Immunohistochemical detection of scrapie prion proteins in clinically normal sheep in Pennsylvania.

Following diagnosis of scrapie in a clinically suspect Suffolk sheep, 7 clinically normal flockmates were purchased by the Pennsylvania Department of Agriculture to determine their scrapie status using an immunohistochemical procedure. Two of the 7 euthanized healthy sheep had positive immunohistochemical staining of the prion protein of scrapie (PrP-Sc) in their brains, nictitating membranes, and tonsils. The PrP-Sc was localized in the areas of the brain where, histopathologically, there was neurodegeneration and astrocytosis. The PrP-Sc occurred within germinal centers of the affected nictitating membranes and tonsils and was located in the cytoplasm of the dendrite-like cells, lymphoid cells, and macrophages. These results confirm that immunohistochemical examination of the nictitating membrane can be used as a screen for the presence of scrapie infection in clinically normal sheep at a capable veterinary diagnostic laboratory. In sheep with a PrP-Sc-positive nictitating membrane, the diagnosis of scrapie should be confirmed by histopathology and immunohistochemical examination of the brain following necropsy. Following full validation, immunohistochemistry assays for detection of PrP-Sc in nictitating membrane lymphoid tissues can improve the effectiveness of the scrapie control and eradication program by allowing diagnosis of the disease in sheep before the appearance of clinical signs.     

Composition and mechanism of antitumor effects of Hericium erinaceus mushroom extracts in tumor-bearing mice

We investigated antitumor effects of the following four extracts of freeze-dried Hericium erinaceus mushrooms in Balb/c mice intracutaneously transplanted on the backs with CT-26 colon cancer cells: HWE, hot water extraction by boiling in water for 3 h; MWE, microwaving in 50% ethanol/water at 60 W for 3 min; and ACE and AKE, boiling in 1% HCl or 3% NaOH for 2 h. HWE and MWE with a higher content of β-glucans, determined by an assay kit, than ACE and MKE were active in all bioassays. Gas chromatography/mass spectrometry analyses showed the presence of 40, 27, 16, and 13 compounds, respectively, in the four extracts. Daily intraperitoneal (ip) injections of HWE and MWE for 2 weeks significantly reduced tumor weights by 38 and 41%. Tumor regressions were associated with changes in the following cancer biomarkers as compared to phosphate buffer (PBS)-treated control mice: 2.7- and 2.4-fold increases in cytolytic activity of splenic natural killer (NK) cells; restored nitric oxide production and phagocytosis in peritoneal macrophages to 95-98% of normal levels; ～2-fold increase in released pro-inflammatory cytokines tumor necrosis factor-α, interleukin-1β, and interleukin-6 from macrophages; and ～56 and ～60% reductions in the number of blood vessels inside the tumor. The pro-angiogenic factors vascular endothelial growth factor (VEGF), cyclooxygenase 2 (COX-2), and 5-lipoxygenase (5-LOX) were also significantly reduced in mRNA and protein expression by tumor genes. Enzyme-linked immunosorbent assay of tumor cells confirmed reduced expression of COX-2 and 5-LOX (32 and 31%). Reduced COX-2 and 5-LOX expression down-regulated VEGF expression, resulting in inhibition of neo-angiogenesis inside the tumors. The results indicate that induction of NK activity, activation of macrophages, and inhibition of angiogenesis all contribute to the mechanism of reduction of tumor size.     

Productive efficiency of the sandfish <Emphasis Type="Italic">Arctoscopus japonicus</Emphasis> coastal gillnet fishery using stochastic frontier analysis

It is important to estimate the productive efficiencies of industries, especially the fishing industry, in order to determine policies that can improve business conditions. In this study, the productive efficiency of the sandfish coastal gillnet fishery on the east coast of Korea has been estimated using stochastic frontier analysis (SFA). A translog production function wherein the inefficiency was represented by a truncated-normal distribution was established; the output variable was the trip production quantity, the input variables were physical production factors directly related to the fishing activities of vessels, such as tonnage, horsepower, and the number of employed fishers. The average productive efficiency of the sample was 0.59 [0.40–0.79], which implied that productive inefficiency occurs in sandfish coastal gillnet vessels. Moreover, it was verified that there are no differences among the average productive efficiencies of fishing vessels of different tonnages.     

