The isolation, characterization and biological activities of the non-glucan polysaccharides from the high-starch-content plant Pueraria mirifica.

The dried root of Pueraria mirifica (P. mirifica) is an edible foodstuff widely used in Asian countries. P. mirifica is known for its high starch content. The isolation of polysaccharides from high-starch plant parts is challenging due to the interference of starch. Therefore, this study aimed to develop a technique for isolating and investigating the structure and activity of non-glucan polysaccharides from P. mirifica (PMP). An effective starch removal process was developed using α-amylase hydrolysis and thorough membrane dialysis. Four non-glucan polysaccharides were isolated, and PMP-2 was subjected to structural elucidation. The results indicated that PMP-2 has a molecular weight of 124.4 kDa and that arabinose and galactose are the main components, accounting for 27.8 % and 58.5 %, respectively. Methylation and NMR analysis suggested that PMP-2 is an Arabinogalactan composed of 1,6-linked Galp and 1,4-linked Galp as the main chain, with arabinan and rhamnose as side chains. Furthermore, PMP-C and PMP-2 exhibited concentration-dependent antioxidant activities against DPPH, ABTS, and hydroxyl radicals and certain immunomodulatory activities related to the release of NO, TNF-α and IL-6. These findings suggest that PMP-2 has potential therapeutically active ingredient in functional foods. The developed method successfully removed starch and isolated non-glucan polysaccharides from the high-starch content plant P. mirifica and can be applied to other high-starch plants.

Enzymatic synthesis of Vaccinium blue using vaccinoside as a bifunctional precursor.

Since Tang dynasty in China, the fresh leaves of Vaccinium bracteatum (VBL) have been applied as natural pigment to produce black rice. However, detailed information on its biosynthetic mechanism still remained unclear. Following rice dyeing capacity assay, vaccinoside, one of iridoid glycosides, was identified as the key active compound. Increased methodical research demonstrated vaccinoside as a distinct bifunctional precursor, which could be catalyzed by polyphenol oxidase or ß-glucosidase independently, followed by reaction with 15 amino acids to give blue pigments (VBPs; λmax 581-590 nm) of different hues. Two synthetic pathways of VBPs were proposed, using multiple techniques such as HPLC, HPSEC, UV-Vis spectrum and colorimeter as analysis tools. Black rice was interpreted to be prepared by cooking, using vaccinoside, intrinsic enzymes from fresh VBL and rice protein in combination. These findings promote the understanding of VBP formation mechanisms and provide an efficient method of producing novel Vaccinium blue pigments.

[Optimization and application of caprylic acid precipitation in the purification of monoclonal antibody].

In response to the market demand for therapeutic antibodies, the upstream cell culture scale and expression titer of antibodies have been significantly improved, while the production efficiency of downstream purification process is relatively fall behind, and the downstream processing capacity has become a bottleneck limiting antibody production throughput. Using monoclonal antibody mab-X as experimental material, we optimized the caprylic acid (CA) precipitation process conditions of cell culture fluid and low pH virus inactivation pool, and studied two applications of using CA treatment to remove aggregates and to inactivate virus. Based on the lab scale study, we carried out a 500 L scale-up study, where CA was added to the low pH virus inactivation pool for precipitation, and the product quality and yield before and after precipitation were detected and compared. We found that CA precipitation significantly reduced HCP residuals and aggregates both before and after protein A affinity chromatography. In the aggregate spike study, CA precipitation removed about 15% of the aggregates. A virus reduction study showed complete clearance of a model retrovirus during CA precipitation of protein A purified antibody. In the scale-up study, the depth filtration harvesting, affinity chromatography, low pH virus inactivation, CA precipitation and depth filtration, and cation exchange chromatography successively carried out. The mixing time and stirring speed in the CA precipitation process significantly affected the CA precipitation effect. After CA precipitation, the HCP residue in the low pH virus inactivation solution decreased 895 times. After precipitation, the product purity and HCP residual meet the quality criteria of monoclonal antibodies. CA precipitation can reduce the chromatography step in the conventional purification process. In conclusion, CA precipitation in the downstream process can simplify the conventional purification process, fully meet the purification quality criterion of mab-X, and improve production efficiency and reduce production costs. The results of this study may promote the application of CA precipitation in the purification of monoclonal antibodies, and provide a reference for solving the bottleneck of the current purification process.

