Diversity and evolution of major Panax species revealed by scanning the entire chloroplast intergenic spacer sequences

Ginseng is an important medicinal plant, but almost no genomic information is known for it. For the primary step to understand the Panax genome, we inspected the chloroplast genome sequence diversity and used that to infer the evolution of Panax species using them. We inspected a total of 101 intergenic spacers (IGS) covering 44,563 bp (96.8 % of the total IGS) from four Panax species. Diversity was inspected by three steps: gel electrophoresis, high resolution melting (HRM) analysis, and further confirmation by sequencing. Even though low levels of InDel polymorphism were detected by gel electrophoresis, high levels of reproducible polymorphisms were identified by HRM analysis. Sequencing of the HRM-polymorphic spaces and pair-wise multiple sequence alignments revealed up to 247 sequence variations between species. We have identified 62 IGS showing polymorphism between species. Among them, the trnE-trnT, trnT-psbD, ndhF-rpl32, and rpl14-rpl16 spaces are more informative for studying the diversity of Panax relatives. Phylogenetic analysis and molecular evolution studies revealed that P. notoginseng is most diverged from the other Panax species, with a nucleotide substitution rate of 0.0039. P. quinquefolius shows a close relationship with P. ginseng and P. japonicus, with a nucleotide substitution rate of 0.0009. Meanwhile, no sequence variation was detected between P. ginseng and P. japonicus. Calculation of molecular clocks revealed that P. notoginseng diverged more than 1.30 million years ago from the other Panax species and then, P. quinquefolius diverged from P. ginseng by migration in the American continent more than 0.29 million years ago.     

His-His-Leu, an angiotensin I converting enzyme inhibitory peptide derived from Korean soybean paste, exerts antihypertensive activity in vivo.

It has been reported that soybean peptide fractions isolated from Korean fermented soybean paste exert angiotensin I converting enzyme (ACE) inhibitory activity in vitro. In this study, further purification and identification of the most active fraction inhibiting ACE activity were performed, and its antihypertensive activity in vivo was confirmed. Subsequently, a novel ACE inhibitory peptide was isolated by preparative HPLC. The amino acid sequence of the isolated peptide was identified as His-His-Leu (HHL) by Edman degradation. The IC(50) value of the HHL for ACE activity was 2.2 microg/mL in vitro. Moreover, the synthetic tripeptide HHL (spHHL) resulted in a significant decrease of ACE activity in the aorta and led to lowered systolic blood pressure (SBP) in spontaneously hypertensive (SH) rats compared to control. Triple injections of spHHL, 5 mg/kg of body weight/injection resulted in a significant decrease of SBP by 61 mmHg (p < 0.01) after the third injection. These results demonstrated that the ACE inhibitory peptide HHL derived from Korean fermented soybean paste exerted antihypertensive activity in vivo.     

Carbon Control on Nitrogen Dynamics in the Forest Floor of an N-Enriched Deciduous Forest Ecosystem

Recent evidence from nitrogen (N) saturation studies indicates that forest floors in moderately impacted forests are the primary sink for atmospheric N inputs. Some researchers have suggested that the sink capacity of organic horizons is dependent on the amount of available carbon (C), which can be used for microbial N assimilation. To test the hypothesis that C limitation in forest floors exposed to chronic N deposition leads to an enhanced N leaching, a field C input manipulation experiment is under way in a deciduous forest. Since September 1999 aboveground C input has been doubled (by doubling litter input or by amending glucose) or excluded in replicated plots. Here we report the short-term response of concentrations of dissolved inorganic N (DIN: NO₃ ^?-N and NH₄ ⁺-N) in forest floor percolate to the C input manipulation. In autumn following the C input manipulation, DIN concentrations in forest floor percolate decreased in all plots except the No Litter plots compared to the pre-treatment summer concentrations. In contrast, the concentrations of DIN in the No Litter plots remained high. A different seasonal pattern of DIN leaching among treatments, along with measurements of microbial biomass C and potential nitrification rates of forest floor samples, indicates that seasonal N dynamics in the forest floor are largely regulated by C availability changes assoicated with litterfall C input.     

Association of antioxidant vitamins and oxidative stress levels in pregnancy with infant growth during the first year of life

OBJECTIVE: Whereas there are numerous reports in the literature relating the impact of maternal nutritional status on subsequent birth outcome, much less is known about the long-term impact on infant growth after birth. Therefore, we conducted a prospective cohort study to investigate the association of maternal micronutrient status (vitamins A, C and E, folate) and oxidative stress status in pregnancy with infant growth during the first year of life. DESIGN: Prospective cohort study. SETTING: Outpatient clinic of obstetrics, Ewha Womans University Hospital, Seoul, South Korea.Subjects and methodsTwo groups were constructed for this study - the Ewha pregnancy cohort (n = 677) and the infant growth cohort comprising follow-up live newborns of all the recruited pregnant women (n = 317). Maternal serum vitamin and urinary oxidative stress levels were collected and infant weights and heights were measured at birth and at 6 and 12 months after birth. RESULTS: Division of the subjects into folate-deficient and normal groups revealed that infant weight and height at 0, 6 and 12 months were adversely affected by folate deficiency. High maternal vitamin C was associated with increased infant weight and height at birth and after birth.ConclusionOur findings indicate the importance of preventing folate deficiency and supplementing vitamin C during pregnancy.     