Assessment of growth and seed oil composition of kenaf (Hibiscus cannabinus L.) germplasm

This study was conducted to evaluate the growth characteristics and fatty acid composition among 15 kenaf mutants derived from the kenaf germplasm C14 and 15 kenaf accessions originating from Russia, India, China, Iran, and Italy. The overall growth performance (plant height, stem diameter, flowering date, leaf, and flower size) of the stem color mutant lines derived from C14 are similar to those of the original variety. However, the flower color mutant lines derived from C14 showed flowering to occur 10 days later when compared with the original variety and showed smaller leaf sizes than the original variety. Late-ripened kenaf accessions (Jinju, Auxu, and Jnagdae) can yield more bio-mass compared with early or medium-maturing germplasm. The late maturity kenaf (Auxu, Jinju, and Jangdae) has a higher oil percentage than the early maturity germplasm. Linoleic, oleic, and palmitic acids were the predominant fatty acids in all kenaf seeds. The stem color mutant lines significantly surpassed the parental means of all saturated fatty acids. In addition, the flower color mutant lines showed broad ranges of variation in oleic acid. The 15 accessions showed a wide range of fatty acid compositions, spanning from 29.75 to 38.30% saturated fatty acids and 61.70 to 70.24% total unsaturated fatty acids, and the late maturity kenaf has a higher linoleic acid percentage than the early maturity germplasm. The flowering period was highly positively (P ≤ 0.01) correlated with the plant height, stem diameter, oil percent, and linolenic acid (C_18:3), and it was significantly negatively (P ≤ 0.01) correlated with stearic acid (C_18:0). These results will provide valuable information to assist the parental selection of kenaf breeding.     

Molecular mapping of a new gene for resistance to rice blast (Pyricularia grisea Sacc.)

A single dominant blast resistance gene conferring resistance to a Korean rice blast isolate was identified in rice variety `Suweon 365'. We report the chromosomal localization and molecular mapping of this blast resistance gene designated as Pi-18, which confers resistance to Korean isolate `KI-313' of the blast pathogen. To know whether there is a relationship among genes conditioning resistance to location-specific isolates of the blast pathogen and thereby to identify linked markers to resistance gene for isolate KI-313 collected in Korea, RFLP markers previously reported to be linked to major blast resistance genes in different rice germplasm and other markers mapped to nearby regions were surveyed for polymorphism between a resistant (`Suweon 365') and a susceptible (`Chucheongbyeo') parent. Linkage associations of the RFLP markers with the resistance gene were verified using an F2 and F3 segregating population of known blast reaction. RFLP analysis showed that Pi-18 was located near the end of chromosome 11, linked to a single copy clone RZ536 at a distance of 5.4 centiMorgans (cM) and that this gene was different from Pi-1(t). An allelism test revealed that this gene was also different from Pi-k. Currently, a combination of RAPD and microsatellite primers is being employed to find additional markers in this region. Tightly linked DNA markers will facilitate selection for resistant genotypes in breeding programs and provide the basis for map based cloning of this new blast resistance gene. This revised version was published online in July 2006 with corrections to the Cover Date.     

Photosynthetic characteristics of highbush blueberry (Vaccinium corymbosum cv. Bluecrop) leaves in response to water stress and subsequent re-irrigation

SUMMARY

Gas exchange and photosystem II (PSII) activities in the leaves of 2-year-old ‘Bluecrop’ highbush blueberry (Vaccinium corymbosum) were monitored during water stress and subsequent re-irrigation to investigate the effects of the intensity of water stress on changes in photosynthetic characteristics. The blueberry shrubs were not irrigated for 3 to 5 weeks, then re-irrigated daily up to 8 weeks. The decrease in soil water potential during water stress caused a progressive decrease in leaf water potential. Soil water potentials decreased to -0.26 MPa and -0.34 MPa at 3 and 5 weeks, respectively, following water stress, but recovered following subsequent re-irrigation, while the soil water potential in daily-irrigated shrubs was maintained at over -0.13 MPa throughout the experiment. Chlorophyll concentrations decreased with an increasing duration of water stress. Chlorophyll concentrations in leaves on shrubs subjected to water stress for 5 weeks did not recover following re-irrigation, unlike those subjected to water stress for 3 weeks. The leaves on shrubs subjected to water stress for 5 weeks maintained lower levels of chlorophyll during reirrigation. The net rate of CO₂ assimilation (A_n) decreased significantly with an increasing duration of water stress. Reirrigation reversed the decrease in A_n in leaves on shrubs subjected to water stress for 3 weeks. Stomatal conductance (g_s) exhibited a similar pattern to A_n. The actual quantum yield of photosystem II (Φ_PSII) and the electron transport rate (ETR) also decreased significantly with an increasing duration of water stress, although the F_v/F_m ratio was not affected. Φ_PPSII and ETR values in the leaves on shrubs subjected to water stress for 5 weeks did not recover after reirrigation, unlike those subjected to water stress for 3 weeks. Non-photochemical quenching increased with an increasing duration of water stress, but subsequent re-irrigation did not reverse the increase. These results indicate that the timing of re-irrigation of water-stressed ‘Bluecrop’ highbush blueberry is critical in order to maintain their photosynthetic capacity. Among the photosynthetic characteristics measured, Φ_PSII and ETR could be used as sensitive indicators to assess the physiological status of leaves of ‘Bluecrop’ highbush blueberry growing under water stress conditions.     