[Effects of simulated rainfall enhancement on sediment CO2 flux in dry lakebed of Barkol Lake, China]. / 模拟增雨对巴里坤湖干涸湖底沉积物CO2通量的影响.

Understanding the responses of lake sediment carbon process to climate change is an important part of a comprehensive understanding of lake carbon budget. To explore the effects of future rainfall increase on sediment carbon flux, undisturbed sediment samples were collected from the bottom of dry lake Barikun in Hami, Xinjiang for the incubation experiment. Based on the increase rate of precipitation (4 mm·10 a-1) and the distribution characteristics of rainfall in the plant growing season in Hami, Xinjiang since 1960, five rainfall treatments were set (86 mm, T0; 94 mm, T1; 102 mm, T2; 110 mm, T3; 126 mm, T4) based on the rainfall in growing season of 2016 (86 mm). We analyzed the effects of rainfall increase on sediment CO2 flux. Results showed that compared with that before rainfall, the sediment CO2 flux increased after 1 day of rainfall in the study area. Compared with that during May to July, the CO2 flux of sediments in August to October decreased. There was no variation of CO2 accumulative emission among the T0-T3 treatments from May to October. However, the average CO2 emission rate under the T3 treatment (0.22 µmol·m-2·s-1) was significantly higher than that under the T4 treatment (0.14 µmol·m-2·s-1). All treatments showed CO2 sink at the first day of rainfall (1 d), with T4 treatment (-0.13 µmol·m-2·s-1) having the highest "carbon sink" capacity. After 1 day, the CO2 sink converted to CO2 source under the five rainfall treatments, with the CO2 emission rate under T3 treatment (0.34 µmol·m-2·s-1) being significantly higher than those under other treatments. Compared with May, the CO2 emission fluxes of T2-T4 treatments were significantly higher than those at the time from August to October. Under the condition with relatively stable temperature, the CO2 flux of sediments was significantly correlated with the sediment moisture and air humidity. In the next 60 years, the continuous increase of future rainfall may be an important factor promoting CO2 emission from lake sediment in arid regions, and thus affecting global warming.

Geniposide attenuates neonatal mouse brain injury after hypoxic-ischemia involving the activation of PI3K/Akt signaling pathway.

Perinatal hypoxic-ischemia (HI) is a leading cause of acute mortality and neurologic complications in newborns. Geniposide, a natural product extracted from the herb Gardenia jasminoides, has been shown to possess neuroprotective effects in neurologic deficits. This study aims to investigate whether Geniposide has therapeutic potential to HI brain injury and the underlying mechanisms. C57/bl6 mice were subjected to HI insult on postnatal day 10. Geniposide (20 mg/kg b.w.) was administered intragastrically every day after HI insult for 7 successional days. Then mice at P18 were sacrificed and brain tissues were collected for further analysis. Geniposide treatment significantly inhibited cell apoptosis, reduced serum IgG leakage into brain tissue, attenuated astrogliosis and microgliosis, prevented loss of pericytes, loss of tight junction and adherens junction proteins. The PI3K/Akt signaling pathway, which related proteins were downregulated after HI insult, was activated by Geniposide treatment. Geniposide treatment after neonatal HI insult attenuated HI-induced cell apoptosis, IgG leakage, microgliosis, astrogliosis, pericytes loss and junction protein degradation. Geniposide could protect against HI-induced brain injury, which might be through the activation of PI3K/Akt signaling pathway.

Thermal-wave resonant cavity signal processing.

