Oral vaccination of mice with lipid-encapsulated Mycobacterium bovis BCG: anatomical sites of bacterial replication and immune activity

Lipid microencapsulation of Mycobacterium bovis bacille Calmette-Guérin (BCG) produces an oral delivery vaccine that can establish systemic cell-mediated immune reactivity and protection against aerosol mycobacterial challenge in mice. Here, we describe the lymphatic and mucosal sites of bacterial replication, and location of Mycobacterium-specific IFN-gamma-secreting cell populations, following oral vaccination of BALB/c mice. Eight weeks following a single oral dose of lipid-encapsulated BCG, viable BCG organisms were recovered from the mesenteric lymph nodes (MLN) of 11/12 mice investigated (93%). Live bacteria were also occasionally recovered from the cervical lymph nodes (17%) and Peyer's patches (8%), but not from homogenates of the lungs or spleen. Strong Mycobacterium-specific IFN-gamma production was recorded among isolated splenocytes, but not among populations of mononuclear cells derived from the MLN or lungs. Oral vaccination of mice with lipid-encapsulated BCG thus appears to promote a state of systemic immunological reactivity more akin to that observed following parenteral rather than conventional oral vaccination, despite the fact that replicating bacilli are restricted to lymphatic tissues of the alimentary tract. Possible patterns of lymphocyte sensitization and trafficking are discussed.     

Biochemical and endocrine responses to impact and collision during elite Rugby League match play

The purpose of this study was to investigate the relationship between the prematch and short-term postmatch biochemical and endocrine responses to the intensity, number, and distribution of impacts associated with collisions during elite Rugby League match play. Seventeen elite male Rugby League players each provided blood and saliva samples 24 hours prematch, 30 minutes prematch, 30 minutes postmatch, and then at 24-hour intervals for a period of 5 days postmatch to determine plasma creatine kinase concentration ([CK]) and salivary cortisol concentration ([sCort]). The intensity, number, and distribution of impact forces experienced by players during match play were recorded using portable global positioning systems (GPSs). The change in the dependent variables at each sample collection time was compared to 24 hours prematch and 30-minute prematch measures. The [CK] and [sCort] increased significantly (p < 0.05) during match play. Significant correlations (p < 0.05) were observed between the number of hit-ups and peak [CK] 24 hours postmatch, 48 hours postmatch, and 72 hours postmatch (p < 0.05). The number of impacts recorded in zone 5 (8.1-10.0G) and zone 6 (>10.1G) during match play was significantly correlated (p < 0.05) to [CK] 30 minutes postmatch, 24 hours post, 48 hours post, and 72 hours postmatch. The GPS was able to provide data on the intensity, number, and distribution of impacts resulting from collisions during match play. Elite Rugby League match play resulted in significant skeletal muscle damage and was highly dependent on the number of heavy collisions >8.1G. [CK] remained elevated 120 hours postmatch identifying that at least 5 days modified activity is required to achieve full recovery after elite Rugby League match play.     

Modelling larval movement data from individual bioassays

We consider modelling the movements of larvae using individual bioassays in which data are collected at a high‐frequency rate of five observations per second. The aim is to characterize the behaviour of the larvae when exposed to attractant and repellent compounds. Mixtures of diffusion processes, as well as Hidden Markov models, are proposed as models of larval movement. These models account for directed and localized movements, and successfully distinguish between the behaviour of larvae exposed to attractant and repellent compounds. A simulation study illustrates the advantage of using a Hidden Markov model rather than a simpler mixture model. Practical aspects of model estimation and inference are considered on extensive data collected in a study of novel approaches for the management of cabbage root fly.     

Morphometric analysis and taxonomic revision of the Vriesea paraibica complex (Bromeliaceae)

The species related to Vriesea paraibica (Bromeliaceae, Tillandsioideae) have controversial taxonomic limits. For several decades, this group has been identified in herbarium collections as V. × morreniana, an artificial hybrid that does not grow in natural habitats. The aim of this study was to assess the morphological variation in the V. paraibica complex through morphometric analyses of natural populations. Two sets of analyses were performed: the first involved six natural populations (G1) and the second was carried out on taxa that emerged from the first analysis, but using material from herbarium collections (G2). Univariate ANOVA was used, as well as discriminant analysis of 16 morphometric variables in G1 and 18 in G2. The results of the analyses of the two groups were similar and led to the selection of diagnostic traits of four species. Lengths of the lower and median floral bracts were significant for the separation of red and yellow floral bracts. Vriesea paraibica and V. interrogatoria have red bracts; these two species are differentiated by the widths of the lower and median portions of the inflorescence and by scape length. These structures are larger in the former and smaller in the latter. Of the species with yellow floral bracts, V. eltoniana is distinguished by longer leaf blades and scapes and V. flava is characterized by its shorter sepal lengths. © 2009 The Linnean Society of London, Botanical Journal of the Linnean Society, 2009, 159 , 163–181.     

