Dynamic Stabilization of DNA Assembly by Using Pyrrole-Imidazole Polyamide

We used N-methylpyrrole (Py)-N-methylimidazole-(Im) polyamide as an exogenous agent to modulate the formation of DNA assemblies at specific double-stranded sequences. The concept was demonstrated on the hybridization chain reaction that forms linear DNA. Through a series of melting curve analyses, we demonstrated that the binding of Py−Im polyamide positively influenced both the HCR initiation and elongation steps. In particular, Py−Im polyamide was found to drastically stabilize the DNA duplex such that its thermal stability approached that of an equivalent hairpin structure. Also, the polyamide served as an anchor between hairpin pairs in the HCR assembly, thus improving the originally weak interstrand stability. We hope that these proof-of-concept results can inspire future use of Py−Im polyamide as a molecular tool to modulate the formation of DNA assemblies.     

Markedly compressible behaviors of gellan hydrogels in a constrained geometry at ultraslow strain rates

The fracture behaviors of gellan hydrogels under compression remarkably depend on the strain rate as well as the boundary conditions for lateral expansion. In the geometry with no constraint for lateral expansion (conventional uniaxial compression), the gels rupture at relatively small strains independently of the compression rates. In contrast, when the gels are compressed extremely slowly (at a strain rate of ca. 10⁻⁵ s⁻¹) in the geometry prohibiting the lateral expansion at their top and bottom surfaces, they are remarkably compressible down to 2% of the initial height without macroscopic fracture and they are accompanied by a large amount of water release. In such markedly compressed gels, many microscopic cracks are formed around the central layer, where strain concentration occurs due to the nonuniform deformation arising from the constrained geometry. In the highly compressible case, the formation of macroscopic cracks is prevented by the localization of microscopic cracks as well as the enhancement in mechanical toughness by a significant increase in polymer concentration due to water release.     

Cytotoxic T lymphocyte activity after intravitreal inoculation of herpes simplex virus type I in mice

After inoculating mice with herpes simplex virus (HSV) by a corneal or intravitreal route, cytotoxic T lymphocyte (CTL) activity in the cervical lymph nodes and spleen was assayed. The spleen and cervical lymph nodes were removed at various points till 2 weeks after inoculation, and CTL activity was assayed in a groups: (A) mice intravitreally inoculated with HSV, and (B) mice with topical application of HSV. The reactivity of delayed type sensitivity was determined by the thickness of the mouse ear pinna on the 6th day in both groups. CTL activity in the spleen was at the same level in both groups. Up to 10 days after inoculation CTL activity in the cervical lymph nodes in group (A) was lower than in group (B). The reactivity of delayed type sensitivity in group (A) was lower than in group (B). These results indicate that an anterior chamber associated immune deviation (ACAID)-like phenomenon occurred after HSV inoculation into the vitreous cavity.     

Slit flow of molten polyethylene filled with plastic flakes

The processability of polyethylene in sheet-extrusion or calendering may be improved by the addition of plastic flakes. The effects of plastic flake parameters and flow conditions on the viscosity and the die swell of the suspension were determined, and the structure in flow (morphology or dispersion state) and applications were discussed. Flakes of biaxially oriented polystyrene, high-impact polystyrene, polypropylene, polyamide or polypropylene spheres were dry-blended with polyethylene. The viscosity was determined by means of a slit or a capillary rheometer. The quenched extrudate was annealed and its die swell was measured. Flakes of biaxially oriented polystyrene shrunk into rod-like form during heating, but other plastics retained the flake-form. The relative viscosity and the swell ratio of the suspensions depended on the type of plastic and operating conditions. The rheological behavior of suspensions qualitatively suggested that polypropylene flakes deformed or crowded (including stacking), polyamide flakes deformed slightly, and polystyrene flakes were difficult to buckle, but crowded slightly. The thickness of extrudates can be controlled easily by flake addition due to decreased die swell, while the output would be reduced to some extent.     

Properties of networks obtained by internal plasticization of epoxy resin with aromatic and aliphatic glycidyl compounds

In order to improve the fracture properties of p, p′-diaminodiphenylmethane-cured epoxy resin, various kinds of aromatic and aliphatic glycidyl compounds were investigated as a modifier at an amount of 30 wt %. Several compounds promoted the fracture toughness. In any glycidyl compounds, however, heat resistance was decreased by the modification. The dynamic mechanical properties of the modified epoxy resins were measured. The crosslinking density ρ was calculated from the theory of rubber elasticity, and the mechanical properties of the resins were discussed in regard to the crosslinking density. Tensile strength was scarcely affected by the crosslinking density. Elongation at break and Izod impact strength increased remarkably with decrease in crosslinking density. The fracture toughness K_Ic- increased with decrease in crosslinking density except at small ρ.     

