Search results for: cell uptake

Authors: Ana Paula Rosifini Alves Claro, André Luiz Reis Rangel, Nathan Trujillo, Ketul C. Popat

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In the present work, in vitro cytotoxicity was evaluated after nanotubes growth on Ti15Mo alloy surface. TiO2 nanotubes were obtained by anodizing technique at room temperature in an electrolyte with 0.25 %NH4F and glycerol at a constant anodic potential of 20 V for 24 hours. The morphology of nanotubes was observed by field emission scanning electron microscopy (FE-SEM; XL 30 FEG, Philips). Crystal structure was analyzed by wide-angle X-ray diffraction. A cell culture model using human fibroblast-like cells was used to study the effect of TiO2 nanotubes growth on the cytotoxicity of the Ti15Mo alloy for 1, 4 and 7 days culture period. The MTT assay was used to evaluate cell viability and cell adhesion was evaluated by scanning electron microscopy. Results show that Ti15Mo alloy with TiO2 nanotubes on surface is nontoxic and exhibit good interaction with surface.

Keywords: titanium alloys, TiO2 nanotubes, cell growth, Ti-15Mo alloy

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Authors: Jie Ding, Yingying Pan, Shammy Raj, Lindy Schaffrick, Jolene Wong, Antoinette Nguyen, Sharada Manchikanti, Larry Unsworth, Peter Kwan, Edward E. Tredget

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Background: Exosomes (EXOs) have been considered a new target that is thought to be involved in and treat wound healing. More research is needed to fully understand the EXO characteristics and mechanisms of EXO-mediated wound healing, especially wound healing after burn injury. Methods: Total EXOs were isolated from 85 serum samples of 29 burn patients and 13 healthy individuals. We characterized the EXOs for morphology and density, serum concentration, protein level, marker expression, size distribution, and cytokine content. After confirmation of EXO uptake by dermal fibroblasts, we also explored functional regulation of primary human normal skin and hypertrophic scar fibroblast cell lines by the EXOs in vitro, including cell proliferation and apoptosis. Results: EXOs dynamically changed their morphology, density, size, and cytokine level during wound healing in burn patients, which were correlated with burn severity and the stages of wound healing. EXOs from both burn patients and healthy individuals stimulated dermal fibroblast proliferation and apoptosis. Conclusion: EXO features may be important signals that influence wound healing after burn injury; however, to understand the mechanisms by which EXOs regulated the fibroblasts in healing wounds, further studies will be required in the future.

Keywords: exosome, burn, wound healing, hypertrophic scarring, cytokines

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Authors: Cheng-Cao Sun, Shu-Jun Li, De-Jia Li

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Through analysis of a published micro-array-based high-throughput assessment, we discovered that miR-1275 was markedly down-regulated in nasopharyngeal carcinoma (NPC) tissues. However, little is known about its effect and mechanism involved in NPC development and progression. In this study, we investigated the role of miR-1275 on the development of NPC. The results indicated that miR-1275 was significantly down-regulated in primary NPC tissues, and very low levels were found in NPC cell lines. Ectopic expression of miR-1275 in NPC cell lines significantly suppressed cell growth as evidenced by cell viability assay and colony formation assay, through inhibition of HOXB5. In addition, miR-1275 suppresses G1/S transition through inhibition of HOXB5. Further, oncogene HOXB5 was revealed to be a putative target of miR-1275, which was inversely correlated with miR-1275 expression in NPC. Collectively, our study demonstrates that as a tumor suppressor, miR-1275 played a pivotal role on NPC through inhibiting cell proliferation, and suppressing G1/S transition by targeting oncogenic HOXB5.

Keywords: microRNA-1275 (miR-1275), HOXB5, nasopharyngeal carcinoma, proliferation

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Authors: Shikha Masand, Sudesh Kumar Yadav

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Certain protein kinases have been shown to be crucial for plant cell signaling pathways associated with plant immune responses. Here we identified a horsegram [Macrotyloma uniflorum (Lam.) Verdc.] malectin-like leucine rich receptor-like protein kinase (RLK) gene MuRLK. The functional MuRLK protein preferentially binds to mannose and N-acetyl glucosamine residues. MuRLK exists in the cytoplasm and also localizes to the plasma membrane of plant cells via its N-terminus. Over-expression of MuRLK in Arabidopsis enhances the basal resistance to infection with Pseudomonas syringae pv. tomato, Alternaria brassicicola and Hyaloperonospora arabidopsidis, are associated with elevated ROS bursts, MAPK activation, thus ultimately leading to hypersensitive cell death. Moreover, salicylic acid-dependent and jasmonic acid-dependent defense responses are also enhanced in the MuRLK-overexpressed plants that lead to HR-induced cell death. Together, these results suggest that MuRLK plays a key role in the regulation of plant cell death, early and late defense responses after the recognition of microbial pathogens.

Keywords: horsegram, Pseudomonas syringae pv. tomato, MuRLK, ROS burst, cell death, plant defense

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Authors: Emrah Ersoy, Yusuf Ozcatalbas

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The aim of this study is to investigate formability of Al based closed cell metallic foams at high temperature. The foam specimens with rectangular section were produced from AlMg1Si0.6TiH20.8 alloy preform material. Bending and free bending tests based on gravity effect were applied to foam specimens at high temperatures. During the tests, the time-angular deformation relationships with various temperatures were determined. Deformation types formed in cell walls were investigated by means of Scanning Electron Microscopy (SEM) and optical microscopy. Bending deformation about 90° was achieved without any defect at high temperatures. The importance of a critical temperature and deformation rate was emphasized in maintaining the deformation. Significant slip lines on surface of cell walls at tensile zones of bending specimen were observed. At high strain rates, the microcrack formation in boundaries of elongated grains was determined.