The thermal-wave resonant cavity (TWRC) technique has been used for thermal diffusivity measurements by many researchers. This study aims to reduce the uncertainty associated with TWRC signal processing (curve fitting) by means of numerical simulation and experimental verification. Simulations show that the plot of signal amplitude versus cavity length can be fitted to a simplified model reported previously when the initial fitting position is at least twice the thermal-wave diffusion length (2 µg), and that the uncertainty caused by different end positions is negligible in the range of 6-10 µg. Upon consideration of the simulation results, signal-to-noise ratio, and clearly defined amplitude curve shape, fitting ranges of about 2.2-8.0 µg and 2.2-8.7 µg were chosen for the experimental data. Thermal diffusivity values (1.438 ± 0.001) × 10-7 and (1.436 ± 0.001) × 10-7 m2 s-1, respectively, were obtained for distilled water, in excellent agreement with the accepted literature value. The ratio of standard deviation to the mean value is smaller than 0.07%, one order of magnitude lower than typical results reported in the literature. Similar simulation results were obtained for air and methanol as intra-cavity samples.

Correlation of liver function with intestinal flora, vitamin deficiency and IL-17A in patients with liver cirrhosis.

The aim of this study was to investigate the correlation of liver function, intestinal flora, vitamin D and interleukin-17A (IL-17A) levels in patients with liver cirrhosis. A total of 52 patients diagnosed with posthepatitic cirrhosis and admitted into Yantai Infectious Disease Hospital (Yantai, China) from January to December in 2012 (liver cirrhosis group), and 52 patients with chronic hepatitis B (hepatitis group), and 40 healthy volunteers receiving physical examination in the hospital (normal control group) were selected into the study. The liver function, hepatitis B virus (HBV) deoxyribonucleic acid (DNA) level, intestinal flora distribution, vitamin D and IL-17A levels of all patients were detected, and the correlation among them was analyzed via Pearson's analysis. The number of Enterobacteriaceae, Enterococcus, Staphylococcus aureus and Saccharomyces in hepatitis and liver cirrhosis groups was significantly greater than in the normal control group (P<0.05), but the number of Lactobacillus, Bacteroides, Bifidobacterium and Clostridium was significantly decreased (P<0.05); the serum IL-17A levels in hepatitis and liver cirrhosis were obviously higher than that in the normal control group (P<0.05), but the serum vitamin D 25(OH) D and 1,25(OH)2D levels were obviously lower than that in the normal control group (P<0.05). In patients with liver cirrhosis, Enterobacteriaceae was positively correlated with prothrombin time (PT), Enterococcus was positively correlated with alanine aminotransferase and aspartate aminotransferase (AST) levels, Bifidobacterium was negatively correlated with AST, alkline phosphatase (AKP) and HBV DNA levels, and Bacteroides was negatively correlated with AST level and PT. There was a significant negative correlation between serum IL-17A and total bilirubin in patients with liver cirrhosis, and 25(OH) D was negatively correlated with AST, AKP and HBV DNA levels. In patients with liver cirrhosis, there was significant positive correlation between Enterococcus and IL-17A, and between Lactobacillus and 25(OH)D, but other bacteria were not obviously associated with IL-17A and vitamin D. Intestinal flora imbalance, vitamin D deficiency and IL-17A imbalance play an important role in the evolution of liver cirrhosis.

Frequency analysis of temperature-dependent interferometric signal for the measurement of the temperature coefficient of refractive index.

A method of frequency analysis for the measurement of the temperature coefficient of refractive index (dn/dT) using a Fabry-Perot interferometer was developed and tested against ethanol and water. The temperature-dependent interferometric signal described by Airy's formula was analyzed in both the temperature and frequency domains. By fast Fourier transform, a low-pass filter was designed and employed to eliminate the noise superimposed on the signal. dn/dT was determined accurately from the noise-removed signal by peak analysis. Furthermore, the signal frequency parameters may be utilized for the material thermophysical property characterization. This method lays the foundation for an online dn/dT instrument for monitoring chemical processes.

A simplified model for thermal-wave cavity self-consistent measurement of thermal diffusivity.