Effects of Enantiomeric Blend of Verbenone on Response ofIps paraconfusus to Naturally Produced Aggregation Pheromone in the Laboratory

The aggregation pheromone produced by maleIps paraconfusus (Coleoptera: Scolytidae) tunneling in Monterey pine,Pinus radiata, logs was trapped on Porapak Q. A concentration of an extract of trapped volatiles that attracted beetles was determined in the laboratory through a concentration–response analysis of walking behavior of males and females. The interruptant effects of four concentrations of verbenone on response to a constant concentration of this naturally produced aggregation pheromone were tested with males and females. Independent of its enantiomeric composition [99.5% (S)-(–), 93.1 % (S)-(–), and 98.3% (R)-(+)], verbenone significantly reduced the percentage of females (but not males) reaching the attractant source. However, when the time required for beetles of both sexes taken together to reach the attractant source was considered, verbenone of higher enantiomeric purity had a greater effect on beetle behavior. Solutions of 99.5% (S)-(–)- and 98.3% (R)-(+)-verbenone increased the time required for beetles to reach the attractive source when compared to 93.1% (S)-(–)-verbenone. When pooled across enantiomeric blends, increasing concentrations of verbenone resulted in slower responses in beetles that reached the attractant source within 2 min. Males and females did not respond to verbenone alone.     

Nondestructive determination of fatty acid composition of husked sunflower (Helianthus annua L.) seeds by near-infrared spectroscopy

Determination of the fatty acid composition of sunflower (Helianthus annua L.) seeds by near-infrared (NIR) spectroscopy was examined. Sunflower seeds were husked (removed from their hulls by a husking machine or manually with a knife). NIR spectra of these seeds were scanned from 1100 to 2500 nm at 2-nm intervals in a whole-grain cell with a wideangle moving drawer for machine-husked seeds or in a single-grain cup for a manually husked single-grain seed. The extracted oils from machine-husked seeds also were scanned by sandwiching them between a pair of slide glasses to create a thin layer and by placing them on a syrup cup. For extracted oil, the absorption band around 1720 nm filled out to the shorter wavelength region in the NIR second-derivative spectra as the percentage of the linoleic acid moiety increased, because linoleic acid absorbs in this region. On the other hand, for husked seeds and for a single-grain seed, as the percentage of linoleic acid increased, the trough at 1724 nm where oleic and saturated acids absorb decreased in the second-derivative NIR spectra. Determination of the fatty acid composition of sunflower seeds could be carried out successfully according to the NIR spectral pattern for both extracted oil (r=−0.989) and kernel seed (r=−0.993). This is important, especially for a manually husked single-grain seed (r=−0.971), because it can still be germinated after such nondestructive analysis.     

Mutational analysis of structure--activity relationships in human tumor necrosis factor-alpha

To determine the region of human tumor necrosis factor-alpha(TNF-), essential for cytotoxic activity against mouse L-M cells,single amino-acid-substituted TNF- mutant proteins (muteins)were produced in Escherichia coli by protein engineering techniques.An expression plasmid for TNF- was mutagenized by passage throughan E.coli mutD5 mutator strain and by oligonucleotide-directedmutagenesis. Approximately 100 single amino-acid-substitutedTNF- muteins were produced and assayed for cytotoxic activity.The cytotoxic activities of purified TNF- muteins, e.g. TNF-31T,-32Y, -82D, -85H, -115L, -141Y, -144K and -146E, were < 1%of that of parent TNF-. These results indicate that the integrityof at least four distinct regions of the TNF- molecule is requiredfor full biological activity. These regions are designated asfollows: region I, from position 30 to 32; region II, from position82 to 89; region III, from position 115 to 117; region FV, fromposition 141 to 146. In addition, TNF-141Y could not completelycompete with parent TNF- for binding to the receptor. This demonstratesthat region IV, and at least aspartk acid at position 141, mustbe involved in the TNF receptor binding site.     

Rinsing effect of alkaline electrolyzed water on nickel surfaces

The improvement in the surface cleanliness of electroplated nickel by rinsing in alkaline electrolyzed water (AEW) was determined. When the nickel plated sample was rinsed with the AEW, it was found that the amount of residual sulfate ion on the surface of a sample decreased approximately by half compared to one rinsed only with de-ionized pure water. Because nanosize hydrogen bubbles are present in the AEW, and the zeta-potential has a negative value, we then surmised that the mechanism of rinsing was as follows: The sulfate ions are selectively absorbed on the nanosize colloidal hydrogen bubbles, or substituted for anions absorbed on the hydrogen bubbles. The sulfate ions absorbed on the nickel surface then become detached. The detached sulfate ions are absorbed on the surface of the hydrogen bubbles, and negatively charge the hydrogen bubbles. It can be considered that any detached sulfate ions do not re-adhere due to the electrical repulsion force of the negatively charged nickel surface. Thus the sample is efficiently rinsed.