Trypsin immobilization in ordered porous polymer membranes for effective protein digestion

Qiao, J.,Kim, J.Y.,Wang, Y.Y.,Qi, L.,Wang, F.Y.,Moon, M.H. Elsevier Pub. Co 2016 Analytica chimica acta Vol.906 No.-

Fast and effective protein digestion is a vital process for mass spectrometry (MS) based protein analysis. This study introduces a porous polymer membrane enzyme reactor (PPMER) coupled to nanoflow liquid chromatography-tandem MS (nLC-ESI-MS/MS) for on-line digestion and analysis of proteins. Poly (styrene-co-maleic anhydride) (PS-co-MAn) was fabricated by the breath figure method to make a porous polymer membrane in which the MAn group was covalently bound to enzyme. Based on this strategy, microscale PPMER (μPPMER) was constructed for on-line connection with the nLC-ESI-MS/MS system. Its capability for enzymatic digestion with bovine serum albumin (BSA) was evaluated with varied digestion periods. The on-line proteolysis of BSA and subsequent analysis with μPPMER-nLC-ESI-MS/MS revealed that peptide sequence coverage increased from 10.3% (digestion time 10 min) to 89.1% (digestion time 30 min). μPPMER can efficiently digest proteins due to the microscopic confinement effect, showing its potential application in fast protein identification and protease immobilization. Applications of on-line digestion using μPPMER with human plasma and urinary proteome samples showed that the developed on-line method yielded equivalent or better performance in protein coverage and identified more membrane proteins than the in-solution method. This may be due to easy accommodation of hydrophobic membrane proteins within membrane pores.

Nutritional Evaluation of Chinese Nonconventional Protein Feedstuffs for Growing-Finishing Pigs - 2. Rapeseed Meal

Li, Defa,Qiao, S.Y.,Yi, G.F.,Jiang, J.Y.,Xu, X.X.,Thacker, P.,Piao, X.S.,Han, In K. Asian Australasian Association of Animal Productio 2000 Animal Bioscience Vol.13 No.1

Two experiments were conducted to determine ileal digestibilities for the amino acids contained in rapeseed meal using the regression technique and then applying the values obtained, in a growth trial, using growing-finishing pigs. For the digestibility trial, four 20 kg crossbred $(Yorkshire{\times}Landrace{\times}Beijing\;Black)$ barrows were fitted with simple T-cannula in the terminal ileum. After recovery, the barrows were fed one of four experimental diets according to a $4{\times}4$ Latin Square design. The pigs were fed corn-soybean meal based diets supplemented with 0, 25, 50 or 75% rapeseed meal. For the growth trial, 80 crossbred $(Yorkshire{\times}Landrace{\times}Beijing\;Black)$ growing pigs $(20{\pm}2.4kg)$ were fed corn-soybean meal diets supplemented with 0, 3, 6, 9 or 12% rapeseed meal. Four pens (2 gilts and 2 castrates) were assigned to each treatment. With the exception of isoleucine and methionine, the digestibility coefficients for the indispensible amino acids declined as the level of rapeseed meal in the diet increased. There was little agreement between the amino acid digestibilities determined with the regression technique and values previously published for rapeseed meal. During the growing (22-42 kg) period, the addition of rapeseed meal had no significant effects on gain, feed intake or feed conversion. During the finishing period (58-91 kg), daily gain was not affected by rapeseed meal inclusion but feed conversion declined (p<0.04) as the level of rapeseed meal in the diet increased.

Effects of Isoleucine Supplementation of a Low Protein, Corn-Soybean Meal Diet on the Performance and Immune Function of Weanling Pigs

Zheng, C.T.,Li, D.F.,Qiao, S.Y.,Gong, L.M.,Zhang, D.F.,Thacker, P.,Han, In K. Asian Australasian Association of Animal Productio 2001 Animal Bioscience Vol.14 No.1