A simplified theoretical model was developed for the thermal-wave cavity (TWC) technique in this study. This model takes thermal radiation into account and can be employed for absolute measurements of the thermal diffusivity of gas and liquid samples without any knowledge of geometrical and thermal parameters of the components of the TWC. Using this model and cavity-length scans, thermal diffusivities of air and distilled water were accurately and precisely measured as (2.191 ± 0.004) × 10(-5) and (1.427 ± 0.009) × 10(-7) m(2) s(-1), respectively, in very good agreement with accepted literature values.

Immobilization of cellulase on a reversibly soluble-insoluble support: properties and application.

Cellulase was coupled to N-succinyl-chitosan (NSC) showing soluble-insoluble characteristics with pH change. Cellulase immobilized on NSC (NSCC) is in a soluble state during the enzyme reaction, yet can be recovered in its insoluble form by lowering the pH of the reaction solution after the reaction. NSCC was obtained under the optimized immobilization conditions of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC) 10 mg, cellulase 15 mg, and pH 5.0. The retention activity of the immobilized cellulase was found to be 48.8%. The effects of pH and temperature on the activity and stability of NSCC were studied and compared with those of free cellulase. The optimum temperature and pH of NSCC was 45 degrees C and 4.0, respectively, which was found unchanged compared with the free one. The stability of cellulase against change in the pH and temperature was improved by the immobilization. The effectiveness of employing NSCC for extracting flavonoids from Ginkgo biloba leaf powder was investigated. Results showed that NSCC enhanced extraction yield up to 2.35-fold when compared with the conventional method. Moreover, NSCC retained 83.5% of its initial activity after five batches of hydrolysis reaction.

Thermal-lens study of photochemical reaction kinetics.

We consider the time dependence of the absorption coefficient due to the photoinduced chemical reaction (PCR) and species diffusion to calculate the temperature rise in the thermal-lens (TL) effect. The TL signal at the detector plane is also calculated. This theoretical approach removes the restriction that the PCR time constant is much greater than the characteristic TL time constant, which was assumed in a previously published model. Hydrocarbon fuel and aqueous Cr(VI) samples are investigated, and quantitative experimental results for the thermal, optical, and PCR properties are obtained. While similar results were obtained for the Cr(VI) solution using the previous and present models, the relative difference between the PCR time constants extracted from the same experimental data for a hydrocarbon fuel sample is found to be more than 220%. This demonstrates the significant difference of the two models.

The effect of relative humidity on binary gas diffusion.

The dependence of diffusion coefficient of O2-N2 mixture in the presence of water vapor was experimentally determined as a function of relative humidity (RH) with different temperatures using an in-house made Loschmidt diffusion cell. The experimental results showed that O2-N2 diffusion coefficient increased more than 17% when RH increased from 0% to 80% at 79 degrees C. In the experiments, the RH in both top and bottom chambers of the diffusion cell were the same, and the pressure inside the diffusion cell was kept as ambient pressure (1 atm.). Maxwell-Stefan theory was employed to analyze the mass transport in the diffusion cell, and found that there was no effective water vapor diffusion taking place, indicating that the gas diffusion in this ternary (O2-N2-water vapor) system could be considered binary gas (O2-N2) diffusion. The Fuller, Schettler, and Giddings (FSG) empirical equation of the kinetic theory of gases was generalized to accommodate the dependence of the binary diffusion coefficient on the RH. The prediction of the generalized equation was found to be consistent with experimental results with the difference of less than 1.5%, showing that the generalized equation could be applied to calculate the diffusion coefficients of the binary gaseous mixture with different temperature and RH values. The effect of water vapor on the increase of O2-N2 diffusion coefficient was discussed using molecule theory.

Time-resolved thermal mirror technique with top-hat cw laser excitation.

