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Lignin in Biological Systems
Published in Severian Dumitriu, Valentin Popa, Polymeric Biomaterials, 2020
In other experiments, a purified fraction EPS4 also obtained from LEM by ethanol precipitation followed by hydrophobic chromatography and gel filtration chromatography completely inhibited the HIV-1-induced cytopathic effect in vitro at concentrations of greater than or equal to 10 μg/mL. Chemical and spectral analysis revealed that EPS4 is composed of water-soluble lignins containing minor amounts of protein (3.2%) and sugars (12.2%). Taken together with the previously reported observation that EPS 4 promotes the activation of macrophages and the proliferation of bone marrow cells, the fraction appears to possess both an immunostimulating activity and anti-HIV effect in vitro (Suzuki et al. 1998).
Applications of Environmental Biotechnology to Bioremediation
Published in Donald L. Wise, Debra J. Trantolo, Remediation of Hazardous Waste Contaminated Soils, 2018
John Sanseverino, James T. Fleming, Armin Heitzer, Bruce M. Applegate, Gary S. Sayler
The rationale of this method is to lyse all the bacterial cells in a soil sample, extract the DNA, and analyze the soil for the degradative function of interest. The advantage of nucleic acid extraction over colony hybridization include (1) cultivation of the sample is not required, and therefore a representative yield of bacterial DNA is achieved;8,9 and (2) sampling, extraction, and analysis can be accomplished in less than five days instead of in over two weeks.10–13 Various methods have been developed to extract bacterial DNA from soils or sediments. Ogram et al.14 developed the direct lysis method to lyse bacterial cells within the soil matrix (Figure 3). Soil bacteria are lysed by mechanical shearing in a bead beater using 100-μm beads. Subsequently, the DNA is separated from the debris through a series of sodium phosphate washes and centrifugations and concentrated by polyethylene glycol or ethanol precipitation. The DNA can be further purified by a series of phenol:chloroform extractions and cesium chloride ultracentrifugation.
Extraction, Isolation and Utilisation of Bioactive Compounds from rice Waste
Published in Quan V. Vuong, Utilisation of Bioactive Compounds from Agricultural and Food Waste, 2017
Alkali solvents can be used to extract DFs by using sodium hydroxide and calcium hydroxide (Daou and Zhang 2013, Wan et al. 2014). Defatted rice bran was treated with 0.15N NaOH for 64.3 minutes in a ratio of 1:5. Starch was hydrolyzed by amylaze and amyl glucosidase. The soluble fiber was collected by ethanol precipitation and centrifugation, with the residue being insoluble fiber. The highest yields of total DFs and soluble DF were 31 per cent and 2.69 per cent, respectively (Daou and Zhang 2013). Soluble DF (7.89 per cent) in rice bran was extracted using a 1:30 ratio of 2 per cent Ca(OH) and defatted rice bran at 60°C over 4 hours (Wan et al. 2014). The starch was removed by amylase at 90°C for 15 minutes before submergence in 2 per cent Ca(OH)2. Another study extracted fiber from rice husk by using NaOH (4 per cent), followed by a bleaching process using NaCl 1.7 wt per cent in acetic acid buffer and an acid hydrolysis (H2SO4 10 M) (Johar et al. 2012). The concentration of final fiber reached 96 per cent, but the severe conditions of this method could limit the potential uses of the extracted fiber. The combination of enzymes and chemicals in DF extraction could increase the extractability and shorten the extraction time (Maphosa and Jideani 2016). This method is more environmental friendly than the chemical method.