Synthesis of conjugated linoleic acid by human-derived Bifidobacterium breve LMC 017: utilization as a functional starter culture for milk fermentation

This study was performed to discover bifidobacteria isolated from human intestines that optimally convert linoleic acid (LA) to conjugated linoleic acid (CLA) and to optimize the culture conditions of milk fermentation. One hundred and fifty neonatal bifidobacteria were screened for CLA-producing ability, and Bifidobacterium breve LMC 017 was selected as it showed about 90% conversion of free LA in MRS broth. The selected strain showed resistance at 0.5% LA in microaerophillic conditions. When monolinolein (LA 90%) was used as a substrate for CLA production, the conversion rate was lower compared to free LA, but the growth rate was unaffected during the milk fermentation. There was no significant difference in CLA production between aerobic and anaerobic conditions, and little decline in CLA was shown after the maximal CLA level had been reached. CLA production increased by 80% with 24 h of incubation in milk containing additional skim milk (5%), where the proteins may have facilitated the production of CLA by enhancing the interaction of substrate with the bacteria. CLA production did not decline after 9 h of fermentation and an additional 12 weeks of storage with other commercial starters. This demonstrates the possibility of using this strain as a costarter in the production of CLA-enriched yogurt.     

Enzyme-linked immunosorbent assay for screening aflatoxin B1 in cottonseed products and mixed feed: collaborative study

A joint AOAC/IUPAC (International Union of Pure and Applied Chemistry) interlaboratory study of an enzyme-linked immunosorbent screening assay (ELISA) for aflatoxins was conducted in laboratories in Canada, France, Japan, South Africa, Switzerland, The Netherlands, Tunisia, and the United States. Twenty-eight samples of raw and roasted peanuts, corn, whole cottonseed, cottonseed meal, ammoniated cottonseed meal, and poultry feed containing various quantities of natural aflatoxins and supplemented when appropriate with aflatoxin B1 were distributed to participating laboratories for testing. The assay is based on conjugation of pure aflatoxin B1 to an enzyme and the competition between this conjugate and (free) aflatoxins in the product for aflatoxin-specific antibodies coated onto microtiter well walls. After a wash step to remove all unbound aflatoxins, a substrate, added to each well, is catalyzed from a colorless to a green solution by any bound enzyme-conjugated aflatoxin B1 present. The intensity of the color decreases as the amount of free aflatoxin B1 in the product increases. Overall correlation was good between ELISA and thin-layer chromatographic (TLC) results for cottonseed products and mixed feed. Variable results were reported for corn and peanut product samples. Although some positive samples (greater than 15 ng/g) of cottonseed products and mixed feed were reported to contain less than 15 ng/g by visual determination, a review of data for absorbance measurements showed that the contamination level was close to the greater than or equal to 15 ng/g standard and would not have been reported as negative under routine screening.(ABSTRACT TRUNCATED AT 250 WORDS)     

Global status of wheat leaf rust caused by <Emphasis Type="Italic">Puccinia triticina</Emphasis>

Leaf rust caused by Puccinia triticina is the most common and widely distributed of the three wheat rusts. Losses from leaf rust are usually less damaging than those from stem rust and stripe rust, but leaf rust causes greater annual losses due to its more frequent and widespread occurrence. Yield losses from leaf rust are mostly due to reductions in kernel weight. Many laboratories worldwide conduct leaf rust surveys and virulence analyses. Most currently important races (pathotypes) have either evolved through mutations in existing populations or migrated from other, often unknown, areas. Several leaf rust resistance genes are cataloged, and high levels of slow rusting adult plant resistance are available in high yielding CIMMYT wheats. This paper summarizes the importance of leaf rust in the main wheat production areas as reflected by yield losses, the complexity of virulence variation in pathogen populations, the role cultivars with race-specific resistance play in pathogen evolution, and the control measures currently practiced in various regions of the world.     