Keywords: Al alloy, Closed cell, Hot deformation, Metallic foam

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Authors: Sana Abbasa, Shahzad Bhattiab, Nadir Khan

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Sargassum siliquastrum is brown seaweed with traditional claims for some medicinal properties. This research was done to investigate the methanol extract of S. siliquastrum for antiproliferative activity against human cervical cancer cell line, HeLa and its mode of cell death. From methylene blue assay, S. siliquastrum exhibited antiproliferative activity on HeLa cells with IC50 of 3.87 µg/ml without affecting non-malignant cells. Phase contrast microscopy indicated the confluency reduction in HeLa cells and changes on the cell shape. Nuclear staining with Hoechst 33258 displayed the formation of apoptotic bodies and fragmented nuclei. S. siliquastrum also induced early apoptosis event in HeLa cells as confirmed by FITC-Annexin V/propidium iodide staining by flow cytometry analysis. Cell cycle analysis indicated growth arrest of HeLa cells at G1/S phase. Protein study by flow cytometry indicated the increment of p53, slight increase of Bax and unchanged level of Bcl-2. In conclusion, S. siliquastrum demonstrated an antiproliferative activity in HeLa cell by inducing G1/S cell cycle arrest via p53-mediated pathway.

Keywords: sargassum siliquastrum, cervical cancer, P53, antiproleferation

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Authors: Drashti G. Daraji, Rosa E. Moo-Puc, Hitesh D. Patel

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Substituted 2-Thioimidazole analogues have been synthesized and confirmed by advanced spectroscopic techniques. Among them, ten compounds have been selected and evaluated for their in vitro anti-cancer activity at the National Cancer Institute (NCI) for testing against a panel of 60 different human tumor cell lines derived from nine neoplastic cancer types. Furthermore, synthesized compounds were tested for their in vitro antiprotozoal activity, and none of them exhibited significant potency against antiprotozoans. It was observed that the tested all compounds seem effective on the UACC-62 melanoma cancer cell line as compared to other cancer cell lines and also exhibited the least potent in the Non-Small Cell Lung Cancer cell line in one-dose screening. In silico studies of these derivatives were carried out by molecular docking techniques and Absorption, Distribution, Metabolism, and Excretion (ADME) using Schrödinger software to find potent B-Raf kinase inhibitor (PDB ID: 3OG7). All the compounds have been performed for docking study; Compound D4 has a good docking score for melanoma cancer as compared with other.

Keywords: anticancer activity, cancer cell line, 2-thio imidazole, one-dose assay, molecular docking

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Authors: Soroosh Torabi, Brad Berron, Ren Xu, Christine Trinkle

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Microfluidics integrated with 3D cell culture is a powerful technology to mimic cellular environment, and can be used to study cell activities such as proliferation, migration and response to drugs. This technology has gained more attention in cancer studies over the past years, and many organ-on-a-chip systems have been developed to study cancer cell behaviors in an ex-vivo tumor microenvironment. However, there are still some barriers to adoption which include low throughput, complexity in 3D cell culture integration and limitations on non-optical analysis of cells. In this study, a user-friendly microfluidic multi-well plate was developed to mimic the in vivo tumor microenvironment. The microfluidic platform feeds multiple 3D cell culture sites at the same time which enhances the throughput of the system. The platform uses hydrophobic Cassie-Baxter surfaces created by microchannels to enable convenient loading of hydrogel/cell suspensions into the device, while providing barrier free placement of the hydrogel and cells adjacent to the fluidic path. The microchannels support convective flow and diffusion of nutrients to the cells and a removable lid is used to enable further chemical and physiological analysis on the cells. Different breast cancer cell lines were cultured in the device and then monitored to characterize nutrient delivery to the cells as well as cell invasion and proliferation. In addition, the drug response of breast cancer cell lines cultured in the device was compared to the response in xenograft models to the same drugs to analyze relevance of this platform for use in future drug-response studies.

Keywords: microfluidics, multi-well 3d cell culture, tumor microenvironment, tumor-on-a-chip

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Authors: Norihiro Kato, Yuriko Takayama

