Identification and structural basis of binding to host lung glycogen by streptococcal virulence factors

The ability of pathogenic bacteria to recognize host glycans is often essential to their virulence. Here we report structure-function studies of previously uncharacterized glycogen-binding modules in the surface-anchored pullulanases from Streptococcus pneumoniae (SpuA) and Streptococcus pyogenes (PulA). Multivalent binding to glycogen leads to a strong interaction with alveolar type II cells in mouse lung tissue. X-ray crystal structures of the binding modules reveal a novel fusion of tandem modules into single, bivalent functional domains. In addition to indicating a structural basis for multivalent attachment, the structure of the SpuA modules in complex with carbohydrate provides insight into the molecular basis for glycogen specificity. This report provides the first evidence that intracellular lung glycogen may be a novel target of pathogenic streptococci and thus provides a rationale for the identification of the streptococcal alpha-glucan-metabolizing machinery as virulence factors.     

Inundation,Hydrodynamics and Vegetation Influence Carbon Dioxide Concentrations in Amazon Floodplain Lakes

Ecosystems - Extensive floodplains and numerous lakes in the Amazon basin are well suited to examine the role of floodable lands within the context of the sources and processing of carbon within...     

Conformational analysis of cyclic hexapeptides designed as constrained ligands for the SH2 domain of the p85 subunit of phosphatidylinositol-3-OH kinase

The structures of the cyclic hexapeptide cyclo(-Gly-Tyr-Val-Pro-Met-Leu-) ( 1 ) and its phosphotyrosyl (pTyr) derivative cyclo[-Gly-Tyr(PO₃H₂)-Val-Pro-Met-Leu-] ( 2 ), designed as constrained models of a sequence that interacts with the src homology 2 (SH2) region of the p85 subunit of phosphatidylinositol-3-OH kinase (PI-3 kinase), were studied in methanol/water solutions by 500 MHz nmr spectroscopy. Compound 1 was found to exist as a 2:1 mixture of isomers about the Val-Pro bond (trans and cis prolyl) between 292–330 K in 75% CD₃O (D,H)/(D,H)₂O solutions. A third species of undetermined structure (ca. 5%) was also observed. Compound 2, a model of phosphorylated peptide ligand that binds to the PI-3 kinase SH2 domain, exhibited similar conformational isomerism. When either compound was dissolved in pure solvent [i.e., 100% CD₃O(H,D) or (H,D)₂O] the ratio of cis to trans isomers was ca 1:1. A battery of one- and two-dimensional nmr experiments at different temperatures and solvent compositions allowed a complete assignment of both the cis and trans forms of 1 and indicated the trans compound to be the major isomer. The spectral properties of the phosphorylated derivative 2 paralleled those of 1 , indicating like conformations for the two compounds. Analysis of rotating frame Overhauser spectroscopy data, coupling constants, amide proton temperature dependence, and amide proton exchange rates generated a set of constraints that were employed in energy minimization and molecular dynamics calculations using the CHARMM force field. The trans isomer exists with the tyrosine and C-terminal Tyr(+3) (Met) residues at opposite corners of the 18-membered ring separated by a distance of 16–18 Å, in contrast with the cis isomer where the side chains of these residues are much closer in space (7–14 Å). It was previously shown that the pTyr and the third amino acid C-terminal to this residue are the critical recognition elements for pTyr-peptide binding to the PI-3 kinase SH2 domain. Such cyclic structures may offer appropriate scaffolding for positioning important amino acid side chains of pTyr-containing peptides as a means of increasing their binding affinities to SH2 domains, and in turn provide a conceptual approach toward the design of SH2 domain directed peptidomimetics. © 1995 John Wiley & Sons, Inc.     

Prenatal exposure to a 50 Hz magnetic field has no effect on spatial learning in adult mice

Male CD1 mice were exposed in utero to a 50 Hz sinusoidal magnetic field at 5 mT (rms) for the period of gestation and were raised subsequently without applied fields. At 82-84 days of age, they began a radial-arm-maze experiment that was designed to test for deficits in spatial learning in memory. Mice exposed in utero and sham-exposed mice exhibited no statistically significant differences in performances. © 1996 Wiley-Liss, Inc.     

