Identification of the epitope for a highly cross-reactive monoclonal antibody on the major sigma factor of bacterial RNA polymerase

A highly cross-reactive monoclonal antibody (MAb), 2G10, was found to react in a conserved region of Escherichia coli RNA polymerase sigma70. The epitope was localized to amino acids 470 to 486, which included part of conserved region 3.1. The epitope for MAb 3D3, a MAb which maps close to the 2G10 epitope, was also determined.     

The de novo design and synthesis of cyclic urea inhibitors of factor Xa: initial SAR studies

In this report we discuss the design, synthesis, and validation of a novel series of cyclic urea inhibitors of the blood coagulation protein Factor Xa. This work culminates in compound 11, a monoamidine inhibitor of fXa employing a new S4 ligand that reduces the cationic character of these analogs. Compound 11 represents a lead for a series of more potent and selective inhibitors.     

Structural organization of virulence-associated plasmids of Yersinia pestis

The complete nucleotide sequence and gene organization of the three virulence plasmids from Yersinia pestis KIM5 were determined. Plasmid pPCP1 (9,610 bp) has a GC content of 45.3% and encodes two previously known virulence factors, an associated protein, and a single copy of IS100. Plasmid pCD1 (70,504 bp) has a GC content of 44.8%. It is known to encode a number of essential virulence determinants, regulatory functions, and a multiprotein secretory system comprising the low-calcium response stimulation that is shared with the other two Yersinia species pathogenic for humans (Y. pseudotuberculosis and Y. enterocolitica). A new pseudogene, which occurs as an intact gene in the Y. enterocolitica and Y. pseudotuberculosis-derived analogues, was found in pCD1. It corresponds to that encoding the lipoprotein YlpA. Several intact and partial insertion sequences and/or transposons were also found in pCD1, as well as six putative structural genes with high homology to proteins of unknown function in other yersiniae. The sequences of the genes involved in the replication of pCD1 are highly homologous to those of the cognate plasmids in Y. pseudotuberculosis and Y. enterocolitica, but their localization within the plasmid differs markedly from those of the latter. Plasmid pMT1 (100,984 bp) has a GC content of 50.2%. It possesses two copies of IS100, which are located 25 kb apart and in opposite orientations. Adjacent to one of these IS100 inserts is a partial copy of IS285. A single copy of an IS200-like element (recently named IS1541) was also located in pMT1. In addition to 5 previously described genes, such as murine toxin, capsule antigen, capsule anchoring protein, etc., 30 homologues to genes of several bacterial species were found in this plasmid, and another 44 open reading frames without homology to any known or hypothetical protein in the databases were predicted.     

Lack of effect of RPE65 removal on the enzymatic processing of all-trans-retinol into 11-cis-retinol in vitro

RPE65 is a major membrane associated protein found in the vertebrate retinal pigment epithelium (RPE). Various studies have shown this protein to be essential for visual function, possibly at the level of the processing of retinoids. The pigment epithelium is the anatomical site in which the visual chromophore 11-cis retinal is generated. The two critical RPE enzymes in the isomerization pathway are lecithin retinol acyl transferase (LRAT) and isomerohydrolase, which processes all-trans-retinyl esters into 11-cis-retinol. Both enzymes are membrane bound. It is shown here that RPE65 can be largely extracted (90-95%) from RPE membranes by 1 M KCl by itself, or with added detergent CHAPS. The almost quantitative extraction of RPE65 from RPE membranes has little or no effect on in vitro LRAT and isomerohydrolase activities in quantitative enzymatic assays using RPE membranes, suggesting that RPE65 may not have an important role to play in the enzymatic processing of all-trans-retinol into 11-cis-retinol in vitro.     

Bilateral malignant carotid chemodectoma

A clinical case of carotid chemodectoma is reported. Unique character of this case consists in combination of rare manifestations of this disease in a patient (malignancy, bilateral location). Summarizing their own experience and analysis of literature data the authors give recommendations for diagnosis and management of such patients.     

Detection of genetically modified soy in processed foods sold commercially in Malaysia by PCR-based method

Regulations for the use and labeling of genetically modified organism (GMO) products and derived ingredients are being implemented worldwide, that demands reliable and accurate methods to detect GMO in raw materials and food products. In this study, polymerase chain reaction (PCR) method was established for monitoring products derived from GMO that are sold in the markets in Malaysia, which specifically amplify the 35S promoter, nos (nopaline synthase-terminator), EPSPS (5-enolpyruvylshikimate-3-phosphate synthase) and RRS (CTP/CP4EPSPS). Using this method, we investigated the incidence of genetically modified soy (GM-soy) and specifically the presence of roundup ready soy (RRS). All the soybean samples were evidenced by presence of the lectin gene. Out of 85 samples examined, the 18 positive GM samples were raw bean (9), tofu (8) and tempe (1) (a traditional Malay food). The results demonstrate for the first time the presence of GM-soy in Malaysian food products, reinforcing the need for the development of accurate quantitative methods for routine analyses.