Surface Chirality of Gly‐Pro Dipeptide Adsorbed on a Cu(110) Surface

The adsorption of chiral Gly‐Pro dipeptide on Cu(110) has been characterized by combining in situ polarization modulation infrared reflection absorption spectroscopy (PM‐RAIRS) and X‐ray photoelectron spectroscopy (XPS). The chemical state of the dipeptide, and its anchoring points and adsorption geometry, were determined at various coverage values. Gly‐Pro molecules are present on Cu(110) in their anionic form (NH₂/COO⁻) and adsorb under a 3‐point binding via both oxygen atoms of the carboxylate group and via the nitrogen atom of the amine group. Low‐energy electron diffraction (LEED) and scanning tunneling microscopy (STM) have shown the presence of an extended 2D chiral array, sustained via intermolecular H‐bonds interactions. Furthermore, due to the particular shape of the molecule, only one homochiral domain is formed, creating thus a truly chiral surface. Chirality 27:411–416, 2015. © 2015 Wiley Periodicals, Inc.     

Lipid nanotechnologies for structural studies of membrane‐associated proteins

We present a methodology of lipid nanotubes (LNT) and nanodisks technologies optimized in our laboratory for structural studies of membrane‐associated proteins at close to physiological conditions. The application of these lipid nanotechnologies for structure determination by cryo‐electron microscopy (cryo‐EM) is fundamental for understanding and modulating their function. The LNTs in our studies are single bilayer galactosylceramide based nanotubes of ～20 nm inner diameter and a few microns in length, that self‐assemble in aqueous solutions. The lipid nanodisks (NDs) are self‐assembled discoid lipid bilayers of ～10 nm diameter, which are stabilized in aqueous solutions by a belt of amphipathic helical scaffold proteins. By combining LNT and ND technologies, we can examine structurally how the membrane curvature and lipid composition modulates the function of the membrane‐associated proteins. As proof of principle, we have engineered these lipid nanotechnologies to mimic the activated platelet's phosphtaidylserine rich membrane and have successfully assembled functional membrane‐bound coagulation factor VIII in vitro for structure determination by cryo‐EM. The macromolecular organization of the proteins bound to ND and LNT are further defined by fitting the known atomic structures within the calculated three‐dimensional maps. The combination of LNT and ND technologies offers a means to control the design and assembly of a wide range of functional membrane‐associated proteins and complexes for structural studies by cryo‐EM. The presented results confirm the suitability of the developed methodology for studying the functional structure of membrane‐associated proteins, such as the coagulation factors, at a close to physiological environment. Proteins 2014; 82:2902–2909. © 2014 Wiley Periodicals, Inc.     

Cloning and restriction fragment length polymorphism analysis of a cDNA for swine PIT-1, a gene controlling growth hormone expression*

A partial swine cDNA which encodes the functional domain of PIT-1 was isolated by the polymerse chain reaction (PCR). The swine PIT-1 cDNA clone is 95% identical at the protein level to the rat Pit-1 gene. Thus, Pit-l's known function in control of rat growth hormone and prolactin expression is likely to be conserved in swine. This swine cDNA clone was used to investigate genetic variability at PIT-1 in several American and Chinese breeds. Polymorphic BamIII fragments were found in pure-bred Meishan animals (n= 13), but only monomorphic fragments in five American breeds (n= 36).     

Long‐distance dispersal of non‐native pine bark beetles from host resources

1. Dispersal and host detection are behaviours promoting the spread of invading populations in a landscape matrix. In fragmented landscapes, the spatial arrangement of habitat structure affects the dispersal success of organisms. 2. The aim of the present study was to determine the long distance dispersal capabilities of two non‐native pine bark beetles (Hylurgus ligniperda and Hylastes ater) in a modified and fragmented landscape with non‐native pine trees. The role of pine density in relation to the abundance of dispersing beetles was also investigated. 3. This study took place in the Southern Alps, New Zealand. A network of insect panel traps was installed in remote valleys at known distances from pine resources (plantations or windbreaks). Beetle abundance was compared with spatially weighted estimates of nearby pine plantations and pine windbreaks. 4. Both beetles were found ≥25 km from the nearest host patch, indicating strong dispersal and host detection capabilities. Small pine patches appear to serve as stepping stones, promoting spread through the landscape. Hylurgus ligniperda (F.) abundance had a strong inverse association with pine plantations and windbreaks, whereas H. ater abundance was not correlated with distance to pine plantations but positively correlated with distance to pine windbreaks, probably reflecting differences in biology and niche preferences. Host availability and dispersed beetle abundance are the proposed limiting factors impeding the spread of these beetles. 5. These mechanistic insights into the spread and persistence of H. ater and H. ligniperda in a fragmented landscape provide ecologists and land managers with a better understanding of factors leading to successful invasion events, particularly in relation to the importance of long‐distance dispersal ability and the distribution and size of host patches.     