A theoretical model was developed for time-resolved thermal mirror spectroscopy under top-hat cw laser excitation that induced a nanoscale surface displacement of a low absorption sample. An additional phase shift to the electrical field of a TEM(00) probe beam reflected from the surface displacement was derived, and Fresnel diffraction theory was used to calculate the propagation of the probe beam. With the theory, optical and thermal properties of three glasses were measured, and found to be consistent with literature values. With a top-hat excitation, an experimental apparatus was developed for either a single thermal mirror or a single thermal lens measurement. Furthermore, the apparatus was used for concurrent measurements of thermal mirror and thermal lens. More physical properties could be measured using the concurrent measurements.

Peripheral arterial occlusive disease: global gene expression analyses suggest a major role for immune and inflammatory responses.

BACKGROUND: Peripheral arterial disease (PAD), a major manifestation of atherosclerosis, is associated with significant cardiovascular morbidity, limb loss and death. However, mechanisms underlying the genesis and progression of the disease are far from clear. Genome-wide gene expression profiling of clinical samples may represent an effective approach to gain relevant information. RESULTS: After histological classification, a total of 30 femoral artery samples, including 11 intermediate lesions, 14 advanced lesions and 5 normal femoral arteries, were profiled using Affymetrix microarray platform. Following real-time RT-PCR validation, different algorithms of gene selection and clustering were applied to identify differentially expressed genes. Under a stringent cutoff, i.e., a false discovery rate (FDR) <0.5%, we found 366 genes were differentially regulated in intermediate lesions and 447 in advanced lesions. Of these, 116 genes were overlapped between intermediate and advanced lesions, including 68 up-regulated genes and 48 down-regulated ones. In these differentially regulated genes, immune/inflammatory genes were significantly up-regulated in different stages of PAD, (85/230 in intermediate lesions, 37/172 in advanced lesions). Through literature mining and pathway analysis using different databases such as Gene Ontology (GO), and the Kyoto Encyclopedia of Gene and Genomics (KEGG), genes involved in immune/inflammatory responses were significantly enriched in up-regulated genes at different stages of PAD(p < 0.05), revealing a significant correlation between immune/inflammatory responses and disease progression. Moreover, immune-related pathways such as Toll-like receptor signaling and natural killer cell mediated cytotoxicity were particularly enriched in intermediate and advanced lesions (P < 0.05), highlighting their pathogenic significance during disease progression. CONCLUSION: Lines of evidence revealed in this study not only support previous hypotheses, primarily based on studies of animal models and other types of arterial disease, that inflammatory responses may influence the development of PAD, but also permit the recognition of a wide spectrum of immune/inflammatory genes that can serve as signatures for disease progression in PAD. Further studies of these signature molecules may eventually allow us to develop more sophisticated protocols for pharmaceutical interventions.

Top-hat cw-laser-induced time-resolved mode-mismatched thermal lens spectroscopy for quantitative analysis of low-absorption materials.

Thermal lens spectroscopy is a highly sensitive and versatile photothermal technique for material analysis, providing optical and thermal properties. To use less expensive multimode non-Gaussian lasers for quantitative analysis of low-absorption materials, this Letter presents a theoretical model for time-resolved mode-mismatched thermal lens spectroscopy induced by a cw laser with a top-hat profile. The temperature profile in a sample was calculated, and the intensity of the probe beam center at the detector plane was also derived using the Fresnel diffraction theory. Experimental validation was performed with glass samples, and the results were found well consistent with literature values of the thermo-optical properties of the samples.

[Research on the orientedly immobilized urease via concanavalin A].

Concanavalin A (ConA) is immobilized on a pre-activated chitosan microspheres, and then oriented immobilization of urease is carried out based on the strong interaction between ConA and glycoprotein. The optimum immobilization conditions are as follows: glutaraldehyde concentration is 3.5%, ConA concentration 1 mg/mL, ConA pH 7.0 and urease concentration 0.4 mg/mL. For orientedly immobilized urease, the highest activity was allowed at pH 5.0-6.0 and temperature 77 degrees C, and the Michaelis constant (Km) was disclosed to be 11.76 mmol/L by Lineweaver-Burk plot. Compared with the free urease and the randomly immobilized urease, the optimum pH of the orientedly immobilized urease becomes smaller and the pH domain wider. Orientedly immobilized urease presents higher temperature resistance, higher affinity to the substrate, and higher stability of operation.

