Determination of usnic acid in lichen toxic to elk by liquid chromatography with ultraviolet and tandem mass spectrometry detection

Usnic acid is unambiguously confirmed by tandem mass spectrometry (MS/MS) in tumbleweed shield lichen, Xanthoparmelia chlorochroa. The lichen contains 2% usnic acid by liquid chromatography with UV quantification at 282 nm. The UV linear range for usnic acid quantification is from its 4 ng limit of detection to 2 microg injected. UV signal saturation is recognized by distortion of the usnic acid UV spectrum. Positive ion electrospray-tandem mass spectrometry offers no similar means to recognize quantification data recorded above the linear range of electrospray. Electrospray ionization capacity and matrix effects limit the reliability of tandem mass spectrometry quantification. The combination of UV quantification and MS confirmation provides a reliable analytical method for measuring usnic acid levels in plant material.     

Antiviral activity of Alpinia katsumadai extracts against rotaviruses

In vitro anti-rotavirus activity of Alpinia katsumadai (AK) extracts were evaluated against bovine G8P[7] and porcine G5P[7] rotaviruses in two different assay strategies, a mixed treatment assay and a post treatment assay. In the mixed treatment assay, six AK extracts [AK-1 (EtOH extract), AK-3 (H(2)O layer), AK-5 (40% methanol fraction), and AK-9-11 (H(2)O extract, polysaccharide fraction, supernatant fraction)] exhibited inhibitory activities against G5P[7] rotavirus with the EC(50) values ranging from 0.7±0.4 to 33.7±6.5 μg/mL. Extracts AK-1, AK-3, and AK-5 inhibited rotavirus infection against G8P[7] rotavirus, the with EC(50) values of 8.4±2.2 μg/mL, 6.5±0.8 μg/mL and 8.4±5.0 μg/mL, respectively. By hemagglutination inhibition (HI) assay, six AK extracts completely inhibited viral adsorption onto human RBCs in both strains of rotaviruses at less than 11 μg/mL. However, in the post treatment assay, there was no anti activity shown against both strains of rotaviruses. As a result, six AK extracts were attributed mainly to having a strong interaction with hemagglutinin protein on the outer surface of rotavirus, resulting to blockage of viral adsorption.     

Development and characterization of mouse monoclonal antibodies reactive with chicken CD83

This study was carried out to develop and characterize mouse monoclonal antibodies (mAbs) against chicken CD83 (chCD83), a membrane-bound glycoprotein belonging to the immunoglobulin superfamily that is primarily expressed on mature dendritic cells (DCs). A recombinant chCD83/IgG4 fusion protein containing the extracellular region of chCD83 was expressed in Chinese Hamster Ovary (CHO) cells and isolated from the spent cell culture medium by protein G affinity chromatography. The extracellular region of the chCD83 protein was purified and used to immunize mice. A cell fusion was performed, from which 342 hybridomas were screened for mAbs to chCD83. Two mAbs, chCD83-159 and chCD83-227, stained the greatest percentage of chCD83-transfected CHO cells and were selected for further characterization. By flow cytometry, both mAbs reacted with a chicken macrophage cell line, HD11. Both mAbs also recognized a single 53 kDa protein on Western blots of lysates from lipopolysaccharide-stimulated spleen mononuclear cells or unstimulated HD11 cells. Immunostaining of chicken secondary lymphoid organs identified chCD83(+) cells with morphologic and subtissue localization properties comparable to mammalian DCs. In vitro stimulation of spleen mononuclear cells with concanavalin A (Con A) decreased the percentage of chCD83(+) cells compared with cells treated with medium alone. Interestingly, spleen cells treated with Con A in the presence of chCD83-227 mAb exhibited decreased percentage of MHCII(+) cells compared with cells treated with an isotype-matched negative control mAb. These chCD83 mAbs may be useful for future investigations of chicken immune cell maturation and mechanisms of action.     

