The Effect of Various Probiotic Strains or Avilamycin Feed Additive on Immune Defense Markers and Acute-Phase Response to Salmonella Infection in Chickens.

Probiotics are a nutritional tool for disease prevention. It has been proposed that stimulation of immune response could affect the growth-promoting properties of antimicrobial growth promoters as well as the control of foodborne pathogens. The current study compares immune response in the blood of 280 non-infected and Salmonella-infected chickens fed either with the growth promoter avilamycin or with one of five probiotic strains of Lactobacillus and Bifidobacterium, which also showed growth-promoting properties. All of the probiotic strains stimulated superoxide anion production and the proliferation of leukocytes, while raising lysozyme and γ-globulin levels (by up to 65%, p < 0.01), which are important factors in native and cell-mediated immune defense against pathogens. In contrast, among the two strains examined, specific Salmonella antibodies were induced only by L. salivarius, and not by B. animalis, as assessed by the ELISA method and confirmed by an agglutination reaction (p < 0.05). In the avilamycin-fed group, both non-infected and infected chickens showed decreased levels of these immune markers (by 30%) and increased levels of ceruloplasmin by up to 35%. In contrast, the probiotics suppressed acute-phase response assessed by ceruloplasmin by up to 32%. This correlation implies that various antimicrobial feed additives have a distinct effect on immunomodulation, which may affect different mechanisms in the nutrition-related metabolism associated with the rate of weight gain in chickens. The data could contribute to the design of innovative antimicrobial feed additives in the food industry and consequently to well-being of humans.

RNA backbone: consensus all-angle conformers and modular string nomenclature (an RNA Ontology Consortium contribution).

A consensus classification and nomenclature are defined for RNA backbone structure using all of the backbone torsion angles. By a consensus of several independent analysis methods, 46 discrete conformers are identified as suitably clustered in a quality-filtered, multidimensional dihedral angle distribution. Most of these conformers represent identifiable features or roles within RNA structures. The conformers are given two-character names that reflect the seven-angle delta epsilon zeta alpha beta gamma delta combinations empirically found favorable for the sugar-to-sugar "suite" unit within which the angle correlations are strongest (e.g., 1a for A-form, 5z for the start of S-motifs). Since the half-nucleotides are specified by a number for delta epsilon zeta and a lowercase letter for alpha beta gamma delta, this modular system can also be parsed to describe traditional nucleotide units (e.g., a1) or the dinucleotides (e.g., a1a1) that are especially useful at the level of crystallographic map fitting. This nomenclature can also be written as a string with two-character suite names between the uppercase letters of the base sequence (N1aG1gN1aR1aA1cN1a for a GNRA tetraloop), facilitating bioinformatic comparisons. Cluster means, standard deviations, coordinates, and examples are made available, as well as the Suitename software that assigns suite conformer names and conformer match quality (suiteness) from atomic coordinates. The RNA Ontology Consortium will combine this new backbone system with others that define base pairs, base-stacking, and hydrogen-bond relationships to provide a full description of RNA structural motifs.

Purification and separation of hydrophobic serum amyloid A precursor isoforms by a one-step preparative method.

The levels of two major serum amyloid A precursor isoforms, SAA1 and SAA2, which are associated with high-density lipoproteins (HDL) are increased during inflammation. The hydrophobic character and the small size difference--corresponding to just 0.8 kDa--between these two members of the SAA family hinder their separation and purification on a large scale by conventional methods. In the current work, both mouse SAA proteins were purified from HDL-SAA and acute-phase serum within 10 h in a one-step procedure using the high-resolution, continuous-elution preparative gel electrophoresis Prep-Cell system in combination with Tris/Glycine SDS-PAGE. Moreover, applying the Tris/Tricine system on the Prep-Cell resulted not only in purification of the SAA proteins, but also in their separation within 16 h. The SAA isoforms were freed from SDS using a Centricon concentrator and were identified using monoclonal antibodies. Optical density profile plots of gel protein or Western blot bands in combination with a colorimetric spectrophotometric protein assay showed that the recovery of the isoforms ranged from 38% to 60%. These results show that the preparative gel electrophoresis system Prep-Cell is a suitable device for separating SAA1 and SAA2 proteins in a simplified, convenient, and fast procedure, which can be applied on a small or large scale.

Cellular defense against oxidized low density lipoproteins and fibrillar amyloid beta in murine cells of monocyte origin with possible susceptibility to the oxidative stress induction.

Reactive oxygen species (ROS) are associated with aging and the correlation between Alzheimer's disease and atherosclerosis is a subject of the discussion. The aim of this study was to determine whether genetic factors affect cellular defense against cytotoxic beta-amyloid (Abeta) which is considered to be the source of ROS. Low levels of Abeta (1-10 microM) led to a significant suppression of redox potential as measured by MTT assay in bone marrow-derived cell lines. The atherosclerosis-resistant cells (GG2EE) were less affected than the susceptible cells (ANA1) in the time-, dose-, and Abeta species-dependent manner. Cell death in amyloid treated resident susceptible macrophages (C57BL/6J), measured by lactate dehydrogenase release, was induced during prolonged incubation and increased when compared with the resistant macrophages (C3H/HeJ, P = 0.005). SDS-PAGE showed that Abeta persisted intracellularly during this period. The cytotoxicity of oxidized low density lipoproteins (oxLDLs) significantly affected only the susceptible cells which actually lowered this cytotoxicity, thus, implying that the harmful effect of the oxLDLs was diminished when compared to that of Abeta. This fact demonstrates that in vitro the defense by cells of monocyte origin against Abeta may be determined in part genetically whereas the reaction to oxLDLs could be fully underlined by genetic susceptibility.

Structural requirements for lysosomal targeting of the prosaposin precursor protein.

Although the Man-6-P-independent lysosomal sorting of prosaposin, a precursor of four saposins (A, B, C, and D) is not understood, a protein/lipid interaction is considered. Immunocytochemical analysis revealed that each single saposin linked to the C-terminus of prosaposin and to secretory albumin, drives the chimeric protein to lysosomes in COS-7 cells. Quantitative image analysis demonstrated that saposins are targeted with different efficiency (P<0.05) and in a less smooth manner than the precursor. Despite a very close homology, the charge distribution at the surface of 3D comparative models between saposins appeared different. Western blotting monitored prosaposin in cells also as a di- or trimeric form, whereas the chimeric saposins as monomeric. This implies that each amphipathic saposin-like motif may be a part of the overall structural requirements for binding of the precursor to the membrane lipids of transport vesicle. The crystal structure of saposin B demonstrating two dimeric units for lipid binding supports current findings.