Improved multiple-locus variable-number tandem-repeat assay for Staphylococcus aureus genotyping, providing a highly informative technique together with strong phylogenetic value.

We describe an improved multiple-locus variable-number tandem-repeat (VNTR) analysis (MLVA) scheme for genotyping Staphylococcus aureus. We compare its performance to those of multilocus sequence typing (MLST) and spa typing in a survey of 309 strains. This collection includes 87 epidemic methicillin-resistant S. aureus (MRSA) strains of the Harmony collection, 75 clinical strains representing the major MLST clonal complexes (CCs) (50 methicillin-sensitive S. aureus [MSSA] and 25 MRSA), 135 nasal carriage strains (133 MSSA and 2 MRSA), and 13 published S. aureus genome sequences. The results show excellent concordance between the techniques' results and demonstrate that the discriminatory power of MLVA is higher than those of both MLST and spa typing. Two hundred forty-two genotypes are discriminated with 14 VNTR loci (diversity index, 0.9965; 95% confidence interval, 0.9947 to 0.9984). Using a cutoff value of 45%, 21 clusters are observed, corresponding to the CCs previously defined by MLST. The variability of the different tandem repeats allows epidemiological studies, as well as follow-up of the evolution of CCs and the identification of potential ancestors. The 14 loci can conveniently be analyzed in two steps, based upon a first-line simplified assay comprising a subset of 10 loci (panel 1) and a second subset of 4 loci (panel 2) that provides higher resolution when needed. In conclusion, the MLVA scheme proposed here, in combination with available on-line genotyping databases (including http://mlva.u-psud.fr/), multiplexing, and automatic sizing, can provide a basis for almost-real-time large-scale population monitoring of S. aureus.

CT angiography helps to differentiate acute from chronic carotid occlusion: "carotid ring sign".

INTRODUCTION: Currently, there is no reliable method to differentiate acute from chronic carotid occlusion. We propose a novel CTA-based method to differentiate acute from chronic carotid occlusions that could potentially aid clinical management of patients. METHODS: We examined 72 patients with 89 spontaneously occluded extracranial internal carotids with CT angiography (CTA). All occlusions were confirmed by another imaging modality and classified as acute (imaging <1 week of presumed occlusion) orchronic (imaging >4 weeks), based on circumstantial clinical and radiological evidence. A neuroradiologist and a neurologist blinded to clinical information determined the site of occlusion on axial sections of CTA. They also looked for (a) hypodensity in the carotid artery (thrombus), (b) contrast within the carotid wall (vasa vasorum), (c) the site of the occluded carotid, and (d) the "carotid ring sign" (defined as presence of a and/or b). RESULTS: Of 89 occluded carotids, 24 were excluded because of insufficient circumstantial evidence to determine timing of occlusion, 4 because of insufficient image quality, and 3 because of subacute timing of occlusion. Among the remaining 45 acute and 13 chronic occlusions, inter-rater agreement (kappa) for the site of proximal occlusion was 0.88, 0.45 for distal occlusion, 0.78 for luminal hypodensity, 0.82 for wall contrast, and 0.90 for carotid ring sign. The carotid ring sign had 88.9% sensitivity, 69.2% specificity, and 84.5% accuracy to diagnose acute occlusion. CONCLUSION: The carotid ring sign helps to differentiate acute from chronic carotid occlusion. If further confirmed, this information may be helpful in studying ischemic symptoms and selecting treatment strategies in patients with carotid occlusions.

Portable School Stop Signs and Other Non-Uniform School Stop Control Devices, HR-190, 1978

Portable (roll-out) stop signs are used at school crossings in over 300 cities in Iowa. Their use conforms to the Code of Iowa, although it is not consistent with the provisions of the Manual on Uniform Traffic Control Devices adopted for nationwide application. A survey indicated that most users in Iowa believe that portable stop signs provide effective protection at school crossings, and favor their continued use. Other non-uniform signs that fold or rotate to display a STOP message only during certain hours are used at school crossings in over 60 cities in Iowa. Their use does not conform to either the Code of Iowa or the Manual on Uniform Traffic Control Devices. Users of these devices also tend to favor their continued use. A survey of other states indicated that use of temporary devices similar to those used in Iowa is not generally sanctioned. Some unsanctioned use apparently occurs in several states, however. A different type of portable stop sign for school crossings is authorized and widely used in one state. Portable stop signs similar to those used in Iowa are authorized in another state, although their use is quite limited. A few reports in the literature reviewed for this research discussed the use of portable stop signs. The authors of these reports uniformly recommended against the use of portable or temporary traffic control devices. Various reasons for this recommendation were given, although data to support the recommendation were not offered. As part of this research, field surveys were conducted at 54 locations in 33 communities where temporary stop control devices were in use at school crossings. Research personnel observed the obedience to stop control and measured the vehicular delay incurred. Stopped delay averaged 1.89 seconds/entering vehicle. Only 36.6 percent of the vehicles were observed to come to a complete stop at the study locations controlled by temporary stop control devices. However, this level of obedience does not differ from that observed at intersections controlled by permanent stop signs. Accident experience was compiled for 76 intersections in 33 communities in Iowa where temporary stop signs were used and, for comparative purposes, at 76 comparable intersections having other forms of control or operating without stop control. There were no significant differences in accident experience An economic analysis of vehicle operating costs, delay costs, and other costs indicated that temporary stop control generated costs only about 12 percent as great as permanent stop control for a street having a school crossing. Midblock pedestrian-actuated signals were shown to be cost effective in comparison with temporary stop signs under the conditions of use assumed. Such signals could be used effectively at a number of locations where temporary stop signs are being used. The results of this research do not provide a basis for recommending that use of portable stop signs be prohibited. However, erratic patterns of use of these devices and inadequate designs suggest that improved standards for their use are needed. Accordingly, nine recommendations are presented to enhance the efficiency of vehicular flow at school crossings, without causing a decline in the level of pedestrian protection being afforded.

CD28-negative cytolytic effector T cells frequently express NK receptors and are present at variable proportions in circulating lymphocytes from healthy donors and melanoma patients.

In humans, NK receptors are expressed by natural killer cells and some T cells, the latter of which are preferentially alphabetaTCR+ CD8+ cytolytic T lymphocytes (CTL). In this study we analyzed the expression of nine NK receptors (p58.1, p58.2, p70, p140, ILT2, NKRP1A, ZIN176, CD94 and CD94/NKG2A) in PBL from both healthy donors and melanoma patients. The percentages of NK receptor-positive T cells (NKT cells) varied strongly, and this variation was more important between individual patients than between individual healthy donors. In all the individuals, the NKT cells were preferentially CD28-, and a significant correlation was found between the percentage of CD28- T cells and the percentage of NK receptor+ T cells. Based on these data and the known activated phenotype of CD28- T cells, we propose that the CD28- CD8+ T cell pool represents or contains the currently active CTL population, and that the frequent expression of NK receptors reflects regulatory mechanisms modulating the extent of CTL effector function. Preliminary results indicate that some tumor antigen-specific T cells may indeed be CD28- and express NK receptors in vivo.