Comparative analysis of skin surface lipids of the labia majora, inner thigh, and forearm.

The purpose of this study was to develop a procedure for the collection of skin surface corneocyte lipids from the semioccluded and intimate regions of the labia majora and inner thigh of women, to evaluate the polar and nonpolar composition, and to compare the distribution of the lipid classes relative to a collection of lipids from the forearm. The solvent system of ethanol-cyclohexane was well tolerated across all sites. While the yield of polar lipids was similar across all 3 sites, there were only marginal differences in the relative abundance of ceramides, a class of lipids closely associated with skin barrier activity. The yield of neutral lipids was significantly less for the labia majora and was associated with a reduced yield of wax esters, triglycerides and free fatty acids, likely associated with reduced sebaceous gland activity. Factors that may contribute to an inferior skin barrier activity for the labia majora are discussed and suggest a possible deficiency of ω-6 fatty acid linked to the sphingosine base of ceramide EOS.

A simple, rapid, and effective method for the extraction of Mycobacterium paratuberculosis DNA from fecal samples for polymerase chain reaction.

Diagnosis of paratuberculosis (Johne's disease) is stymied by the lack of 1 diagnostic tool that can be used to detect both subclinically and clinically infected animals. At present, fecal culture remains the single diagnostic test that can detect infection in both disease states provided the animals actively shed Mycobacterium paratuberculosis in their feces. Yet, fecal culture has a disadvantage associated with the protracted incubation period of 8-16 weeks before results are available. Detection of nucleic acids specific to M. paratuberculosis in fecal samples is a technique that can circumvent the culture method. This study describes a rapid, simple, and effective method to extract DNA from fecal samples and modification of a polymerase chain reaction assay for optimal sensitivity of detection. An evaluation of 1,000 well-characterized fecal samples was performed by the Colorado Department of Agriculture (Denver, CO) and the National Animal Disease Center (Ames, IA) to determine the sensitivity, specificity, and reproducibility of the new method. Results from this study show that the sensitivity of detection was highly dependent on the load of bacteria in the fecal sample with 81% detection of samples containing >70 colony-forming units (cfu)/g of feces and a 45% detection rate for samples containing less than 1 cfu/g. Similarly, reproducibility of the technique between the 2 laboratories (n = 250 samples) was much higher (75%) for the fecal samples containing high levels of M. paratuberculosis and reduced to 25% for samples with less than 1 cfu/g. An overall specificity of 83% was obtained for known negative samples. The method described here is rapid, simple, and inexpensive compared with other techniques. In addition, this method can detect animals that are shedding less than 1 cfu/g.