Genetic variants within the COL5A1 gene are associated with ligament injuries in physically active populations from Australia, South Africa, and Japan
Published in European Journal of Sport Science, 2023
Javier Alvarez-Romero, Mary-Jessica N. Laguette, Kirsten Seale, Macsue Jacques, Sarah Voisin, Danielle Hiam, Julian A. Feller, Oren Tirosh, Eri Miyamoto-Mikami, Hiroshi Kumagai, Naoki Kikuchi, Nobuhiro Kamiya, Noriyuki Fuku, Malcolm Collins, Alison V. September, Nir Eynon
Saliva was collected using an Oragene DNA collection kit (DNA Genotek, ON, Canada). DNA was extracted using an ethanol precipitation and the PrepIT®-L2P reagent (DNA Genotek, ON, Canada) in accordance with the manufacturer’s instructions. The rs12722 and rs10628678 variants were genotyped by TaqMan® SNP Genotyping Assay (assay ID: C____370252_20 [rs12722] and AH20W1K [rs10628678]) and using real-time PCR systems LightCycler 480 (Roche Applied Science, Mannheim, Germany), or QuantStudio 5 (Thermo Fisher Scientific). Genotypes were called based on the TaqMan® assay results using LightCycler® 480 SW software (version 1.5, Roche Molecular Systems) or QuantStudioTM Design & Analysis software (version 1.4.3, Thermo Fisher Scientific). Similarly, duplicates, positive and negative controls were included in each experiment.
Electrospun polysaccharide scaffolds: wound healing and stem cell differentiation
Published in Journal of Biomaterials Science, Polymer Edition, 2022
Preethi G. U., Unnikrishnan B. S., Sreekutty J., Archana M. G., Deepa Mohan, Raveendran Pillai K., Sreelekha T. T.
The polysaccharide was isolated as per previously reported procedures [8]. Briefly, Tamarind seed kernels were obtained, shade dried, and powdered for the extraction of polysaccharides. Seed kernel powder was treated with petroleum ether (Merck Emsure, boiling point, 60 °C − 80 °C) for 72 hrs which initiated the removal of fat content. The treated powder was later boiled in distilled water for 4 hrs, cooled to room temperature, and centrifuged at 20000Xg for 15 min. Ethanol precipitation was done to the supernatant and centrifuged to collect the residue. The residue was dissolved in distilled water treated with chloroform to remove denatured protein. Ethanol precipitation was repeated on the aqueous portion, centrifuged and the residue dissolved in distilled water followed by dialysis across distilled water for about 48 hrs and concentrated. These steps were repeated, and the final sample was analyzed for its concentration using high-performance liquid chromatography (HPLC, Shimadzu). Later, the sample was lyophilized using CHRIST ALPHA 2-4 LD PLUS (Germany) and stored at 4 °C [11].
Different influence of sulfated chitosan with different sulfonic acid group sites on HUVECs behaviors
Published in Journal of Biomaterials Science, Polymer Edition, 2020
Guijuan Han, Xiaohui Xia, Zhicheng Pan, Yucheng Lin, Lihua Li, Yanpeng Jiao, Changren Zhou, Shan Ding
The gene expression levels of angiogenesis-related genes affected by different SCS or heparin were measured via real-time polymerase chain reaction (RT-PCR) assay. HUVECs cells were cultured on 6-well plates and incubation with different SCS and heparin at a concentration of 500 μg/mL in culture medium for 7 and 14 days. The total RNA was extracted using TRIzol reagent (Invitrogen) according to the manufacturer’s instructions and collected by ethanol precipitation. Subsequently, RNA was reverse transcribed to complementary DNA (cDNA) using the PrimeScript RT reagent kit (Invitrogen) in a 20 μL of reaction mixture. The expression levels of angiogenesis-related genes, including von Willebrand factor (vWF), CD31 and vascular endothelial growth factor (VEGF) were quantified using real-time PCR (ABI PRISM®7500 Sequence Detection System) with SYBR PremixEx Taq II (Invitrogen). The primers sequence for the target genes are listed in Table 1. The relative expression level of each gene of interest was normalized to that of the housekeeping gene β-actin. Normalized relative mRNA quantities were calculated by the 2−ΔΔCT method and reported as fold induction. Experiments were performed independently in triplicate.