Comparative analysis of pathogenesis-related protein 10 (<Emphasis Type="Italic">PR10</Emphasis>) genes between fungal resistant and susceptible peppers

To elucidate the functional roles of PR10 genes from two pepper species during plant-pathogen interactions, PR10 genes were isolated from fungal-resistant (Capsicum baccatum var. PBC80) and fungal-susceptible (C. annuum var. Yeoju) pepper fruits infected with anthracnose fungus (Colletotrichum acutatum). Despite strong nucleotide sequence identity, there were significant differences in the patterns of gene expression and protein accumulation between the genes from the two host species. Induced expression of the PR10 mRNA in PBC80 (bacPR10) was highly maintained from 24 h after infection (HAI) rather than that in Yeoju (annPR10). These mRNA expression patterns were correlated with the level of respective protein that was detected as two or three bands in each species. Substantial induction of bacPR10 proteins was confirmed by 2D-gel analysis followed by immunoblotting. Immunolocalization study showed that deposition of bacPR10 was exclusively observed in the pericarp of PBC80 fruits after fungal infection, suggesting functional significance in defence. Additionally, in vitro analysis of the enzymatic properties of PR10 proteins revealed that recombinant bacPR10 had higher ribonucleolytic activity and exhibited less sensitivity to proteinase treatment than did annPR10. Taken together, these results support the idea that relative abundance and prolonged longevity of bacPR10 in PBC80 fruits may contribute to their increased resistance in response to the anthracnose fungus, as compared with Yeoju fruit.     

Expression of secreted and membrane-bound mucins in the airways of piglets experimentally infected with Mycoplasma hyopneumoniae

Immunohistochemistry was used to demonstrate secreted mucins MUC2, MUC5AC and MUC5B and membrane-bound mucin MUC4 in the pulmonary bronchioles of piglets experimentally infected with Mycoplasma hyopneumoniae. Conventional status, Landrace-Duroc cross-bred piglets, 13 days of age, were randomised to two groups. One group (n=20) was infected by the intra-tracheal route with the SNU98703 strain of M. hyopneumoniae, and a group of 12 animals acted as uninfected controls. Five infected and three uninfected piglets were euthanased on the day of infection and at 7, 21, and 35 days post-inoculation (PI). Membrane-bound MUC4 and secreted MUC5AC were the predominant mucins produced in the bronchioles of the piglets in response to M. hyopneumoniae infection, but by day 35 PI, all labelled mucins had returned to pre-infection levels, contemporaneous with reduced pulmonary lesion scores. The increased mucin production may result from direct stimulation of the epithelium by mycoplasmal infection, or may arise indirectly following M. hyopneumoniae-induced ciliostasis.     

Effects of dietary electrolyte balance on the chemistry of blood and urine in lactating sows and sow litter performance

One hundred fifty-three sows (average parity of 2.2) were used to determine the effects of dietary electrolyte balance (calculated as mEq/kg of diet for Na + K - Cl) on sows and their litters during lactation. The sows were fed corn-soybean meal-based diets (1.0% lysine, 1.0% valine, 0.95% Ca, and 0.80% P; as-fed basis) starting on d 109 of gestation and throughout the 21-d lactation experiment. Dietary electrolyte balance (dEB) was 0, 100, 200, 350, and 500 mEq/kg (as-fed basis), well above and below the dEB of 185 mEq/kg found in a simple corn-soybean meal-based lactation diet. To achieve the desired dEB, diets had the following: 1) 1.8% HCl (6 N) and 1.06% CaCl2, 2) 1.0% CaCl2, 3) 0.04% NaHCO3, 4) 1.29% NaHCO3, and 5) 2.54% NaHCO3 (as-fed basis). Increasing dEB increased blood pH (linear and quadratic effects, P < 0.001), partial pressure of carbon dioxide (linear effect, P < 0.001), HCO3- concentration (linear and quadratic effects, P < 0.001), and blood base excess (linear and quadratic effects, P < 0.001). However, increased dEB resulted in lower blood concentrations of K (linear and quadratic effects, P < 0.04), Cl (linear and quadratic effects, P < 0.001), and ionized Ca (linear and quadratic effects, P < 0.001). Changing dEB did not affect ADFI; water usage, litter weight gain; sow weight change; sow backfat change; percentages of CP, lactose, and fat in the milk; percentage of sows returning to estrus; days to estrus; and number of pigs born alive in the subsequent litter (P = 0.06). However, piglet survivability to d 10 and overall was greatest with the lower dEB treatments (linear effect, P < 0.05). The pH (linear and quadratic effects, P < 0.001) and colony forming units of total bacteria (linear effect, P < 0.03) in the urine increased as dEB of the diet was increased. In conclusion, dEB had pronounced effects on the physiological status of sows and decreasing dEB below that in a simple corn-soybean meal-based diet decreased bacterial counts in the urine and increased piglet survivability. However, milk composition, sow and litter weights at weaning, and subsequent rebreeding performance of the sows were not affected by dEB.