluated the toxicities of the B. tinctoria root extracts at 10 to 100 mg mL-1 concentrations in raw 264.7 cells and observed 80% cell viability. The anti-inflammatory effect of B. tinctoria root extract in lipopolysaccharide (LPS)-stimulated Raw 264.7 cells were observed with concentrations at 10, 30, and 50 μg mL-1. The results showed that 77.27-66.82% of nitric oxide (NO) production was inhibited by 50 μg mL-1 B. tinctoria root extract. The protein expression of Inducible NO synthase (iNOS) expression dramatically decreased by 93.14% and 52.65% in raw 264.7 cells treated with water and ethanol extracts of B. tinctoria root, respectively. Moreover, cyclooxygenase-2 (COX-2) protein expression decreased by 42.85% and 69.70% in raw 264.7 cells treated with water and ethanol extracts of B. tinctoria root, respectively. Furthermore, the mRNA expression of pro-inflammatory markers, such as tumor necrosis factor-alpha, interleukin-1β, interleukin-6, monocyte chemoattractant protein-1, and prostaglandin E synthase 2, was significantly suppressed in a concentration-dependent manner. Additionally, the B. tinctoria root extracts effectively inhibited enzymes involved in physiological activities. The B. tinctoria root extracts showed excellent anti-inflammatory effects and can be used as a functional material for biological activities.

Keywords: cytokine, macrophage, pro-inflammatory, protein expression, real-time PCR

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Authors: Mansi Patel, Priti Mehta

Abstract:

Background: Ionizing radiations have detrimental effects on humans, and the growing technological encroachment has increased human exposure to it enormously. So, the safety issues have emphasized researchers to develop radioprotector from natural resources having minimal toxicity. A substance having anti-inflammatory, antioxidant, and immunomodulatory activity can be a potential candidate for radioprotection. One such plant with immense potential i.e. Bamboo was selected for the present study. Purpose: The study aims to evaluate the potential of Indian bamboo leaves for protection against the clastogenic effect of gamma radiation. Methods: The protective effect of bamboo leaf extract against gamma radiation-induced genetic damage in human peripheral blood lymphocytes (HPBLs) was evaluated in vitro using Cytokinesis blocked micronuclei assay (CBMN). The blood samples were pretreated with varying concentration of extract 30 min before the radiation exposure (4Gy & 6Gy). The reduction in the frequency of micronuclei was observed for the irradiated and control groups. The effect of various concentration of bamboo leaf extract (400,600,800 mg/kg) on the development of radiation induced sickness and altered mortality in mice exposed to 8 Gy of whole-body gamma radiation was studied. The developed symptoms were clinically scored by multiple endpoints for 30 days. Results: Treatment of HPBLs with varying concentration of extract before exposure to a different dose of γ- radiation resulted in significant (P < 0.0001) decline of radiation induced micronuclei. It showed dose dependent and concentration driven activity. The maximum protection ~ 70% was achieved at nine µg/ml concentration. Extract treated whole body irradiated mice showed 50%, 83.3% and 100% survival for 400, 600, and 800mg/kg with 1.05, 0.43 and 0 clinical score respectively when compared to Irradiated mice having 6.03 clinical score and 0% survival. Conclusion: Our findings indicate bamboo leaf extract reduced the radiation induced cytogenetic damage. It has also increased the survival ratio and reduced the radiation induced sickness and mortality when exposed to a lethal dose of gamma radiation.

Keywords: bamboo leaf extract, Cytokinesis blocked micronuclei (CBMN) assay, ionizing radiation, radio protector

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Authors: Wenxin Mei, Jinhua Sun, Qingsong Wang

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The volume expansion of lithium-ion batteries is mainly induced by intercalation induced stress within the negative electrode, resulting in capacity degradation and even battery failure. Stress generation due to lithium intercalation into graphite particles is investigated based on an electrochemical-mechanical model in this work. The two-dimensional model presented is fully coupled, inclusive of the impacts of intercalation-induced stress, stress-induced intercalation, to evaluate the lithium concentration, stress generation, and displacement intra and inter-particle. The results show that the distribution of lithium concentration and stress exhibits an analogous pattern, which reflects the relation between lithium diffusion and stress. The results of inter-particle stress indicate that larger Von-Mises stress is displayed where the two particles are in contact with each other, and deformation at the edge of particles is also observed, predicting fracture. Additionally, the maximum inter-particle stress at the end of lithium intercalation is nearly ten times the intraparticle stress. And the maximum inter-particle displacement is increased by 24% compared to the single-particle. Finally, the effect of graphite particle arrangement on inter-particle stress is studied. It is found that inter-particle stress with tighter arrangement exhibits lower stress. This work can provide guidance for predicting the intra and inter-particle stress to take measures to avoid cracking of electrode material.