The Antitumor Activity of the Novel Compound Jesridonin on Human Esophageal Carcinoma Cells

Jesridonin, a small molecule obtained through the structural modification of Oridonin, has extensive antitumor activity. In this study, we evaluated both its in vitro activity in the cancer cell line EC109 and its in vivo effect on tumor xenografts in nude mice. Apoptosis induced by Jesridonin was determined using an MTT assay, Annexin-V FITC assay and Hoechest 33258 staining. Apoptosis via mitochondrial and death receptor pathways were confirmed by detecting the regulation of MDM2, p53, and Bcl-2 family members and by activation of caspase-3/-8/-9. In addition, vena caudalis injection of Jesridonin showed significant inhibition of tumor growth in the xenograft model, and Jesridonin-induced cell apoptosis in tumor tissues was determined using TUNEL. Biochemical serum analysis of alkaline phosphatase (ALP), alanine transaminase (ALT), aspartate transaminase (AST), gamma-glutamyl transferase (GGT), total protein (TP) and albumin (ALB) indicated no obvious effects on liver function. Histopathological examination of the liver, kidney, lung, heart and spleen revealed no signs of JD-induced toxicity. Taken together, these results demonstrated that Jesridonin exhibits antitumor activity in human esophageal carcinomas EC109 cells both in vitro and in vivo and demonstrated no adverse effects on major organs in nude mice. These studies provide support for new drug development.     

Nickel chloride inhibits the DNA repair of UV-treated but not methyl methanesulfonate-treated chinese hamster ovary cells

Nickel, a human carcinogen, has been shown to enhance the cytotoxicity, mutagenicity, and sister-chromatid exchanges (SCE) induced by ultraviolet (UV) light but not by methyl methanesulfonate (MMS). To verify that the cocytotoxicity and cogenotoxicity of nickel are correlated with its inhibition on DNA repair, the effects of nickel on the DNA repair induced by UV and by MMS have been investigated. Our analyses of DNA repair of single-strand breaks by alkaline elution and alkaline sucrose sedimentation indicate that nickel inhibited the DNA repair in UV-treated, but not in MMS-treated cells. Therefore, the inhibition of DNA repair seems to play an important role in the cocytotoxicity and comutagenicity of nickel. However, the inhibition of DNA repair seems not to play a decisive role in enhancing SCE, because we have previously shown that arsenite inhibits the UV-induced DNA repair, but has no enhancing effect on the UV-induced SCE. Our results also show that nickel had obvious inhibitory effects on DNA ligation and postreplication repair, but had no apparent effect on nucleotide excision and DNA polymerization in the UV repair. The results of the DNA ligation inhibition by nickel in UV but not in MMS repair suggest that different ligases are used in the DNA repair of UV- and MMS-induced damages.     

DIPTEREX RESISTANCE IN THE HOUSEFLY AND THE SYNERGIC MECHANISM OF SV1 IN RELATION TO PENETRATION THROUGH CUTICLE

Abstract  SV₁ was observed to have obvious synergism and could delay housefly ( Musca domestica vicina ) resistance development to Dipterex. The penetration rates of Dipterex through housefly cuticle were determined in a susceptible and two resistant strains. The results indicated that the penetration in the resistant housefly strains was obviously slower than in the susceptible one. The penetrating rate of SV₁+ Dipterex (in mixture) was higher than that of Dipterex. The penetration reduction in resistant houseflies may be an important factor in bringing forth resistance. The increase of the penetrating rate of Dipterex and the decrease of its metabolic rate are regarded as the important mechanisms of SV₁ synergism to Dipterex.