Quantitative analysis of anticancer 3-deoxyanthocyanidins in infected sorghum seedlings

3-Deoxyanthocyanidins are structurally related to the anthocyanin pigments, which are popular as health-promoting phytochemicals. Here, it is demonstrated that the 3-deoxyanthocyanidins are more cytotoxic on human cancer cells than the 3-hydroxylated anthocyanidin analogues. At 200 microM concentration, luteolinidin reduced the viability of HL-60 and HepG2 cells by 90 and 50%, respectively. Sorghum is a major source of 3-deoxyanthocyanidins, which are present as seed pigments and as phytoalexins responding to pathogen attack. On the basis of the collision-induced dissociation spectra of luteolinidin and apigeninidin, an LC-MS/MS method, operating in multiple-reaction monitoring mode, was developed for the specific detection and accurate quantification of these compounds in complex mixtures, which may be difficult to analyze using absorbance measurements. The results demonstrated that inoculated sorghum seedlings could be utilized for convenient and large-scale production of 3-deoxyanthocyanidins. A quantity of almost 270 microg/g (fresh weight) of luteolinidin was produced 72 h after fungal inoculation of 1-week-old seedlings.     

台灣入侵紅火蟻之遺傳種群研究

In the case of recently established populations of Solenopsis invicta in Taiwan, analysis of genetic variation may provide various dimensions regarding the historical demographic events of this invasive species, which represents a novel opportunity to study the genetic consequences of invasiveness over an extremely short time period. Here we describe genetic structure of two introduced S. invicta populations,Taoyuan and Chiayi, in Taiwan using two classes of markers, nuclear (microsatellites) and mitochondrial (mtDNA) genomes. Pronounced regional differentiation inferred from either AMOVA or Bayesian clustering approach at both genomes suggests that the two populations most likely originate from separate introductions.Furthermore, significant mtDNA but not nuclear differentiation between sympatric social forms suggests two scenarios that interform gene flow is mainly mediated by males, and queens of each form commonly fail to become egg-layers of alternate form. Isolation by distance (IBD) obtained from microsatellites is absent in monogynes, indicating sexuals of this form retain the superior dispersal ability to homogenize the nuclear signature among spatially isolated areas; however, lack of IBD in less vagile polygyne may result from frequent human mediated jump dispersal that erodes the geographical restrictions of genetic exchange. The patterns observed here not only provide insights into how social organization influences the interform gene flow but also reinforce different breeding strategies pursued by two forms in the character of shaping the genetic variation at two levels of genomes.     

Release of nutrients dissolved organic carbon during decomposition of Chamaecyparis obtusa var. formosana leaves in a mountain forest in Taiwan

Forest ecosystems in Taiwan are periodically influenced by typhoons that cause large amounts of litter input to the soil. The potential rapid decomposition of such litter under the warm and moist climatic conditions in Taiwan may lead to nutrient losses via seepage. The goal of this study was to investigate the dynamics of C, N, K, Ca, Mg, and dissolved organic carbon (DOC) during decomposition of Chamaecyparis obtusa var. formosana leaves in a field study at the Yuanyang Lake site in N Taiwan. We simulated the effect of a typhoon by adding about three times the annual aboveground litterfall (totally 13,900 kg ha^–1) as fresh leaves. Litterbags were taken at 7 dates over 16 months, followed by detection of mass loss and element composition in the remaining litter. Aqueous extracts of the remaining litter were analyzed for DOC and major elements. The properties of DOC were characterized by fluorescence spectra and by its stability against microbial decomposition. The litter mass loss was 35% after 16 months. The losses of Ca after 16 months from the litter bags were about equivalent to mass loss (39%), while those of K and Mg reached 86% and 60% of the initial amount, respectively. From the 13,900 kg ha^–1 of litter applied in total, 59 kg K ha^–1 and 12 kg Mg ha^–1 were released in the 16 months decomposition period, most of it in the first 4 months. The total release of Ca amounted to 69 kg ha^–1 but was more evenly distributed throughout the 16 months of observation. The absolute amount of N in the decomposing litter increased by 37% while the C : N decreased from 69 to 34. Extrapolated to the manipulation treatment, this resulted in a N gain of 36 kg N ha^–1 within 16 months. The leaching of K and DOC in laboratory extractions followed an asymptotic function with highest leaching from the initial litter and subsequent decrease with time of decomposition. On the contrary, the leaching of Ca and Mg reached a maximum after 2–4 months of incubation. About 2% of the C was extractable with water from the initially incubated leaves. The bioavailability of the extracted DOC decreased with litter age. Our results indicate that the decomposition of large amounts of litter induces a high risk of K and Mg losses with seepage, but the risk for N losses is low. The sources of N accumulation in decomposing litter at this site require further studies. In the initial phase of litter decomposition, the release of DOC seems to be an important contribution to mass loss.     

Extracellular production of a functional soy cystatin by Bacillus subtilis

A recombinant Bacillus subtilis producing soy cystatin was developed by subcloning with a soy cystatin gene cloned in Escherichia coli. An active form of cystatin against the cysteine protease from Pacific whiting fillets contaminated with Myxosporidia parasite was constitutively expressed and secreted extracelluarly into the medium. Two gene fragments of signal peptides from kerA and sacB were introduced and compared for secretion efficiency of cystatin. The secretion level of active cystatin improved with the signal peptide of kerA when compared to that of sacB. Inhibitor activity was reduced rapidly after peak expression of the target protein at 36 h of fermentation. The addition of 1% glucose, a suppressor of protease, into the medium sustained the increase of the cystatin activity during fermentation. This study introduced a potential new method for fermentation production of cystatin.     