Keywords: electrochemical-mechanical model, graphite particle, lithium concentration, lithium ion battery, stress

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Authors: C. Rattanakawin, S. Vasailor

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Acid leaching of flotation concentrate of oxide copper ore containing mainly of malachite was performed in a standard agitation tank with various parameters. The effects of solid to liquid ratio, sulfuric acid concentration, agitation speed, leaching temperature and time were examined to get proper conditions. The best conditions are 1:8 solid to liquid ratio, 10% concentration by weight, 250 rev/min, 30 ^oC and 5-min leaching time in respect. About 20% Cu grade assayed by atomic absorption technique with 98% copper recovery was obtained from these combined optimum conditions. Dissolution kinetics of the concentrate was approximated as a logarithmic function. As a result, the first-order reaction rate is suggested from this leaching study.

Keywords: agitation leaching, dissolution kinetics, flotation concentrate, oxide copper ore, sulfuric acid

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Authors: Lila A. Alkhattaby, Norah A. Alsayegh, Mohammad S. Ansari, Mohammad O. Ansari

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In this work, we doped zinc oxide ZnO with potassium K we have synthesized using the sol-gel method. Structural properties were depicted by X-ray diffractometer (XRD) and energy distribution spectroscopy, X-ray diffraction studies confirm the nanosized of the particles and favored orientations along the (100), (002), (101), (102), (110), (103), (200), and (112) planes confirm the hexagonal wurtzite structure of ZnO NPs. The optical properties study using the UV-Vis spectroscopy. The band gap decreases from 4.05 eV to 3.88 eV, the lowest band gap at 10% doped concentration. The photoluminescence (PL) spectroscopy results show two main peaks, a sharp peak at ≈ 384 nm in the UV region and a broad peak around 479 nm in the visible region. The highest intensity of the band-edge luminescence was for 2% doped concentration because of the combined effect of the decreased probability of nonradiative recombination and has better crystallinity.

Keywords: K doped ZnO, photoluminescence spectroscopy, UV-Vis spectroscopy, x-ray spectroscopy

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Authors: Muhammad Muzamil Khan, Asadullah Madni, Nina Filipczek, Jiayi Pan, Nayab Tahir, Hassan Shah, Vladimir Torchilin

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Lipid-polymer hybrid nanoparticles (LPHNP) are delivery systems for controlled drug delivery at tumor sites. The superior biocompatible properties of lipid and structural advantages of polymer can be obtained via this system for controlled drug delivery. In the present study, cisplatin-loaded lipid-chitosan hybrid nanoparticles were formulated by the single step ionic gelation method based on ionic interaction of positively charged chitosan and negatively charged lipid. Formulations with various chitosan to lipid ratio were investigated to obtain the optimal particle size, encapsulation efficiency, and controlled release pattern. Transmission electron microscope and dynamic light scattering analysis demonstrated a size range of 181-245 nm and a zeta potential range of 20-30 mV. Compatibility among the components and the stability of formulation were demonstrated with FTIR analysis and thermal studies, respectively. The therapeutic efficacy and cellular interaction of cisplatin-loaded LPHNP were investigated using in vitro cell-based assays in A2780/ADR ovarian carcinoma cell line. Additionally, the cisplatin loaded LPHNP exhibited a low toxicity profile in rats. The in-vivo pharmacokinetics study also proved a controlled delivery of cisplatin with enhanced mean residual time and half-life. Our studies suggested that the cisplatin-loaded LPHNP being a promising platform for controlled delivery of cisplatin in cancer therapy.

Keywords: cisplatin, lipid-polymer hybrid nanoparticle, chitosan, in vitro cell line study

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Authors: Imen Hamed, Burcu Ak, Oya Işık, Leyla Uslu, Kubilay Kazım Vursavuş

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In this study, 3 species of Dunaliella (Dunaliella sp. Salt Lake isoalte (Tuz Gölü), Dunaliella salina CCAP19/18, and Dunaliella bardawil LB 2538) and their optical density, dry matter, chlorophyll a, total carotenoids, and β-carotene production were investigated in a batch system. The aim of this research was to compare carotenoids, and β-carotene production were investigated in a batch those 3 species. Therefore 2 stress factors were used: 2 different temperatures (20°C and 30°C) and 2 different salinities (30‰, and 60‰) were tested over a 17-day study. The highest growth and chlorophyll a was reported for Dunaliella sp. under 20°C/30‰ and 20°C/60‰ conditions respectively followed by D. bardawil and D. salina. Significant differences were noticed (p<0.05) for the other 3 species. The growth decreased as temperature and salinity increased since the lowest growth was noticed for the 30°C/60‰ group. The chlorophyll a content decreased also as temperature increased however when the NaCl concentration increased an augmentation of the content was noticed . In the 17^th day of experiment the highest carotenoids concentration was reported for D. bardawil 20°C/30‰ (65,639±0,400 μg.mL⁻¹) and the most important β carotene concentration was for D. salina 20°C/60‰ (8,98E-07±0,013 mol/L).

