Polycistronic lentivirus induced pluripotent stem cells from skin biopsies after long term storage,blood outgrowth endothelial cells and cells from milk teeth

The generation of human induced pluripotent stem cells (hiPSCs) requires the collection of donor tissue, but clinical circumstances in which the interests of patients have highest priority may compromise the quality and availability of cells that are eventually used for reprogramming. Here we compared (i) skin biopsies stored in standard physiological salt solution for up to two weeks (ii) blood outgrowth endothelial cells (BOECs) isolated from fresh peripheral blood and (iii) children's milk teeth lost during normal replacement for their ability to form somatic cell cultures suitable for reprogramming to hiPSCs. We derived all hiPSC lines using the same reprogramming method (a conditional (FLPe) polycistronic lentivirus) and under similar conditions (same batch of virus, fetal calf serum and feeder cells). Skin fibroblasts could be reprogrammed robustly even after long-term biopsy storage. Generation of hiPSCs from juvenile dental pulp cells gave similar high efficiencies, but that of BOECs was lower. In terms of invasiveness of biopsy sampling, biopsy storage and reprogramming efficiencies skin fibroblasts appeared best for the generation of hiPSCs, but where non-invasive procedures are required (e.g. for children and minors) dental pulp cells from milk teeth represent a valuable alternative.     

Fertilisation practice changes rhizosphere microbial community structure in the agroecosystem

Rhizosphere microbial community is important for the acquisition of soil nutrients and closely related to plant species. Fertilisation practice changed soil quality. With the hypothesis of stronger rhizosphere effect of plant on rhizosphere microbial community than fertilisation management, we designed this research based on a long‐term field experiment (1982–present). This study consists of no fertilisation (NF), mineral fertilisers (NPK), mineral fertilisers plus 7,500 kg/ha of wheat straw addition (WS) and mineral fertilisers plus 30,000 kg/ha of cow manure (CM). After analysing, we found that fertilisation management not only elevated crop yield but also affected crop rhizosphere microbial community structure. The influence of fertilisation practice on wheat rhizosphere microbial structure was stronger than that of wheat. For wheat rhizosphere bacterial community, it was significantly affected by soil water content (SWC), nitrogen (TN), phosphorus (TP), pH, available phosphorus (AVP) and nitrogen (AVN), dissolved organic nitrogen (DON) and carbon (DOC). Besides SWC, pH, AVP, AVN, TN, TP and DOC, the wheat rhizosphere fungi community was also significantly affected by soil organic matter (SOM) and available potassium (AVK). Moreover, compared to rhizosphere bacterial community, the influences of soil physiochemical properties on rhizosphere fungal community was stronger. In conclusion, fertilisation practice was the primary factor structuring rhizosphere microbial community by changing soil nutrients availabilities in the agroecosystem.     

Social media user’s knowledge on soil plant relationship in Saudi Arabia

The study aimed to investigate the soil-plant relationship by looking at knowledge levels of social media users. The study examined the relationship between the users’ characteristics and their knowledge on soil-plant relationship. Online survey was designed and distributed to gather the data. The number of response was received from 383 respondents where 375 participants provided completed information and the remaining eight responses were eliminated due to quality standards. The results show most of the participants (68.8%) are moderately depend on social media to acquire knowledge on soil-plant relationship and indicated “Twitter” as the highly utilized platform followed by the “Youtube”. Meanwhile, 48.8% of the participants indicated that social media have a high impact on their knowledge-based information on soil-plant relationship and agricultural contents. Inferential analysis shows there was a significant positive relationship (p < 0.01) between independent variables; Twitter, Youtube, Instagram, and Snapchat and the degree of trust on these platforms, and the level’s knowledge on soil-plant relationship (dependent variable). Only 13% of the variance accounted for the impact of social media on participants’ agricultural knowledge-base can be predicted from the combination of participants’ reliance on a set of social media platforms (Twitter, Instagram, Youtube, and Snapchat). The study revealed the trust and confidence of the users using social media on agricultural information/soil plant relationship had a medium effect in social and educational research.     

Characterization of CIM monoliths as enzyme reactors

The immobilization of the enzymes citrate lyase, malate dehydrogenase, isocitrate dehydrogenase and lactate dehydrogenase to CIM monolithic supports was performed. The long-term stability, reproducibility, and linear response range of the immobilized enzyme reactors were investigated along with the determination of the kinetic behavior of the enzymes immobilized on the CIM monoliths. The Michaelis-Menten constant K(m) and the turnover number k(3) of the immobilized enzymes were found to be flow-unaffected. Furthermore, the K(m) values of the soluble and immobilized enzyme were found to be comparable. Both facts indicate the absence of a diffusional limitation in immobilized CIM enzyme reactors.     