This experiment was conducted to investigate the effects of crystalline isoleucine supplementation of a low protein, corn-soybean meal diet on the performance and immune function of weanling pigs. Forty-five crossbred ($Duroc{\times}Landrace{\times}Large\;White$) piglets, weighing an average of $11.00{\pm}0.07kg$, were assigned to either a control diet containing 20% crude protein (0.64% isoleucine), a 16% crude protein diet without isoleucine supplementation (0.41% isoleucine) or a 16% crude protein diet supplemented with isoleucine (0.64% isoleucine). Reducing the crude protein content of the diet from 20 to 16% significantly (p<0.05) reduced both average daily gain and feed intake. Feed conversion also tended (p=0.07) to be poorer for a low protein diet without isoleucine supplementation. Isoleucine supplementation of the 16% crude protein diet increased both gain and feed intake to a level similar to that obtained by pigs fed the 20% crude protein diet (p>0.05). Blood urea nitrogen, serum total protein and serum globulin were significantly (p<0.05) higher for pigs fed the unsupplemented 16% crude protein diet than for pigs fed the isoleucine-supplemented diet or the control. Egg albumin antibody titre decreased significantly (p<0.05) in pigs fed the diet with isoleucine supplementation, whereas the antibody titre of pigs fed the low protein and low isoleucine diet was similar to that of pigs fed the diet containing 20% crude protein and 0.64% isoleucine. It was suggested that crystalline isoleucine supplementation of a low protein and low isoleucine diet improved pig performance but suppressed humoral immune function.

Use of Chinese Sunflower Meal as a Nonconventional Protein Feedstuff for Growing-Finishing Pigs

Li, Defa,Yi, G.F.,Qiao, S.Y.,Zheng, C.T.,Xu, X.X.,Piao, X.S.,Han, In K.,Thacker, P. Asian Australasian Association of Animal Productio 2000 Animal Bioscience Vol.13 No.5

Two experiments were conducted to determine the ileal digestibility of the amino acids contained in sunflower meal using the regression technique and then applying the values obtained, in a growth trial, using growing-finishing pigs. For the digestibility trial, four 20 kg crossbred ($Yorkshire{\times}Landrace{\times}Beijing$ Black) barrows were fitted with simple Tcannula in the terminal ileum. After recovery, the barrows were fed one of four experimental diets according to a $4{\times}4$ Latin Square design. The pigs were fed corn-soybean meal based diets supplemented with 0, 25, 50 or 75% sunflower meal. For the growth trial, 80 crossbred ($Yorkshire{\times}Landrace{\times}Beijing$ Black) growing pigs (21.5 kg) were fed corn-soybean meal diets supplemented with 0, 5, 10 or 15% sunflower meal. Five pens (2 gilts and 2 castrates) were assigned to each treatment. With the exception of arginine and valine, the digestibility coefficients for the indispensible amino acids declined as the level of sunflower meal in the diet increased. During both the growing (21.5-49.1 kg) and finishing (49.1-90.3 kg) periods and over the entire experiment (21.5-90.3 kg), average daily gain declined in a linear manner (p<0.05) with increasing amounts of sunflower meal. Feed intake was not significantly altered while feed conversion declined in a linear manner (p<0.05) during the grower period only. When the entire experimental period was taken into account, there would appear to be little penalty in either growth or feed conversion for including sunflower meal at levels up to 10% of the diet. Therefore, the price relationship between sunflower meal and other high-protein feedstuffs may provide an excellant opportunity for pork producers to use sunflower meal in order to reduce feed costs.

Effects of Replacing Spray Dried Porcine Plasma with Solpro500 on Performance, Nutrient Digestibility and Intestinal Morphology of Starter Pigs

Lai, C.H.,Qiao, S.Y.,Li, Defa,Piao, X.S.,Bai, L.,Mao, X.F. Asian Australasian Association of Animal Productio 2004 Animal Bioscience Vol.17 No.2

The objective of this study was to evaluate Solpro500 (a wheat hydrolysate containing a high level of glutamine) as a replacement for spray dried porcine plasma (SDPP) in diets fed to nursery pigs. One hundred and eight pigs (Dalland, $5.39{\pm}0.80$ kg BW) weaned at 21 days were assigned to one of three treatment groups for a 28 day feeding trial. The experimental diets were based on corn and soybean meal and were supplemented with either 8% SDPP, 4% SDPP plus 4% Solpro500 or 8% Solpro500. Each treatment was fed to six pens with six pigs per pen (4 barrows and 2 gilts). The experimental results indicated no significant difference (p>0.05) in daily gain, feed intake or feed efficiency for pigs fed the three experimental diets. However, the diarrhea index for pigs fed either 4% SDPP and 4% Solpro500 or 8% Solpro500 was lower (p<0.05) than that for pigs fed 8% SDPP. No differences (p>0.05) were found in the apparent fecal digestibility of dry matter, organic matter or crude protein between pigs fed the three diets. The intestinal morphology (villous height, villous width and crypt depth) was not affected by diet treatments (p>0.05). In conclusion, Solpro500 SDPP can replace SDPP without any negative effects on nursery pig performance.