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Some gram-negative bacteria enable the simultaneous activation of gene expression involved in N-acylhomoserine lactone (AHL) dependent cell-to-cell communication system. Such regulatory system for the bacterial group behavior is termed as quorum sensing (QS) because a diffusible AHL signal can accumulate around the cell during the increase of the cell density and trigger activation of the sequential QS process. By blocking the QS, the expression of diverse genes related to infection, antibiotic production, and biofilm formation is inhibited. Conditioning of QS by regulation of the DNA-receptor-AHL interaction is a potential target for enhancing host defenses against pathogenicity. We focused on engineered application of transcriptional regulator SpnR produced in opportunistic human pathogen Serratia marcescens. The SpnR can interact with AHL signals at an N-terminal domain and also with a promoter region of a QS target gene at a C-terminal domain. As the initial process of the QS activation, the SpnR forms a complex with the AHL to enhance the expression of pig cluster; the SpnR normally acts as an activator for the expression of the QS-dependent gene. In this research, we attempt to artificially control QS by changing the role of SpnR. The QS-dependent prodigiosin production is expected to inhibit by externally added SpnR in the culture broth of AS-1 strain because the AHL concentration was kept below the threshold by AHL-SpnR complex formation. Maltose-binding protein (MBP)-tagged SpnR (MBP-SpnR) was overexpressed in Escherichia coli and purified using an affinity chromatography equipped with an amylose resin column. The specific interaction between AHL and MBP-SpnR was demonstrated by quartz crystal microbalance (QCM) sensor. AHL with amino end-group was coupled with COOH-terminated self-assembled monolayer prepared on a gold electrode of 27-MHz quartz crystal sensor using water-soluble carbodiimide. After the injection of MBP-SpnR into a cup-type sensor cell filled with the buffer solution, time course of resonant frequency change (ΔFs) was determined. A decrease of ΔFs clearly showed the uptake of MBP-SpnR onto the AHL-immobilized electrode. Furthermore, no binding affinity was observed after the heat-inactivation of MBP-SpnR at 80ºC. These results suggest that MBP-SpnR possesses a specific affinity for AHL. MBP-SpnR was added to the culture medium as an AHL trap to study inhibitory effects on intracellularly accumulated prodigiosin. With approximately 2 µM MBP-SpnR, the amount of prodigiosin induced was half that of the control without any additives. In conclusion, the function of SpnR could be switched by adding it to the cell culture. Exogenously added MBP-SpnR possesses high affinity for AHL derived from cells and acts as an inhibitor of AHL-mediated QS.

Keywords: intracellular signaling, microbial biotechnology, quorum sensing, transcriptional regulator

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Authors: Remzi Şahin, Sadık Ata, Kevser Dincer

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In this study, performance of proton exchange membrane (PEM) fuel cell was experimentally investigated. The efficiency of energy conversion in PEM fuel cells is dependent on the catalytic activities of the catalysts used in the cathode and anode of membrane electrode assemblies. Membrane is considered the heart of PEM fuel cells without which they cannot produce electricity. PEM fuel cell performance increased with coating carbon nanotube (CNT). CNT show a unique combination of stiffness, strength, and tenacity compared to other fiber materials which usually lack one or more of these properties. Two different experiments were performed and the membrane performance has been determined by repeating the two experiments that were done before coating. The purposes of these experiments are the observation of power change due to a temperature change in the same voltage value.

Keywords: carbon nanotube (CNT), proton exchange membrane (PEM), fuel cell, spin method

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Authors: Mahfoodha Al-Kitani, Dylan Thompson, Keith Stokes

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Sickle cell disease (SCD) and sickle cell trait (SCT) are the most common hematological diseases in Oman according to the national survey of genetic blood disorders. The aim of this study was to determine markers of physical fitness and anthropometrics indices in children with sickle cell disease and children with sickle cell trait and compare them with normal healthy children of the same age. One hundred and twenty male children participated in the present study divided to three groups: 40 with sickle disease (SCD; age, 13.3(.80), height, 131.9(3.5), mass, 29.2(3.1)); 40 with sickle cell trait (SCT; age, 12.2(.80), height, 141.0(9.9), mass, 38.0(4.4)); and 40 controls with normal hemoglobin (Con; age, 12.8(.80), height, 139.4(8.7), mass, 37.2(4.3)). All children completed a 5-min running exercise test on a treadmill at speed corresponding to 5 km/hr. Heart rate and was recorded during exercise and during 10-min of recovery. Blood lactate was measured before and 5 min after the completion of exercise. Children with SCD exhibited a higher mean value (P < 0.05) for percent body fat and fat mass than the normal healthy subjects and SCT subjects. Resting values of hemoglobin were similar in SCT (11.04(.78)) and control (10.8(94)) groups, and lower in SCD (8.89(.54); P < 0.05). There was a strong correlation between peak heart rate and resting hemoglobin levels for the three groups (r= -.472. n= 120, p < .0005).The SCD group (175.2(10.3)) exhibited higher mean heart rate during exercise than those observed in the SCT (143.7(9.5)) and normal control children (144.5(22.4); P < 0.05). Additionally, SCD children showed higher serum lactate values before and after treadmill exercise compared to the other groups (P < 0.05). Children with sickle cell trait demonstrate similar physical fitness level and similar exercise responses to treadmill stress test to normal children. In contrast, SCD children have lower body mass, higher fat mass and lower physical fitness than children with SCT and healthy controls.

Keywords: sickle cell disease, sickle cell trait, children, exercise

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Authors: Tan Yong Sheng Edgar, Yeong Wai Yee

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Age-related macular degeneration (AMD) is one of the leading cause of blindness. It can cause severe visual loss due to damaged retinal pigment epithelium (RPE). RPE is an important component of the retinal tissue. It functions as a transducing boundary for visual perception making it an essential factor for sight. The RPE also functions as a metabolically complex and functional cell layer that is responsible for the local homeostasis and maintenance of the extra photoreceptor environment. Thus one of the suggested method of treating such diseases would be regenerating these RPE cells. As such, we intend to grow these cells using a synthetic scaffold to provide a stable environment that reduces the batch effects found in natural scaffolds. Stiffness of the scaffold will also be investigated to determine the optimal Young’s modulus for cultivating these cells. The cells will be generated into a monolayer cell sheet and their functions such as formation of tight junctions and gene expression patterns will be assessed to evaluate the cell sheet quality compared to a native RPE tissue.

