Mild and transient heat shock enhances DNA integration following lipofection of recombinant plasmids in 4T1 cells

316-320Cancer cells having stably integrated genes encoding tumor-associated antigens could be utilized as a vaccine, in-vitro stimulators of antigen-primed T-cells, and target for cytotoxicity assay, etc. Lipofection is a simple and safer technique for stable transfection of plasmid DNA. However, the poor rate of genomic integration has limited its application. In the current study, the effect of mild and transient heat shock following lipofection on the improvement of genomic integration was evaluated. The cDNA fragments encoding chicken MMP-11peptide (V32-K365) and the immunoglobulin-like domain 2 of chicken VEGFR-2 were cloned separately into pcDNA3.1 vector. Lipofection was carried out using Lipofectamine® 2000 (Life Technologies, USA) in 4T1 cells followed by a heat shock at 42°C for 10 min. Transfected cells were selected for a period of four weeks against 500 µg/mL G418 in RPMI 1640 media supplemented with 10% fetal bovine serum. Distinct G418-resistant colonies appeared after 14 days of selection. Heat shock significantly (P <0.05) increased the number of viable colonies following antibiotic selection. The immunofluorescent study confirmed the stable integration of the target DNAs into the cells. It is concluded that mild and brief heat shock following lipofection improves the stable integration of recombinant pcDNA3.1 plasmids into 4T1 cells

Mild and transient heat shock enhances DNA integration following lipofection of recombinant plasmids in 4T1 cells

Cancer cells having stably integrated genes encoding tumor-associated antigens could be utilized as a vaccine, in-vitro stimulators of antigen-primed T-cells, and target for cytotoxicity assay, etc. Lipofection is a simple and safer technique for stable transfection of plasmid DNA. However, the poor rate of genomic integration has limited its application. In the current study, the effect of mild and transient heat shock following lipofection on the improvement of genomic integration was evaluated. The cDNA fragments encoding chicken MMP-11peptide (V32-K365) and the immunoglobulin-like domain 2 of chicken VEGFR-2 were cloned separately into pcDNA3.1 vector. Lipofection was carried out using Lipofectamine® 2000 (Life Technologies, USA) in 4T1 cells followed by a heat shock at 42°C for 10 min. Transfected cells were selected for a period of four weeks against 500 µg/mL G418 in RPMI 1640 media supplemented with 10% fetal bovine serum. Distinct G418-resistant colonies appeared after 14 days of selection. Heat shock significantly (P &lt;0.05) increased the number of viable colonies following antibiotic selection. The immunofluorescent study confirmed the stable integration of the target DNAs into the cells. It is concluded that mild and brief heat shock following lipofection improves the stable integration of recombinant pcDNA3.1 plasmids into 4T1 cells

Linearno i nelinearno sirenje ionsko-zvučnih valova u plazmi u posudi koja sadrži i negativne ione

Linear and nonlinear propagation of ion-acoustic waves are theoretically investigated in a multicomponent plasma consisting of electrons, positive ions and negative ions bounded in a cylindrical waveguide. The stability of the ion-acoustic wave is discussed taking into account the role of finite geometry and the concentration of negative ions of the plasma. The effect of nonlinearity on the ion-acoustic wave is investigated through the derivation of the effective potential (Sagdeev potential) and the results are discussed graphically with the variation of ion-streaming and the geometry of the bounded plasma.Teorijski istražujemo linearno i nelinearno širenje ionsko-zvučnih valova u višekomponentnoj plazmi koja se sastoji od elektrona, te pozitivnih i negativnih iona i nalazi se u valjkastom valovodu. Raspravljamo stabilnost ionsko-zvučnih valova uzimajućci u obzir utjecaj ograničenog prostora i koncentracije negativnih iona u plazmi. Istražujemo učinke nelinearnosti na ionsko-zvučni val putem izvođenja efektivnog (Sagdeevog) potencijala a ishodi numeričkih računa predočuju se grafički za različita strujanja iona i oblike valovoda

Problem samofokusiranja i poprečne stabilnosti u magnetiziranoj plazmi s ionskom strujom

