Effect of Synbiotic and Probiotic Supplementation on Serum Levels of Endothelial Cell Adhesion Molecules in Hemodialysis Patients: a Randomized Control Study

Probiotics and Antimicrobial Proteins - The aim of this study was to investigate the effect of synbiotic and probiotic supplementation on serum vascular dysfunction and necrosis markers in...     

Design and Synthesis of Heterocyclic Conjugated Peptides as Novel Antimicrobial Agents

Antimicrobial peptides have been recognized as a novel class of antibiotics and several candidates are currently in clinical trials. In the present study, new antimicrobial compounds were synthesized by coupling quinazolinone moiety with the fragments of elastin sequences VP, GVP, VGVP and GVGVP. They were evaluated for their antibacterial activity against both gram positive and gram negative bacterial strains. We are here reporting that heterocyclic conjugated tetra peptide and penta peptide showed enhanced antibacterial activity compare to the conventional antimicrobial drugs.     

Proline accumulation and the expression of Δ1-pyrroline-5-carboxylate synthetase in two safflower cultivars

Proline (Pro) accumulation under water stress was measured in safflower (Carthamus tinctorius L.) drought tolerant cv. A1 and sensitive cv. Nira. Activities of pyrroline-5-carboxylate reductase (P5C reductase) and pyrroline-5-carboxylate synthetase (P5C synthetase), two enzymes involved in the Pro biosynthetic pathway were also estimated. Water stress resulted in a reduction in the leaf dry mass and chlorophyll content along with a gradual accumulation of Pro. RT-PCR results show higher expression of Δ¹-pyrroline-5-carboxylate synthetase (p5cs) gene in correlation with up-regulated Pro accumulation in cv. A1. P5C reductase was found to be the Pro synthesis rate limiting whereas P5C synthetase did not show any specific response to the drought stress in both cultivars.     

USP32 is an active, membrane-bound ubiquitin protease overexpressed in breast cancers

USP32, on chromosomal band 17q23.1-17q23.2, is a highly conserved but uncharacterized gene that gave rise during evolution to a well-known hominoid-specific proto-oncogene, USP6. We investigated the expression profile of USP32 in human tissues and examined its functions to gain insight into this novel member of the well-conserved ubiquitination system. We detected ubiquitous USP32 expression across tissues and confirmed the predicted deubiquitination function owing to the presence of conserved peptidase signature aspargine, cysteine, histidine, and aspartic acid domains of ubiquitin-specific proteases. A Golgi localization of GFP-fused USP32 was detected by fluorescent protection assay and BODIPY-TR staining. In addition, stable silencing of USP32 caused a significant decrease in the proliferation and migration rate of cells. Based on these and the fact that USP32 maps to 17q23, which is commonly amplified in breast cancers, we analyzed USP32 expression in breast cancer cells. We detected high expression of USP32 in 50% (9 of 18) of breast cancer cell lines and 22% (9 of 41) of primary breast tumors compared to mammary epithelial cells. In summary, we report the preliminary characterization of this novel deubiquitinating enzyme on 17q23 and demonstrate its functional role in the ubiquitin system and its potential involvement in tumorigenesis.     