Contrasting Effects of Farmyard Manure (FYM) and Compost for Remediation of Metal Contaminated Soil

We investigated effect of farm yard manure (FYM) and compost applied to metal contaminated soil at rate of 1% (FYM-1, compost-1), 2% (FYM-2, compost-2), and 3% (FYM-3, compost-3). FYM significantly (P < 0.001) increased dry weights of shoots and roots while compost increased root dry weight compared to control. Amendments significantly increased nickel (Ni) in shoots and roots of maize except compost applied at 1%. FYM-3 and -1 caused maximum Ni in shoots (11.42 mg kg^?1) and roots (80.92 mg kg^?1), respectively while compost-2 caused maximum Ni (14.08 mg kg^?1) and (163.87 mg kg^?1) in shoots and roots, respectively. Plants grown in pots amended with FYM-2 and compost-1 contained minimum Cu (30.12 and 30.11 mg kg^?1) in shoots, respectively. FYM-2 and compost-2 caused minimum zinc (Zn) (59.08 and 66.0 mg kg^?1) in maize shoots, respectively. FYM-2 caused minimum Mn in maize shoots while compost increased Mn in shoots and roots compared to control. FYM and compost increased the ammonium bicarbonate diethylene triamine penta acetic acid (AB-DTPA) extractable Ni and Mn in the soil and decreased Cu and Zn. Lower remediation factors for all metals with compost indicated that compost was effective to stabilize the metals in soil compared to FYM.     

Nutrient Mineralization and Leaf Litter Preference by the Earthworm Pontoscolex corethrurus on Iron Ore Mine Wastes

Abstract The effects of the earthworm Pontoscolex corethrurus (Muller) on the rate of mineralization of cattle dung‐amended iron (Fe^{2 +} ) ore mine wastes and its preference for partially decomposed leaf litter with contrasting chemical composition were studied in pot trials. The growth and survival rates of earthworms showed significant positive correlations with percent of organic matter. During 96 days of exposure, the earthworms significantly increased exchangeable Ca^{2 +} , Mg^{2 +} , PO₄^{3 ?} and NH₄‐N. Iron ore mine wastes amended with 5–10% organic matter supported earthworm fauna better than mine wastes amended with 0–3% organic matter. The leaf litter preference shown by the earthworm was, in descending order, Phyllanthus reticulatus, Tamarindus indica, Anacardium occidentale, Casuarina equisetifolia, Acacia auriculiformis, and Eucalyptus camaldulensis. A significant positive correlation was observed between the survival and growth rates of earthworms and the nutrient contents of partially decomposed leaf litter. The first three plant species were significantly richer in nutrients, mainly organic carbon, calcium, phosphorus, and nitrogen, than the other two plant species. Acacia auriculiformis and E. camaldulensis litter were preferred less because of their high lignin and polyphenolic compounds, despite being rich in other macronutrients like nitrogen and phosphorus. It is concluded that the introduction of P. corethrurus to cattle dung‐amended (5–10%) iron ore mine wastes or revegetation of the sites with P. reticulatus, T. indica, and A. occidentale plant species should be attempted before earthworm introduction. The litter from these species acts as a source of food for earthworms, thereby hastening the process of restoration of abandoned iron ore mines of Goa, India.     

Recombinant dengue virus type 3 envelope domain III protein from Escherichia coli

Dengue is a public health problem of global significance for which there is neither an effective antiviral therapy nor a preventive vaccine. The envelope protein of dengue virus is the major antigen to elicit neutralizing antibody response and protective immunity in hosts. Optimization of culture media was carried out for enhanced production of recombinant dengue virus type 3 envelope domain III (rDen 3 EDIII) protein in E. coli. Further, batch and fed-batch cultivation process were also developed in optimized medium. After fed-batch cultivation, the dry cell weight was about 22.80 g/L of culture. The rDen 3 EDIII protein was purified using immobilized metal affinity chromatography. This process produced ～649 mg of purified rDen 3 EDIII protein per liter of culture. The purity of the protein was determined by SDS-PAGE analysis and the reactivity was checked by Western blotting as well as ELISA. These results show that the purified protein may be used for the dengue diagnosis or further prophylactic studies for dengue infection.     

Rapid cryopreservation of five mammalian and one mosquito cell line at −80°C while attached to flasks in a serum free cryopreservative

Cell culturing, and the requisite storage of cell lines at ultra-low temperatures, is used in most laboratories studying or using eukaryotic proteomics, genomics, microarray, and RNA technologies. In this study we have observed that A72(dog), CRFK(cat), NB324K(human), MCF7(human), WI38(human), and C636(mosquito) cells were effectively cryopreserved at −80°C while attached to the substratum of 25cm² tissue culture flasks. This was accomplished using a serum free crypreservative recently developed by Corsini and co-workers. The technique allows for significant savings of time and money in laboratories that rapidly process numerous cell lines.     

Enhancement of Hydrocarbon Removal in a Clay and Drilling-Waste Polluted Soil

The application of biological processes in restoring oil polluted sites is growing due to their efficiency in removing different classes of pollutants. The aim of this study was to determine the ability of microorganisms present in a drilling-waste polluted soil (36,200 mg TPH kg^?1 soil) to remove weathered hydrocarbons under stimulated and non-stimulated soil conditions. The hypothesis under study was whether petroleum hydrocarbons removal could be enhanced by manipulating C/N ratio, water content and addition of three agroindustrial wastes. A Box-Behnken design was employed to evaluate the effect of each variable. Results demonstrated that, for orange peels and banana trunk treatments, the variable with the largest effect (p < 0.01) on hydrocarbon removal was the C/N ratio, indicating that higher ratio (100/3) improved removal (79.5–82%). The largest effect (p < 0.001) on hydrocarbon removal for pineapple wastes was observed with higher water content (60%) achieving the highest removal (89%). After 90 days of experimentation, the type of agricultural waste and the agricultural waste/soil ratio were not statistically significant in any treatment. However, their addition was important relative to non-stimulated soil, which showed a hydrocarbon removal of 17%. Data reported in this study showed the application of bioremediation in clay and drilling waste-polluted soils.