Transferrin Receptor Targeted Delivery Of Sirna For Gene Therapy

Gene therapy is thought to be a solution for various difficult to treat diseases such as cancer. Small interfering RNA (siRNA) as a promising anti-sense molecule can specifically silence disease related gene have been exploit in different diseases. However, lack of safe and efficient siRNA delivery systems limits the application of siRNA therapy in clinic. Transferrin receptor (TfR) is an essential transmembrane receptor involved in iron uptake. TfR universally express in most cells/ tissues but upregulated in certain cells, for example, many cancer cells and activated T cells (ATCs). To overcome the biological barriers and increase siRNA delivery efficiency, in this dissertation, various TfR targeted siRNA delivery systems have been developed. Holo-Tf and TfR binding peptide were conjugated to polyethylenimine (PEI), an efficient gene delivery polymer, via different crosslinkers. Tf-PEI was evaluated in asthma model and breast cancer model. Tf-PEI polyplexes demonstrated satisfactory physicochemical properties and selectivity to cells overexpressing TfR (e.g. basal like breast cancer cells and ATCs). Tf-PEI can achieve efficient gene knockdown ex vivo and targeted delivery to ATCs in asthma model via pulmonary delivery. Peptide-PEI was evaluated in lung cancer model. Peptide-PEI can selectively deliver siRNA to lung cancer cells overexpressing TfR and achieve efficient GAPDH gene knockdown. In conclusion, the present study described the feasibility to efficiently deliver siRNA to ATCs in asthma and cancer cells in various cancer model via targeting TfR. This results could help to develop an efficient delivery system of siRNA for asthma therapy and cancer therapy

Characterization of Dopaminergic System in the Striatum of Young Adult Park2(-/-) Knockout Rats

Mutations in parkin gene (Park2) are linked to early-onset autosomal recessive Parkinson's disease (PD) and young-onset sporadic PD. Park2 knockout (PKO) rodents;however, do not display neurodegeneration of the nigrostriatal pathway, suggesting age-dependent compensatory changes. Our goal was to examine dopaminergic (DAergic) system in the striatum of 2 month-old PKO rats in order to characterize compensatory mechanisms that may have occurred within the system. The striata form wild type (WT) and PKO Long Evans male rats were assessed for the levels of DAergic markers, for monoamine oxidase (MAO) A and B activities and levels, and for the levels of their respective preferred substrates, serotonin (5-HT) and beta-phenylethylamine (beta-PEA). The PKO rats displayed lower activities of MAOs and higher levels of beta-PEA in the striatum than their WT counterparts. Decreased levels of beta-PEA receptor, trace amine-associated receptor 1 (TAAR-1), and postsynaptic DA D2 (D2L) receptor accompanied these alterations. Drug-naive PKO rats displayed normal locomotor activity;however, they displayed decreased locomotor response to a low dose of psychostimulant methamphetamine, suggesting altered DAergic neurotransmission in the striatum when challenged with an indirect agonist. Altogether, our findings suggest that 2 month-old PKO male rats have altered DAergic and trace aminergic signaling

High Relaxivity Gadolinium-Polydopamine Nanoparticles

AbstractThis study reports the preparation of a series of gadolinium‐polydopamine nanoparticles (GdPD‐NPs) with tunable metal loadings. GdPD‐NPs are analyzed by nuclear magnetic relaxation dispersion and with a 7‐tesla (T) magnetic resonance imaging (MRI) scanner. A relaxivity of 75 and 10.3 mM−1 s−1 at 1.4 and 7 T is observed, respectively. Furthermore, superconducting quantum interference device magnetometry is used to study intraparticle magnetic interactions and determine the GdPD‐NPs consist of isolated metal ions even at maximum metal loadings. From these data, it is concluded that the observed high relaxivities arise from a high hydration state of the Gd(III) at the particle surface, fast rate of water exchange, and negligible antiferromagnetic coupling between Gd(III) centers throughout the particles. This study highlights design parameters and a robust synthetic approach that aid in the development of this scaffold for T1‐weighted, high relaxivity MRI contrast agents

Structure and Function of Iron-Loaded Synthetic Melanin

We describe a synthetic method for increasing and controlling the iron loading of synthetic melanin nanoparticles and use the resulting materials to perform a systematic quantitative investigation on their structure 12property relationship. A comprehensive analysis by magnetometry, electron paramagnetic resonance, and nuclear magnetic relaxation dispersion reveals the complexities of their magnetic behavior and how these intraparticle magnetic interactions manifest in useful material properties such as their performance as MRI contrast agents. This analysis allows predictions of the optimal iron loading through a quantitative modeling of antiferromagnetic coupling that arises from proximal iron ions. This study provides a detailed understanding of this complex class of synthetic biomaterials and gives insight into interactions and structures prevalent in naturally occurring melanins

Targeted Delivery of siRNA to Transferrin Receptor Overexpressing Tumor Cells via Peptide Modified Polyethylenimine

The use of small interference RNA (siRNA) to target oncogenes is a promising treatment approach for cancer. However, siRNA cancer therapies are hindered by poor delivery of siRNA to cancer cells. Transferrin receptor (TfR) is overexpressed in many types of tumor cells and therefore is a potential target for the selective delivery of siRNA to cancer cells. Here, we used the TfR binding peptide HAIYPRH (HAI peptide) conjugated to cationic polymer branched polyethylenimine (bPEI), optimized the coupling strategy, and the TfR selective delivery of siRNA was evaluated in cells with high (H1299) and low TfR expression (A549 and H460). The HAI-bPEI conjugate exhibited chemico-physical properties in terms of size, zeta-potential, and siRNA condensation efficiency similar to unmodified bPEI. Confocal microscopy and flow cytometry results revealed that HAI-bPEI selectively delivered siRNA to H1299 cells compared with A549 or H460 cells. Moreover, HAI-bPEI achieved more efficient glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene knockdown in H1299 cells compared with bPEI alone. However, despite optimization of the targeting peptide and coupling strategy, HAI-bPEI can only silence reporter gene enhanced green fluorescent protein (eGFP) at the protein level when chloroquine is present, indicating that further optimization of the conjugate is required. In conclusion, the HAI peptide may be useful to target TfR overexpressing tumors in targeted gene and siRNA delivery approaches

