In vitro transfection of HeLa cells with temperature sensitive polycationic copolymers

Cataloged from PDF version of article.In this study, we investigated different types of polyethyleneimine (PEI) and their block copolymers with N-isopropylacrylamide (NIPA) as temperature-sensitive polycationic non-viral vectors for transfection of HeLa cells in cell culture media. First carboxyl-terminated poly(NIPA) was synthesized and then copolymerized with PEIs branched or linear and with two different molecular weights (2 and 25 kDa). Addition of PEI units to the poly(NIPA) chains increased the LCST values up to body temperature. Zeta potentials of the copolymers were significantly lower than the corresponding PEI homopolymers. A green fluorescent protein expressing plasmid was used as a model. Complexes of this plasmid both with PEIs and their copolymers were formed. The zeta potentials of these complexes were between -3.1 and +21.3. Higher values were observed for the complexes prepared with branched and higher molecular weight PEIs. Copolymerization caused a profound decrease in the positive charges. Particle sizes of the complexes were in the range of 190-1235 nm. Using high polymer/plasmid ratios caused aggregation. The smallest complexes were obtained with the copolymer prepared with branched PEI with 25-kDa molecular weight. Copolymers were able to squeeze plasmid DNA more at the body temperature. Cytotoxicity was observed with PEIs especially with the branched higher molecular weights. Copolymerization reduced the cytotoxicity. The best in vitro DNA uptake efficiency (70%) was achieved with the complex prepared with poly(NIPA)/PEI25B. However, poly(NIPA)/PEI25L was the most successful vector for an effective gene expression without any significant toxicity. © 2004 Elsevier B.V. All rights reserved

Wind, jet, hybrid corona and hard X-ray flares: multiwavelength evolution of GRO J1655-40 during the 2005 outburst rise

We have investigated the complex multiwavelength evolution of GRO J1655-40 during the rise of its 2005 outburst. We detected two hard X-ray flares, the first one during the transition from the soft state to the ultra-soft state, and the second one in the ultra-soft state. The first X-ray flare coincided with an optically thin radio flare. We also observed a hint of increased radio emission during the second X-ray flare. To explain the hard flares without invoking a secondary emission component, we fit the entire data set with the eqpair model. This single, hybrid Comptonization model sufficiently fits the data even during the hard X-ray flares if we allow reflection fractions greater than unity. In this case, the hard X-ray flares correspond to a Comptonizing corona dominated by non-thermal electrons. The fits also require absorption features in the soft and ultra-soft state which are likely due to a wind. In this work we show that the wind and the optically thin radio flare co-exist. Finally, we have also investigated the radio to optical spectral energy distribution, tracking the radio spectral evolution through the quenching of the compact jet and rise of the optically thin flare, and interpreted all data using state transition models.Comment: 16 pages, 11 figure

Toward Continuous Monitoring of Breath Biochemistry: A Paper-Based Wearable Sensor for Real-Time Hydrogen Peroxide Measurement in Simulated Breath

Exhaled breath contains a large amount of biochemical and physiological information concerning one’s health and provides an alternative route to noninvasive medical diagnosis of diseases. In the case of lung diseases, hydrogen peroxide (H2O2) is an important biomarker associated with asthma, chronic obstructive pulmonary disease, and lung cancer and can be detected in exhaled breath. The current method of breath analysis involves condensation of exhaled breath, is not continuous or real time, and requires two separate and bulky devices, complicating the periodic or long-term monitoring of a patient. We report the first disposable paper-based electrochemical wearable sensor that can monitor exhaled H2O2 in artificial breath calibration-free and continuously, in real time, and can be integrated into a commercial respiratory mask for on-site testing of exhaled breath. To improve precision for sensing H2O2, we perform differential electrochemical measurement by amperometry in which screen-printed Prussian Blue-mediated and nonmediated carbon electrodes are used for differential analysis. We were able to measure H2O2 in simulated breath in a concentration-dependent manner in real time, confirming its functionality. This proposed system is versatile, and by modifying the chemistry of the sensing electrodes, our method of differential sensing can be extended to continuous monitoring of other analytes in exhaled breath

