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Feeding legume grains to pigs usually increases losses of endogenous proteins at the terminal ileum. However, the identity of such proteins is largely unknown. This study was undertaken to determine the ileal flow and identity of soluble proteins present in large concentrations in ileal digesta of young pigs fed soybean meal (SBM), peas (P), faba beans (FB), or blue lupin (L) in expt. 1, and white (WPC) or black (BPC) chickpeas in expt. 2. Protein in the control diet (C) was provided by casein. Ileal digesta proteins were analyzed using sodium dodecyl sulfate polyacrylamide gel electrophoresis, Coomassie blue staining, densitometry and N-terminal amino acid sequencing. Three protein bands at molecular masses of 25, 27, and 30 kDa had a higher ileal flow (P < 0.05) in the pigs fed the legume-based diets compared to those fed the control diet in expt. 2. This was true for the 25- and 30-kDa proteins (P < 0.05) and the 27-kDa protein (P < 0.10) in pigs fed the legume-containing diets in expt. 1. These proteins shared N-terminal amino acid sequences with enzymes of the serine protease family including pig trypsin (25 kDa) and blood coagulation factor IX or chymotrypsin (27 and 30 kDa). KEY WORDS: * piglets * endogenous proteins * serine-protease * proteins * densitometry * sequence analysis

Abstract Background RNA quality and quantity are important factors for ensuring the accuracy of gene expression analysis and other RNA-based downstream applications. Extraction of high quality nucleic acids is difficult from neuronal cells and brain tissues as they are particularly rich in lipids. In addition, most common RNA extraction methods are phenol-based, resulting in RNA that may be incompatible with downstream applications such as gene expression. Findings In this work, a comparative analysis of the RNA quality obtained from SK-N-MC cells was performed using six commonly used RNA isolation kits: two phenol-based kits and four non-phenol based kits. The non-phenol based kits tested AxyPrep Multisource Total RNA Miniprep, RNeasy® Mini, EasySpin and Ilustra RNAspin Mini RNA Isolation, all performed well and resulted in the isolation of high quality RNA, as evaluated by A260/A280. The RNA extracted with AxyPrep Multisource Total RNA Miniprep, RNeasy® Mini and EasySpin provided the highest RNA yields. In particular, the RNA isolated by AxyPrep Multisource Total RNA Miniprep Kit did not show any detectable genomic DNA contamination even without previous DNase treatment or after RNA direct PCR amplification using universal 18S primers. Conclusions The RNA extracted from SK-N-MC cells with AxyPrep Multisource Total RNA Miniprep Kit was superior with respect to the RNA quality and concentration. This kit does not use aggressive organic solvents and RNA free of genomic DNA was isolated without the need for DNase treatment.

Reaction of the tri(μ-sulfido)triiron(III) tris(β-diketiminate) cyclophane complex, Fe3S3LEt/Me (1), or of the di(μ-sulfido)diiron(III) complex Fe2S2HLEt/Me (5), with the related tri(bromide)triiron(II) complex Fe3Br3LEt/Me (2) results in electron and ligand redistribution to yield the mixed-ligand multiiron complexes, including Fe3Br2SLEt/Me (3) and Fe2Br2SHLEt/Me (4). The cleavage and redistribution observed in these complexes is reminiscent of necessary Fe–S bond cleavage for substrate activation in nitrogenase enzymes, and provides a new perspective on the lability of Fe–S bonds in FeS clusters. [ABSTRACT FROM AUTHOR]

In this study, a single, improved methodology was used to extract, fractionate and purify the 11S (legumin-type or related to the [Alpha]-conglutin from Lupinus albus L.), 7S (vicilin-type or related to the [Beta]-conglutin from L. albus) and 2S (related to the [Gamma]-conglutin from L. albus) families of proteins from eight legume species: L. albus, Glycine max (L.) Merr., Pisum sativum L., Vicia faba L., Cicer arietinum L., Phaseolus vulgaris L., Lens culinaris Med. and Arachis hypogaea L. The sedimentation coefficients obtained varied from 1.9 to 8.1 for the [Gamma]-conglutin-related proteins, from 5.1 to 10.5 for the [Beta]-conglutin-related proteins and from 12.0 to 14.9 for the [Alpha]-conglutin-related globulins. The [Gamma]-conglutin-related proteins is the most heterogenous group. Antibodies produced against each type of [Gamma]-conglutin polypeptide chain recognize the other polypeptide chain as well as other polypeptides in the corresponding globulins from all species examined. The 7S globulins are typically composed of a large number of polypeptides, covering a wide range of molecular masses (10 to 70 kD). The presence of disulphide bonds is apparently absent and the occurrence of glycopolypeptides is not widespread. Finally, the 11S globulins are characteristically formed by a limited number of polypeptides that may be divided into a lighter group (20-25 kD) and a heavier group (35-50 kD). The presence of disulphide bonds is apparently widespread but the occurrence of glycopolypeptides seems to be relatively rare. Both the 7S family and the 11S globulins studied by immunoblotting exhibit a low level of structural similarity.