Keywords: RPE, scaffold, characterization, biomaterials, colloids and nanomedicine

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Authors: Dong-An Wang

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All kinds of microspheres have been extensively employed as carriers for drug, gene and therapeutic cell delivery. Most therapeutic cell delivery microspheres rely on a two-step methodology: fabrication of microspheres and subsequent seeding of cells onto them. In this study, we have developed a novel one-step cell encapsulation technique using a convenient and instant water-in-oil single emulsion approach to form cell-encapsulated gelatin microspheres. This technology is adopted for hyaline cartilage tissue engineering, in which autologous chondrocytes are used as therapeutic cells. Cell viability was maintained throughout and after the microsphere formation (75-100 µm diameters) process that avoids involvement of any covalent bonding reactions or exposure to any further chemicals. Further encapsulation of cell-laden microspheres in alginate gels were performed under 4°C via a prompt process. Upon the formation of alginate constructs, they were immediately relocated into CO2 incubator where the temperature was maintained at 37°C; under this temperature, the cell-laden gelatin microspheres dissolved within hours to yield similarly sized cavities and the chondrocytes were therefore suspended within the cavities inside the alginate gel bulk. Hence, the gelatin cell-laden microspheres served two roles: as cell delivery vehicles which can be removable through temperature curing, and as porogens within an alginate hydrogel construct to provide living space for cell growth and tissue development as well as better permeability for mutual diffusions. These cell-laden microspheres, namely “temperature-cured dissolvable gelatin microsphere based cell carriers” (tDGMCs), were further encapsulated in a chondrocyte-laden alginate scaffold system and analyzed by WST-1, gene expression analyses, biochemical assays, histology and immunochemistry stains. The positive results consistently demonstrated the promise of tDGMC technology in delivering these non-anchorage dependent cells (chondrocytes). It can be further conveniently translated into delivery of other non-anchorage dependent cell species, including stem cells, progenitors or iPS cells, for regeneration of tissues in internal organs, such as engineered hepatogenesis or pancreatic regeneration.

Keywords: biomaterials, tissue engineering, microsphere, hydrogel, porogen, anchorage dependence

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Authors: Violina R. Angelova, Venelina T. Popova, Radka V. Ivanova, Givko T. Ivanov, Krasimir I. Ivanov

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A comparative research on the impact of compost on uptake and allocation of nutrients and heavy metals and quality of Oriental tobacco Krumovgrad 90 has been carried out. The experiment was performed on an agricultural field contaminated by the lead zinc smelter near the town of Kardzali, Bulgaria, after closing the lead production. The compost treatments had significant effects on the uptake and allocation of plant nutrients and heavy metals. The incorporation of compost leads to decrease in the amount of heavy metals present in the tobacco leaves, with Cd, Pb and Zn having values of 36%, 12% and 6%, respectively. Application of the compost leads to increased content of potassium, calcium and magnesium in the leaves of tobacco, and therefore, may favorably affect the burning properties of tobacco. The incorporation of compost in the soil has a negative impact on the quality and typicality of the oriental tobacco variety of Krumovgrad 90. The incorporation of compost leads to an increase in the size of the tobacco plant leaves, the leaves become darker in colour, less fleshy and undergo a change in form, becoming (much) broader in the second, third and fourth stalk position. This is accompanied by a decrease in the quality of the tobacco. The incorporation of compost also results in an increase in the mineral substances (pure ash), total nicotine and nitrogen, and a reduction in the amount of reducing sugars, which causes the quality of the tobacco leaves to deteriorate (particularly in the third and fourth harvests).

Keywords: chemical composition, compost, heavy metals, oriental tobacco, quality

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Authors: Folajinmi Oluwasina, Magbagbeola Dairo, Ikeoluwapo Ajayi

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Contraception has proven to be an effective way of controlling fertility and spacing births. Studies have shown that contraception can avert the high-risk pregnancies and consequently reduce maternal deaths up to 32%. Uptake of modern contraception is promoted as a mechanism to address the reproductive health needs of men and women, as well as the crucial challenge of rapid population increase. A cross- sectional descriptive study using a two- stage systematic sampling technique was used to select 530 women of reproductive age out of 20,000 households. Respondents were interviewed using a semi-structured questionnaire. Knowledge was assessed on a 5 point score in which a score of ≤ 2 rated poor while perception was scored on 36 points score in which a score of ≤ 18 was rated low. Data were analyzed using descriptive statistics, Chi-square test and logistic regression at p< 0.05. There were 530 respondents. Age of respondents was 30.3 ±7.8 years, and 73.0% were married. About 90% had good knowledge of contraception while 60.8% had used contraceptives. The commonest source of information about contraception was mass media (72.8%). Minority (26.1%) obtained husbands approval before using contraceptive while 20.0% had used modern contraceptives before the first birth. Many (54.5%) of the respondents agreed that contraception helps in improving standard of living and 64.7% had good perception about contraception. Factors that hindered effective uptake of contraception services included poor service provider’s attitude (33.3%) and congestion at the service centers (4.5%). Respondents with nonuse of contraceptive before first birth are less likely to subsequently use contraceptives (OR= 0.324, 95% CI= 0.1-0.5). Husband approval of contraceptives use was the major determinant of women’s contraceptive use (OR = 3.4, 95% CI = 1.3-8.7). Respondents who had family planning centers not more than 5 kilometers walking distance to their residence did not significantly use contraception services (41.5%) more than 21.1% of those who had to take means of transportation to the service venues. This study showed that majority of the respondents were knowledgeable and aware of contraception services, but husband’s agreement on the use of modern contraceptives remains poor. Programmes that enhances husbands approval of modern contraception is thus recommended.