Synthesis,biological evaluation,and molecular docking of new series of antitumor and apoptosis inducers designed as VEGFR-2 inhibitors

Based on quinazoline, quinoxaline, and nitrobenzene scaffolds and on pharmacophoric features of VEGFR-2 inhibitors, 17 novel compounds were designed and synthesised. VEGFR-2 IC₅₀ values ranged from 60.00 to 123.85 nM for the new derivatives compared to 54.00 nM for sorafenib. Compounds 15_a, 15_b, and 15_d showed IC₅₀ from 17.39 to 47.10 µM against human cancer cell lines; hepatocellular carcinoma (HepG2), prostate cancer (PC3), and breast cancer (MCF-7). Meanwhile, the first in terms of VEGFR-2 inhibition was compound 15_d which came second with regard to antitumor assay with IC₅₀ = 24.10, 40.90, and 33.40 µM against aforementioned cell lines, respectively. Furthermore, Compound 15_d increased apoptosis rate of HepG2 from 1.20 to 12.46% as it significantly increased levels of Caspase-3, BAX, and P53 from 49.6274, 40.62, and 42.84 to 561.427, 395.04, and 415.027 pg/mL, respectively. Moreover, 15_d showed IC₅₀ of 253 and 381 nM against HER2 and FGFR, respectively.     

Thyroid hormone-regulated enhancer blocking: cooperation of CTCF and thyroid hormone receptor

The highly conserved, ubiquitously expressed, zinc finger protein CTCF is involved in enhancer blocking, a mechanism crucial for shielding genes from illegitimate enhancer effects. Interestingly, CTCF-binding sites are often flanked by thyroid hormone response elements (TREs), as at the chicken lysozyme upstream silencer. Here we identify a similar composite site positioned upstream of the human c-myc gene. For both elements, we demonstrate that thyroid hormone abrogates enhancer blocking. Relief of enhancer blocking occurs even though CTCF remains bound to the lysozyme chromatin. Furthermore, chromatin immunoprecipitation analysis of the lysozyme upstream region revealed that histone H4 is acetylated at the CTCF-binding site. Loss of enhancer blocking by the addition of T3 led to increased histone acetylation, not only at the CTCF site, but also at the enhancer and the promoter. Thus, when TREs are adjacent to CTCF-binding sites, thyroid hormone can regulate enhancer blocking, thereby providing a new property for what was previously thought to be constitutive enhancer shielding by CTCF.     

Personality influences tetanus-specific antibody response in adult male rhesus macaques after removal from natal group and housing relocation

Previous research has suggested that personality is related to immune function in macaques. Using a prospective design, we examined whether variation in the personality dimension "Sociability" in adult male rhesus macaques (Macaca mulatta) was related to the in vivo secondary antibody response to a tetanus toxoid booster immunization following removal from natal groups and relocation to individual housing. We also explored whether the timing of the immunization following relocation had an impact on the immune response. Blood was sampled at the time of booster immunization, at 14 and 28 days post-immunization, and approximately 9 months post-immunization. Plasma was assayed for tetanus-specific IgG by enzyme-linked immunoassay (ELISA). There was no difference between High- and Low-Sociable animals in antibody levels at the time of the booster immunization. Multivariate analysis of variance (MANOVA) revealed that High-Sociable animals had a significantly higher antibody response following relocation and immunization compared to Low-Sociable animals. There was no effect of timing of the immunization on the immune response. The results confirm that personality factors can affect animals' immune responses, and that the dimension Sociability may be influential in a male's response to social separation and relocation.     

The quest for the holy "G" of chromosomal passengers

Twelve years after reporting a novel chromosomal passenger protein, Mollinari et al. in this issue of Developmental Cell finally report its molecular identification, and the results are stunning. TD-60 is a member of the RCC1 family of guanine nucleotide exchange factors implicating small GTPases in chromosome congression and cytokinesis.