Evidence of Long-Range Transport of Pollutants from the Size-Fractionated Ionic Composition of Aerosols in the Jeju Island of Korea

The ionic composition of total suspended particulate (TSP) and fine (PM_2.5) fractions was investigated from an 1,100 site in the middle of Mt. Halla in Jeju Island, Korea from March to November 2006. The sum concentrations of cation and anion species in TSP fraction were 205 ± 170 and 183 ± 164 neq m^?3, respectively, while those for PM_2.5 as 118 ± 129 and 88.5 ± 89.3 neq m^?3, respectively. In TSP, the concentration of the major ions changed in the order of SO₄ ^2? > NH₄ ⁺ > Ca²⁺ > Na⁺ > NO₃ ^? > Mg²⁺ > K⁺ > Cl^?, while its PM_2.5 counterpart as NH₄ ⁺ > SO₄ ^2? > Ca²⁺ > NO₃ ^? > Na⁺ > Mg²⁺ > K⁺ > Cl^?. Inspection of the temporal variabilities of ionic components indicated that most ions peaked in spring or fall months. The back trajectory analysis showed that the atmospheric composition of the major ionic species was affected fairly sensitively by long-range transport from China under the favorable meteorological conditions. In contrast, the lowest ionic concentration levels were seen most abundantly, when air masses passed from South Sea. Hence, the analysis of ionic concentration data suggests that their distributions are controlled by the combined effects of various source processes including the most prominent Chinese origin and the meteorological condition favorable for such transport.     

Identification and structure elucidation of a p-phenoxybenzaldehyde in bamboo shoots by HPLC-ESI/MS/MS and NMR

In this study, a derivative of p-phenoxybenzaldehyde in bamboo shoots was investigated. Bamboo shoots were ground and extracted with water, and an aqueous suspension was purified by SPE using Oasis HLB cartridges. After the SPE procedure, the analytes were analyzed by HPLC with refractive index detection (HPLC-RI). In the HPLC-RI analysis for sucralose, a putative sucralose was detected. In the subsequent HPLC-PDA analysis, the suspicious peak showed a unique UV absorption spectrum with the maximum wavelength at 285 nm indicating the existence of an aromatic ring. The contents of the unknown compound in bamboo shoot products ranged from 0.01 to 0.15 mg/g. The identity of the unknown compound was further confirmed by HPLC-ESI/MS/MS. The molecular weight of the unknown compound was determined to be 244. The chemical structure of the unknown compound was elucidated on the basis of NMR spectroscopic analyses ((1)H, (13)C, DEPT, COSY, HMQC, and HMBC). Finally, the structure of the unknown compound was characterized as 4-(4-dihydroxymethylphenoxy)benzaldehyde.     

Direct inhibition of maintenance respiration in western hemlock roots exposed to ambient soil carbon dioxide concentrations

Root respiration often exhibits a direct and immediate decline with increasing concentrations of ambient soil carbon dioxide concentration ([CO(2)]), and recent evidence suggests this decline may be attributable to a decline in maintenance respiration within the root. If true, this concept could provide a clue to the biochemical process underlying respiratory inhibition as well as improve our knowledge of the timing and degree to which this inhibition occurs in nature. To test the hypothesis that maintenance respiration exhibits a direct, negative response to increasing [CO(2)], we measured total respiration in intact root systems of western hemlock (Tsuga heterophylla (Raf.) Sarg.) seedlings grown at different relative growth rates and exposed to soil [CO(2)]s ranging from 91 to 7008 &mgr;mol mol(-1). Analysis of covariance was used to separate maintenance from total respiration. Total respiration declined exponentially with increasing [CO(2)]. Maintenance respiration, which comprised 85% of total respiration over all treatments, also declined exponentially with increasing [CO(2)]. Growth respiration was not inhibited at any [CO(2)]. These findings may explain why roots of some fast-growing species do not show [CO(2)] inhibition.