Longitudinal assessment of antisaccades in patients with multiple sclerosis

We have previously demonstrated that assessment of antisaccades (AS) provides not only measures of motor function in multiple sclerosis (MS), but measures of cognitive control processes in particular, attention and working memory. This study sought to demonstrate the potential for AS measures to sensitively reflect change in functional status in MS. Twenty-four patients with relapsing-remitting MS and 12 age-matched controls were evaluated longitudinally using an AS saccade task. Compared to control subjects, a number of saccade parameters changed significantly over a two year period for MS patients. These included saccade error rates, latencies, and accuracy measures. Further, for MS patients, correlations were retained between OM measures and scores on the PASAT, which is considered the reference task for the cognitive evaluation of MS patients. Notably, EDSS scores for these patients did not change significantly over this period. These results demonstrate that OM measures may reflect disease evolution in MS, in the absence of clinically evident changes as measured using conventional techniques. With replication, these markers could ultimately be developed into a cost-effective, non-invasive, and well tolerated assessment tool to assist in confirming progression early in the disease process, and in measuring and predicting response to therapy.     

HD exchange and PLIMSTEX determine the affinities and order of binding of Ca2+ with troponin C

Troponin C (TnC), present in all striated muscle, is the Ca(2+)-activated trigger that initiates myocyte contraction. The binding of Ca(2+) to TnC initiates a cascade of conformational changes involving the constituent proteins of the thin filament. The functional properties of TnC and its ability to bind Ca(2+) have significant regulatory influence on the contractile reaction of muscle. Changes in TnC may also correlate with cardiac and various other muscle-related diseases. We report here the implementation of the PLIMSTEX strategy (protein ligand interaction by mass spectrometry, titration, and H/D exchange) to elucidate the binding affinity of TnC with Ca(2+) and, more importantly, to determine the order of Ca(2+) binding of the four EF hands of the protein. The four equilibrium constants, K(1) = (5 ± 5) × 10(7) M(-1), K(2) = (1.8 ± 0.8) × 10(7) M(-1), K(3) = (4.2 ± 0.9) × 10(6) M(-1), and K(4) = (1.6 ± 0.6) × 10(6) M(-1), agree well with determinations by other methods and serve to increase our confidence in the PLIMSTEX approach. We determined the order of binding to the four EF hands to be III, IV, II, and I by extracting from the H/DX results the deuterium patterns for each EF hand for each state of the protein (apo through fully Ca(2+) bound). This approach, demonstrated for the first time, may be general for determining binding orders of metal ions and other ligands to proteins.     

Patterns of cell signaling pathway activation that characterize mammary development

Previous work has detailed the histological and biochemical changes associated with mammary development and remodeling. We have now made use of gene expression profiling, and in particular of the previously described signatures of cell signaling pathway activation, to explore the events associated with mammary gland development. We find that there is elevated E2F-specific pathway activity prior to lactation and relatively low levels of other important signaling pathways, such as RAS, MYC and SRC. Upon lactation and continuing into the involution phase, these patterns reverse with a dramatic increase in RAS, SRC and MYC pathway activity and a decline in E2F activity. At the end of involution, these patterns return to that of the adult non-lactating mammary gland. The importance of the changes in E2F pathway activity, particularly during the proliferative phase of mammary development, was confirmed through the analysis of mice deficient for various E2F proteins. Taken together, these results reveal a complex pattern of pathway activity in relation to the various phases of mammary gland development.     

Genital stimulation facilitates maternal behavior in estrous ewes

Only a small proportion of ewes at estrus have been found to respond maternally to newborn lambs, and this low maternal responsiveness may be partially attributable to the absence of the genital stimulation which occurs at parturition. Therefore, the effect of artificial genital stimulation on maternal behavior of estrous ewes was investigated. Estrus was synchronized in 33 ewes by placement and withdrawal of progestin-saturated vaginal sponges. Estrous ewes were divided into two groups, a control group and a group receiving 5 min of artificial genital stimulation, and observed following presentation of newborn lambs. Significantly more stimulated ewes licked the lamb and emitted low-pitched bleats in a 30-min test. When genital stimulation was subsequently administered to control ewes, more of them also became maternal so that the two groups were no longer significantly different. These results indicate that absence of genital stimulation is one of the factors contributing to the low maternal responsiveness of estrous ewes. They also demonstrate for the first time that artificial genital stimulation is effective in eliciting maternal behavior in nonpregnant ewes even at physiological concentrations of estradiol.     

Linking root production to aboveground plant characteristics and water table in a temperate bog

To develop agronomic application rates for alum-amended poultry litter, nutrient and Al availability of this particular organic fertilizer were examined using long-term field weathering studies. It was revealed that concentrations of N, P, K⁺, and Al³⁺ in the poultry litter leachate ranged from 5 to 6,503 mg L^-1, 2 to 84 mg L^-1, 2 to 5135 mg L^-1, and 0.05 to 4.6 mg L^-1, respectively. Release of nutrients occurred principally in the first growing season and would last for years. During 19 months of field weathering, 10.5 kg N, 1.1 kg P, 34.5 kg K⁺, and 0.031 kg Al³⁺ were released per ton of the poultry litter and recovered in the leachate. The results suggest that no Al toxicity would be generated to crops if alum-amended poultry litter is used as a fertilizer. If applied at 13.3 ton ha^-1, the poultry litter will supply 150.0 kg ha^-1 N, 15.2 kg ha^-1 P, and 456.2 kg ha^-1 K⁺ to seasonal crops. Application of alum-amended poultry litter at available N-based rates will meet crop nutrient requirements while minimizing nutrient runoff losses.     

Distribution of antimicrobial resistance and virulence genes in Enterococcus spp. and characterization of isolates from broiler chickens

Enterococci are now frequent causative agents of nosocomial infections. In this study, we analyzed the frequency and distribution of antibiotic resistance and virulence genotypes of Enterococcus isolates from broiler chickens. Fecal and cecal samples from nine commercial poultry farms were collected to quantify total enterococci. Sixty-nine presumptive enterococci were isolated and identified by API 20 Strep, and their susceptibilities to antibiotics were determined. Genotypes were assessed through the use of a novel DNA microarray carrying 70 taxonomic, 17 virulence, and 174 antibiotic resistance gene probes. Total enterococcal counts were different from farm to farm and between sample sources (P < 0.01). Fifty-one (74%) of the isolates were identified as E. faecium, whereas nine (13%), seven (10%), and two (3%) isolates were identified as E. hirae, E. faecalis, and E. gallinarum, respectively. Multiple-antibiotic resistance was evident in E. faecium and E. faecalis isolates. The most common multiple-antibiotic resistance phenotype was Bac Ery Tyl Lin Str Gen Tet Cip. Genes conferring resistance to aminoglycoside (aac, aacA-aphD, aadB, aphA, sat4), macrolide (ermA, ermB, ermAM, msrC), tetracycline (tetL, tetM, tetO), streptogramin (satG_vatE8), bacitracin (bcrR), and lincosamide (linB) antibiotics were detected in corresponding phenotypes. A range of 9 to 12 different virulence genes was found in E. faecalis, including ace, agg, agrB(Efs) (agrB gene of E. faecalis), cad1, the cAM373 and cCF10 genes, cob, cpd1, cylAB, efaA(Efs), and gelE. All seven E. faecalis isolates were found to carry the gelE gene and to hydrolize gelatin and bile salts. Results from this study showed the presence of enterococci of public and environmental health concerns in broiler chicken farms and demonstrated the utility of a microarray to quickly and reliably analyze resistance and virulence genotypes of Enterococcus spp.