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Biomedical subjects

A H Kennedy

Publications and source records attributed to A H Kennedy.

10 recordsLinked to original sources

Stratum corneum lipids of human epidermal keratinocyte air-liquid cultures: implications for barrier function.

PURPOSE: The purpose of this study is to investigate the permeability barrier, i.e., the stratum corneum (SC) lipids, of human epidermal keratinocyte air-liquid cultures and compare them with those of human SC. METHOD: The SC lipids composition was analyzed by TLC technique, the organization by electron microscopic procedure, and the phase transition temperature by infrared spectroscopic method. RESULTS: Electron microscopy demonstrated that The SC lipids of cultures were largely retained inside the comeocytes, and that the intercellular lipids lack both the basic unit repetition (i.e., broad: narrow: broad: broad: narrow: broad of electron lucent bands) and the covalently-bound lipid envelope normally found in human SC. These characteristics are similar to those found in SC from patients with atopic dermatitis or psoriasis, or from animals with essential fatty acid deficiency, suggesting that the cultures may be hyperproliferative. In addition, the high free sterol content and the altered fatty acid/ceramide composition of these cultures argue that the compromised barrier function is linked to hyperproliferation and lipid synthesis, or vice versa. Infrared spectroscopic analyses confirm that there are major conformational differences between the lipids of human and cultured SC. CONCLUSIONS: The profound differences in SC lipid composition, organization and conformational properties attest that permeability alone is not a sufficiently sensitive marker to define barrier equivalence between cultures and human skin.

Air↗

Accuracy and reliability of temperature measurement by instrument and site.

The accuracy and reliability of three instruments (IVAC, [IVAC Corp., San Diego, CA] TempaDOT [PyMaH Corp., Somerville, NJ], off-the-shelf glass) were determined at axillary, oral and rectal sites taken on children by experienced RNs given one of three levels of inservice education. TempaDOT was found to be the most clinically valid temperature measurement instrument for 502 children in this acute care setting.

Adolescent↗

Barrier function of human keratinocyte cultures grown at the air-liquid interface.

Stratum corneum (SC), the outermost and least permeable layer of skin, is the major barrier to passive transepidermal water loss. In the research described in this paper, we have used human keratinocyte cultures, grown at the air-liquid (A/L) interface, to examine the relationship between epidermal differentiation (including SC formation) and barrier function. Histologically, the A/L culture showed several markers of complete differentiation, including the presence of well-organized and defined epidermal cell layers, keratohyalin granules, and a multilayered SC. The permeability of tritiated water through epidermal cultures, which had grown for 3 weeks at the A/L interface, was measured with a microdiffusion apparatus. The results of these experiments demonstrated that: a) the human keratinocyte cultures developed a substantial barrier (i.e., a multilayered SC) to water diffusion across the entire surface. If the relative humidity of the culturing environment was lowered from 100% to around 75%, the barrier was significantly improved; b) the differentiation promoter, 1.25-dihydroxy-vitamin-D3, increased the number of SC layers and reduced water permeation through the culture; c) the nature of the keratinocyte support matrix could be altered to improve the morphology as well as the barrier function of the epidermal cultures. Overall, the observations are consistent with the relationship that is believed to exist between SC intercellular lipid content and percutaneous penetration. Confirmation of this hypothesis will further the considerable potential of human keratinocyte A/L cultures as a valuable and relevant model in which to study drug absorption and metabolism.

Air↗

Transdermal drug transport and metabolism. I. Comparison of in vitro and in vivo results.

Using excised human skin and tissue grafted to athymic mice, the in vitro and in vivo delivery and metabolism of a salicylate diester were compared. Concentration profiles of this drug and its metabolites were obtained for the outer several hundred microns of the skin. These results show significant differences in the extent of enzymatic cleavage and distribution of metabolites between in vitro and in vivo studies. Furthermore, these data suggest that in vitro results may overestimate metabolism because of increased enzymatic activity and/or decreased capillary removal.

Administration, Cutaneous↗

The water permeability of primary mouse keratinocyte cultures grown at the air-liquid interface.

In order to study the development of the epidermal permeability barrier in vitro, tritiated water (HTO) flux was measured across murine keratinocytes cultured at the air-liquid interface. Using a micro-diffusion technique, it was shown that air-liquid cultures form areas where the water diffusion is comparable to that of intact neonatal mouse skin. When water permeability is measured over a large area of the culture surface, however, significantly higher flux is obtained. These results show that under the culture conditions used, areas of water barrier comparable to intact neonatal mouse skin coexist with regions of less complete barrier formation.

Animals↗

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↗

An education program for nurses from referring hospitals in a perinatal regionalization system.

Administering high-quality perinatal care at a regional center involves more than providing in-center service. An outreach education program was developed for nurses in referring hospitals. The authors believe that through the carefully phased development of an outreach education program, quality perinatal care on all levels can be realized.

Education, Nursing, Continuing↗