Novel conantokins from Conus parius venom are specific antagonists of N-methyl-D-aspartate receptors

We report the discovery and characterization of three conantokin peptides from the venom of Conus parius. Each peptide (conantokin-Pr1, -Pr2, and -Pr3) contains 19 amino acids with three gamma-carboxyglutamate (Gla) residues, a post-translationally modified amino acid characteristic of conantokins. The new peptides contain several amino acid residues that differ from previous conantokin consensus sequences. Notably, the new conantokins lack Gla at the 3rd position from the N terminus, where the Gla residue is replaced by either aspartate or by another post-translationally modified residue, 4-trans-hydroxyproline. Conantokin-Pr3 is the first conantokin peptide to have three different post-translational modifications. Conantokins-Pr1 and -Pr2 adopt alpha-helical conformations in the presence of divalent cations (Mg2+ and Ca2+) but are generally unstructured in the absence of divalent cations. Conantokin-Pr3 adopts an alpha-helical conformation even in the absence of divalent cations. Like other conantokins, the new peptides induced sleep in young mice and hyperactivity in older mice upon intracranial injection. Electrophysiological assays confirmed that conantokins-Pr1, -Pr2, and -Pr3 are N-methyl-d-aspartate (NMDA) receptor antagonists, with highest potency for NR2B-containing NMDA receptors. Conantokin-Pr3 demonstrated approximately 10-fold selectivity for NR2B-containing NMDA receptors. However, conantokin-Pr2 showed minimal differences in potency between NR2B and NR2D. Conantokins-Pr1, -Pr2, and -Pr3 all demonstrated high specificity of block for NMDA receptors, when tested against various ligand-gated ion channels. Conus parius conantokins allow for a better definition of structural and functional features of conantokins as ligands targeting NMDA receptors.     

Similarity of the CDC and WHO weight-for-length growth charts in predicting risk of obesity at age 5 years

The World Health Organization (WHO) 2006 weight-for-length (WFL) or BMI growth charts are now recommended as the new standard for children under 24 months. The objective of this study was to examine associations of ever being overweight during 1-24 months, based on the older Centers for Disease Control and Prevention (CDC) and WHO cutpoints, with risk of obesity at age 5 years. From well-child visits to a Massachusetts multi-site group practice during 1980-2008, we studied 15,488 children with length/height and weight measurements at 1, 6, 12, 18, and 24 months, and at 5 years. The main exposures were ever being overweight during 1-24 months using each of three cutpoints: CDC WFL ≥ 95th percentile, WHO WFL or BMI ≥ 97.7th percentile. The main outcome was obesity at 5 years (CDC BMI ≥ 95th percentile). We calculated multivariable odds ratios (ORs), adjusted for age, sex, race/ethnicity, and year. At 5 years, 10.8% of participants were obese. During 1-24 months, 21.3, 18.3, and 20.2% were ever overweight using CDC WFL, WHO WFL, and WHO BMI cutpoints, respectively. ORs (95% confidence interval (CI)) for associations of ever being overweight during 1-24 months with obesity at 5 years were 6.0 (5.4, 6.6), 6.3 (5.7, 7.0), and 6.0 (5.4, 6.7), respectively. Ever being overweight in the first 2 years of life is a strong predictor of obesity at 5 years. CDC WFL, WHO WFL, and WHO BMI cutpoints for overweight in early childhood provided similar estimates of later obesity risk.     

Taxonomy and phylogeny of new wood- and soil-inhabiting Sporothrix species in the Ophiostoma stenoceras-Sporothrix schenckii complex

Sporothrix, one of the anamorph genera of Ophiostoma, includes the important human pathogen S. schenckii and various fungi associated with insects and sap stain of wood. A survey of fungi from wood utility poles in South Africa yielded two distinct groups of Sporothrix isolates from different geographical areas. DNA sequence and morphological data derived in this study showed that isolates in these groups represent two novel species in the S. schenckii-O. stenoceras species complex. A new species isolated from pine poles and rosebush wood and phylogenetically closely related to S. pallida is described here as Sporothrix stylites. Phylogenetic analyses also confirmed the synonymy of S. albicans and S. nivea with S. pallida. Sporothrix stylites and S. pallida also are related closely to the isolates from soil, previously treated as "environmental" isolates of S. schenckii. Soil isolates are clearly distinct from human isolates of S. schenckii. We describe the former here as Sporothrix humicola. The isolates from eucalypt poles group peripheral to most other species in the S. schenckii-O. stenoceras complex and are newly described as Sporothrix lignivora. Phylogenetic analyses of sequences of isolates from soil and wood together with those of clinical isolates showed that the human-pathogenic strains form an aggregate of several cryptic species.     

Trypanocidal tetrahydrofuran lignans from Peperomia blanda

Five tetrahydrofuran lignans and two known flavones were isolated from the aerial parts of Peperomia blanda. The structures of the isolated lignans were elucidated by interpretation of their spectroscopic data, including by gHMQC and gHMBC. The relative and absolute configurations of the isolates were determined from NOESY interactions and optical properties, respectively. Four of the lignans were diastereomeric whilst one was of mixed biosynthetic origin. All but one of the lignans exhibited high in vitro trypanocidal activity when assayed against epimastigotes of Trypanosoma cruzi strain Y.     