A model to assess fundamental and realized carrying capacities of island ecosystem: A case study in the southern Miaodao Archipelago of China

Shi, H.,Shen, C.,Zheng, W.,Li, F.,Wang, X.,Chi, Y.,Guo, Z.,Qiao, M.,Gao, L.,Ding, D. Springer Science + Business Media 2016 Acta oceanologica Sinica Vol.35 No.2

Manganese-cobalt hexacyanoferrate cathodes for sodium-ion batteries

Pasta, Mauro,Wang, Richard Y.,Ruffo, Riccardo,Qiao, Ruimin,Lee, Hyun-Wook,Shyam, Badri,Guo, Minghua,Wang, Yayu,Wray, L. Andrew,Yang, Wanli,Toney, Michael F.,Cui, Yi The Royal Society of Chemistry 2016 Journal of materials chemistry. A, Materials for e Vol.4 No.11

<P>Prussian Blue analogues (PBAs) have shown promise as electrode materials for grid-scale batteries because of their high cycle life and rapid kinetics in aqueous-based electrolytes. However, these materials suffer from relatively low specific capacity, which may limit their practical applications. Here, we investigate strategies to improve the specific capacity of these materials while maintaining their cycling stability and elucidate mechanisms that enhance their electrochemical properties. In particular, we have studied the electrochemical and structural properties of manganese hexacyanoferrate (MnHCFe) and cobalt hexacyanoferrate (CoHCFe) in an aqueous, sodium-ion electrolyte. We also studied manganese-cobalt hexacyanoferrate (Mn-CoHCFe) solid solutions with different Mn/Co ratios that combine properties of both MnHCFe and CoHCFe. The materials have the characteristic open-framework crystal structure of PBAs, and their specific capacities can be significantly improved by electrochemically cycling (oxidizing and reducing) both the carbon-coordinated Fe and the nitrogen-coordinated Co or Mn ions.<I>In situ</I>synchrotron X-ray diffraction studies and<I>ex situ</I>soft X-ray absorption spectroscopy combined with an in-depth electrochemical characterization provide insight into the different electrochemical properties associated with the Fe, Co, and Mn redox couples. We show that cycling the C-coordinated Fe preserves the crystal structure and enables the outstanding kinetics and cycle life previously displayed by PBAs in aqueous electrolytes. On the other hand, the N-coordinated Co and Mn ions exhibit a slower kinetic regime due to structural distortions resulting from the weak N-coordinated crystal field, but they still contribute significantly towards increasing the specific capacity of the materials. These results provide the understanding needed to drive future development of PBAs for grid-scale applications that require extremely high cycle life and kinetics.</P>

Effect of Feeding Organic Acid With or Without Enzyme on Intestinal Microflora, Intestinal Enzyme Activity and Performance of Weaned Pigs

Li, Defa,Liu, S.D.,Qiao, S.Y.,Yi, G.F.,Liang, C.,Thacker, P. Asian Australasian Association of Animal Productio 1999 Animal Bioscience Vol.12 No.3

Ninety-six, 35 day old, crossbred pigs, were fed either a basal diet based on com, soybean meal, fish meal and whey or one of three similar diets supplemented with either 0.5% organic acid or enzyme both alone and in combination. Neither organic acid nor enzyme produced any significant (p<0.05) improvements in daily gain or feed conversion. Organic acid, both alone and in combination with enzyme, significantly (p=0.04) decreased the pH in the lower colon. None of the treatments produced any effects on E. Coli or Lactobacillus numbers in any part of the gastrointestinal tract. Feeding enzyme increased trypsin (p=0.01), chymotrypsin (p=0.03) and amylase (p=0.08) levels in the jejunum. Chymotrypsin levels were higher (p=0.04) in the ileum of pigs fed enzyme. Serum glucose levels were lower (p=0.01) on day 14 in pigs fed enzyme either alone or in combination with acid. Under the conditions of this experiment (10% dietary whey, pigs weaned at 35 days of age), neither organic acid nor enzyme were effective in improving starter pig performance. Therefore, there would appear to be little justification for the routine inclusion of these products in diets fed to pigs weaned at 35 days or later.