Glycine transport in mouse eggs and preimplantation conceptuses

At least two Na+-dependent systems for glycine transport became detectable, while another became undetectable during preimplantation development of mouse conceptuses. Glycine was taken up by a process in eggs and cleavage-stage conceptuses which closely resembles system Gly. Mediated transport at these stages was more rapid at higher Cl- concentrations, sigmoidally related to the exogenous Na+ concentration, and strongly inhibited by sarcosine but not by amino acids with larger side chains. Moreover, neither Li+ nor choline could substitute for Na+ in stimulating glycine transport. System Gly was the only mediated process detected for glycine uptake in unfertilized and fertilized eggs and two-cell conceptuses, but two, less conspicuous, sarcosine-resistant, Na+-dependent components of transport also appeared to be present in eight-cell conceptuses. One of the latter components seemed to remain relatively inconspicuous when conceptuses formed blastocysts, while system Gly became undetectable. In contrast, the other less conspicuous component in eight-cell conceptuses appeared to become the most conspicuous transport process in blastocysts. The latter process, previously designated system B0,+, was shown here also to interact strongly with a broad scope of zwitterionic and cationic amino acid structures. Moreover, transport of glycine via system B0,+ was more rapid at higher Cl- concentrations, and this Na+-dependent process as well as Na+-independent leucine uptake were inhibited by choline. Furthermore, Na+-dependent amino acid transport in two-cell conceptuses and blastocysts was inhibited by 1.0 or 10 mM ouabain, but the inhibition was incomplete at both concentrations. Since Na+/K+-ATPase has not been detected in two-cell conceptuses, inhibition of amino acid transport by ouabain may not have been due solely to an effect on this enzyme. The level of system Gly activity decreased during the development of eight-cell conceptuses from eggs, and this decrease could contribute to an associated decline in intracellular glycine. Since other amino acids begin to compete strongly with glycine for transport when system B0,+ replaces system Gly in conceptuses, this qualitative change in transport activity may help account for a further decrease in the glycine content of conceptuses, reported elsewhere to occur after they form blastocysts.     

Repression of cap-dependent translation by 4E-binding protein 1: competition with p220 for binding to eukaryotic initiation factor-4E.

An important aspect of the regulation of gene expression is the modulation of translation rates in response to growth factors, hormones and mitogens. Most of this control is at the level of translation initiation. Recent studies have implicated the MAP kinase pathway in the regulation of translation by insulin and growth factors. MAP kinase phosphorylates a repressor of translation initiation [4E-binding protein (BP) 1] that binds to the mRNA 5' cap binding protein eukaryotic initiation factor (eIF)-4E and inhibits cap-dependent translation. Phosphorylation of the repressor decreases its affinity for eIF-4E, and thus relieves translational inhibition. eIF-4E forms a complex with two other polypeptides, eIF-4A and p220, that promote 40S ribosome binding to mRNA. Here, we have studied the mechanism by which 4E-BP1 inhibits translation. We show that 4E-BP1 inhibits 48S pre-initiation complex formation. Furthermore, we demonstrate that 4E-BP1 competes with p220 for binding to eIF-4E. Mutants of 4E-BP1 that are deficient in their binding to eIF-4E do not inhibit the interaction between p220 and eIF-4E, and do not repress translation. Thus, translational control by growth factors, insulin and mitogens is affected by changes in the relative affinities of 4E-BP1 and p220 for eIF-4E.     

A quantitative genetic analysis of tissue-specific catalase activity inMus musculus

Tissue-specific catalase activity in 3-week-old animals from inbred mouse strains 129/ReJ, BALB/c, C3H/HeAnl/Cas-1^b, C3H/HeSnJ, C3H/S, C57BL/6J, and Swiss-Webster was found to be highly variable by analysis of variance (P=0.01). Appropriate crosses were made among strains which were classified as normal (BALB/c, C3H/HeSnJ, C3H/S), hypocatalasemic (129/ReJ, C57BL/6J), and acatalasemic (C3H/HeAnl/Cas-1^b) with respect to blood catalase activity to study the inheritance of the blood, kidney, liver, and lung catalase activity levels in a number of generations (reciprocal F₁'s, F₂, two backcrosses —BC₁ and BC₂— and some RI lines). Segregation analysis and statistical methods which tested different models of inheritance as well as calculations of heritability were used in an effort to assess and evaluate genetic parameters that affect catalase activity. Results indicate that the inheritance of blood catalase activity in the cross involving acatalasemic and normal (BALB/c, C3H/HeSnJ) strains is compatible with the single-locus difference between the parental strains; however, the difference between the acatalasemic and the hypocatalasemic strain (C57BL/6J) would require additional genetic interaction for a satisfactory explanation. A similar pattern of generalization also applies to the inheritance of kidney catalase activity. The segregation pattern for the liver and lung catalase activity in most crosses is significantly different from the expectations of the single locus model. These results are compatible with the concept that a number of genes must affect tissue-specific catalase activity in mice. These may include previously described (e.g., Ce-1 and Ce-2) or novel genetic regulators/modifiers which interact with a single structural gene (Cas-1) or its product to produce the catalase phenotype characteristic of specific tissues in each strain.This investigation was supported by a Natural Sciences and Engineering Research Council of Canada operating grant to S.M.S.     

