24 research outputs found
3D interconnect technology based on low temperature copper nanoparticle sintering
No full textWe explore a methodology for patterned copper nanoparticle paste for 3D interconnect applications in wafer to wafer (W2W) bonding. A novel fine pitch thermal compression bonding process (sintering) with coated copper nanoparticle paste was developed. Most of the particle size is between 10-30 nm. Lithographically defined stencil printing using photoresist and lift-off was used to apply and pattern the paste. Variations in sintering process parameters, such as: pressure, geometry and ambient atmosphere, were studied. Compared to Sn-Ag-Cu (SAC) microsolder bumps, we achieved better interconnect resistivity after sintering at 260 °C for 10 min, in a 700 mBar hydrogen forming gas (H2/N2) environment. The electrical resistivity was 7.84 ± 1.45 μΩ·cm, which is about 4.6 times that of bulk copper. In addition, metallic nanoparticle interconnect porosity can influence the electrical properties of the interconnect. Consequently, we investigated the porosity effect on conductivity using finite element simulation. A linear relationship between the equivalent conductivity and particle overlapping ratio was found.Accepted Author ManuscriptElectronic Components, Technology and MaterialsEKL Processin
497. Central CART Gene Delivery by Recombinant AAV Vector Decreases Lean Body Mass in Diet-Induced Obese Male Rats
No full textThe Research of Introducing Project Practice Training Mode for Computer Professional Talent
No full textAbstractIt is an essential problem must to face how to raises the computer major student's project design and development ability to meet the stern employment competition situation. The paper analyzed the shortcoming of existing computer practice teaching system, proposed a novel education pattern with project practice, taken the corresponding reform measures. It was emphasized for the characteristics of project practice-- the planning, the authenticity and the affectivity. The practice result indicated that, student's practice ability, independent study ability and unity cooperation ability obtains enhanced greatly after the project practice
Infection of Purified Nuclei by Adeno-associated Virus 2
No full textAdeno-associated virus 2 (AAV), a nonpathogenic human parvovirus, requires co-infection with a helper virus for its optimal replication. Although AAV possesses a broad host range, certain cell types lack the machinery necessary for efficient entry into the cell and intracellular trafficking of AAV into the nucleus, where the viral second-strand DNA synthesis must occur before gene expression. We have demonstrated that in less-permissive mouse fibroblasts, the virus fails to transport to the nucleus due to altered endocytic processing. However, relatively little is known about the intracellular site of viral uncoating and transport of the virion across the nuclear envelope. Here, we provide evidence that AAV can efficiently enter intact nuclei purified from both permissive and less-permissive cell types. Furthermore, entry into the nucleus is time- and temperature-dependent, but is not saturable and seems to occur independently of the nuclear pore complex. We also demonstrate that purified nuclei contain all of the machinery necessary for uncoating and viral second-strand DNA synthesis even in the absence of a helper virus. These studies provide new insights into the basic biology of AAV and may also have implications for the optimal use of AAV vectors in human gene therapy
Homology modeling of a Class A GPCR in the inactive conformation: A quantitative analysis of the correlation between model/template sequence identity and model accuracy
No full textPublished in final edited form as: J Mol Graph Model. 2016 November ; 70: 140–152. doi:10.1016/j.jmgm.2016.10.004.With the present work we quantitatively studied the modellability of the inactive state of Class A G protein-coupled receptors (GPCRs). Specifically, we constructed models of one of the Class A GPCRs for which structures solved in the inactive state are available, namely the β2 AR, using as templates each of the other class members for which structures solved in the inactive state are also available. Our results showed a detectable linear correlation between model accuracy and model/template sequence identity. This suggests that the likely accuracy of the homology models that can be built for a given receptor can be generally forecasted on the basis of the available templates. We also probed whether sequence alignments that allow for the presence of gaps within the transmembrane domains to account for structural irregularities afford better models than the classical alignment procedures that do not allow for the presence of gaps within such domains. As our results indicated, although the overall differences are very subtle, the inclusion of internal gaps within the transmembrane domains has a noticeable a beneficial effect on the local structural accuracy of the domain in question