Self-focusing in a magnetized plasma consisting of cold electrons and ions has been studied in a situation when ions have a streaming velocity and the external magnetic field is at an angle µ with the streaming direction. By the use of a modified scaling, we have deduced an analogue of a non-linear Schrodinger equation involving only the space variables, which actually controls the transversal stability of the system, thus signaling the possibility of self-focusing. The various situations arising due to the variation of the propagation angle µ and the streaming velocity u0 are discussed in detail for the specific cases of D2 and N2 ions.Proučavamo samofokusiranje u magnetiziranoj plazmi koja se sastoji of hladnih elektrona i iona u uvjetima kada ioni struje a magnetsko je polje pod nekim kutom prema smjeru njihovog strujanja. Primjenom modificirane promjene mjera, izveli smo jednadžbu koja je analogna Schrodingerovoj jednadžbi s prostornim varijablama koja opisuje poprečnu stabilnost sustava i tako najavljuje mogućnost samofokusiranja. Raspravljaju se podrobno razni uvjeti koji nastaju zbog promjena smjera strujanja iona (kuta θ) i brzine strujanja (u0) za posebne slučajeve D2 i N2 iona

Management of the Power Output of Photovoltaic Array and Fuel Cell

This paper comprises with the hybrid model of photovoltaic array (PV) and fuel cell (FC) for maximizing and managing the power generation in the system. In this model two different power sources had been used one is photovoltaic array and another one is fuel cell; both the sources are independent of their individual�s working and can be used as per their requirement. The output generated by photovoltaic array and fuel cell is connected to the Cuk converter which regulates the voltage and providing constant dc supply at the output end of Cuk converter. Use of fuel cell in the model helps to compensate the photovoltaic array output during night time or cloudy weather. If demand is less than power supply than surplus energy is used to generate hydrogen from pure water which get stored in storage tank for future generation in fuel cell. Controllers are used to reduce steady state error, harmonics and output impedances

A Review on Speech Recognition Methods

Voice recognition is the identification of a speaker on the basis of the characteristics of voices. For this, features of speech patterns that differ between individuals are used to achieve the objective. In this paper speaker recognition system are discussed. Implementation of speaker's voice recognition system with MATLAB makes possible use of voice for real life applications. This paper provides a brief review of different DSP based techniques applied for speech recognition

Manas Kr. Mondal

ABSTRACT: A total of 18 lactating multiparous cows (4-6 years aged) were selected from the out patients at the Addl. Block Animal Health Center, Matiali Block, Jalpaiguri, West Bengal, India and divided into three group, i.e normally cyclic (C), repeat breeder (RB) and post partum anoestrous (A). Blood samples were collected from all these cows for haematological and biochemical parameters. Erythrocyte sedimentation rate (ESR) and total leukocyte count (TEC) count were higher (P&lt;0.05) in repeat breeder and anoestrous cows compared to cyclic ones; however the Hb and PCV values were low (P&lt;0.05). Serum glucose and protein levels were low (P&lt;0.05) repeat breeding cows than the normally cyclic cows. The results indicate that there may not be any specific haemato-biochemical marker for common reproductive disorders in cows

Net-charge particle ratio fluctuations in pppp collisions at several LHC energies

Event-by-event particle ratio fluctuations for simulated data sets of three different models named UrQMD, AMPT, and Pythia are studied using the fluctuation variable $\nu_{dyn}$. The simulated data sets produced in $pp$ collisions at four different LHC energies $\sqrt{s} = 2.76, 5.02, 7$ and $13$ TeV are generated and considered for this analysis. The variation of fluctuation parameter $\nu_{dyn}$ for accepted pair of meson and baryon combination $i.e.$ $[\pi, K]$, $[\pi, p]$ and $[p, K]$ with the increasing value of the mean multiplicity of charged particles ($\langle N_{ch} \rangle$) are investigated. It has been observed that the correlation between the particle pair $[\pi, K]$ is more than that of the two other particle pairs $[\pi, p]$ and $[p, K]$. However, the energy-wise inspection of the fluctuation variable $\nu_{dyn}$ for $0-10\%$ centrality data shows the increase in the correlation between the particles in each pair for all three models considered.Comment: 16 pages, 18 figures, Submitted to Journal for peer-revie