Longitudinal study of ventilation threshold and maximal O2 uptake in athletic boys

Ventilation threshold (VET) and peak O2 uptake (VO2max) were determined annually from ages 11 to 15 yr in 18 athletic boys. The treadmill protocol consisted of a constant-run speed with grade increments every second minute. Ventilation, VO2, and CO2 production were measured using online open-circuit spirometry. Coefficients of variation for determination of VO2max and VET were 3.4 and 5.6%, respectively. VO2max increased across age 11-15 yr, from 60.8 to 68.0 ml X kg-1 X min-1. VET at 11 yr was 34.4 and at 15 yr 41.9 ml X kg-1 X min-1, thus increasing from 56 to 62% of VO2max. Previous studies of children have shown a decline of VET relative to VO2max across age; however, in the present study the increase may have been due to the training of the boys in competitive athletics. However, the trained youth did not achieve the high relative threshold of trained adults. Across age, both VO2max and VET scaled to weight to the power 1 (in a log-log transformation). The increase in VO2max (l/min) showed greatest increments corresponding to gains in size (a growth curve), whereas increases of VET were consistent year to year. Thus VET was altered independently of VO2max. Factors other than size (and presumably muscle mass) such as the maturation of an enzymatic profile of fast glycolytic fibers might have an important influence on the threshold during youth.     

Comparison of Sewage and Animal Fecal Microbiomes by Using Oligotyping Reveals Potential Human Fecal Indicators in Multiple Taxonomic Groups

Most DNA-based microbial source tracking (MST) approaches target host-associated organisms within the order Bacteroidales, but the gut microbiota of humans and other animals contain organisms from an array of other taxonomic groups that might provide indicators of fecal pollution sources. To discern between human and nonhuman fecal sources, we compared the V6 regions of the 16S rRNA genes detected in fecal samples from six animal hosts to those found in sewage (as a proxy for humans). We focused on 10 abundant genera and used oligotyping, which can detect subtle differences between rRNA gene sequences from ecologically distinct organisms. Our analysis showed clear patterns of differential oligotype distributions between sewage and animal samples. Over 100 oligotypes of human origin occurred preferentially in sewage samples, and 99 human oligotypes were sewage specific. Sequences represented by the sewage-specific oligotypes can be used individually for development of PCR-based assays or together with the oligotypes preferentially associated with sewage to implement a signature-based approach. Analysis of sewage from Spain and Brazil showed that the sewage-specific oligotypes identified in U.S. sewage have the potential to be used as global alternative indicators of human fecal pollution. Environmental samples with evidence of prior human fecal contamination had consistent ratios of sewage signature oligotypes that corresponded to the trends observed for sewage. Our methodology represents a promising approach to identifying new bacterial taxa for MST applications and further highlights the potential of the family Lachnospiraceae to provide human-specific markers. In addition to source tracking applications, the patterns of the fine-scale population structure within fecal taxa suggest a fundamental relationship between bacteria and their hosts.     

Structure and mechanism of the farnesyl diphosphate synthase from Trypanosoma cruzi: implications for drug design

Typanosoma cruzi, the causative agent of Chagas disease, has recently been shown to be sensitive to the action of the bisphosphonates currently used in bone resorption therapy. These compounds target the mevalonate pathway by inhibiting farnesyl diphosphate synthase (farnesyl pyrophosphate synthase, FPPS), the enzyme that condenses the diphosphates of C5 alcohols (isopentenyl and dimethylallyl) to form C10 and C15 diphosphates (geranyl and farnesyl). The structures of the T. cruzi FPPS (TcFPPS) alone and in two complexes with substrates and inhibitors reveal that following binding of the two substrates and three Mg2+ ions, the enzyme undergoes a conformational change consisting of a hinge-like closure of the binding site. In this conformation, it would be possible for the enzyme to bind a bisphosphonate inhibitor that spans the sites usually occupied by dimethylallyl diphosphate (DMAPP) and the homoallyl moiety of isopentenyl diphosphate. This observation may lead to the design of new, more potent anti-trypanosomal bisphosphonates, because existing FPPS inhibitors occupy only the DMAPP site. In addition, the structures provide an important mechanistic insight: after its formation, geranyl diphosphate can swing without leaving the enzyme, from the product site to the substrate site to participate in the synthesis of farnesyl diphosphate.