5,399 research outputs found

    FLASH: ultra-fast protocol to identify RNA-protein interactions in cells

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    Determination of the in vivo binding sites of RNA-binding proteins (RBPs) is paramount to understanding their function and how they affect different aspects of gene regulation. With hundreds of RNA-binding proteins identified in human cells, a flexible, high-resolution, high-throughput, highly multiplexible and radioactivity-free method to determine their binding sites has not been described to date. Here we report FLASH (Fast Ligation of RNA after some sort of Affinity Purification for High-throughput Sequencing), which uses a special adapter design and an optimized protocol to determine protein-RNA interactions in living cells. The entire FLASH protocol, starting from cells on plates to a sequencing library, takes 1.5 days. We demonstrate the flexibility, speed and versatility of FLASH by using it to determine RNA targets of both tagged and endogenously expressed proteins under diverse conditions in vivo

    Bifenthrin toxicity, inheritance of resistance, cross-resistance to insecticides in Helicoverpa armigera

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    Aim of study: It is first report to sort out resistance development; its mode and inheritance in Helicoverpa armigera against bifenthrin till several generations using progeny reciprocal crosses and back crosses, combined with observing the cross resistance of bifenthrin against pyrethroid, organophosphate, pyrazole and new chemistry insecticides.Area of study: This study was conducted at agriculture fields of University of Agriculture, Faisalabad, Pakistan.Material and methods: Bifenthrin selected strain of H. armigera was reciprocally crossed to bifenthrin susceptible strains. Resulting F1 progeny was back-crossed to resistant strain. Cross resistance of bifenthrin to six insecticides (cypermethrin, triazophos, emamectin benzoate, fipronil, lambda-cyhalothrin, profenofos) was observed.Main results: Resistance ratio was higher in bifenthrin selected strain. h value showed that resistance was autosomal with incomplete dominance. Polygenic mode of resistance; resistance controlled by more than one gene; was found against bifenthrin in H. armigera. Cross resistance of bifenthrin selected strain against different insecticides was found higher.Research highlights: Reciprocal crosses of F1 progeny combined with LC50 exhibits that resistance can be controlled using multiple insecticides at different intervals against H. armigera. These results can be implicated to develop an integrated pest management strategy to control H. armigera

    Covalently cross-linked proteins & polysaccharides: Formation, characterisation and potential applications

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    This review presents recent research conducted on the development of various protein-polysaccharide conjugates, their functional properties and industrial applications. These conjugates are formed by the glycosylation of food proteins with carbohydrates via the Maillard reaction and are capable of improving the functional properties of proteins. The Maillard reaction facilitates covalent bonding between a reducing group of a carbohydrate and an amino group of a protein under controlled conditions of temperature, time, pH, and relative humidity. There is a great deal of interest in modifying the functional properties of proteins and in the use of novel conjugates for various industrial applications. This review discusses various methods and their implications for preparing and characterising these conjugates. Furthermore, the physicochemical properties of conjugates such as solubility, thermal stability, emulsifying activity, emulsion stabilising properties, gelling and foaming properties are also analysed. A novel processing technology, spinning disc reactor, could be an alternative process for the production of protein-polysaccharide conjugates, with desirable functionality in different food systems

    Mass transfer efficiency of a tall and low plate free area liquid pulsed sieve-plate extraction column

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    Acknowledgements The authors would like to acknowledge Chakwal group of industries for funding the project. Ms. Madiha, Ms. Zona, Mr. Sohaib, Mr. Abdullah, Mr. Mudassar, and Mr. Salahuddin also deserve our acknowledgements for their assistance in different ways.Peer reviewedPublisher PD

    Development of an improved oxygen electrode for use in alkaline H2-O2 fuel cells Quarterly report, Apr. 1 - Jun. 30, 1967

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    Preparation of institial compounds of transition metals for hydrogen oxygen fuel cell cathode

    Improved enzymatic accessibility of peanut protein isolate pre-treated using thermosonication

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    Thermosonication pre-treatment was used to enhance the pancreatin-induced proteolysis of peanut protein isolate (PPI). Response surface methodology was applied to optimize the thermosonication conditions (including power-output and temperature), and the highest degree of hydrolysis (7.16%) was obtained at 475.0 W, 72 °C. SDS-PAGE analysis showed that at this optimized condition, the enzymatic accessibility of the major constitutive protein arachin in thermosonicated PPI (TS-PPI) was substantially improved compared to that in untreated PPI or sonicated PPI (475 W, 30°C; S-PPI), resulting in a remarkable increase in protein solubility for the hydrolysates. Protein denaturation and conformation profiles of untreated PPI, S-PPI and TS-PPI were investigated using differential scanning calorimetry, intrinsic fluorescence emission spectroscopy, Fourier transform infra-red spectroscopy and thioflavin-T (ThT) fluorescence assay. It was found that heat could present a markedly additive effect to ultrasound on denaturing peanut proteins, leading to significant changes in protein conformation. TS-PPI was characterized by the appearance of high proportion of parallel intermolecular β-sheets and a strong fluorescence enhancement upon binding to ThT, suggesting that the protein unfolding and aggregation induced by thermosonication probably resulted in the formation of fibril protein aggregates in TS-PPI rather than spherical protein aggregates formed in S-PPI. As a result, the protein conformation of TS-PPI appeared to be more unfolded and flexible than that of untreated PPI or S-PPI, and therefore was more easily accessible to protease. This study shows that thermosonication pre-treatment could be a highly effective and feasible technique to improve the enzymatic accessibility of globular proteins, producing prominent functional benefits for the protein hydrolysates

    Dichromacy: Color Vision Impairment and Consanguinity in Heterogenous Population of Pakistan

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    Background and Objectives: Dichromacy, an X-linked recessive disorder is identified worldwide, more in males than females. In European Caucasians, its incidence is 8% in males and 0.5% in females. In India, it is 8.73% in males and 1.69% in females, and in Iran, it is 8.18% in males and 0.43% in females. Population based epidemiological data about dichromacy in different ethnic groups in Pakistan is not available. The aim of this study was to find out the population prevalence of inherited red-green dichromacy in a heterogenous population of the district of Chiniot, Punjab, Pakistan, and to determine the impact of consanguinity and ethnicity. Methods: In this cross-sectional study, boys and girls of the higher secondary schools were examined in the three tehsils of district Chiniot. Pseudoisochromatic Ishihara Test has been employed for detection of dichromacy in the study population. The sample size was calculated statistically as 260, which was expanded to 705 and divided by population density of the three tehsils. Results: Screening of 359 males and 346 females revealed 19 (5.29%) dichromat males and only 2 (0.58%) females. The study population belonged to 23 castes / isonym groups. The consanguinity found in the district of Chiniot is 84.82% and in the dichromat families, it is 85.71%, of which 52.37% are first cousin. Interpretation & Conclusion: The study has shown that the incidence of dichromacy could be reduced through genetic counselin
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