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Biomedical subjects

J E McKie

Publications and source records attributed to J E McKie.

8 recordsLinked to original sources

Stratum corneum lipid phase transitions and water barrier properties.

In mammals, the outer skin layer, the stratum corneum, is the ultimate barrier to water loss. In order to relate barrier function to stratum corneum structure, samples from porcine skin were investigated by using differential scanning calorimetry (DSC), infrared (IR) spectroscopy, and water permeability techniques. Results of DSC and IR studies show that stratum corneum lipids undergo thermal transitions between 60 and 80 degrees C similar to lipid thermotropic transitions seen in a variety of synthetic and biological membranes. Results of water flux experiments performed under conditions similar to those of the DSC and IR studies show an abrupt change in permeability at about 70 degrees C. At low temperatures, water flux values are similar to those obtained for human skin in vivo, yielding an activation energy of 17 kcal/mol, in excellent agreement with values obtained for water flux through a variety of lipid biomembranes. In contrast, at temperatures above about 70 degrees C, water flux is characterized by an activation energy only slightly higher than that of free diffusion, suggesting that the stratum corneum offers little diffusional resistance under these conditions. These combined results suggest that increased disorder in stratum corneum lipid structure, brought about by thermotropic transitions, results in dramatically altered diffusional resistance of this tissue to water flux. Thus, as found for numerous biological membranes, water flux and lipid order in porcine stratum corneum are inversely related.

Animals↗

Role of stratum corneum lipid fluidity in transdermal drug flux.

Fatty acids are effective penetration enhancers for the transdermal delivery of certain co-applied drugs. In order to assess the mechanism of enhancement, spectrometric, calorimetric, and flux techniques were used to study porcine stratum corneum following treatment with a series of cis- and trans-positional isomers of octadecenoic acid. Results obtained from spectrometric and calorimetric measurements show increased lipid fluidity following treatment of the stratum corneum with the cis monoenoic acids which have the site of unsaturation centrally located. Under similar conditions, these same cis monounsaturated acids resulted in enhancement of salicyclic acid flux through porcine skin. The striking parallelism between flux and fluidity measurements suggests that transdermal drug flux may be ultimately related to stratum corneum lipid structure.

Animals↗

Lipid thermotropic transitions in human stratum corneum.

The techniques of differential scanning calorimetry (DSC) and thermal perturbation infrared (IR) spectrometry were used to investigate thermal transitions in intact, fractionated, and lipid-extracted human stratum corneum. The DSC results show 3 major and one minor thermal transition in the range of 30-120 degrees C. Of particular interest to this study are 2 transitions seen near 65 degrees and 75 degrees C in intact stratum corneum and a stratum corneum membrane preparation, but absent from lipid-extracted samples. Results of IR spectrometry show that thermally induced spectral changes related to enhanced motion of the lipid acyl chains also occur in the region of about 60-80 degrees C. The combined DSC and IR results show that the thermal transitions near 65 degrees and 75 degrees C depend on water concentration in a manner identical to that seen for a variety of lipid-water systems. Taken together, these results suggest that thermal transitions occurring near 65 degrees and 85 degrees C involve increased acyl chain motion of stratum corneum lipids.

Calorimetry, Differential Scanning↗

A noninvasive, in vivo technique to quantitatively measure water concentration of the stratum corneum using attenuated total-reflectance infrared spectroscopy.

In order to noninvasively measure water concentration in the stratum corneum, infrared spectra were obtained using an attenuated total-reflectance technique in conjunction with Fourier transform spectroscopy. A weak water-absorbance band near 2,100 cm-1 was detected in both in vivo and in vitro spectra. The significance of this band is that it occurs in a region of the mid-infrared where the stratum corneum and most topically applied substances show no absorbance. In vitro spectra obtained as a function of ambient relative humidity showed an increase in the absorbance near 2,100 cm-1 with increasing water concentration in the stratum corneum. The combined in vivo and in vitro results lead to a quantitative assessment of water concentration in the uppermost layers of the stratum corneum.

Animals↗

Novel approach to bacterial identification that uses the autobac system.

A new system for the rapid identification of gram-negative bacilli on the Autobac system is described. This system utilizes growth inhibition profiles to a panel of differentially inhibitory chemical agents. These profiles are analyzed with a two-stage quadratic discriminant analysis to arrive at the organism identification. The system identifies 30 different groups of gram-negative bacilli, including the most clinically significant Enterobacteriaceae and glucose nonfermenters. A total of 3,726 strains, distributed among the 30 groups, was tested. The Autobac system agreed with the conventional biochemical identification 88.4% of the time. When the individual group results were weighted to reflect clinical frequency, the result was a 93.1% agreement.

Anti-Bacterial Agents↗

Rapid screening for bacteriuria by light scatter photometry (Autobac): a collaborative study.

A total of 2,720 urine specimens from three laboratories were evaluated by Autobac (Pfizer Diagnostics) and were compared with simultaneous colony counts for evidence of bacteriuria. Of 599 specimens with a colony count of greater than or equal to 10(5) colony-forming units per ml, 93.8% were detected within 6 h. This detection rate increased to 97% of 447 positive urine specimens when only specimens from patients not on antimicrobials were evaluated. The majority (77.9%) of positive specimens were detected as early as 3 h. Those specimens with greater than or equal to 10(5) colony-forming units per ml, which were negative by Autobac at 6 h, included organisms which are frequently considered to be contaminants (diphtheroids, lactobacilli, alpha and gamma streptococci, yeasts, and Staphylococcus epidermidis), or were from patients who were being treated with antimicrobial agents. Of 2,121 urine specimens with colony counts of less than 10(5), 98.1% were correctly determined to be negative by Autobac at 3 h. This percentage decreased to 86.0 at 6 h. The majority of these false-positive specimens were those with colony counts of 10(4) to 10(5) colony-forming units per ml. There appeared to be no major difference in results from the three laboratories or among the four lots of broth used in this study.

Anti-Bacterial Agents↗

Rapid determination of minimum inhibitory concentrations of antimicrobial agents by regression analysis of light scattering data.

A novel, rapid, automated method for determining the minimum inhibitory concentrations of antimicrobial drugs within ranges acceptable for therapeutic application is described and validated in this report. By employing a simple modification of the Autobac photometer currently in use and in optically clear, modified Mueller-Hinton broth, the method utilizes forward light scattering data measured in the presence of two concentrations of the antimicrobial agent to compute the minimum inhibitory concentrations. Empirically derived regression equations which simultaneously use scattering data from two drug concentrations are employed in this computation, rather than simple breakpoint analysis in which the light scattering measured in the presence of each drug concentration is referred to a threshold level. The minimum inhibitory concentrations obtained with this new method were highly reproducible and, as shown by side by side comparisons were in excellent agreement with the minimum inhibitory cencentrations obtained with the International Collaborative Study broth dilution method.

Anti-Bacterial Agents↗