Gli effetti squelch binaurale e ombra nei bambini con impianto cocleare monolaterale e protesi acustica controlaterale

Lo scopo di questo studio è stato quello di indagare il grado di effetto squelch binaurale (BSE) e di effetto ombra (HSE) nei bambini che fanno uso di impianto cocleare (CI) e protesi acustica controlaterale (HA). Sono stati arruolati 19 bambini con CI che indossavano regolarmente una HA. Per valutare la performance vocale in presenza di rumore è stato utilizzato un test di identificazione vocalica. Il test è stato eseguito in tre condizioni di ascolto: (1) ascolto bimodale con sorgente di rumore sul lato HA; (2) ascolto con CI con sorgente di rumore sul lato HA; (3) ascolto con CI con sorgente di rumore sul lato CI. I valori medi di BSE e di HSE osservati sono 11.8% e 17.1% rispettivamente. L’analisi statistica condotta ha evidenziato una differenza significativa sia nel confronto tra le tre condizioni di ascolto che per l’analisi post-hoc (p < 0.001). In conclusione, la maggioranza dei bambini con CI hanno mostrato BSE ed HSE con un miglioramento significativo nella percezione vocale in presenza di rumore

Autocatalytic metallization of fabrics using Si ink, for biosensors, batteries and energy harvesting

Commercially available metal inks are mainly designed for planar substrates (for example, polyethylene terephthalate foils or ceramics), and they contain hydrophobic polymer binders that fill the pores in fabrics when printed, thus resulting in hydrophobic electrodes. Here, a low‐cost binder‐free method for the metallization of woven and nonwoven fabrics is presented that preserves the 3D structure and hydrophilicity of the substrate. Metals such as Au, Ag, and Pt are grown autocatalytically, using metal salts, inside the fibrous network of fabrics at room temperature in a two‐step process, with a water‐based silicon particle ink acting as precursor. Using this method, (patterned) metallized fabrics are being enabled to be produced with low electrical resistance (less than 3.5 Ω sq−1). In addition to fabrics, the method is also compatible with other 3D hydrophilic substrates such as nitrocellulose membranes. The versatility of this method is demonstrated by producing coil antennas for wireless energy harvesting, Ag–Zn batteries for energy storage, electrochemical biosensors for the detection of DNA/proteins, and as a substrate for optical sensing by surface enhanced Raman spectroscopy. In the future, this method of metallization may pave the way for new classes of high‐performance devices using low‐cost fabrics

BRCA1 and BRCA2 mutations in Turkish breast/ovarian families and young breast cancer patients

To date, BRCA1 and BRCA2 mutations in breast and/or ovarian patients have not been characterized in the Turkish population. We investigated the presence of BRCA mutations in 53 individuals with a personal and family history of breast and/or ovarian cancer, and 52 individuals with a personal history of breast cancer diagnosed below age 50 without additional family history. We have identified 11 mutations (nine BRCA1 and two BRCA2) using combined techniques involving protein truncation test, direct sequencing and heteroduplex analysis. We found eight out of 53 patients (15.1%) with a family history to carry BRCA gene mutations (seven BRCA1 and one BRCA2). Of these, four were found in 43 families presenting only breast cancer histories, and four were found in families presenting ovarian cancer with or without breast cancer. We also demonstrated two BRCA1 and one BRCA2 mutations in three out of 52 (5.8%) early-onset breast cancer cases without additional family history. Three of nine BRCA1 and both BRCA2 mutations detected in this study were not reported previously. These mutations may be specific to the Turkish population. The BRCA1 5382insC mutation, specific to Ashkenazi and Russian populations, was found twice in our study group, representing a possible founder mutation in the Turkish population. © 2000 Cancer Research Campaig

