Synchrotron X-ray reflectivity studies of nanoporous organosilicate thin films with low dielectric constants

Quantitative, non-destructive X-ray reflectivity analysis using synchrotron radiation sources was successfully performed on nanoporous dielectric thin films prepared by thermal processing of blend films of a thermally curable polymethylsilsesquioxane dielectric precursor and a thermally labile triethoxy-silyl-terminated six-arm poly(epsilon-caprolactone) porogen in various compositions. In addition, thermogravimetric analysis and transmission electron microscopy analysis were carried out. These measurements provided important structural information about the nanoporous films. The thermal process used in this study was found to cause the porogen molecules to undergo efficiently sacrificial thermal degradation, generating closed, spherical nanopores in the dielectric film. The resultant nanoporous films exhibited a homogeneous, well defined structure with a thin skin layer and low surface roughness. In particular, no skin layer was formed in the porous film imprinted using a porogen loading of 30 wt%. The film porosities ranged from 0 to 33.8% over the porogen loading range of 0-30 wt%open131

Functional and structural characteristics of anticancer peptide Pep27 analogues

BACKGROUND: A secreted peptide Pep27 initiates the cell death program in S. pneumoniae through signal transduction. This study was undertaken to evaluate the relation between the structure and cytotoxic activity of Pep27 and its analogues on cancer cells. RESULTS: Pep27anal2 characterized substituting ((2)R→W), ((4)E→W), ((11)S→W) and ((13)Q→W) in native Pep27, exhibited greater hydrophobicity and anticancer activity than Pep27 and other analogues. The IC(50 )values of Pep27anal2 were approximately 10 – 30 μM in a number of cell lines (AML-2, HL-60, Jurkat, MCF-7 and SNU-601). Confocal microscopy showed that Pep27anal2-FITC was localized in the plasma membrane, and then moving from the membrane to subcellular compartments with the initiation of membrane blebbing. Flow cytometric analysis using propidium iodide and Annexin V also revealed that Pep27anal2 induced apoptosis with minor membrane damage. Electron microscopy revealed that Pep27 induced apoptosis in Jurkat cells. The anticancer activity of Pep27anal2 was neither abrogated by pan-caspase inhibitor (Z-VAD-fmk) nor related to cytochrome c release from mitochondria. The 3D solution structures of these two Pep27 peptides revealed that both form a random coil conformation in water; however, they adopted stable α-helical conformations in solutions. CONCLUSION: The results indicate that Pep27anal2 can penetrate the plasma membrane, and then induce apoptosis in both caspase-and cytochrome c-independent manner. The hydrophobicity of Pep27anal2 appears to play an important role in membrane permeabilization and/or anticancer properties. The structure-functional relationships of these peptides are also discussed. It is proposed that Pep27anal2 is a potential candidate for anticancer therapeutic agents

Plasma modification of poly(2-heptadecyl-4-vinylthieno[3,4-d]thiazole) low bandgap polymer and its application in solar cells

For the first time, we here propose a green methodology to modify a low bandgap polymer for highly efficient solar cells using atmospheric pressure plasma jet or soft plasma operating on different feeding gases (air, Ar and N-2). The physical properties of the modified polymer were investigated using conductivity measurements, UV-visible spectroscopy, photoluminescence spectroscopy, X-ray photoelectron spectroscopy, cyclic voltammograms, atomic force microscopy, cathodoluminescence and confocal Raman spectroscopy. Further, we examined the variation of the work function of the polymer before and after plasma treatment using a gamma-focused ion beam. Additionally, photovoltaic cells based on the plasma-modified polymer having ITO/PEDOT:PSS/PHVTT (with or without plasma modification): PC71BM/LiF/Al configuration were fabricated and then characterized. We found that the power conversion efficiency (PCE) of the plasma-modified polymer increased dramatically as compared to the control polymer (without plasma treatment). PCE of the control polymer was found to be 4.11%, while after air, Ar and N-2 gas plasma treatment the polymer showed PCEs of 4.85%, 4.87% and 5.14% respectively. Thus, plasma treatment not only alters the surface properties, but also modifies the bulk properties (changes in HOMO and LUMO bandgap level). Hence, this work provides new dimensions to explore more about plasma and polymer chemistry

