Validating and corroborating the deposition of two annual growth zones in asteriscus otoliths of common carp Cyprinus carpio from South Africa's largest impoundment

A total of 816 common carp Cyprinus carpio asteriscus otolith pairs were collected from Lake Gariep, South Africa. Otoliths were interpreted whole, submerged in methyl salicylate and viewed under transmitted light. The precision of growth zone counts of the primary reader was estimated at 5·54 and 7·03% using the average per cent error method and the coefficient of variation, respectively. Age-bias plots indicated no systematic bias between the primary reader and the three secondary readers for up to nine growth zones (95% of the sample). Growth zone deposition rate was validated using a mark-recapture experiment of chemically tagged C. carpio (n = 21) conducted in a large earthen pond under ambient conditions in the vicinity of Lake Gariep. The validation results were corroborated for the wild population by edge analysis and a length-based age-structured model. All three methods suggest that growth zone formation occurred biannually, exemplifying the importance of age validation as a prerequisite for understanding the life history of C. carpio.     

RecA and RadA proteins of Brucella abortus do not perform overlapping protective DNA repair functions following oxidative burst

Very little is known about the role of DNA repair networks in Brucella abortus and its role in pathogenesis. We investigated the roles of RecA protein, DNA repair, and SOS regulation in B. abortus. While recA mutants in most bacterial species are hypersensitive to UV damage, surprisingly a B. abortus recA null mutant conferred only modest sensitivity. We considered the presence of a second RecA protein to account for this modest UV sensitivity. Analyses of the Brucella spp. genomes and our molecular studies documented the presence of only one recA gene, suggesting a RecA-independent repair process. Searches of the available Brucella genomes revealed some homology between RecA and RadA, a protein implicated in E. coli DNA repair. We considered the possibility that B. abortus RadA might be compensating for the loss of RecA by promoting similar repair activities. We present functional analyses that demonstrated that B. abortus RadA complements a radA defect in E. coli but could not act in place of the B. abortus RecA. We show that RecA but not RadA was required for survival in macrophages. We also discovered that recA was expressed at high constitutive levels, due to constitutive LexA cleavage by RecA, with little induction following DNA damage. Higher basal levels of RecA and its SOS-regulated gene products might protect against DNA damage experienced following the oxidative burst within macrophages.     

Kinetics of an individual transmembrane helix during bacteriorhodopsin folding

The kinetics of an individual helix of bacteriorhodopsin have been monitored during folding of the protein into lipid bilayer vesicles. A fluorescence probe was introduced at individual sites throughout helix D of bacteriorhodopsin and the changes in the fluorescence of the label were time-resolved. Partially denatured, labelled bacteriorhodopsin in SDS was folded directly into phosphatidylcholine lipid vesicles. Stopped-flow mixing of the reactants allowed the folding kinetics to be monitored with millisecond time resolution by time-resolving changes in the label fluorescence, intrinsic protein fluorescence as well as in the absorption of the retinal chromophore. Monitoring specific positions on helix D showed that two kinetic phases were altered compared to those determined by monitoring the average protein behaviour. These two phases, of 6.7 s(-1) and 0.33 s(-1), were previously assigned to formation of a key apoprotein intermediate during bacteriorhodopsin folding. The faster 6.7s(-1) phase was missing when time-resolving fluorescence changes of labels attached to the middle of helix D. The amplitude of the 0.33 s(-1) phase increased along the helix, as single labels were attached in turn from the cytoplasmic to the extracellular side. An interpretation of these results is that the 6.7 s(-1) phase involves partitioning of helix D within the lipid headgroups of the bilayer vesicle, while the 0.33 s(-1) phase could reflect transmembrane insertion of this helix. In addition, a single site on helix G was monitored during folding. The results indicate that, unlike helix D, the insertion of helix G cannot be differentiated from the average protein behaviour. The data show that, while folding of bacteriorhodopsin from SDS into lipids is a co-operative process, it is nevertheless possible to obtain information on specific regions of a membrane protein during folding in vitro.     