Optimization of lipase-catalyzed synthesis of octyl hydroxyphenylpropionate by response surface methodology

The ability of immobilized lipase Candida antarctica (Novozyme 435) to catalyze the direct esterification of hydroxyphenylpropionic acid and octanol in a solvent-free system was investigated in this study. Response surface methodology (RSM) and five-level-four-factor central composite rotatable design (CCRD) were employed to evaluate the effects of synthesis parameters, such as reaction time, temperature, enzyme amount, and pH memory, on percentage molar conversion of phenolic acid esters. Reaction time, temperature, and enzyme amount were the most important variables. On the basis of canonical analysis and ridge max analysis, the optimum synthesis conditions with 95.9% molar conversion were reaction time of 58.2 h, temperature of 52.9 degrees C, enzyme amount of 37.8% (w/w), and pH memory of pH 7.     

Fungichromin: a substance from Streptomyces padanus with inhibitory effects on Rhizoctonia solani

Streptomyces padanus strain PMS-702 is an antagonist of Rhizoctonia solani AG-4, the causal agent of damping-off of cabbage. Treatment of cabbage seeds with the culture filtrate of S. padanus strain PMS-702 was effective in reducing the incidence of damping-off of cabbage. The major active ingredient from the culture filtrate of S. padanus strain PMS-702 was purified by silica gel column chromatography and identified as the polyene macrolide, fungichromin, by NMR and mass spectral data. Bioassay studies showed that fungichromin had a strong antifungal activity against R. solani AG-4, and its minimum inhibitory concentration (over 90% inhibition) was found to be 72 microg/mL. This is the first report of fungichromin from S. padanus as an active ingredient for the control of Rhizoctonia damping-off of cabbage.     

selective production and characterization of levan by Bacillus subtilis (Natto) Takahashi

To meet the industrial need of an efficient microbial method for increased levan production, Bacillus subtilis (natto) Takahashi, a commercial natto starter for preparing fermented soybeans (natto), was used to produce levan. After cultivation for 21 h, 40-50 mg of levan mL(-1) was produced in medium containing 20% (w/w) sucrose, which was approximately 50% yield on available fructose. The product consisted of two fractions with different molecular masses (1794 and 11 kDa), which were easily separated by fractionation using an ethanol gradient. The products were well characterized by GPC, 13C NMR, and 1H NMR. The various sugars and concentrations, initial pH, fermentation temperature, and agitation speed affected the levan production by B. subtilis (natto) Takahashi. Takahashi strain is the most efficient levan-producing strain among all of the B. subtilis strains tested and, as previously reported, it produced the highest yield of levan in the least time (21 h) under the common cultivation condition.     

Biological treatment of potato processing wastewaters

A bench-scale laboratory study was conducted to determine the performance of the single-stage treatment system of powdered activated carbon activated sludge reactors and the two-stage treatment system using activated sludge reactors followed by the biological activated carbon columns. Two activated sludge reactor runs and two biological activated carbon (BAC) column runs were conducted in this study. Hydraulic detention time in the aeration tank was 8, 16, and 32 hours for the activated sludge reactor runs. In run 2,4 g/1 of powdered activated carbon (PAC) were added to the activated sludge reactors to improve organic pollutant removal. The feed COD (chemical oxygen demand) was from 2668 to 3309 mg/1. For the BAC runs, the hydraulic detention time varied from 0.288 to 4.034 hours. Results indicate that 92% of COD was removed in the non-PAC activated sludge reactors, while 96% of COD was removed in the PAC activated sludge reactors. Biomass accumulation in the BAC columns improved the COD removal. Effluents from BAC columns contained 34 to 95 mg/1 COD. The two-stage treatment system of activated sludge followed by BAC process had better treatment performance than the single-stage treatment system of PAC activated sludge process.     

Assessment of cardiac output measurement in dogs by transpulmonary pulse contour analysis

Objective – To determine if metatarsal artery pressure (COmet) is comparable to femoral artery pressure (COfem) as the input for transpulmonary pulse contour analysis (PiCCO) in anesthetized dogs, using the lithium dilution method (LiDCO) as a standard for cardiac output (CO) measurement. Design – Prospective randomized study. Setting – University research laboratory. Animals – Ten healthy purpose‐bred mixed breed dogs were anesthetized and instrumented to measure direct blood pressure, heart rate, arterial blood gases, and CO. Interventions – The CO was measured using LiDCO and PiCCO techniques. Animals had their right femoral and left distal metatarsal artery catheterized for proximal (COfem) and distal (COmet) PiCCO analysis, respectively. Measurements were obtained from each animal during low, normal, and high CO states by changing amount of inhalant anesthetics and heart rate. Measurements were converted to CO indexed to body weigh (CI_BW=CO/kg) for statistical analysis. Agreement was determined using Bland and Altman analysis and concordance correlation coefficients. Measurements and Main Results – Thirty paired measurements were taken. The LiDCO CI_BW (± SD) was 68.7 ± 30.3, 176.0 ± 53.0, and 211.1 ± 76.5 mL/kg/min during low, normal, and high CO states, respectively. There was a significant effect of CI_BW state on bias and relative bias with COmet (P<0.001 and P=0.003, respectively). Bias of the COmet method (± SD) was ?116.6 (70.5), 20.1(76.4), and 91.3 (92.0) mL/kg/min at low, normal, and high CI_BW, respectively. Bias of the COfem (± SD) was ?20.3 (19.0), 8.6 (70.9), and ?2.9 (83.0) mL/kg/min at low, normal, and high CI_BW, respectively. The mean relative bias for COfem was ?6.7 ± 44% (limits of agreements: ?81.2 to 67.9%). Conclusion – Compared with lithium dilution, the pulse contour analysis provides a good estimation of CO, but requires femoral artery catheterization in anesthetized dogs.