Growth and wood basic density of acacia hybrid clones at three locations in Vietnam

Field trials testing a total of 27 clones of the interspecific hybrid Acacia mangium × A. auriculiformis and seedling controls of the parental species were established at Ba Vi and Yen Thanh in the north of Vietnam and Long Thanh in the south. At both Ba Vi and Yen Thanh there were significant (P < 0.001) differences in height and diameter at breast height (DBH) among 22 tested clones at 4 years. At Long Thanh, twelve hybrid clones did not differ significantly in DBH at age 3 years, but did (P < 0.001) at age 5 years. At the two northern sites the acacia hybrid clones had significantly greater DBH than control seedlots of the parental species. At Long Thanh, DBH of the hybrid clones and A. mangium was similar, with a genetically improved seedlot of A. mangium displaying the best DBH. Mean wood basic density at breast height of the acacia hybrid clones was 539 kg m⁻³ at Yen Thanh at age 8 years, and 473 kg m⁻³ at Long Thanh at age 5 years; density for A. mangium at Long Thanh was only slightly lower than the hybrid clones at 461 kg m⁻³. Linear regressions of Pilodyn penetration (PP) at breast height on wood basic density explained 60% of the variance in density of treatments (clones and control seedlots) at Yen Thanh and 36% at Long Thanh. There were significant differences between hybrid clones in PP at all three trial sites. Clonal DBH performance was not strongly correlated across the three trial sites; Pearson correlations of clone mean DBH between pairs of sites ranged from −0.47 to 0.20. Clonal rankings for PP were more stable, with Pearson correlations between pairs of sites ranging from r = 0.71 to 0.78.     

Detection and molecular characterization of porcine type 3 orthoreoviruses circulating in South Korea

Orthoreoviruses infect virtually all mammalian species, causing systemic infections including mild gastrointestinal and respiratory illnesses. However, little is known about the prevalence or genetic diversity of porcine orthoreoviruses in South Korea. We examined 237 diarrheic fecal samples collected from 78 pig farms around the country. RT-PCR utilizing primers specific for the L1 gene of mammalian orthoreoviruses showed that 45 (19.0%) samples were positive. The 10 strains isolated from orthoreovirus-positive samples formed typical perinuclear cytoplasmic inclusion bodies and had an atypical hemagglutination pattern; these are characteristics of type 3 orthoreovirus. Phylogenetic analysis of the S1 gene in these 10 Korean and other strains showed that type 3 orthoreoviruses could be divided into four lineages; the 10 Korean strains were included in porcine lineage IV, along with T3/porcine/Sichuan/2006. Sequence analysis showed that strains in lineage IV had nucleotide identities of 97.0-98.1% and deduced amino acid identities of 96.4-98.2%. Sequence analysis of the σ1 protein, a viral attachment protein, revealed that the amino acid sequences associated with neurotropism (amino acids 198-204, 249I, 350D, and 419E) were highly conserved among the Korean strains, confirming that neural tropism was present. In conclusion, our findings suggest that porcine orthoreovirus infections are endemic in pig farms in South Korea and that the 10 novel Korean porcine orthoreoviruses belong to porcine lineage IV of type 3 orthoreovirus. In addition, sequence analysis of S1 genes encoding the σ1 protein showed that the 9 of 10 Korean porcine orthoreoviruses exhibited neural tropism.     

Comparing the osteogenic potential of canine mesenchymal stem cells derived from adipose tissues, bone marrow, umbilical cord blood, and Wharton's jelly for treating bone defects

Alternative sources of mesenchymal stem cells (MSCs) for replacing bone marrow (BM) have been extensively investigated in the field of bone tissue engineering. The purpose of this study was to compare the osteogenic potential of canine MSCs derived from adipose tissue (AT), BM, umbilical cord blood (UCB), and Wharton''s jelly (WJ) using in vitro culture techniques and in vivo orthotopic implantation assays. After canine MSCs were isolated from various tissues, the proliferation and osteogenic potential along with vascular endothelial growth factor (VEGF) production were measured and compared in vitro. For the in vivo assay, MSCs derived from each type of tissue were mixed with β-tricalcium phosphate and implanted into segmental bone defects in dogs. Among the different types of MSCs, AT-MSCs had a higher proliferation potential and BM-MSCs produced the most VEGF. AT-MSCs and UCB-MSCs showed greater in vitro osteogenic potential compared to the other cells. Radiographic and histological analyses showed that all tested MSCs had similar osteogenic capacities, and the level of new bone formation was much higher with implants containing MSCs than cell-free implants. These results indicate that AT-MSCs, UCB-MSCs, and WJ-MSCs can potentially be used in place of BM-MSCs for clinical bone engineering procedures.     