Keywords: Dunaliella sp., Dunaliella salina, Dunaliella bardawil, growth, pigments, stress factors

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Authors: H. Boulahyaoui, S. Alaoui Amine, C. Loutfi, H. El Annaz, N. Touil, El M. El Fahim, S. Mrani

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Three Moroccan children fully vaccinated with Rotarix™ have been hospitalized for Rotavirus Gastroenteritis (RVGE) in the pediatric division of the Farabi Hospital, Oujda. Rotavirus G/P genotypes could not be typed because of their delayed crossing threshold (Ct) resolute with a group A rotavirus (RVA) real time RT-PCR. These strains were adapted to cell culture. All viruses replicated and caused extensive cytopathic effects after four or five passages in MA104 cell lines. Significant improvements have been obtained in the amount of viral particles. Each virus multiplied to a high titer (7.5 TCID50/ml). VP7 and VP4 partial gene sequencing revealed distinct genotypes compared to the Rotarix(®) vaccine strain. Two strains were of G2P[4] genotype whereas the third was G9P[8] genotype. Virus isolation while labor intensive, is recommended as a second test, especially when higher sensitivity for conventional RVA genotyping RT-PCR is needed. VP7 antigenic similarities between these strains and Rotarix were determined.

Keywords: esacpe-vaccine, Morocco, Rotarix, G2P[4], G9P[8]

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Authors: Shenko Chura Aredo, Hailu Belay, Kevin T. Kornegay

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In light of Adama City’s smart city development vision, this study thoroughly investigates the performance of smart security systems with Fifth Generation (5G) network capabilities. It can be logistically difficult to install a lot of cabling, particularly in big or dynamic settings. Moreover, latency issues might affect linked systems, making it difficult for them to monitor in real time. Through a focused analysis that employs Adama City as a case study, the performance has been evaluated in terms of spectrum and energy efficiency using empirical data and basic signal processing formulations at different frequency resources. The findings also demonstrate that cameras working at higher 5G frequencies have more capacity than those operating at sub-6 GHz, notwithstanding frequency-related issues. It has also been noted that when the beams of such cameras are adaptively focussed based on the distance of the last cell edge user rather than the maximum cell radius, less energy is required than with conventional fixed power ramping.

Keywords: 5G, energy efficiency, safety, smart security, spectral efficiency

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Authors: Ehiedu Philomina Kika, Jessica Chinonso Ehilegbu

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The exploration and production of fossil fuel particularly crude oil has led to some serious environmental damage in some oil producing communities like the Bodo Community who rely heavily on their aquatic environment for food and water. This study was therefore carried out to investigate the level of some heavy metals in water and Callinectes amnicola (Swimming Crab) in the month of August, September and October from Bodo creek, Rivers State, Nigeria. The physico-chemical parameters of the water were also analyzed in-situ. The levels of heavy metals, Lead (Pb), Cadmium (Cd), Chromium (Cr), Zinc (Zn), Copper (Cu) were analyzed in water and in Callinectes amnicola (Swimming Crab), using Atomic Absorption Spectrophotometer (AAS) after acid digestion. For the concentration of heavy metals in water, Pb ranged from 0.103 - 0.791 mg/l, Zn 0.0025 - 0.342 mg/l, Cr < 0.001 - 0.304 mg/l, Cd 0.011 - 0.116 mg/l and Cu <0.001 - 0.079 mg/l. For the concentration of heavy metals in Callinectes amnicola (Swimming Crab), the level of Pb ranged from 0.359 - 0.849 mg/l, Zn 0.134 - 0.342 mg/l, Cd 0.053 - 0.103 mg/l, Cr < 0.001 - <0.001 mg/l, Cu < 0.001 - 0.131 mg/l. The concentrations of Pb, Cd and Cr for all water and crab samples collected from the various stations were higher than permissible level suggesting serious anthropogenic influence. Thus, precaution needs to be taken to prevent further contamination and adequate purification measures need to be put in place. Therefore, there should be periodic environmental pollution monitoring, for assessment and awareness especially with regards heavy metal.