Keywords: contraception services, service provider’s attitude, uptake, husbands approval

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Authors: Grecco M. Ante, Princess Rochelle O. Gan

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Plants that can absorb greater than 10,000 µg Ni/g dry mass in their stems and leaves are termed as ‘hypernickelophores’. Chelators are chemicals that make the metals in the soil more soluble, making them a potential enhancer for phytoextraction. This study aims to observe the effect of different concentrations of the chelating agent ethylene diamine tetraacetate (EDTA) on the metal uptake (or rate of phytoextraction) of Nickel by Phyllanthus sp. nov. The plant is found to be a hyperickelophore in normal conditions. The addition of EDTA increased the metal uptake of the plant. The increasing amount of the chelating agent causes a decrease in the phytoextraction of the plant but moves the onset of its peak of maximum nickel content in its tissue to an earlier time. The chelator-assisted phytoextraction of nickel by Phyllanthus sp. nov. is proven to be an efficient auxiliary mining operation for nickel laterite mines.

Keywords: phytomining, Phyllanthus sp. nov., EDTA, nickel, laterite

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Authors: Cheng-Cao Sun, Shu-Jun Li, De-Jia Li

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Determinants of growth and metastasis in cancer remain of great interest to define. MicroRNAs (miRNAs) have frequently emerged as tumor metastatic regulator by acting on multiple signaling pathways. Here, we report the definition of miR-346 as an oncogenic microRNA that facilitates non-small cell lung cancer (NSCLC) cell growth and metastasis. XPC, an important DNA damage recognition factor in nucleotide excision repair was defined as a target for down-regulation by miR-346, functioning through direct interaction with the 3'-UTR of XPC mRNA. Blocking miR-346 by an antagomiR was sufficient to inhibit NSCLC cell growth and metastasis, an effect that could be phenol-copied by RNAi-mediated silencing of XPC. In vivo studies established that miR-346 overexpression was sufficient to promote tumor growth by A549 cells in xenografts mice, relative to control cells. Overall, our results defined miR-346 as an oncogenic miRNA in NSCLC, the levels of which contributed to tumor growth and invasive aggressiveness.

Keywords: microRNA-346, miR-346, XPC, non-small cell lung cancer, oncogenesis

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Authors: Shafieeh Mansoori, Fatemeh Yazdian, Ashrafsadat Hatamian, Majid Azizi

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Monascus purpureus, which has a special metabolite with many therapeutic and medicinal properties including antioxidant, antibiotic, anti-hypercholesterolemia, and immunosuppressive properties, is a traditional Chinese fermentation fungus and is used as a natural dietary supplement. Production of desired metabolites actually determined by optimized growth which is supported by some factors such as substrates and Monascus strains type, moisture content of the fermentation mixture, aeration, and control of contamination issues. In this experiment, M. purpureus PTCC5305 was cultured in both the liquid and solid culture medium. The former medium contain YMP (yeast extract, maltose and peptone), PGC (peptone, glucose complex), and GYP (glucose, yeast extract and peptone) medium. After 8 days, the best medium for the cell production was PGC agar medium on solid culture with 0.28 g dry weight of cell mass whereas the best liquid culture was GYP medium with 3.5 g/l dry weight of cell mass. The lowest cell production was on YMP agar with 0.1 g dry weight of cell mass and then YMP medium with 2.5 g/l dry cell weight.

Keywords: Monascus purpureus, solid state fermentation, submerged culture, Chinese fermentation fungus

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Authors: Armin Khaghani Boroujeni, Leila Dehghani, Parham Talebi Boroujeni, Sahar Rostamian, Ali Asilian

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Background: Hair loss is a common complaint observed in both genders. Androgenetic alopecia is known pattern for hair loss. To assess new regenerative strategies (PRP, A-SC-BT, conditioned media, exosome-based treatments) compared to conventional therapies for hair loss or hair regeneration, an updated review was undertaken. To address this issue, we carried out this systematic review to comprehensively evaluate the efficacy of cell-based therapies on hair loss. Methods: The available online databases, including ISI Web of Science, Scopus, and PubMed, were searched systematically up to February 2022. The quality assessment of included studies was done using the Cochrane Collaboration's tool. Results: As a result, a total of 90 studies involving 2345 participants were included in the present study. The enrolled studies were conducted between 2010 and 2022. The subjects’ mean age ranged from 19 to 55.11 years old. Approaches using platelet rich plasma (PRP) provide a beneficial impact on hair regrowth. However, other cell-based therapies, including stem cell transplant, stem cell-derived conditioned medium, and stem cell-derived exosomes, revealed conflicting evidence. Conclusion: However, cell-based therapies for hair loss are still in their infancy, and more robust clinical studies are needed to better evaluate their mechanisms of action, efficacy, safety, benefits, and limitations. In this review, we provide the resources to the latest clinical studies and a more detailed description of the latest clinical studies concerning cell-based therapies in hair loss.