Lepton polarization asymmetry in radiative dileptonic B-meson decays in MSSM

In this paper we study the polarization asymmetries of the final state lepton in the radiative dileptonic decay of B meson (\bsllg) in the framework of Minimal Supersymmetric Standard Model (MSSM) and various other unified models within the framework of MSSM e.g. mSUGRA, SUGRA (where condition of universality of scalar masses is relaxed) etc. Lepton polarization, in addition of having a longitudinal component (\pl), can have two other components, \pt and \pn, lying in and perpendicular to the decay plane, which are proportional to \ml and hence are significant for final state being $\mu^+ ~ \mu^-$ or $\tau^+ \~\tau^-$. We analyse the dependence of these polarization asymmetries on the parameters of the various models.Comment: typos corrected to match with published versio

Analysis of Various Polarization Asymmetries In The Inclusive b→sℓ+ℓ−b\to s \ell^+ \ell^- Decay In The Fourth-Generation Standard Model

In this study a systematical analysis of various polarization asymmetries in inclusive b \rar s \ell^+ \ell^- decay in the standard model (SM) with four generation of quarks is carried out. We found that the various asymmetries are sensitive to the new mixing and quark masses for both of the $\mu$ and $\tau$ channels. Sizeable deviations from the SM values are obtained. Hence, b \rar s \ell^+ \ell^- decay is a valuable tool for searching physics beyond the SM, especially in the indirect searches for the fourth-generation of quarks ($t', b')$.Comment: 19 Pages, 10 Figures, 3 Table

Structure-based design and synthesis of antiparasitic pyrrolopyrimidines targeting pteridine reductase 1

The treatment of Human African Trypanosomiasis remains a major unmet health need in sub-Saharan Africa. Approaches involving new molecular targets are important and pteridine reductase 1 (PTR1), an enzyme that reduces dihydrobiopterin in Trypanosoma spp. has been identified as a candidate target and it has been shown previously that substituted pyrrolo[2,3-d]pyrimidines are inhibitors of PTR1 from T. brucei (J. Med. Chem. 2010, 53, 221-229). In this study, 61 new pyrrolo[2,3-d]pyrimidines have been prepared, designed with input from new crystal structures of 23 of these compounds complexed with PTR1, and evaluated in screens for enzyme inhibitory activity against PTR1 and in vitro antitrypanosomal activity. 8 compounds were sufficiently active in both screens to take forward to in vivo evaluation. Thus although evidence for trypanocidal activity in a stage I disease model in mice was obtained, the compounds were too toxic to mice for further development

Chromatin profiling of cortical neurons identifies individual epigenetic signatures in schizophrenia

Both heritability and environment contribute to risk for schizophrenia. However, the molecular mechanisms of interactions between genetic and non-genetic factors remain unclear. Epigenetic regulation of neuronal genome may be a presumable mechanism in pathogenesis of schizophrenia. Here, we performed analysis of open chromatin landscape of gene promoters in prefrontal cortical (PFC) neurons from schizophrenic patients. We cataloged cell-type-based epigenetic signals of transcriptional start sites (TSS) marked by histone H3-K4 trimethylation (H3K4me3) across the genome in PFC from multiple schizophrenia subjects and age-matched control individuals. One of the top-ranked chromatin alterations was found in the major histocompatibility (MHC) locus on chromosome 6 highlighting the overlap between genetic and epigenetic risk factors in schizophrenia. The chromosome conformation capture (3C) analysis in human brain cells revealed the architecture of multipoint chromatin interactions between the schizophrenia-associated genetic and epigenetic polymorphic sites and distantly located HLA-DRB5 and BTNL2 genes. In addition, schizophrenia-specific chromatin modifications in neurons were particularly prominent for non-coding RNA genes, including an uncharacterized LINC01115 gene and recently identified BNRNA_052780. Notably, protein-coding genes with altered epigenetic state in schizophrenia are enriched for oxidative stress and cell motility pathways. Our results imply the rare individual epigenetic alterations in brain neurons are involved in the pathogenesis of schizophrenia