Dimeric C34 Derivatives Linked through Disulfide Bridges as New HIV-1 Fusion Inhibitors

C34, a 34-mer fragment peptide, is contained in the HIV-1 envelope protein gp41. A dimeric derivative of C34 linked through a disulfide bridge at its C terminus was synthesized and found to display potent anti-HIV activity, comparable with that of a previously reported PEGylated dimer of C34REG. The reduction in the size of the linker moiety for dimerization was thus successful, and this result might shed some light on the mechanism of the suppression of six-helix bundle formation by these C34 dimeric derivatives. Addition of a Gly-Cys(CH₂CONH₂)-Gly-Gly motif at the N-terminal position of a C34 monomeric derivative significantly increased the anti-HIV-1 activity. This moiety functions as a new pharmacophore, and this might provide a useful insight into the design of potent HIV-1 fusion inhibitors.     

Development of Laser Ionization Techniques for Evaluation of the Effect of Cancer Drugs Using Imaging Mass Spectrometry

Recently, combined therapy using chemotherapy and photodynamic therapy (PDT) has been proposed as a means of improving treatment outcomes. In order to evaluate the efficacy of combined therapy, it is necessary to determine the distribution of the anticancer drug and the photosensitizer. We investigated the use of imaging mass spectrometry (IMS) to simultaneously observe the distributions of an anticancer drug and photosensitizer administered to cancer cells. In particular, we sought to increase the sensitivity of detection of the anticancer drug docetaxel and the photosensitizer protoporphyrin IX (PpIX) by optimizing the ionization-assisting reagents. When we used a matrix consisting of equal weights of a zeolite (NaY5.6) and a conventional organic matrix (6-aza-2-thiothymine) in matrix-assisted laser desorption/ionization, the signal intensity of the sodium-adducted ion of docetaxel (administered at 100 μM) increased about 13-fold. Moreover, we detected docetaxel with the zeolite matrix using the droplet method, and detected PpIX by fluorescence and IMS with α-cyano-4-hydroxycinnamic acid (CHCA) using the spray method.     

Characterization of Acetylxylan Esterase from White-Rot Fungus Irpex lacteus

The carbohydrate esterase family 1 (CE1) in CAZy contains acetylxylan esterases (AXEs) and feruloyl esterases (FAEs). Here we cloned a gene coding for an AXE belonging to CE1 from Irpex lacteus (IlAXE1). IlAXE1 was heterologously expressed in Pichia pastoris, and the recombinant enzyme was purified and characterized. IlAXE1 hydrolyzed p-nitrophenyl acetate, α-naphthyl acetate and 4-methylumbelliferyl acetate, however, it did not show any activity on ethyl ferulate and methyl p-coumarate. We also examined the activity on partially acetylated and feruloylated xylan extracted from corncob by hydrothermal reaction. Similarly, ferulic and p-coumaric acids were not liberated, and acetic acid was only detected in the reaction mixture. The results indicated that IlAXE1 is an acetylxylan esterase actually reacted to acetyl xylan. However, since IlAXE1 was unable to completely release acetic acid esterifying xylopyranosyl residues, it is assumed that acetyl groups exhibiting resistance to deacetylation by IlAXE1 are present in corn cob xylan.     

Exploring the Minimal RNA Substrate of Flexizymes

Flexizymes are tRNA acylation ribozymes that have been successfully used to facilitate genetic code reprogramming. They are capable of charging acid substrates onto various tRNAs and tRNA analogues. However, their minimal RNA substrate has not been investigated. Here we have designed fluorescently labeled short RNAs corresponding to the four, three, and two bases (4bRNA, 3bRNA, 2bRNA) at the tRNA 3′-end and explored the minimal RNA substrate of flexizymes, dFx and eFx. 3bRNA was the observed minimal RNA substrate of the flexizymes, but the efficiency of acylation of this short RNA was two to three times lower than that of 4bRNA. The efficiency of acylation of 4bRNA was comparable with that of the microhelix, a 22-base RNA conventionally used as a tRNA analogue for analyzing acylation efficiency. We also compared the efficiencies of acylation of the microhelix and 4bRNA with various acid substrates. Thanks to the short length of 4bRNA, its acyl-4bRNA products exhibited larger mobility shifts in gel electrophoresis than those exhibited by acyl-microhelix products with every substrate tested. This indicated that 4bRNA was an ideal RNA substrate for analyzing the efficiency of acylation by flexizymes.     

