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Objective To prepare PCM modified liposome (PCM-LIP) containing enhanced green fluorescent protein expression plasmid (pEGFP) and to evaluate its myocardial targeting ability. Methods Liposome was prepared by film-ultrasonic, with PCM used as ligand and DOTAP as cationic lipid material. PCM-LIP containing pEGFP was prepared by incubating liposome with pEGFP at room temperature. The connecting method of PCM was optimized and the connection rate of PCM was determined. The characteristics of liposomes including encapsulation ability, morphology, particle size, zeta potential and stability in phosphate buffer solution (PBS) were observed. The transfection efficiency of liposomes into H9c2 cells was evaluated by inverted fluorescence microscopy and flow cytometry, so as to characterize their myocardial targeting ability and to determine the optimum dosage of PCM. Results PCM-LIP was prepared by insertion method and the amount of PCM accounted for 3% of the lipid. After incubation with pEGFP, PCM-LIP was spherical in shape, with the particle size being (261. 9±2. 2) nm, zeta potential being (-5. 0±0. 6) mV, and PCM-LIP was stable in PBS at -4℃ for 30 d. The transfection efficiency of PCM-LIP was significantly higher than that of unmodified liposome (P<0. 05). Conclusion PCM can improve the transfection efficiency of liposome into cardiomyocytes and PCM-LIP shows a satisfactory myocardial targeting ability.
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ABSTRACT Thiophene oligomer has been investigated using DFT/TDDFT calculations with an aim to check its suitability for opto electronic applications and also to analyse the influence of π-bridge. Our results revealed that thiophene oligomers have excellent π-conjugation throughout. FMO analysis give an estimate of band gap of thiophene oligomer and further revealed HOMO are localized on π - bridge, donor group and LUMO are localized on π - bridge and acceptor group. A TDDFT calculation has been performed to understand the absorption properties of them in gas phase and solvent phase. PCM calculations convey that absorption maxima show positive solvatochromism. Among the designed candidates, the one with more π - bridge show higher wavelength of absorption maxima and would be a choice for better optoelectronic materials. NBO analysis provides support for complete delocalization in these systems. It is interesting to note that oligomer with more π-bridge display an enhanced optoelectronic properties than with less π - bridge.
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The CTLA-4 protein is expressed in activated T cells and plays an essential role in the immune response through its regulatory effect on T cell activation. Polymorphisms of the CTLA-4 gene have been correlated with autoimmune, neoplastic and infectious illnesses. This work aimed to verify possible associations between single nucleotide polymorphisms (SNPs) in CTLA-4, -318C/T in the promoter and +49A/G in exon 1 and paracoccidioidomycosis (PCM) caused by Paracoccidioides brasiliensis. For this purpose, 66 chronic form PCM patients and 76 healthy controls had their allele, genotype and haplotype frequencies determined. The genetic admixture structure of the patients and controls was evaluated to eliminate ancestral bias. The comparison of frequencies indicated no significant differences between patients and controls that could link the SNPs to PCM. Groups were admixture matched with no difference observed in population ancestry inference, indicating that the absence of association between CTLA-4 polymorphisms and PCM could not be attributed to ancestral bias. This study showed that there was no association between the CTLA-4 SNPs -318 and +49 and the resistance or susceptibility to PCM.