Identification of Domains Directing Specificity of Coupling to G-proteins for the Melanocortin MC3 and MC4 Receptors

The melanocortin receptors, MC3R and MC4R, are G protein-coupled receptors that are involved in regulating energy homeostasis. Using a luciferase reporter gene under the transcriptional control of a cAMP-responsive element (CRE), the coupling efficiency of the MC4R and MC3R to G-proteins was previously shown to be different. MC4R exhibited only 30-50% of the maximum activity induced by MC3R. To assess the role of the different MC3R and MC4R domains in G-protein coupling, several chimeric MC3R/MC4R receptors were constructed. The relative luciferase activities, which were assessed after transfecting the chimeric receptors into HEK 293T cells, showed that the i3 (3rd intracellular) loop domain has an essential role in the differential signaling of MC3R and MC4R. To reveal which amino acid residue was involved in the MC4R-specific signaling in the i3 loop, a series of mutant MC4Rs was constructed. Reporter gene analysis showed that single mutations of Arg220 to Ala and Thr232 to either Val or Ala increased the relative luciferase activities, which suggests that these specific amino acids, Arg220 and Thr232, in the i3 loop of MC4R play crucial roles in G-protein coupling and the subtype-specific signaling pathways. An examination of the inositol phosphate (IP) levels in the cells transfected with either MC3R or MC4R after being exposed to the melanocortin peptides revealed significant stimulation of IP production by MC3R but no detectable increase in IP production was observed by MC4R. Furthermore, none of the MC4R mutants displayed melanocortin peptide-stimulated IP production. Overall, this study demonstrated that MC3R and MC4R have distinct signaling in either the cAMP- or the inositol phospholipid-mediated pathway with different conformational requirements

Structural insights into the HBV receptor and bile acid transporter NTCP

B型肝炎ウイルスの受容体“胆汁酸輸送体”の立体構造を解明. 京都大学プレスリリース. 2022-05-18.Roughly 250 million people are infected with hepatitis B virus (HBV) worldwide, and perhaps 15 million also carry the satellite virus HDV, which confers even greater risk of severe liver disease. Almost ten years ago the HBV receptor was identified as NTCP (sodium taurocholate co-transporting polypeptide), which interacts directly with the first 48 amino acid residues of the N-myristoylated N-terminal preS1 domain of the viral large (L) protein. Despite the pressing need for therapeutic agents to counter HBV, the structure of NTCP remains unsolved. This 349-residue protein is closely related to human apical sodium-dependent bile acid transporter (ASBT), another member of the solute carrier family SLC10. Crystal structures have been reported of similar bile acid transporters from bacteria, and these models with ten transmembrane helices are believed to resemble strongly both NTCP and ASBT. Using cryo-electron microscopy we have solved the structure of NTCP bound to an antibody, clearly showing the transporter has no equivalent to the first transmembrane helix of other SLC10 models, leaving the N-terminus exposed on the extracellular face. Comparison of the different structures indicates a common mechanism of bile acid transport, but the NTCP structure also displays a pocket formed by residues known to interact with preS1, presenting new and enticing opportunities for structure-based drug design

Characteristic Analysis of Homo- and Heterodimeric Complexes of Human Mitochondrial Pyruvate Carrier Related to Metabolic Diseases

Human mitochondrial pyruvate carriers (hMPCs), which are required for the uptake of pyruvate into mitochondria, are associated with several metabolic diseases, including type 2 diabetes and various cancers. Yeast MPC was recently demonstrated to form a functional unit of heterodimers. However, human MPC-1 (hMPC-1) and MPC-2 (hMPC-2) have not yet been individually isolated for their detailed characterization, in particular in terms of their structural and functional properties, namely, whether they exist as homo- or heterodimers. In this study, hMPC-1 and hMPC-2 were successfully isolated in micelles and they formed stable homodimers. However, the heterodimer state was found to be dominant when both hMPC-1 and hMPC-2 were present. In addition, as heterodimers, the molecules exhibited a higher binding capacity to both substrates and inhibitors, together with a larger structural stability than when they existed as homodimers. Taken together, our results demonstrated that the hetero-dimerization of hMPCs is the main functional unit of the pyruvate metabolism, providing a structural insight into the transport mechanisms of hMPCs