Sequence variations of the bovine prion protein gene (PRNP) in native Korean Hanwoo cattle

Bovine spongiform encephalopathy (BSE) is one of the fatal neurodegenerative diseases known as transmissible spongiform encephalopathies (TSEs) caused by infectious prion proteins. Genetic variations correlated with susceptibility or resistance to TSE in humans and sheep have not been reported for bovine strains including those from Holstein, Jersey, and Japanese Black cattle. Here, we investigated bovine prion protein gene (PRNP) variations in Hanwoo cattle [Bos (B.) taurus coreanae], a native breed in Korea. We identified mutations and polymorphisms in the coding region of PRNP, determined their frequency, and evaluated their significance. We identified four synonymous polymorphisms and two non-synonymous mutations in PRNP, but found no novel polymorphisms. The sequence and number of octapeptide repeats were completely conserved, and the haplotype frequency of the coding region was similar to that of other B. taurus strains. When we examined the 23-bp and 12-bp insertion/deletion (indel) polymorphisms in the non-coding region of PRNP, Hanwoo cattle had a lower deletion allele and 23-bp del/12-bp del haplotype frequency than healthy and BSE-affected animals of other strains. Thus, Hanwoo are seemingly less susceptible to BSE than other strains due to the 23-bp and 12-bp indel polymorphisms.     

Chemerin analog regulates energy metabolism in sheep

Accumulating data suggest a relationship between chemerin and energy metabolism. Our group previously described gene cloning, expression analysis and the regulatory mechanism of chemerin and its own receptor in mice and cattle. The objective of the present study was to investigate the physiological effect of chemerin on endocrine changes and energy metabolism in sheep using a biologically stable chemerin analog. The chemerin analog was intravenously administrated (100 or 500 µg/head) to sheep, and plasma insulin and metabolites (glucose, nonesterified fatty acids (NEFA), triglyceride, total cholesterol and high‐density lipoprotein (HDL) cholesterol) were analyzed. The chemerin analog dramatically increased the insulin levels, and glucose levels were decreased. NEFA levels were slightly decreased at 20 min but then increased gradually from 60 to 180 min after analog administration. In addition, injection of the chemerin analog immediately increased triglyceride and total cholesterol but not HDL levels. These results suggested that chemerin analog regulated insulin secretion related to glucose metabolism and the release of triglycerides in sheep in vivo. This study provides new information about endocrine and metabolic changes in response to chemerin in sheep.     

Comparative analysis of physicochemicals and antioxidative properties in new red rice (Oryza sativa L. cv. Gunganghongmi)

The main objectives of this study were to investigate physicochemicals and antioxidant activities of new red rice (Oryza sativa cv. Gunganghongmi (GH)) by comparing normal brown (Nampyeongbyeo, NB) and reported red rice (Jukjinjubyeo, JB) in Korea. The nutritional constituents, including protein, oil, sugar, fatty acid, GABA, and γ-oryzanol were not significantly different between normal brown and colored rice. However, the ethanol extract of GH showed the highest phenolic content (24.7 ± 1.3 mg g^?1). The ethanol extracts of GH showed higher scavenging activities against DPPH (0.2 mg mL^?1 = 62.1 ± 2.5%) and ABTS (0.2 mg mL^?1 = 63.2 ± 3.5%) radicals. Moreover, GH more inhibited LPS-induced nitric oxide (NO) production (13.2 ± 1.4 μM) than JB (18.3 ± 2.3 μM) and NB (22.1 ± 1.4 μM) at the same concentration (0.2 mg mL^?1) without cytotoxicity. These results suggest that new red rice (GH) would be considered to be new functional rice due to its anti-oxidative effect and high nutrition.     

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.