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Stimulation of skin sympathetic nerve discharge by central command. Differential control of sympathetic outflow to skin and skeletal muscle during static exercise.

Microneurographic measurements of muscle sympathetic nerve activity (SNA) have suggested that, during static exercise, central command is much less important than skeletal muscle afferents in causing sympathetic neural activation. The possibility remains, however, that the sympathetic discharge produced by central command is targeted mainly to tissues other than skeletal muscle. To examine this possibility, we recorded SNA with microelectrodes placed selectively in skin, as well as in muscle, nerve fascicles of the peroneal nerve during static handgrip maneuvers designed to separate the effects of central command from those of muscle afferents. To study the relative effects of cutaneous sympathetic activation on sudomotor versus vasomotor function, we simultaneously estimated changes in skin blood flow (laser Doppler velocimetry) and in sudomotor (electrodermal) activation in the region of skin innervated by the impaled nerve fascicle. Two minutes of static handgrip at 10%, 20%, and 30% of maximal voluntary contraction caused large and intensity-dependent increases in skin SNA. These increases in SNA immediately preceded the onset of muscle tension, accelerated progressively during sustained handgrip, and resolved promptly with the cessation of motor effort. The handgrip-induced increases in skin SNA were not maintained when handgrip was followed by arrest of the forearm circulation, a maneuver that maintains the stimulation of chemically sensitive muscle afferents while eliminating the influences of central command and mechanically sensitive muscle afferents. During normothermia, static handgrip at 30% maximal voluntary contraction caused sustained increases in skin SNA (+400 +/- 83%, mean +/- SEM, p less than 0.05) and in electrodermal activity (+276 +/- 56%, p less than 0.05) but only transient increases in estimated skin vascular resistance (+11 +/- 2%, p less than 0.05). When skin temperature was increased or decreased to a new stable baseline level, subsequent increases in skin SNA during handgrip were accompanied by sustained but directionally opposite changes in estimated skin vascular resistance, with exercise-induced vasodilation during hyperthermia but exercise-induced vasoconstriction during hypothermia. From these observations, we conclude the following: 1) static exercise markedly increases sympathetic outflow to skin as well as to skeletal muscle; 2) the increases in skin SNA, unlike muscle SNA, appear to be caused mainly by central command rather than by muscle afferent reflexes; and 3) this cutaneous sympathetic activation appears to be targeted both to sweat glands and to vascular smooth muscle, with the relative targeting being temperature dependent.(ABSTRACT TRUNCATED AT 400 WORDS)

Adult↗

Mechanistic relationship among mutagenicity, skin sensitization, and skin carcinogenicity.

Twenty organic Salmonella mutagens, seven of which (including benzo[a]pyrene) are established skin carcinogens, and one of which (2-chloroethanol) is a well-defined noncarcinogen to skin, have been evaluated for skin-sensitizing activity using the local lymph node assay. The relative mutagenicity of the agents to Salmonella was also established. Fourteen of the chemicals were positive in the local lymph node assay, including the seven skin carcinogens. 2-Chloroethanol was inactive as a sensitizing agent. We suggest that a variety of factors contributes to the lack of sensitizing activity of the remaining six bacterial mutagens: extremes of intrinsic chemical reactivity, high water solubility reducing dermal translocation, and inappropriate dermal metabolism. Two reference skin-sensitizing agents (an oxazolinone and fluorescein isothiocyanate) were established as in vitro clastogens after their recognition as nonmutagens to Salmonella. These data imply that mutagenicity, rather than simply activity in the Salmonella assay, is a primary stimulus for electrophilic sensitization and carcinogenic initiation in the skin. We conclude that genotoxicity data for an agent can provide indications of the agent's potential to induce skin sensitization and that genotoxins which are skin-sensitizing agents have an enhanced potential to initiate skin carcinogenesis. We suggest that common, albeit individually distinct, structure-activity relationships underpin genotoxicity, skin sensitization, and the initiation of skin carcinogenesis. These relationships should simplify the hazard evaluation of chemicals and contribute to a reduction in animal usage. Several predictions of skin carcinogenicity are made based on the data presented.

Animals↗

Protoporphyrin IX fluorescence kinetics and localization after topical application of ALA pentyl ester and ALA on hairless mouse skin with UVB-induced early skin cancer.

In order to improve the efficacy of 5-aminolevulinic acid-based (ALA) photodynamic therapy (PDT), different ALA derivatives are presently being investigated. ALA esters are more lipophilic and therefore may have better skin penetration properties than ALA, possibly resulting in enhanced protoporphyrin IX (PpIX) production. In previous studies it was shown that ALA pentyl ester (ALAPE) does considerably enhance the PpIX production in cells in vitro compared with ALA. We investigated the in vivo PpIX fluorescence kinetics after application of ALA and ALAPE to hairless mice with and without UVB-induced early skin cancer. ALA and ALAPE (20% wt/wt) were applied topically to the mouse skin and after 30 min, the solvent was wiped off and PpIX fluorescence was followed in time with in vivo fluorescence spectroscopy and imaging. At 6 and 12 h after the 30 min application, skin samples of visible lesions and adjacent altered skin (UVB-exposed mouse skin) and normal mouse skin were collected for fluorescence microscopy. From each sample, frozen sections were made and phase contrast images and fluorescence images were recorded. The in vivo fluorescence kinetics showed that ALAPE induced more PpIX in visible lesions and altered skin of the UVB-exposed mouse skin, but not in the normal mouse skin. In the microscopic fluorescence images, higher ALAPE-induced PpIX levels were measured in the stratum corneum, but not in the dysplastic layer of the epidermis. In deeper layers of the skin, PpIX levels were the same after ALA and ALAPE application. In conclusion, ALAPE does induce higher PpIX fluorescence levels in vivo in our early skin cancer model, but these higher PpIX levels are not located in the dysplastic layer of the epidermis.