Conventional and EMG telemetry studies of upstream migration and tailrace attraction of adult Atlantic salmon at a hydroelectric installation on the Exploits River,Newfoundland, Canada

Conventional and electromyogram (EMG) radio telemetry studies have documented occurrence of tailrace attraction and residency, and associated energy cost, for migratory wild Atlantic at a power plant on the Exploits River, insular Newfoundland, Canada. All fish demonstrated some degree of tailrace attraction and turbine discharge was the primary factor resulting in ‘false attraction’. In 2002, residency times were generally less than 1 h, although some fish demonstrated residency between 71 and 118 h. In 2004, after plant refurbishing, fish took from 6 to 11 days to reach the power plant and remained in the tailrace area from 3 to 12 days. Frequent entrances into the tailraces indicated fish were demonstrating a searching behaviour to find the upstream migration route. In 2003 and 2004, EMG data were collected from 3 fish in each year migrating to, and past, the tailraces. An Energy Index was calculated integrating EMG signal and time spent at each location to estimate potential energetic cost. Relatively high-energy expenditure was associated with tailrace attraction and residency in both years. In 2003, fish spent highest energy at the lowest tailrace (first encountered in their migration) while in 2004, there were also high-energy costs associated with a new generating unit. In both years, high EMGs both just below and above the power plant suggested these reaches were arduous and difficult to pass.     

The MS Risk Allele of CD40 Is Associated with Reduced Cell-Membrane Bound Expression in Antigen Presenting Cells: Implications for Gene Function

Human genetic and animal studies have implicated the costimulatory molecule CD40 in the development of multiple sclerosis (MS). We investigated the cell specific gene and protein expression variation controlled by the CD40 genetic variant(s) associated with MS, i.e. the T-allele at rs1883832. Previously we had shown that the risk allele is expressed at a lower level in whole blood, especially in people with MS. Here, we have defined the immune cell subsets responsible for genotype and disease effects on CD40 expression at the mRNA and protein level. In cell subsets in which CD40 is most highly expressed, B lymphocytes and dendritic cells, the MS-associated risk variant is associated with reduced CD40 cell-surface protein expression. In monocytes and dendritic cells, the risk allele additionally reduces the ratio of expression of full-length versus truncated CD40 mRNA, the latter encoding secreted CD40. We additionally show that MS patients, regardless of genotype, express significantly lower levels of CD40 cell-surface protein compared to unaffected controls in B lymphocytes. Thus, both genotype-dependent and independent down-regulation of cell-surface CD40 is a feature of MS. Lower expression of a co-stimulator of T cell activation, CD40, is therefore associated with increased MS risk despite the same CD40 variant being associated with reduced risk of other inflammatory autoimmune diseases. Our results highlight the complexity and likely individuality of autoimmune pathogenesis, and could be consistent with antiviral and/or immunoregulatory functions of CD40 playing an important role in protection from MS.     

Non-Invasive Assessment of the Interrelationships of Diet,Pregnancy Rate,Group Composition,and Physiological and Nutritional Stress of Barren-Ground Caribou in Late Winter

The winter diet of barren-ground caribou may affect adult survival, timing of parturition, neonatal survival, and postpartum mass. We used microhistological analyses and hormone levels in feces to determine sex-specific late-winter diets, pregnancy rates, group composition, and endocrine-based measures of physiological and nutritional stress. Lichens, which are highly digestible but contain little protein, dominated the diet (> 68%) but were less prevalent in the diets of pregnant females as compared to non-pregnant females and males. The amount of lichens in the diets of pregnant females decreased at higher latitudes and as winter progressed. Pregnancy rates (82.1%, 95% CI = 76.0 – 88.1%) of adult cows were within the expected range for a declining herd, while pregnancy status was not associated with lichen abundance in the diet. Most groups (80%) were of mixed sex. Male: female ratios (62:100) were not skewed enough to affect the decline. Levels of hormones indicating nutritional stress were detected in areas of low habitat quality and at higher latitudes. Levels of hormones indicated that physiological stress was greatest for pregnant cows, which faced the increasing demands of gestation in late winter. These fecal-based measures of diet and stress provided contextual information for the potential mechanisms of the ongoing decline. Non-invasive techniques, such as monitoring diets, pregnancy rates, sex ratios and stress levels from fecal samples, will become increasingly important as monitoring tools as the industrial footprint continues to expand in the Arctic.     

The brachial plexus in the vervet monkey (Cercopithecus pygerythrus)

The brachial plexus in ten embalmed, mature vervet monkeys was dissected to document the structure and branching pattern of this nerve plexus in this frequently used research animal. In general, the brachial plexus in the vervet monkey was similar to the plexus of other Old World monkeys. However, several aspects were comparable to those observed in domestic animals. Thus the quadrupedal and bipedal abilities of the vervet monkey was reflected in the structure of its brachial plexus.