Targeted Delivery of siRNA to Transferrin Receptor Overexpressing Tumor Cells via Peptide Modified Polyethylenimine

The use of small interference RNA (siRNA) to target oncogenes is a promising treatment approach for cancer. However, siRNA cancer therapies are hindered by poor delivery of siRNA to cancer cells. Transferrin receptor (TfR) is overexpressed in many types of tumor cells and therefore is a potential target for the selective delivery of siRNA to cancer cells. Here, we used the TfR binding peptide HAIYPRH (HAI peptide) conjugated to cationic polymer branched polyethylenimine (bPEI), optimized the coupling strategy, and the TfR selective delivery of siRNA was evaluated in cells with high (H1299) and low TfR expression (A549 and H460). The HAI-bPEI conjugate exhibited chemico-physical properties in terms of size, zeta-potential, and siRNA condensation efficiency similar to unmodified bPEI. Confocal microscopy and flow cytometry results revealed that HAI-bPEI selectively delivered siRNA to H1299 cells compared with A549 or H460 cells. Moreover, HAI-bPEI achieved more efficient glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene knockdown in H1299 cells compared with bPEI alone. However, despite optimization of the targeting peptide and coupling strategy, HAI-bPEI can only silence reporter gene enhanced green fluorescent protein (eGFP) at the protein level when chloroquine is present, indicating that further optimization of the conjugate is required. In conclusion, the HAI peptide may be useful to target TfR overexpressing tumors in targeted gene and siRNA delivery approaches

Effect of the location pattern of rural residential buildings on natural ventilation in mountainous terrain of central China

The use of natural ventilation in buildings to reduce the energy consumption and CO2 emission has been widely investigated and practiced, but few existing studies have considered the exploration and assessment of natural ventilation in different location patterns of rural residential buildings in the mountainous terrain of China. In this paper, the representative rural residential buildings are firstly selected in Huarong, Pingjiang and Liuyang regions of northern Hunan Province to carry out on-site survey works to determine building types, physical parameters and layout forms. Then, the wind tunnel experiments are carried out to investigate the effectiveness of natural ventilation under different location patterns, and the monitored results are compared with simulated data. The results show that the experiments and simulations are in satisfactory agreement. The experimental data also indicate that when the modelled distance of 120 mm (i.e. 12 m between the building and hilly terrain in practical application) is the best option for building natural ventilation. Based on the investigation and statistical data, the natural ventilation effectiveness under different location patterns and operational conditions is simulated using CFD methods, and it is obtained the most favourable location pattern for natural ventilation. The results show that the winter ventilation of buildings in the existing location pattern is significantly obstructed in the hilly terrain, which is favourable to the indoor thermal environment, however, the natural ventilation is compromised to a certain extent in summer. Furthermore, the findings also show that, regardless of the hilly terrain's height at 50 m or 150 m, the buildings are able to avoid natural ventilation in winter to the maximum extent when the distance between the buildings and the frontier of the hilly terrain is double that of the building height (i.e. 12 m). This study could contribute to theoretical instructions for optimum design of natural ventilation of rural residential buildings in the mountainous terrain of central China.Design & Construction Managemen

Phylogenomics reveal Populus gonggaensis as a hybrid between P. lasiocarpa and P. cathayana (Salicaceae)

High levels of intra-specific polymorphism and frequent hybridisation make it difficult to define species and correctly apply their scientific names. Populus L. is a challenging genus with plentiful natural and artificial hybrids. This study is a part of the project ‘Flora of Pan-Himalaya’ and aims to determine the taxonomic identity of P. gonggaensis N. Chao & J.R. He and to find out whether it is of hybrid origin. Whole-genome sequencing data were obtained from 57 samples. The SNP matrix was developed for phylogenetic reconstruction, ABBA-BABA statistics, PCA and ADMIXTURE analysis. The results indicate that P. gonggaensis is a spontaneous hybrid between P. lasiocarpa and P. cathayana. This study points out the importance of SNP data and comprehensive analyses for discovering the potential interspecific hybridisation and clarifies the usage of the name. In addition, the lectotype of P. gonggaensis was designated

Selective comminution and grinding mechanisms of spent carbon anode from aluminum electrolysis using ball and rod mills

The recovery of spent carbon anode (SCA) materials plays important roles in environment protection and resources recycling, while this cannot be efficiently achieved without liberation. In this study, grinding characteristics of spent carbon anode from aluminum electrolysis in both ball mill and rod mill were analyzed, and compared based on the utilization of the selective Fuerstenau upgrading curves. In addition, the different grinding mechanisms of ball and rod milling were evaluated by analyzing the shape factor and surface roughness of the ground samples. Results of mineralogical characterizations indicated that carbon particles (d50 = 46.86 μm) presented in the SCA were closely associated with cryolite particles. At 5 min grinding time, the maximum selective comminution factor (β) values of ball milling and rod milling were 2.00 and 1.63, respectively, indicating a higher degree of selective comminution of SCA was achieved from ball milling. Comparisons of the valuable component content (cv, cum) of -125 μm ground particles and the shape characterizations of 74–125 μm ground particles generated from ball and rod milling manifest that a direct relationship exists between the degree of selective comminution and the grinding mechanism