Administration, Topical↗

Comparison of the percutaneous absorption of hydrophilic and lipophilic compounds in shed snake skin and human skin.

The in vitro transdermal permeation of eight hydrophilic drugs (antipyrine, L-dopa, dopamine hydrochloride, diclofenac sodium, 5-fluorouracil, isoprenaline hydrochloride, nicorandil and morphine hydrochloride) and eight lipophilic drugs (aminopyrine, cyclobarbital, ibuprofen, indomethacin, isosorbide dinitrate, flurbiprofen, ketoprofen and lignocaine) was determined using shed snake skin of Elaphae obsoleta and human skin. The permeation parameters and physiological characteristics of the skin, e.g. the water and lipid content, and the thickness of shed snake skin and human skin were evaluated and compared. In shed snake skin, the permeability coefficients (P) of lipophilic drugs were in the same range as those through the human skin (0.9 to 1.8-times); whereas those of hydrophilic drugs were remarkably lower (3.3 to 6.1-times). The thickness and lipid content of shed snake skin and human stratum corneum were not significantly different (P > 0.05), whereas the water content of shed snake skin was significantly lower than that of human stratum corneum (P < 0.05). The lower permeability of shed snake skin for hydrophilic compounds might be caused by the lower porosity of skin strata. The results suggested a potential use of shed snake skin as barrier membrane for lipophilic compounds percutaneous absorption studies in vitro.

Animals↗

Comparison of bovine udder skin with human and porcine skin in percutaneous permeation experiments.

Rat and pig animal skin has been the most common replacement material for human skin for use in in vitro permeability experiments. Unfortunately, the permeability barrier of skin from laboratory animals is known to be relatively weak, due to significant follicular transport. Pig skin has been shown to be a suitable model for human skin. Unfortunately, it cannot be gathered from the regular slaughtering process, which makes it unsuitable for permeation experiments. We therefore studied the suitability of bovine udder skin, an untreated waste material of the butchering process, as a possible replacement material for use in in vitro permeability tests. We investigated the barrier strength of bovine udder skin against four different substances, and its histology and lipid profile, in comparison with pig skin and heat separated human epidermis. Pig and human skin were found to be equally permeable, whilst bovine udder skin seemed to exhibit a weaker, but less variable, barrier against caffeine, benzoic acid, testosterone, and flufenamic acid. The skin of all three species contained variable contents of the major lipid classes: cholesterol, ceramides, cholesterol ester, fatty acids and triglycerides. Morphological differences mainly comprised variations in the density of hair follicles. Based on these results, the amount of free fatty acids and triglycerides and the density of hair follicles seem to be important factors in the differences between the skin barriers in the three species.

Animals↗

[Evaluation of the skin sympathetic function by the decrease of skin temperature at the finger tip immersed into cold water].

The change of the skin temperature at the finger tip immersed into cold water was measured in normal volunteers and in patients with disturbances of various portions of sympathetic nervous system with copper-constant thermocouple. The skin temperature of the normal subjects decreased soon after the immersion, partly due to cooling by the cold water, and partly due to the decrease of the skin blood flow. The skin sympathetic activity recorded from three normal subjects using the technique of microneurography presented an increase during the decreasing phase of the skin temperature. The skin temperature of the patients who received stellate ganglion block did not fall and maintained higher level compared to that of normal subjects. These data indicated that the decreasing phase of the skin temperature of the immersed finger is related to the increase of skin sympathetic activity. Cases of vascular neuropathy (lesion in post-ganglionic fiber), Shy-Drager syndrome and pure autonomic failure (lesion in pre-ganglionic fiber), cervical spondylosis and cervical disc herniation (lesion in descending tract of sympathetic system in spinal cord), and bleeding in pons tegmentum (lesion in descending tract in brain stem) were examined and all of them presented poor decrease of the skin temperature during the immersion. The skin temperature decreased, but incompletely in the patients, who had suffered from thalamic infarct and post-central gyrus infarct, and did not feel any sense on the immersion. These results indicate that the decreasing phase of the skin temperature of the finger immersed into cold water depends more upon the efferent than upon the afferent system, and this test is useful for evaluation of the skin sympathetic function.

Adult↗

In vitro skin penetration of griseofulvin in rat and human skin from an ointment dosage form.

Penetration and permeation of griseofulvin into and across the rat skin after application of three ointment formulations containing either dimethylacetamide (DMAC) or diethylene glycol monoethylether (DGME) or the ointment base alone (control--without DMAC or DGME) were studied, in vitro. Penetration and permeation of griseofulvin into and across the human skin after application of DGME ointment was also studied. Permeation of griseofulvin across the rat skin was highest for the DMAC ointment, followed by the DGME ointment and lowest for the control. Concentration of griseofulvin in the upper layers of the skin (i.e., surface to 100 micron depth) was also highest for DMAC and lowest for the control. However, the skin levels from the DGME ointment could not be distinguished (statistically) from the other two formulations. In comparison to rat skin, human skin is much less permeable. Amount of griseofulvin (from DGME ointment) that permeated through the rat skin was 14 times the amount that permeated through the human skin. Concentration of griseofulvin in the upper layers of the rat skin were 4 times the concentration of the drug in the upper layers of the human skin. The concentration of griseofulvin in the various layers of the human skin after one topical application were far greater than those reported after prolonged peroral administration.

Administration, Topical↗