Anti-Mycobacterial Nucleoside Antibiotics from a Marine-Derived Streptomyces sp. TPU1236A

Five new nucleoside antibiotics, named streptcytosines A–E (1–5), and six known compounds, de-amosaminyl-cytosamine (6), plicacetin (7), bamicetin (8), amicetin (9), collismycin B (10), and SF2738 C (11), were isolated from a culture broth of Streptomyces sp. TPU1236A collected in Okinawa, Japan. The structures of new compounds were elucidated on the basis of their spectroscopic data (HRFABMS, IR, UV, and 2D NMR experiments including ¹H-¹H COSY, HMQC, HMBC, and NOESY spectra). Streptcytosine A (1) belonged to the amicetin group antibiotics, and streptcytosines B–E (2–5) were derivatives of de-amosaminyl-cytosamine (6), 2,3,6-trideoxyglucopyranosyl cytosine. Compound 1 inhibited the growth of Mycobacterium smegmatis (MIC = 32 µg/mL), while compounds 2–5 were not active at 50 µg/disc. Bamicetin (8) and amicetin (9) showed the MICs of 16 and 8 µg/mL, respectively.     

β 1-4 mannobiose enhances Salmonella-killing activity and activates innate immune responses in chicken macrophages

Salmonella spp. is one of the major causes of food-borne illness in humans, and Salmonella enteritidis (SE) infection in commercial poultry is a world-wide problem. Here we have investigated the in vitro immune-modulating effects of β 1-4 mannobiose (MNB), which was previously found to prevent SE infection in vivo in chickens, using chicken macrophage (MQ-MCSU) cells. Treatment of MQ-NCSU cells with MNB dose-dependently increased both phagocytic activity and Salmonella-killing activity of macrophages, with the highest reduction in SE viability observed at a concentration of 40 μg/ml at 48 h post-infection. Likewise, both hydrogen peroxide (H(2)O(2)) and nitric oxide (NO) production were increased in a dose-dependent manner by MNB. Gene expression analysis of MNB-treated macrophages revealed significant increases in the expression of iNOS, NOX-1, IFN-γ, NRAMP1, and LITAF, genes critical for host defense and antimicrobial activity, when compared to untreated cells. This data confirms that MNB possesses potent innate immune-modulating activities and can up-regulate antibacterial defenses in chicken macrophages.     

Endocrine status and development of porcine testicular tissues in host mice

Clarification of the endocrine status of host mice provides us with basic knowledge with which we can manipulate the growth and function of xenografted testicular tissues. We investigated the hormonal profiles of castrated mice grafted with porcine immature testicular tissues from 30 to 210 or more days after grafting (day 0=castration and grafting). The serum follicle stimulating hormone (FSH) concentrations of the host mice declined (P<0.05) from day 60 compared with those of the castrated, ungrafted mice. The serum inhibin and testosterone levels were higher (P<0.05) than those in the castrated, ungrafted mice from days 30 and 90 days, respectively. The inhibin levels further increased (P<0.05) from day 90, during which time the levels were higher (P<0.05) than those in the intact male mice. In the grafts, formation of lumens occurred in the seminiferous cords on day 90 and spermatozoa appeared in the lumens from day 120. However, spermatogenesis in the grafts did not reach the qualitatively normal levels observed in adult boars. The intensity of the immune reaction to inhibin alpha subunits in the Sertoli cells of the grafts decreased with differentiation of the seminiferous tubules. The present findings indicate that a feedback loop was established between the mouse hypothalamo-pituitary axis and the grafted porcine tissues from 60 days post-grafting. The results also indicate that the serum inhibin levels in the host mice remained high even after the appearance of lumens in the seminiferous tubules of the grafted tissues; this is strikingly different to the situation in normal male animals, in which the serum inhibin levels decline at around the time of tubular differentiation. The lack of efferent ducts in the tubules of the grafted tissues probably caused the accumulation of inhibin to be released into the lumens, resulting in high concentrations of circulating inhibin. These high levels of inhibin may directly affect spermatogenic activity and suppress FSH secretion.     