Traditional Japanese architecture blends beauty and rationale[lessons for software engineering]

It's hard to find a good instance of harmonization of beauty and rationale in the field of software engineering. We can, however, find it in ancient Japanese architecture which is where we might find the clues we need to achieve our goal. Unfortunately, the software community is relatively young, so there are few such secrets or literatures to use as a basis for our communications. Moreover, because the structure of software is not obvious yet architectural structures are clearly visible-the difficulty we face might be larger and deeper than that of other disciplinary fields. However, by carefully examining ancient architectural methods, we believe that we are stepping in the right direction in our pursuit of the patterns that will realize the harmony of beauty and rationale     

Possibility of Thermal Protection in Earth Re-entry Flight by MHD Flow Control with Air-Core Circular Magnet

The possibility of thermal protection in an earth re-entry flight by the magnetohydrodynamic (MHD) flow control with an air-core circular magnet producing about 1 T or less on the wall surface of a space vehicle is examined by r-z two-dimensional numerical simulations. The present study adopts a hemisphere with a radius of 1.0 m as the nose shape of a space vehicle. The numerical results show that aerodynamic heating can be reduced by the MHD flow control with an air-core circular magnet, and also that the maximum wall heat flux and the effect of the MHD flow control become prominent with the magnitude of coil current and the flight altitude. Furthermore, it is found that there is an optimum magnet size to maximize the effect of MHD flow control under constant exciting magnetic energy, and also the optimum size gets reduced with the decrease in the flight altitude. © 2009 Institute of Electrical Engineers of Japan. Published by John Wiley & Sons, Inc.     

Increased leukocyte aggregates are associated with atherosclerosis in patients with hemodialysis

Little data are available on the role of blood rheology in atherosclerosis in hemodialysis (HD) patients. This study sought to assess the relationship between leukocytes conjugated with platelets (leukocyte aggregates [LA]) and atherosclerosis in patients with HD. The present study included 118 patients on HD. As surrogate markers of atherosclerosis, aortic stiffness measured by brachial-ankle pulse wave velocity, and carotid intima-media thickness (IMT) were measured. As an assessment of LA, a method, microchannel array flow analyzer, which makes it possible to directly observe the flow of blood cell elements through the microchannel, was used. We measured a number of LA during 50 μL flow of whole blood through microchannels. In 12 age-matched healthy individuals, a number of LA during 50 μL flow of whole blood was 25.7±5.4, whereas in HD patients it was significantly increased up to 48.2±16.4 (P<0.001). Flow cytometry demonstrated that LA were predominantly monocytes. Leukocyte aggregates were positively associated with plasma levels of fibrinogen (P<0.01), or serum high-sensitive C-reactive protein (P<0.01). Moreover, LA had highly significant associations with brachial-ankle pulse wave velocity (P<0.001) and IMT (P<0.001). In conclusion, we demonstrated hemorheologically that monocyte-platelet conjugates play an important role in aortic stiffness and IMT in HD patients.     

Fabrication of a working Bi-2223 superconducting magnet cooled by liquid nitrogen

A practical Bi-2223 superconducting magnet, working in liquid nitrogen (L.N₂), was designed and fabricated. Bi-2223 tape with a critical current of 147 A was prepared by a controlled overpressure (CT-OP) process at 77.3 K in self-field. Ten double-pancake coils were resistively connected by copper terminals. The bore diameter was 54 mm?, the magnet outer diameter was 122 mm?, the height of the magnet was 124 mm, and the weight of the magnet was about 3 kg. The maximum magnetic field at the center of the bore was 0.48 T with an operating current of 50 A. The experimental results agree well with design predictions calculated by finite element method. AC operation was also performed, and no distortion of the voltage waveform was observed. Therefore, this Bi-2223 superconducting magnet is a suitable replacement for copper magnets designed for applications in science and technology.     

Characterization of acylmono-, mono-, di-, tri- and tetraglycosylsterol and saponin in Adzuki bean (Vigna angularis) seeds

Five sterylglycosides (acylmono-, mono-, di tri- and tetraglycosylsterol) and a saponin were isolated from Adzuki beans and characterized. In the glycosylsterols, the principal component sterols were sitosterol and stigmasterol; the major sugar component was glucose. The glucose units were shown to be linked by β1,6-bonds. The three oligoglycosylsterols were shown to be gentiobiosylsterol, gentiotriosylsterol and gentiotetraosylsterol; the latter two are novel sterylglycosides. The saponin was identified as glucopyranosyl-(β1→2)-glucopyranurosyl-(β1→3′)-soyasapogenol B (Azukisaponin I) which had previously been found in Adzuki beans.     

Enhancement of low density lipoprotein binding to both low density lipoprotein receptor-positive and- negative cells by tetracycline antibiotics

Some tetracycline (TC) antibiotics, including TC and anhydrotetracycline, have been found to enhance specific binding of low density lipoprotein (LDL) to both LDL receptor-positive and-negative cells at relatively higher concentrations. When incubated at 37°C, the ability of LDL receptor-negative human fibroblasts to bind ¹²⁵I-LDL was increased from<2 to 45 ng/mg by 170 μM TC. In normal human fibroblasts and Hep G2 cells, ¹²⁵I-LDL binding was elevated 1.4- to 2-fold by 113 μM TC. The ¹²⁵I-LDL binding in the presence of TC was diminished by both heparin and EDTA. The enhancement by TC was not observed when ¹²⁵I-LDL binding was assayed at 4°C. TC enhanced LDL binding to paraformaldehyde-fixed Hep G2 cells, excluding LDL receptor induction in the mechanism. These results demonstrated that TC enhanced cellular LDL binding through a process not involving functional LDL receptors.