Cytomegalovirus infection via mother's milk: could distinct virus strains determine different disease patterns in preterm twins?

We describe a case of cytomegalovirus infection via the mother's milk in preterm twins who showed a different disease pattern. Molecular analyses to genotype the viral genome disclosed a mixed population of virus strains in the milk. These isolates were differentially transmitted to each of the twins and this may be responsible of the different patterns of disease.     

CTAB–Urea Method Purifies RNA from Melanin for cDNA Microarray Analysis

Melanin represents a major problem for the study of melanoma by microarrays since it is retained during RNA extraction and inhibits the enzymatic reactions used for probe preparation. Here we report a new method for cleaning RNA from melanin, based on the use of the cationic detergent cetyl‐trimethylammonium bromide (CTAB)–urea for RNA precipitation. This method is easy to perform and has a low cost. Purified RNA is recovered with high quality and good yield. CTAB–urea treated RNA from highly pigmented melanoma cells can be successfully reverse transcribed and labeled to obtain probes which can be subsequently used in cDNA microarray experiments, giving consistent and reproducible results.     

Linear basic peptides for targeting interferon-gamma-glycosaminoglycan interactions: synthesis and inhibitory properties.

Glycosaminoglycans (GAGs) play an important role in inflammatory responses due to their ability to interact with cytokines and chemokines, resulting in the localization of these mediators to specific anatomical sites, where they function to direct leukocyte recruitment and activation. Targeting GAG-cytokine/chemokine interactions might may thus have therapeutic applications as anti-inflammatory or immunomodulatory therapy in vivo. Peptides that mimic the heparin-binding domains of cytokines may have a potential use as inhibitors of GAG-cytokine interactions. A linear octapeptide (MC-2) derived from the conserved heparin-binding region of interferon-gamma (IFN-gamma) was synthesized along with four analogs featuring a substitution of Phe for Leu in position 1 and varying number of positive charges on the octapeptide molecule. The relative abilities of the synthesized peptides to inhibit the interactions between IFN-gamma and GAGs were compared. From the results, it follows that the inhibitory potency of the octapeptide analogs was related to the number of positive charges in the molecule, while increased hydrophobicity had no significant effect.     

Novel methylene-linked heterocyclic EP1 receptor antagonists

We describe the SAR, in terms of heterocyclic replacements, for a series of pyrazole EP(1) receptor antagonists. This study led to the identification of several aromatic heterocyclic replacements for the pyrazole in the original compound. Investigation of replacements for the methylene linker uncovered disparate SAR in the thiazole and pyridine series.     

Structure-activity study of 2,3-benzodiazepin-4-ones noncompetitive AMPAR antagonists: identification of the 1-(4-amino-3-methylphenyl)-3,5-dihydro-7,8-ethylenedioxy-4H-2,3-benzodiazepin-4-one as neuroprotective agent

In the search for AMPA receptor (AMPAR) antagonists, 2,3-benzodiazepines represent a family of specific noncompetitive antagonists with anticonvulsant and neuroprotective properties. We have previously shown that 2,3-benzodiazepin-4-ones possess marked anticonvulsant properties and high affinity for the noncompetitive binding site of the AMPAR complex. In this paper, we report the synthesis and pharmacological characterization of a full set of 2,3-benzodiazepin-4-ones in order to better define the structure-activity relationship (SAR) of this class of compounds. Binding assays and functional tests were performed to evaluate the antagonistic activity at the AMPARs. Through these results we have identified a potent AMPAR antagonist, 1-(4-amino-3-methylphenyl)-3,5-dihydro-7,8-ethylenedioxy-4H-2,3-benzodiazepin-4-one (5c). This compound noncompetitively inhibited AMPAR-mediated toxicity in primary mouse hippocampal cultures with an IC(50) of 1.6muM and blocked kainate-induced calcium influx in rat cerebellar granule cells with an IC(50) of 6.4muM. Thus, 5c has the in vitro potential as therapeutic drug in the treatment of various neurological disorders.     

Selection and characterization of a promoter for expression of single-copy recombinant genes in Gram-positive bacteria

Background  

In the past ten years there has been a growing interest in engineering Gram-positive bacteria for biotechnological applications, including vaccine delivery and production of recombinant proteins. Usually, bacteria are manipulated using plasmid expression vectors. The major limitation of this approach is due to the fact that recombinant plasmids are often lost from the bacterial culture upon removal of antibiotic selection. We have developed a genetic system based on suicide vectors on conjugative transposons allowing stable integration of recombinant DNA into the chromosome of transformable and non-transformable Gram-positive bacteria.