Performance of Growing-Finishing Pigs Fed Sesame Meal Supplemented Diets Formulated Using Amino Acid Digestibilities Determined by the Regression Technique

Li, Defa,Qiao, S.Y.,Yi, G.F.,Jiang, J.Y.,Xu, X.X.,Piao, X.S.,Han, In K.,Thacker, P. Asian Australasian Association of Animal Productio 2000 Animal Bioscience Vol.13 No.2

Two experiments were conducted to determine ileal digestibilities for the amino acids contained in sesame meal using the regression technique and then applying the values obtained in a growth trial using growing-finishing pigs. For the digestibility trial, four, 20 kg crossbred (Yorkshire $\times$ Landrace $\times$ Beijing Black) barrows were fitted with a simple T-cannula in the terminal ileum. After recovery, the barrows were fed one of four experimental diets according to a $4{\times}4$ Latin Square design. The pigs were fed corn-soybean meal based diets supplemented with 0, 25, 50 or 75 percent sesame meal. For the growth trial, 210 crossbred (Yorkshire $\times$ Landrace $\times$ Henan Min) growing pigs ($21.8{\pm}1.4kg$), were fed corn-soybean meal based diets supplemented with 0, 3, 6, 9, or 12% sesame meal. Three pens (7 gilts and 7 castrates) were assigned to each treatment. With the exception of arginine and phenylalanine, the digestibility coefficients for the indispensible amino acids declined as the level of sesame meal in the diet increased. There was little agreement between the amino acid digestibilities determined with the regression technique and values previously published for sesame meal determined with the direct method. Daily gain and feed conversion both declined (linear effect p=0.02 and 0.06 respectively) as the level of sesame meal in the diet increased.

Branched-chain Amino Acids are Beneficial to Maintain Growth Performance and Intestinal Immune-related Function in Weaned Piglets Fed Protein Restricted Diet

Ren, M.,Zhang, S.H.,Zeng, X.F.,Liu, H.,Qiao, S.Y. Asian Australasian Association of Animal Productio 2015 Animal Bioscience Vol.28 No.12

As a novel approach for disease control and prevention, nutritional modulation of the intestinal health has been proved. However, It is still unknown whether branched-chain amino acid (BCAA) is needed to maintain intestinal immune-related function. The objective of this study was to determine whether BCAA supplementation in protein restricted diet affects growth performance, intestinal barrier function and modulates post-weaning gut disorders. One hundred and eight weaned piglets ($7.96{\pm}0.26kg$) were randomly fed one of the three diets including a control diet (21% crude protein [CP], CON), a protein restricted diet (17% CP, PR) and a BCAA diet (BCAA supplementation in the PR diet) for 14 d. The growth performance, plasma amino acid concentrations, small intestinal morphology and intestinal immunoglobulins were tested. First, average daily gain (ADG) (p<0.05) and average daily feed intake (ADFI) (p<0.05) of weaned pigs in PR group were lower, while gain:feed ratio was lower than the CON group (p<0.05). Compared with PR group, BCAA group improved ADG (p<0.05), ADFI (p<0.05) and feed:gain ratio (p<0.05) of piglets. The growth performance data between CON and BCAA groups was not different (p>0.05). The PR and BCAA treatments had a higher (p<0.05) plasma concentration of methionine and threonine than the CON treatment. The level of some essential and functional amino acids (such as arginine, phenylalanine, histidine, glutamine etc.) in plasma of the PR group was lower (p<0.05) than that of the CON group. Compared with CON group, BCAA supplementation significantly increased BCAA concentrations (p<0.01) and decreased urea concentration (p<0.01) in pig plasma indicating that the efficiency of dietary nitrogen utilization was increased. Compared with CON group, the small intestine of piglets fed PR diet showed villous atrophy, increasing of intra-epithelial lymphocytes (IELs) number (p<0.05) and declining of the immunoglobulin concentration, including jejunal immunoglobulin A (IgA) (p = 0.04), secreted IgA (sIgA) (p = 0.03) and immunoglobulin M (p = 0.08), and ileal IgA (p = 0.01) and immunoglobulin G (p = 0.08). The BCAA supplementation increased villous height in the duodenum (p<0.01), reversed the trend of an increasing IELs number. Notably, BCAA supplementation increased levels of jejunal and ileal immunoglobulin mentioned above. In conclusion, BCAA supplementation to protein restricted diet improved intestinal immune defense function by protecting villous morphology and by increasing levels of intestinal immunoglobulins in weaned piglets. Our finding has the important implication that BCAA may be used to reduce the negative effects of a protein restricted diet on growth performance and intestinal immunity in weaned piglets.