NOTES Adeno-Associated Virus Type 2-Mediated Gene Transfer: Correlation of Tyrosine Phosphorylation of the Cellular Single-Stranded D Sequence-Binding Protein with Transgene Expression in Human Cells In Vitro and Murine Tissues In Vivo
No full textAlthough the adeno-associated virus type 2 (AAV)-based vector system has gained attention as a potentially useful alternative to the more commonly used retroviral and adenoviral vectors for human gene therapy, the single-stranded nature of the viral genome, and consequently the rate-limiting second-strand viral DNA synthesis, significantly affect its transduction efficiency. We have identified a cellular tyrosine phosphoprotein, designated the single-stranded D sequence-binding protein (ssD-BP), which interacts specifically with the D sequence at the 3 * end of the AAV genome and may prevent viral second-strand DNA synthesis in HeLa cells (K. Y. Qing et al., Proc. Natl. Acad. Sci. USA 94:10879–10884, 1997). In the present studies, we examined whether the phosphorylation state of the ssD-BP correlates with the ability of AAV to transduce various established and primary cells in vitro and murine tissues in vivo. The efficiencies of transduction of established human cells by a recombinant AAV vector containing the b-galactosidase reporter gene were 293> KB> HeLa, which did not correlate with the levels of AAV infectivity. However, the amounts of dephosphorylated ssD-BP which interacted with the minus-strand D probe were also as follows: 293> KB> HeLa. Predomi-nantly the phosphorylated form of the ssD-BP was detected in cells of the K562 line, a human erythroleukemi
Fabrication of a Micro-Needle Array Electrode by Thermal Drawing for Bio-Signals Monitoring
No full textA novel micro-needle array electrode (MAE) fabricated by thermal drawing and coated with Ti/Au film was proposed for bio-signals monitoring. A simple and effective setup was employed to form glassy-state poly (lactic-co-glycolic acid) (PLGA) into a micro-needle array (MA) by the thermal drawing method. The MA was composed of 6 × 6 micro-needles with an average height of about 500 μm. Electrode-skin interface impedance (EII) was recorded as the insertion force was applied on the MAE. The insertion process of the MAE was also simulated by the finite element method. Results showed that MAE could insert into skin with a relatively low compression force and maintain stable contact impedance between the MAE and skin. Bio-signals, including electromyography (EMG), electrocardiography (ECG), and electroencephalograph (EEG) were also collected. Test results showed that the MAE could record EMG, ECG, and EEG signals with good fidelity in shape and amplitude in comparison with the commercial Ag/AgCl electrodes, which proves that MAE is an alternative electrode for bio-signals monitoring
Flexible microneedle array electrode using magnetorheological drawing lithography for bio-signal monitoring
No full textAdeno-associated Virus 2-Mediated Transduction and Erythroid Lineage-Restricted Long-Term Expression of the Human β-Globin Gene in Hematopoietic Cells from Homozygous β-Thalassemic Mice
No full textAdeno-associated virus 2 (AAV), a nonpathogenic human parvovirus, has gained attention as a potentially useful vector for human gene therapy. Here, we report successful AAV-mediated stable transduction and high-efficiency, long-term, erythroid lineage-restricted expression of a human β-globin gene in primary murine hematopoietic stem cells in vivo. Bone marrow-derived primitive Sca-1+, lin− hematopoietic stem cells from homozygous β-thalassemic mice were transduced ex vivo with a recombinant AAV vector containing a normal human β-globin gene followed by transplantation into low-dose-irradiated B6.c-kitW41/41 anemic recipient mice. Six months posttransplantation, tail-vein blood samples were analyzed by PCR amplification to document the presence of the transduced human β-globin gene sequences in the peripheral blood cells. Semiquantitative PCR analyses revealed that the transduced human β-globin gene sequences were present at ∼1 copy per cell. The efficiency of the human β-globin gene expression was determined to be up to 35% compared with the murine endogenous β-globin gene by semiquantitative RT-PCR analyses. Peripheral blood samples from several positive recipient mice obtained 10 months posttransplantation were fractionated to obtain enriched populations of granulocytes, lymphocytes, and erythroid cells. PCR analyses revealed the presence of the human β-globin gene sequences in granulocytes and lymphocytes, indicating multilineage reconstitution. However, only the erythroid population was positive following RT-PCR analyses, suggesting lineage-restricted expression of the transduced human β-globin gene. Southern blot analyses of total genomic DNA samples isolated from bone marrow cells from transplanted mice also documented proviral integration. These results provide further support for the potential use of recombinant AAV vectors in gene therapy of β-thalassemia and sickle-cell disease