Chemomicin A, a new angucyclinone antibiotic produced by Nocardia mediterranei subsp. kanglensis 1747-64.

A new angucyclinone antibiotic, chemomicin A was isolated from cultured broth of Nocardia mediterranei subsp. kanglensis 1747-64. Its chemical structure was determined to be 1,2,3,4a,5,6,6a,12a,12b-nonahydro-1,2,3,8,12,12b-hexahydroxy-3-methyl-6a,12a-epoxybenz[a]anthracen-4,7(12H)-dione by a detailed spectroscopic analysis. Chemomicin A had antimicrobial activity against Bacillus subtilis and Enterococcus faecium with MIC values of 10.2 and 20.4 microM, respectively, and showed cytotoxicity against human colorectal cancer HCT116 cells and human esophageal carcinoma YES-2 cells with IC50 values of 127 and 153 microM, respectively.

Deinococcus yunweiensis sp. nov., a gamma- and UV-radiation-resistant bacterium from China.

A Gram-negative, non-spore-forming, non-motile, rod-shaped, red-pigmented strain, designated YIM 007T, was found as a contaminant on an agar plate in the laboratory of Yunnan Institute of Microbiology, China. The optimum growth pH and temperature for the isolate were 7.0-7.5 and 30 degrees C, respectively. The predominant respiratory quinone was MK-8. The polar lipid profile consisted mainly of various unknown phosphoglycolipids and glycolipids. The major cellular fatty acids were C(16 : 1)omega7c, C(16 : 0), C(17 : 0) and C(17 : 1)omega8c. l-Ornithine was detected in its peptidoglycan. The DNA G+C content was 64.1 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain YIM 007(T) showed 16S rRNA gene sequence similarity levels of 86.8-92.1 % to the other described Deinococcus species. Based on the high 16S rRNA gene sequence divergence and phenotypic differences, it is proposed that the unknown strain should be classified as a novel species in the genus Deinococcus with the name Deinococcus yunweiensis sp. nov. The type strain is YIM 007T (=KCTC 3962T=DSM 17005T).

Preliminary report on the biological effects of space flight on the producing strain of a new immunosuppressant, Kanglemycin C.

Kanglemycin C (K-C) is a new immunosuppressant isolated from the culture broth of Nocardia mediterranei var. kanglensis 1747-64. To improve the productivity of K-C and to study the biological effects of space flight on its producing strain, spores from five K-C producing strains (U-10, U-15, U-7, M-13, gamma-33) mutated from the wild strain N. mediterranei var. kanglensis 1747-64 were carried into space by an unmanned spaceship, "Shenzhou III" (Divine Vessel III) on March 25, 2002. Comparatively, the strain U-7 was the highest K-C producing strain among the above five starting strains when cultivated in 500-ml Erlenmeyer flasks. After a 6 day and 18 h flight, the treated spores went through serial screening processes to screen for high-yield K-C mutant strains, using thin layer chromatography and high performance liquid chromatography (HPLC). The K-C yield produced by one mutant strain, designated as F-16, derived from the starting strain U-7 was increased by up to 200% when compared to that produced by the starting strain U-7 in 500-ml Erlenmeyer flasks after careful postflight HPLC analysis. Another mutant strain, designated as F-210, derived from the starting strain M-13 showed reduced productivity of K-C as well as exhibited changes in some morphological and physiological characteristics. For example, the broth color of the strain F-210 changed from yellow to purple after 96 h of culture, but that of the ground control strain M-13 remained yellow. Similarly, the mycelium morphological change from filamentous to coccoid of F-210 occurred later than that of ground control M-13. Examination of the survivability of postflight spores indicated that exposure to radiation, during the 162 h of space flight, plays a critical role in the survival rates of spores such that spores exposed to strong radiation exhibited lower survival rates than spores exposed to weak radiation.