Compressive deformation of wood impregnated with low molecular weight phenol formaldehyde (PF) resin II: effects of processing parameters

To obtain high-strength phenol formaldehyde (PF) resin-impregnated compressed wood at low pressing pressure, the effects of resin content, preheating temperature, pressing temperature, and pressing speed on the compressive deformation of oven-dried low molecular weight PF resin-impregnated wood was investigated. With an increase of PF resin content, the Youngs modulus of the cell wall perpendicular to the fiber direction decreases, and collapse-initiating pressure decreases linearly with the Youngs modulus. This indicates that the occurrence of cell wall collapse is strain-dependent. By increasing preheating temperatures, the collapse-initiating pressure increases due to the increment of the Youngs modulus of the cell wall. An increase in pressing temperature results in the thermal softening of the cell wall and causes collapse at a lower pressure. The wood is compressed effectively despite accelerated resin curing. The pressing speed significantly affects the viscoelastic deformation of the cell wall and the wood is well deformed with decreasing pressing speed, although the differences in density and mechanical properties are relatively small after a pressure-holding period of 30min. In all the parameters examined in this study, the Youngs modulus and bending strength increase with increasing density.     

Identification of a novel cytotoxic protein, Cry45Aa, from Bacillus thuringiensis A1470 and its selective cytotoxic activity against various mammalian cell lines

Parasporal inclusion proteins produced by Bacillus thuringiensis strain A1470 exhibit strong cytotoxicity against human leukemic T cells when activated by protease treatment. One of the cytotoxic proteins was separated by anion exchange chromatography and gel filtration chromatography and designated Cry45Aa. Its gene was then expressed in recombinant Escherichia coli, in which the Cry45Aa precursor was accumulated in an inclusion body. It was solubilized in sodium carbonate buffer and processed with proteinase K, and cytotoxic activities of the protein against various mammalian cell lines were evaluated using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide assay. The protein exhibited high cytotoxic activity against CACO-2, Sawano, MOLT-4, TCS, and HL60 cells and moderate activity against U-937 DE-4, PC12, and HepG2 cells. On the other hand, the EC50 values against Jurkat, K562, HeLa, A549, Vero, COS-7, NIH3T3, CHO, and four normal tissue cells (human primary hepatocyte cells, UtSMC, MRC-5, and normal T cells) were >2 microg/mL.     

Durability evaluation of particleboards by accelerated aging tests

Summary Specimens of four commercially produced particleboards were exposed to various accelerated aging treatments: 1) 6-cycle ASTM D-1037 exposure, 2) 6-cycle WCAMA exposure, 3) BS 5669 test, 4) cyclic soak-dry test, and 5) cyclic boil-dry test. Thickness swelling (TS) and internal bond strength (IB) were measured after each step of each cycle. Phenol and phenol (face)/isocyanate (core)bonded boards are more durable than urea and melamine urea bonded boards. The effect of treatment 1) is most severe on TS and IB retention, followed in order by treatments 2), 5), 4) and 3). When the value of springback reached about 35%, IB became equal to zero.     

The influences of boiling and drying treatments on the behaviors of tension wood with gelatinous layers in <Emphasis Type="Italic">Zelkova serrata</Emphasis>

This study examined how boiling and drying treatments influenced various physical properties of the tension wood with gelatinous fibers (G-fibers) of a 29-yearold Zelkova branch. By boiling treatment, tension wood with numerous G-fibers contracted considerably in the longitudinal direction and the longitudinal Young’s modulus decreased in spite of the water-saturated condition. The drying treatment caused green tension wood and boiled tension wood with numerous G-fibers to shrink longitudinally and increased their longitudinal Young’s moduli. These specific behaviors in tension wood were highly correlated with the proportion of G-fibers in a specimen and were probably caused by the microscopic behavior of cellulose microfibril (CMF) in the gelatinous layers (G-layers). The longitudinal shrinkage of tension wood due to drying suggests the existence of a hygro-sensible, noncrystalline region in the CMF, which is abundant in the G-layer. Furthermore, the noncrystalline region in the CMF softens during boiling treatment, resulting in the reduction of the longitudinal Young’s modulus in tension wood. The longitudinal contraction of tension wood with G-fibers by boiling might be caused by the tensile growth stress remaining in green G-layers. However, no changes were detected in the 004 d-spacing of cellulose crystal in tension wood from the boiling and drying treatments, regardless of the proportion of G-fibers.