Developmental origins of childhood overweight: potential public health impact

Several modifiable pre- and postnatal determinants of childhood overweight are known, but no one has examined how they influence risk of overweight in combination. We estimated the risk of overweight at age 3 years according to levels of maternal smoking during pregnancy, gestational weight gain, breastfeeding duration, and infant sleep duration. We studied 1,110 mother-child pairs in Project Viva, a prospective prebirth cohort study. The main outcome measure was child overweight (BMI for age and sex >or=95th percentile) at age 3. We ran logistic regression models with all four modifiable risk factors as well as the covariates maternal BMI and education, child race/ethnicity, and household income. From the model, we obtained the estimated probability of overweight for each of the 16 combinations of the four risk factors. During pregnancy, 9.8% of mothers smoked and 50% gained excessive weight. In infancy, 73% mother-child pairs breastfed for <12 m, and 31% of infants slept <12 h/day. Among the 3-year-old children in the cohort, 9.5% were overweight. In the prediction model, the estimated probability of overweight ranged from 0.06 among children exposed to favorable levels of all four risk factors, to 0.29 with adverse levels of all four. Healthful levels of four behaviors during early development predicted much lower probability of overweight at age 3 than adverse levels. Interventions to modify several factors during pregnancy and infancy could have substantial impact on prevention of childhood overweight.     

Drought timing differentially affects above- and belowground productivity in a mesic grassland

Climate models forecast an intensification of the global hydrological cycle with droughts becoming more frequent and severe, and shifting to times when they have been historically uncommon. Droughts, or prolonged periods of precipitation deficiency, are characteristic of most temperate grasslands, yet few experiments have explored how variation in the seasonal timing of drought may impact ecosystem function. We investigated the response of above- and belowground net primary production (ANPP & BNPP) to altered drought timing in a mesic grassland in NE Kansas. Moderate drought treatments (25% reduction from the mean growing season precipitation [GSP]) were imposed by erecting rainout shelters in late spring (LSP), early summer (ESM), and mid-summer (MSM, n = 10 plots/treatment). These treatments were compared to two controls (long-term average GSP [LTA] and ambient GSP [AMB]) and a wet treatment (+30% of the long-term average GSP [WET]). We found that LSP drought did not significantly reduce ANPP relative to control plots while the ESM and MSM drought did despite equivalent reductions in soil moisture. In contrast, the WET treatment did not affect ANPP. Neither the WET nor the drought treatments altered BNPP as compared to the controls. Our results suggest that forecasts of ecosystem responses to climate change will be improved if both the seasonal timing of alterations in precipitation as well as differential responses of above- and belowground productivity to drought are incorporated into models.     

Discovery of renin inhibitors containing a simple aspartate binding moiety that imparts reduced P450 inhibition

Discovery of potent renin inhibitors which contain a simplified alkylamino Asp-binding group and exhibit improved selectivity for renin over Cyp3A4 is described. Structure-function results in this series are rationalized based on analysis of selected compounds bound to renin, and the contribution of each molecular feature leading to the reduced P450 inhibition is quantified.     

Selective inhibition of uracil tRNA methylases of E. coli by ethionine.

L-ethionine has been found to inhibit uracil tRNA methylating enzymes in vitro under conditions where methylation of other tRNA bases is unaffected. No selective inhibitor for uracil tRNA methylases has been identified previously. 15 mM L-ethionine or 30 mM D,L-ethionine caused about 40% inhibition of tRNA methylation catalyzed by enzyme extracts from E. coli B or E. coli M3S (mixtures of methylases for uracil, guanine, cytosine, and adenine) but did not inhibit the activity of preparations from an E. coli mutant that lacks uracil tRNA methylase. Analysis of the 14CH3 bases in methyl-deficient E. coli tRNA after its in vitro methylation with E. coli B3 enzymes in the presence or absence of ethionine showed that ethionine inhibited 14CH3 transfer to uracil in tRNA, but did not diminish significantly the 14CH3 transfer to other tRNA bases. Under similar conditions 0.6 mM S-adenosylethionine and 0.2 mM ethylthioadenosine inhibited the overall tRNA base methylating activity of E. coli B preparations about 50% but neither of these ethionine metabolites preferentially inhibited uracil methylation. Ethionine was not competitive with S-adenosyl methionine. Uracil methylation was not inhibited by alanine, valine, or ethionine sulfoxide. It is suggested that the thymine deficiency that we found earlier in tRNA from ethionine-treated E. coli B cells, resulted from base specific inhibition by the amino acid, ethionine, of uracil tRNA methylation in vivo.     

Immunization with modified vaccinia virus Ankara-based recombinant vaccine against severe acute respiratory syndrome is associated with enhanced hepatitis in ferrets

Severe acute respiratory syndrome (SARS) caused by a newly identified coronavirus (SARS-CoV) is a serious emerging human infectious disease. In this report, we immunized ferrets (Mustela putorius furo) with recombinant modified vaccinia virus Ankara (rMVA) expressing the SARS-CoV spike (S) protein. Immunized ferrets developed a more rapid and vigorous neutralizing antibody response than control animals after challenge with SARS-CoV; however, they also exhibited strong inflammatory responses in liver tissue. Inflammation in control animals exposed to SARS-CoV was relatively mild. Thus, our data suggest that vaccination with rMVA expressing SARS-CoV S protein is associated with enhanced hepatitis.