Keywords: cell-based therapy, exosome, hair restoration, systematic review

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Authors: Qi Liu, Jiqin Yao, Xiaohu Zhou, Bo Zheng

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The first artificial cell was produced by Thomas Chang in the 1950s when he was trying to make a mimic of red blood cells. Since then, many different types of artificial cells have been constructed from one of the two approaches: a so-called bottom-up approach, which aims to create a cell from scratch, and a top-down approach, in which genes are sequentially knocked out from organisms until only the minimal genome required for sustaining life remains. In this project, bottom-up approach was used to build a new cell-free expression system which mimics artificial cell that capable of protein expression and communicate with each other. The artificial cells constructed from the bottom-up approach are usually lipid vesicles, polymersomes, hydrogels or aqueous droplets containing the nucleic acids and transcription-translation machinery. However, lipid vesicles based artificial cells capable of communication present several issues in the cell communication research: (1) The lipid vesicles normally lose the important functions such as protein expression within a few hours. (2) The lipid membrane allows the permeation of only small molecules and limits the types of molecules that can be sensed and released to the surrounding environment for chemical communication; (3) The lipid vesicles are prone to rupture due to the imbalance of the osmotic pressure. To address these issues, the hydrogel-based artificial cells were constructed in this work. To construct the artificial cell, polyacrylamide hydrogel was functionalized with Acrylate PEG Succinimidyl Carboxymethyl Ester (ACLT-PEG2000-SCM) moiety on the polymer backbone. The proteinaceous factors can then be immobilized on the polymer backbone by the reaction between primary amines of proteins and N-hydroxysuccinimide esters (NHS esters) of ACLT-PEG2000-SCM, the plasmid template and ribosome were encapsulated inside the hydrogel particles. Because the artificial cell could continuously express protein with the supply of nutrients and energy, the artificial cell-artificial cell communication and artificial cell-natural cell communication could be achieved by combining the artificial cell vector with designed plasmids. The plasmids were designed referring to the quorum sensing (QS) system of bacteria, which largely relied on cognate acyl-homoserine lactone (AHL) / transcription pairs. In one communication pair, “sender” is the artificial cell or natural cell that can produce AHL signal molecule by synthesizing the corresponding signal synthase that catalyzed the conversion of S-adenosyl-L-methionine (SAM) into AHL, while the “receiver” is the artificial cell or natural cell that can sense the quorum sensing signaling molecule form “sender” and in turn express the gene of interest. In the experiment, GFP was first immobilized inside the hydrogel particle to prove that the functionalized hydrogel particles could be used for protein binding. After that, the successful communication between artificial cell-artificial cell and artificial cell-natural cell was demonstrated, the successful signal between artificial cell-artificial cell or artificial cell-natural cell could be observed by recording the fluorescence signal increase. The hydrogel-based artificial cell designed in this work can help to study the complex communication system in bacteria, it can also be further developed for therapeutic applications.

Keywords: artificial cell, cell-free system, gene circuit, synthetic biology

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Authors: Mojdeh Mohseni

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In the context of tissue engineering, the effect of microtopography as afforded by scaffold morphology is an important design parameter. Since the morphology of pores can effect on cell behavior, in this study, porous Chitosan (CHIT) - Gelatin (GEL)- Alginate (ALG) scaffolds with microtubule orientation structure were manufactured by unidirectional freeze-drying method and the effect of pore morphology on differentiation of Mesenchymal Stem Cells (MSCs) was investigated. This study showed that, the provided scaffold with natural polymer had good properties for cell behavior and the pores with highest orientation rate have produced appropriate substrate for the differentiation of stem cells.

Keywords: Chitosan, gelatin, Alginate, pore morphology, stem cell differentiation

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Authors: Zahra Shokrolahi, Mohammad Reza Atashzar

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The goals of treating patients with breast cancer are to cure the disease, prolong survival, and improve quality of life. Immune cells in the tumor microenvironment have an important role in regulating tumor progression. The term of cellular immunotherapy refers to the administration of living cells to a patient; this type of immunotherapy can be active, such as a dendritic cell (DC) vaccine, in that the cells can stimulate an anti-tumour response in the patient, or the therapy can be passive, whereby the cells have intrinsic anti-tumour activity; this is known as adoptive cell transfer (ACT) and includes the use of autologous or allogeneic lymphocytes that may, or may not, be modified. The most important breast cancer cellular immunotherapies involving the use of T cells and natural killer (NK) cells in adoptive cell transfer, as well as dendritic cells vaccines. T cell-based therapies including tumour-infiltrating lymphocytes (TILs), engineered TCR-T cells, chimeric antigen receptor (CAR T cell), Gamma-delta (γδ) T cells, natural killer T (NKT) cells. NK cell-based therapies including lymphokine-activated killers (LAK), cytokine-induced killer (CIK) cells, CAR-NK cells. Adoptive cell therapy has some advantages and disadvantages some. TILs cell strictly directed against tumor-specific antigens but are inactive against tumor changes due to immunoediting. CIK cell have MHC-independent cytotoxic effect and also need concurrent high dose IL-2 administration. CAR T cell are MHC-independent; overcome tumor MHC molecule downregulation; potent in recognizing any cell surface antigen (protein, carbohydrate or glycolipid); applicable to a broad range of patients and T cell populations; production of large numbers of tumor-specific cells in a moderately short period of time. Meanwhile CAR T cells capable of targeting only cell surface antigens; lethal toxicity due to cytokine storm reported. Here we present the most popular cancer cellular immunotherapy approaches and discuss their clinical relevance referring to data acquired from clinical trials .To date, clinical experience and efficacy suggest that combining more than one immunotherapy interventions, in conjunction with other treatment options like chemotherapy, radiotherapy and targeted or epigenetic therapy, should guide the way to cancer cure.