Keywords: Bodo creek, crude oil, heavy metal, swimming crab

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Authors: Guadalupe Stefanny Aguilar-Moreno, Miguel Angel Aguilar-Mendez, Teodoro Espinosa-Solares

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In the agricultural sector, one of the main emitters of greenhouse gases is manure management, which has been increased considerably in recent years. Biogas is an energy source that can be produced from different organic materials through anaerobic digestion (AD); however, production efficiency is still low. Several techniques have been studied to increase its performance, such as co-digestion, the variation of digestion conditions, and nanomaterials used. Therefore, the aim of this investigation was to evaluate the effect of magnetite nanoparticles (NPs) concentration, synthesized by co-precipitation, on the biogas and methane production in AD using chicken litter as a substrate. Synthesis of NPs was performed according to the co-precipitation method, for which a fractional factorial experimental design 25⁻² with two replications was used. The study factors were concentrations (precursors and passivating), time of sonication and dissolution temperatures, and the response variables were size, hydrodynamic diameter (HD) and zeta potential. Subsequently, the treatment that presented the smallest NPs was chosen for their use on AD. The AD was established in serological bottles with a working volume of 250 mL, incubated at 36 ± 1 °C for 80 days. The treatments consisted of the addition of different concentrations of NPs in the microcosms: chicken litter only (control), 20 mg∙L⁻¹ of NPs + chicken litter, 40 mg∙L⁻¹ of NPs + chicken litter and 60 mg∙L⁻¹ of NPs + chicken litter, all by triplicate. Methane and biogas production were evaluated daily. The smallest HD (49.5 nm) and the most stable NPs (21.22 mV) were obtained with the highest passivating concentration and the lower precursors dissolution temperature, which were the only factors that had a significant effect on the HD. In the transmission electron microscopy performed to these NPs, an average size of 4.2 ± 0.73 nm was observed. The highest biogas and methane production was obtained with the treatment that had 20 mg∙L⁻¹ of NPs, being 29.5 and 73.9%, respectively, higher than the control, while the treatment with the highest concentration of NPs was not statistically different from the control. From the above, it can be concluded that the magnetite NPs promote the biogas and methane production in AD; however, high concentrations may cause inhibitory effects among methanogenic microorganisms.

Keywords: agricultural sector, anaerobic digestion, nanotechnology, waste management

Procedia PDF Downloads 139

Authors: Wen-Yang Hu, Ranli Lu, Mark Maienschein-Cline, Danping Hu, Larisa Nonn, Toshi Shioda, Gail S. Prins

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Background: Genetic mutations are highly associated with increased prostate cancer risk. In addition to whole genome sequencing, somatic mutations can be identified by aligning transcriptome sequences to the human genome. Here we analyzed bulk RNAseq and single cell RNAseq data of human prostate cancer cells and their matched non-cancer cells in benign regions from 4 individual patients. Methods: Sequencing raw reads were aligned to the reference genome hg38 using STAR. Variants were annotated using Annovar with respect to overlap gene annotation information, effect on gene and protein sequence, and SIFT annotation of nonsynonymous variant effect. We determined cancer-specific novel alleles by comparing variant calls in cancer cells to matched benign cells from the same individual by selecting unique alleles that were only detected in the cancer samples. Results: In bulk RNAseq data from 3 patients, the most common variants were the noncoding mutations at UTR3/UTR5, and the major variant types were single-nucleotide polymorphisms (SNP) including frameshift mutations. C>T transversion is the most frequently presented substitution of SNP. A total of 222 genes carrying unique exonic or UTR variants were revealed in cancer cells across 3 patients but not in benign cells. Among them, transcriptome levels of 7 genes (CITED2, YOD1, MCM4, HNRNPA2B1, KIF20B, DPYSL2, NR4A1) were significantly up or down regulated in cancer stem cells. Out of the 222 commonly mutated genes in cancer, 19 have nonsynonymous variants and 11 are damaged genes with variants including SIFT, frameshifts, stop gain/loss, and insertions/deletions (indels). Two damaged genes, activating transcription factor 6 (ATF6) and histone demethylase KDM3A are of particular interest; the former is a survival factor for certain cancer cells while the later positively activates androgen receptor target genes in prostate cancer. Further, single cell RNAseq data of cancer cells and their matched non-cancer benign cells from both primary 2D and 3D tumoroid cultures were analyzed. Similar to the bulk RNAseq data, single cell RNAseq in cancer demonstrated that the exonic mutations are less common than noncoding variants, with SNPs including frameshift mutations the most frequently presented types in cancer. Compared to cancer stem cell enriched-3D tumoroids, 2D cancer cells carried 3-times higher variants, 8-times more coding mutations and 10-times more nonsynonymous SNP. Finally, in both 2D primary and 3D tumoroid cultures, cancer stem cells exhibited fewer coding mutations and noncoding SNP or insertions/deletions than non-stem cancer cells. Summary: Our study demonstrates the usefulness of bulk and single cell RNAseaq data in identifying somatic mutations in prostate cancer, providing an alternative method in screening candidate genes for prostate cancer diagnosis and potential therapeutic targets. Cancer stem cells carry fewer somatic mutations than non-stem cancer cells due to their inherited immortal stand DNA from parental stem cells that explains their long-lived characteristics.

Keywords: prostate cancer, stem cell, genomic mutation, RNAseq

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