Unusual composition of peptidoglycan in Bordetella pertussis

The composition of the peptidoglycan of Bordetella pertussis and the nature of its turnover products was determined by a new combination of analytical techniques: high performance liquid chromatography of an enzymatic peptidoglycan hydrolysate and fast atom bombardment mass spectrometry and fast atom bombardment collision-activated dissociation tandem mass spectrometry. Sixteen major components of the peptidoglycan were purified, and assignment of complete or partial chemical structures was achieved for nine and seven species, respectively. At this level of resolution, a previously unrecognized heterogeneity of monomeric (five new species; nine total) and dimeric species (five new species; five total) was detected. No species containing diaminopimelyl-diaminopimelic acid cross-links or lysyl-arginine substitutions were found. Previous estimates of total cross-linkage and average chain length were revised downward to 32% and 21 disaccharide residues, respectively. Detection of a chemically novel species, a disaccharide octapeptide monomer, in both the peptidoglycan hydrolysate and culture supernatant fluid, suggests that an N-acetyl-muramyl-L-alanine amidase acts on the intact peptidoglycan of Bordetella and participates in cell wall turnover. Five peptidoglycan turnover products were identified in the supernatant fluid of late logarithmic phase cultures, including the 1,6-anhydro monomeric species known as tracheal cytotoxin. Peptidoglycan turnover was detected at a low rate of approximately 10%/generation, a value sufficient to account for the generation of all tracheal cytotoxin found in culture supernatant fluids.     

Further studies on the rabbit erythroid cell plasma membrane transferrin receptor

Studies reproted here indicate that a major cell surface glycoprotein of the rabbit bone marrow erythroid cell may be involved in binding transferrin. The glycoprotein has an apparent molecular weight of 18,000. It is suggested that bone marrow erythroid cells may provide an invaluable source of the red cell membrane transferrin receptor for future studies.     

Benzimidazole-based DGAT1 inhibitors with a [3.1.0] bicyclohexane carboxylic acid moiety

Previously disclosed benzimidazole-based DGAT1 inhibitors containing a cyclohexane carboxylic acid moiety suffer from isomerization at the alpha position of the carboxylic acid group, generating active metabolites which exhibit DGAT1 inhibition comparable to the corresponding parent compounds. In this report, we describe the design, synthesis and profiling of benzimidazole-based DGAT1 inhibitors with a [3.1.0] bicyclohexane carboxylic acid moiety. Our results show that single isomer 3A maintains in vitro and in vivo inhibition against DGAT1. In contrast to previous lead compounds, 3A does not undergo isomerization during in vitro hepatocyte incubation study or in vivo mouse study.     

Meteorological factors and ambient bacterial levels in a subtropical urban environment

We conducted a study to investigate the characteristics and determinants of ambient bacteria in Taipei, Taiwan from August 2004 to March 2005. We monitored ambient culturable bacteria in Shin-Jhuang City, an urban area in the Taipei metropolitan areas, using duplicate Burkard Portable Air Samplers with R2A agar. The average concentration of total bacteria was 1,986 colony-forming units per cubic meter of air (CFU/m³) (median?=?780 CFU/m³) over the study period, with the highest level in autumn. Most bacterial taxa had similar seasonal variation, with higher concentrations in autumn and winter. During the study period, Gram negative rods and cocci were predominant. Multivariate analyses indicated that wind speed and wind direction significantly influenced ambient bacterial distribution. Temperature and relative humidity were also important environmental factors positively associated with ambient bacterial concentrations. We observed statistically significant relationships between ambient bacteria and air pollutants, including sulfur dioxide (SO₂), ozone, particulate matter (aerodynamic diameter ≤10 μm (PM₁₀)), methane and total hydrocarbons. The concentrations of methane and total hydrocarbons during the previous day were positively associated with total bacteria and Gram negative rods, respectively. Ozone level on the previous day had a negative relationship with Gram negative cocci. SO₂ level with a 3-day lag was positively correlated with concentrations of both total bacteria and Gram negative cocci. In the future, more longitudinal studies are needed to confirm the relationships and possible mechanisms between ambient bacteria and meteorological factors, as well as to evaluate the ecological and health impacts of ambient bacteria.     

Blocking HIV-1 entry by a gp120 surface binding inhibitor

We report the mode of action of a proteomimetic compound that binds to the exterior surface of gp120 and blocks HIV-1 entry into cells. Using a one cycle time-of-addition study and antibody competition binding studies, we have determined that the compound blocks HIV-1 entry through modulation of key protein-protein interactions mediated by gp120. The compound exhibits anti-HIV-1 replication activities against several pseudotype viruses derived from primary isolates and the resistant strains isolated from existing drug candidates with equal potency. Together, these data provide evidence that the proteomimetic compound represents a novel class of HIV-1 viral entry inhibitor that functions through protein surface recognition in analogy to an antibody.     

Evidence for competition between Ixodes scapularis and Dermacentor albipictus feeding concurrently on white-tailed deer

Competition among ticks, and among ectoparasites generally, has rarely been demonstrated. Ixodes scapularis and Dermacentor albipictus are both hard ticks commonly found feeding on deer harvested at Letterkenny Army Depot, in south central Pennsylvania, USA. The two species have contrasting life histories resulting in D. albipictus spending notably more time on the shared host. We hypothesized that this would give D. albipictus an advantage in locating and occupying optimal attachment sites (highly vascularized areas like the head and ears). Ticks were collected from 224 hunter-killed deer in December 2005 and November 2006 to determine if there is evidence of competition for attachment sites when these two species concurrently infest deer. A timed sample (3?min per region) of representative ticks was collected from the head (ears, face and neck regions) and body (axillae regions). Ixodes scapularis was more abundant and prevalent overall than D. albipictus. Dermacentor albipictus was found almost exclusively on the head, whereas I. scapularis was more evenly distributed, but somewhat more abundant on the body than on the head. The proportion of I. scapularis on the head was reduced at high D. albipictus abundances, but I. scapularis abundance did not alter the distribution of D. albipictus. This study supports the hypothesis of competition for preferred attachment sites between these two species of ticks, and suggests that D. albipictus may be competitively dominant over I. scapularis on the head region of concurrently infested white-tailed deer.     

Structural analysis of a 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase with an N-terminal chorismate mutase-like regulatory domain

3-Deoxy-D-arabino-heptulosonate 7-phosphate synthase (DAHPS) catalyzes the first step in the biosynthesis of a number of aromatic metabolites. Likely because this reaction is situated at a pivotal biosynthetic gateway, several DAHPS classes distinguished by distinct mechanisms of allosteric regulation have independently evolved. One class of DAHPSs contains a regulatory domain with sequence homology to chorismate mutase-an enzyme further downstream of DAHPS that catalyzes the first committed step in tyrosine/phenylalanine biosynthesis-and is inhibited by chorismate mutase substrate (chorismate) and product (prephenate). Described in this work, structures of the Listeria monocytogenes chorismate/prephenate regulated DAHPS in complex with Mn(2+) and Mn(2+) + phosphoenolpyruvate reveal an unusual quaternary architecture: DAHPS domains assemble as a tetramer, from either side of which chorismate mutase-like (CML) regulatory domains asymmetrically emerge to form a pair of dimers. This domain organization suggests that chorismate/prephenate binding promotes a stable interaction between the discrete regulatory and catalytic domains and supports a mechanism of allosteric inhibition similar to tyrosine/phenylalanine control of a related DAHPS class. We argue that the structural similarity of chorismate mutase enzyme and CML regulatory domain provides a unique opportunity for the design of a multitarget antibacterial.