Keywords: breast cancer , cell therapy , CAR T cell , CIK cells

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Authors: Violina Angelova

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A comparative study has been carried out into the impact of organic meliorants on the uptake of heavy metals, micro and macroelements and the phytoremediation potential of enhanced tobacco BAC F3. The soil used as part of this experiment was sampled from the vicinity of the Non-Ferrous-Metal Works near Plovdiv, Bulgaria. The pot experiment carried out consisted of a randomized, complete block design containing nine treatments and three replications (27 pots). The treatments consisted of a control (with no organic meliorants) and compost and vermicompost meliorants (added at 5%, 10%, 15%, and 30%, and recalculated based on their dry soil weight). Upon reaching commercial ripeness, the tobacco plants were gathered. Heavy metals, micro and macroelement contents in roots, stems, and leaves of tobacco were analyzed by the method of the microwave mineralization. To determine the elements in the samples, inductively coupled emission spectrometry (Jobin Yvon Emission - JY 38 S, France) was used. The distribution of the heavy metals, micro, and macroelements in the organs of the enhanced tobacco has a selective character and depended above all on the parts of the plants and the element that was examined. Pb, Zn, Cu, Fe, Mn, P and Mg distribution in tobacco decreases in the following order: roots > leaves > stems, and for Cd, K, and Ca - leaves > roots > stems. The high concentration of Cd in the leaves and the high translocation factor indicate the possibility of enhanced tobacco to be used in phytoextraction. Tested organic amendments significantly influenced the uptake of heavy metals, micro and macroelements by the roots, stems, and leaves of tobacco. A correlation was found between the quantity of the mobile forms and the uptake of Pb, Zn, and Cd by the enhanced tobacco. The compost and vermicompost treatments significantly reduced heavy metals concentration in leaves and increased uptake of K, Ca and Mg. The 30% compost and 30% vermicompost treatments led to the maximal reduction of heavy metals in enhanced tobacco BAC F3. The addition of compost and vermicompost further reduces the ability to digest the heavy metals in the leaves, and phytoremediation potential of enhanced tobacco BAC F3. Acknowledgment: The financial support by the Bulgarian National Science Fund Project DFNI Н04/9 is greatly appreciated.

Keywords: heavy metals, micro and macroelements, enhanced tobacco BAC F3, phytoremediation, organic meliorants

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Authors: Suhadiyana Hanapi, Alhassan Salami Tijani, W. A. N Wan Mohamed

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In this paper, a prototype PEM fuel cell vehicle integrated with a 1 kW air-blowing proton exchange membrane fuel cell (PEMFC) stack as a main power sources has been developed for a lightweight cruising vehicle. The test vehicle is equipped with a PEM fuel cell system that provides electric power to a brushed DC motor. This vehicle was designed to compete with industrial lightweight vehicle with the target of consuming least amount of energy and high performance. Individual variations in driving style have a significant impact on vehicle energy efficiency and it is well established from the literature. The primary aim of this study was to assesses the power and fuel consumption of a hydrogen fuel cell vehicle operating at three difference driving technique (i.e. 25 km/h constant speed, 22-28 km/h speed range, 20-30 km/h speed range). The goal is to develop the best driving strategy to maximize performance and minimize fuel consumption for the vehicle system. The relationship between power demand and hydrogen consumption has also been discussed. All the techniques can be evaluated and compared on broadly similar terms. Automatic intelligent controller for driving prototype fuel cell vehicle on different obstacle while maintaining all systems at maximum efficiency was used. The result showed that 25 km/h constant speed was identified for optimal driving with less fuel consumption.

Keywords: prototype fuel cell electric vehicles, energy efficient, control/driving technique, fuel economy

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Authors: Isobel Hambleton, Mark Carrington

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Trypanosoma brucei, the causal agent of a range of diseases in humans and livestock, evades the mammalian immune system through a population survival strategy based on the expression of a series of antigenically distinct variant surface glycoproteins (VSGs). RNAi mediated knockdown of the active VSG gene triggers a precytokinesis cell cycle arrest. To determine whether this phenotype is the result of reduced VSG transcript or depleted VSG protein, we used morpholino antisense oligonucleotides to block translation of VSG mRNA. The same precytokinesis cell cycle arrest was observed, suggesting that VSG protein abundance is monitored closely throughout the cell cycle. An inducible expression system has been developed to test various GPI-anchored proteins for their ability to rescue this cell cycle arrest. This system has been used to demonstrate that wild-type VSG expressed from a T7 promoter rescues this phenotype. This indicates that VSG expression from one of the specialised bloodstream expression sites (BES) is not essential for cell division. The same approach has been used to define the minimum essential features of a VSG necessary for function.

Keywords: bloodstream expression site, morpholino, precytokinesis cell cycle arrest, variant surface glycoprotein

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Authors: Sanjay Tushar Choudhary

Abstract:

This paper focuses on the thermodynamic analysis of Solid Oxide Fuel Cell (SOFC). In the present work the SOFC has been modeled to work with internal reforming of fuel which takes place at high temperature and direct energy conversion from chemical energy to electrical energy takes place. The fuel-cell effluent is a high-temperature steam which can be used for co-generation purposes. Syn-gas has been used here as fuel which is essentially produced by steam reforming of methane in the internal reformer of the SOFC. A thermodynamic model of SOFC has been developed for planar cell configuration to evaluate various losses in the energy conversion process within the fuel cell. Cycle parameters like fuel utilization ratio and the air-recirculation ratio have been varied to evaluate the thermodynamic performance of the fuel cell. Output performance parameters like terminal voltage, cell-efficiency and power output have been evaluated for various values of current densities. It has been observed that a combination of a lower value of air-circulation ratio and higher values of fuel utilization efficiency gives a better overall thermodynamic performance.

