Successful closure of feline axillary wounds by reconstruction of the elbow skin fold.

This report describes the successful closure of five chronic feline axillary wounds. The aetiology was known to be forelimb entrapment in a neck collar in three cases and was suspected in the others. Each cat underwent a single surgical procedure during which the wound was debrided, the normal structure of the elbow skin fold was restored and the remaining skin defect was closed primarily. None of the cats had undergone any previous reconstruction attempts. No postoperative complications were observed and the wounds healed uneventfully. Cats have well-developed elbow skin folds, allowing a wide range of limb motion to occur. Having a forelimb trapped in a neck collar not only creates a wound in the axilla but also disrupts the normal anatomy of the skin fold. This report demonstrates that restoring the elbow skin fold before closing the wound may improve the chances of a successful reconstruction at the first surgical intervention.

Comparative analyses of multi-species sequences from targeted genomic regions.

The systematic comparison of genomic sequences from different organisms represents a central focus of contemporary genome analysis. Comparative analyses of vertebrate sequences can identify coding and conserved non-coding regions, including regulatory elements, and provide insight into the forces that have rendered modern-day genomes. As a complement to whole-genome sequencing efforts, we are sequencing and comparing targeted genomic regions in multiple, evolutionarily diverse vertebrates. Here we report the generation and analysis of over 12 megabases (Mb) of sequence from 12 species, all derived from the genomic region orthologous to a segment of about 1.8 Mb on human chromosome 7 containing ten genes, including the gene mutated in cystic fibrosis. These sequences show conservation reflecting both functional constraints and the neutral mutational events that shaped this genomic region. In particular, we identify substantial numbers of conserved non-coding segments beyond those previously identified experimentally, most of which are not detectable by pair-wise sequence comparisons alone. Analysis of transposable element insertions highlights the variation in genome dynamics among these species and confirms the placement of rodents as a sister group to the primates.

An activator of glutamate decarboxylase genes regulates the expression of enteropathogenic Escherichia coli virulence genes through control of the plasmid-encoded regulator, Per.

Enteropathogenic Escherichia coli (EPEC) is a major cause of infantile diarrhoea in a number of developing countries and is the prototype of pathogenic bacteria that cause attaching and effacing (A/E) intestinal lesions. A chromosomal pathogenicity island, termed the locus of enterocyte effacement (LEE), contains all the genes necessary for the A/E phenotype as well as genes for a type III secretion system and intimate adhesion. Genes in the LEE and genes involved in the synthesis of bundle-forming pili (BFP) are positively regulated by the plasmid-encoded regulator (Per) and comprise the per regulon. In order to identify factors that control the per regulon, we screened an EPEC genomic library for clones that modulate the expression of per. A plasmid clone that decreased the expression of per was isolated using a lacZ reporter gene fused to the per promoter. Subcloning revealed that YhiX, a putative AraC/XylR family transcriptional regulator, was the effector of per repression. Through downregulation of per, a plasmid overproducing YhiX reduced the synthesis of intimin, BfpA, Tir, and CesT, factors important for EPEC virulence. yhiX is located downstream of gadA, which encodes glutamate decarboxylase, an enzyme involved in acid resistance of E. coli. YhiX was found to be an activator of gadA, and the cloned yhiX gene increased production of glutamate decarboxylases (GAD) and activated the transcription of the gadA and gadB promoters. Therefore, yhiX was renamed gadX. Analysis of a gadX mutant grown in the different culture media with acidic and alkaline pH showed that regulation of perA, gadA and gadB by GadX was altered by the external pH and the culture media condition. Under conditions in which EPEC infects cultured epithelial cells, GadX negatively regulated perA expression, and the derepression in the gadX mutant increased translocation of Tir into epithelial cells relative to wild-type EPEC. DNA mobility shift experiments showed that purified GadX protein bound to the perA, gadA and gadB promoter regions in vitro, indicating that GadX is a transcriptional regulator of these genes. On the basis of these results, we propose that GadX may be involved in the appropriate expression of genes required for acid resistance and virulence of EPEC. Our data are consistent with a model in which environmental changes resulting from passage from the stomach to the proximal small intestine induce the functional effect of GadX on per and GAD expression in order to prevent inappropriate expression of the products of these two systems.

Testing Damasio's somatic marker hypothesis with psychopathic individuals: risk takers or risk averse?

Damasio and colleagues (A. R. Damasio, 1994; A. R. Damasio, D. Tranel, & H. Damasio, 1990) have theorized about a possible relationship between somatic markers and the behavior of psychopathic individuals (Ps), but, to date, there are no published data regarding the proposed relationship. The authors assessed 86 Caucasian and 71 African American male offenders using R. D. Hare's (1991) Psychopathy Checklist--Revised and used a modified version of Bechara and colleagues' (A. Bechara, A. R. Damasio, H. Damasio, & S. W. Anderson, 1994; A. Bechara, H. Damasio, D. Tranel, & A. R. Damasio, 1997) gambling task to test the hypothesis that Ps would, consistent with the somatic marker hypothesis, fail to become risk averse. Results indicated that level of anxiety, but not psychopathy, was predictive of response choices. Several limitations and implications of the study are noted.

A method for assessing the migratory response of Naegleria fowleri utilizing [3H]uridine-labeled amoebae.

A new procedure is described to assay the migratory response of Naegleria fowleri (ATCC 30894) amoebae to potential chemoattractants. The method utilizes a blind-well Boyden chemotaxis chamber, two micropore filters of different construction, and amoebae-labeled with [3H]uridine. The technique was standardized by determining the influence of incubation time, filter construction, filter pore size and geometry, amoebae to filter pore ratio, and chemoattractant concentration. Radiolabeled amoebae were placed in Boyden chambers that contained the combination of an upper polycarbonate filter with distinct pores with a diameter of 8 microns and a lower filter of nitrocellulose with a 150-micron depth to separate the wells. A ratio of two amoebae to one filter pore and a 2-h incubation period were chosen to obtain optimal migration conditions. Nerve cell extract was used as the chemoattractant. The migratory responses of both highly pathogenic and weakly pathogenic strains of N. fowleri to nerve cell extract were compared using either the radiolabel procedure or the conventional single filter, leading-front method. Using either method, a highly pathogenic cloned strain of N. fowleri amoebae moved in a directional manner (chemotactically) in vitro to B103 rat neuroblastoma cell extract. In contrast, a weakly pathogenic strain of amoebae responded in a nondirectional manner (chemokinetically) to nerve cell extract. While both the leading-front assay and the radiolabel assay give accurate results, the measurement of radiolabeled cells allows one to test a greater number of attractants in one assay and the procedure eliminates observer bias.