Regeneration of pigeonpea (Cajanus cajan) from cotyledonary node via multiple shoot formation

Summary Plant regeneration, which is the major limiting factor for transformation of Cajanus cajan, has been obtained via multiple shoot formation from the cotyledonary node region of seedlings germinated on MS medium containing 2 mgl^–1 6-benzylaminopurine. A mass of multiple shoot-initials formed at the axillary bud region of the cotyledonary node of the seedlings within two weeks. The cotyledonary node along with the mass of shoot-initials excised from the seedling, continued to form new shoot-initials on MS medium containing 6-benzylaminopurine (2 mgl^–1) and supplemented topically with indole-3-acetic acid. The formation of new shoot-initials was also observed from the cotyledonary nodal explant, after cutting off its surface layers to completely remove the pre-existing shoot-initials and culturing it on 6-benzylaminopurine (2 mgl^–1) containing medium. The shoots elongated rapidly on basal MS medium and rooted efficiently in MS medium supplemented with indole-3-butyric acid (0.5 mgl^–1). The procedure described is efficient, and highly reproducible and a common response was observed for all the six varieties tested.Abbreviations BAP 6-benzylaminopurine - IAA indole-3-acetic acid - IBA indole-3-butyric acid - NAA -naphthaleneacetic acid - MS medium, Murashige and Skoog (1962) medium - CN cotyledonary node     

Non‐enzymatic aqueous peroxyoxalate chemiluminescence immune detection using a CCD camera and a CMOS device

A new method for non‐enzymatic aqueous peroxyoxalate chemiluminescence (POCL) biomolecular detection using imaging chip‐based devices has been developed. A water‐soluble amide of oxalic acid was synthesized and used in the investigation and characterization of POCL immunodetection in an aqueous environment. Six fluorescent dyes commonly used in biological detection were tested, and the intensity of light generated from the aqueous POCL reactions was characterized in the liquid phase. Direct detection sensitivity comparisons between a standard fluorescent method and this POCL method were performed in both liquid and solid phases. Results showed that detection sensitivity using the POCL method is comparable to that of the fluorescent method. POCL biomolecular detection on a nitrocellulose membrane was also investigated using a charge‐coupled device (CCD) camera. Again, POCL detection sensitivity proved to be comparable to that using the fluorescent detection method. In an application of aqueous POCL biomolecular detection, Staphylococcus aureus enterotoxin B (SEB) and its antibody were used to demonstrate immuno‐ and affinity detection. For further applications, such as DNA and protein arrays, simultaneous detection of biomolecules labelled with different fluorescent labels was investigated, using a complementary metal oxide semiconductor (CMOS) colour imaging chip. Copyright © 2008 John Wiley & Sons, Ltd.     

Bag culture: a method for root-root co-culture

A method named "bag culture" was developed for coculturing of Linum persicum (section Syllinum) and L. austriacum (section Linum) hairy roots. For this propose L. austriacum and L. persicum hairy root cultures were established using Agrobacterium rhizogenes in McCown medium. L. persicum hairy roots in bags (1 mm2 mesh) were successfully grown together with L. austriacum hairy roots. The amounts of podophyllotoxin (PTOX) and 6-methoxypodophyllotoxin (MPTOX) produced by L. persicum hairy root cultures were detected using HPLC. The results indicated that the amounts of both lignans and growth indexes of the two hairy roots decreased, that may be partly due to a competition between the two types of culture in using precursors of biosynthetic metabolites and the amount of culture medium which is available for each hairy root. However, MPTOX (0.17 g/100 g DW) and PTOX (0.02 g/100 g DW) levels of the L. persicum single culture in bag were significantly higher than of the other cultures which may be due to the immobilization effect of the bag.