Keywords: current density, SOFC, suel utilization factor, recirculation ratio

Procedia PDF Downloads 477

Authors: Young Cheol Choi, Eun-Jin Moon, Sung-Won Yoo, Sang-Hwa Jung, In-Hwan Yang

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Stainless steel slag (STS) is a by-product generated from the stainless steel refining process. The recycling of STS produced in Korea for construction applications is limited due to its poor hydraulic properties. On the other hand, STS has high carbonation reactivity to CO2 as it contains gamma-C2S content. This material is ideal for mineral carbonation which is one of the techniques proposed for carbon emission reduction. The objective of this study is to investigate the feasibility of developing a zero-cement STS binder activated by carbonation as alternative cementitious material. The quantitative analyses for CO2 uptake of STS powder and STS blended cement were investigated using thermogravimetric analysis (TGA), X-ray diffraction (XRD). In addition, the compressive strength and microstructure of STS pastes after CO2 curing were evaluated. Test results showed that STS can be activated by carbonation to gain a sufficient strength as alternative cementitious material.

Keywords: gamma-C2S, CO2 uptake, carbonation, stainless steel slag

Procedia PDF Downloads 442

Authors: Hamed Karimian, Noraziah Nordin, Mohamad Ibrahim Noordin, Syam Mohan, Mahboubeh Razavi, Najihah Mohd Hashim, Happipah Mohd Ali

Abstract:

Satureja bachtiarica Bunge is a perennial medicinal plant belonging to the Lamiaceae family and endemic species in Iran. Satureja bachtiarica Bunge with the local name of Marzeh koohi is edible vegetable use as flavoring agent, anti-bacterial and to relieve cough and indigestion. In this study, the anti-cancer effect of Satureja bachtiarica Bunge on the MDA-MB-231 cell line as an Breast cancer cell model has been analyzed for the first time. Satureja bachtiarica Bunge was extracted using different solvents in the order of increasing polarity. Cytotoxicity activity of hexane extract of Satureja bachtiarica Bunge (SBHE) was observed using MTT assay. Acridine orange/Propidium iodide staining was used to detect early apoptosis; Annexin-V-FITC assay was carried out to observe the detection of cell-surface Phosphatidylserine (PS), with Annexin-Vserving as a marker for apoptotic cells. Caspase 3/7, 8 and-9 assays showed significantly activation of caspase-9 where lead intrinsic mitochondrial pathway. Bcl-2/Bax expressions and cell cycle arrest were also investigated. SBHE had exhibited significantly higher cytotoxicity against MDA-MB-231 Cell line compare to other cell lines. A significant increase in chromatin condensation in the cell nucleus was observed by fluorescence analysis. Treatment of MDA-MB-231 cells with SBHE encouraged apoptosis, by down-regulating Bcl-2 and up-regulating Bax, which lead the activation of caspase 9. Moreover, SBHE treatment significantly arrested MDA-MB-231 cells in the G1 phase. Together, the results presented in this study demonstrated that SBHE inhibited the proliferation of MDA-MB-231 cells, leading cell cycle arrest and programmed cell death, which was confirmed to be through the mitochondrial pathway.

Keywords: Satureja bachtiarica Bunge, MDA-MB-231, apoptosis, annexin-V, cell cycle

Procedia PDF Downloads 314

Authors: Raed Kouta

Abstract:

A fuel cell is a multi-physical system. Its electrical performance depends on chemical, electrochemical, fluid, and mechanical parameters. Many studies focus on physical and chemical aspects. Our study contributes to the evaluation of the influence of mechanical aspects on the performance of a fuel cell. This study is carried out as part of a reliability approach. Reliability modeling allows to consider the uncertainties of the incoming parameters and the probabilistic modeling of the outgoing parameters. The fuel cell studied is the one often used in land, sea, or air transport. This is the Low-Temperature Proton Exchange Membrane Fuel Cell (PEMFC). This battery can provide the required power level. One of the main scientific and technical challenges in mastering the design and production of a fuel cell is to know its behavior in its actual operating environment. The study proposes to highlight the influence on the production of electrical energy: Mechanical design and manufacturing parameters and their uncertainties (Young module, GDL porosity, permeability, etc.). The influence of the geometry of the bipolar plates is also considered. An experimental design is proposed with two types of materials as well as three geometric shapes for three joining pressures. Other experimental designs are also proposed for studying the influence of uncertainties of mechanical parameters on cell performance. - Mechanical (static, dynamic) and thermal (tightening - compression, vibrations (road rolling and tests on vibration-climatic bench, etc.) loads. This study is also carried out according to an experimental scheme on a fuel cell system for vibration loads recorded on a vehicle test track with three temperatures and three expected performance levels. The work will improve the coupling between mechanical, physical, and chemical phenomena.

Keywords: fuel cell, mechanic, reliability, uncertainties

Procedia PDF Downloads 158

Authors: O. S. Adeyemi, C. G. Whiteley

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The present study determined the toxic potential of metal nanoparticles in cell culture system. Silver and gold nanoparticles were synthesized and characterized following established "green" protocols. The synthesized nanoparticles, in varying concentrations ranging from 0.1–100 µM were evaluated for toxicity in metastatic MDA-MB-231 cells. The nanoparticles promoted a generation of reactive oxygen species and reduced cell viability to less than 50% in the demonstration of cellular toxicity. The nanoparticles; gold and the silver-gold mixture had IC50 values of 56.65 and 18.44 µM respectively. The IC50 concentration for silver nanoparticles could not be determined. Furthermore, the probe of the cell death using flow cytometry and confocal microscopy revealed the partial involvement of apoptosis as well as necrosis. Our results revealed cellular toxicity caused by the nanoparticles but the mechanism remains yet undefined.

Keywords: cell death, nanomedicine, nanotoxicology, toxicity

Procedia PDF Downloads 355