Phonophoretic delivery of 10% hydrocortisone through the epidermis of humans as determined by serum cortisol concentrations.

BACKGROUND AND PURPOSE: The purpose of this investigation was to determine whether application of hydrocortisone phonophoresis enhances transcutaneous delivery of topically applied hydrocortisone in humans, as determined by blood cortisol levels. SUBJECTS: The subjects were 16 men and women, between the ages of 18 and 33 years (X = 25, SD = 2.74), without symptoms of any ongoing inflammatory condition. METHODS: A gel coupling medium containing 10% hydrocortisone acetate was used. Ultrasound was delivered over a 50-cm2 area for 5 minutes at an intensity of 1.0 W/cm2 and a frequency of 1.0 MHz. Each subject received a control treatment (ultrasound alone) and an experimental treatment (hydrocortisone phonophoresis) on the volar aspect of the forearm 1 week apart. Blood was drawn, under both control and experimental conditions, from a cubital vein just proximal to the treatment site prior to each treatment and 0,5, and 15 minutes posttreatment. Serum cortisol concentrations were measured using a fluorescence polarization immunoassay. RESULTS: No rise in serum cortisol concentrations following hydrocortisone phonophoresis was detected. CONCLUSION AND DISCUSSION: These findings suggest that there was no penetration of hydrocortisone through the epidermis and into the underlying vasculature. Clinical implications regarding hydrocortisone levels within the subcutaneous tissues are discussed, and further research is suggested.

Analysis of the development of cellular subsets present in the neural crest using cell sorting and cell culture.

We have tested the hypothesis that developmentally significant cellular subsets are present in the early stages of neural crest ontogenesis. Cultured quail trunk neural crest cells probed with the monoclonal antibodies HNK-1 and R24 exhibited heterogeneous staining patterns. Fluorescence-activated cell sorting was used to isolate the HNK-1+ and HNK-1- cell populations at 2 days in vitro. When these cell populations were cultured, the HNK-1+ sorted cells differentiated into melanocytes, unpigmented cells, and numerous catecholamine-positive (CA+) cells. In contrast, the HNK-1- sorted cells gave rise to melanocytes and unpigmented cells, but few, if any, CA+ cells. When neural crest cells at 2 days in vitro were labeled with R24 and sorted, both the R24+ the R24- sorted cell populations produced numerous CA+ cell, melanocytes, and unpigmented cells. These results provide evidence for the existence of developmental preferences in some subsets of neural crest cells early in embryogenesis.

Phenotypic properties of catecholamine-positive cells that differentiate in avian neural crest cultures.

We have investigated several phenotypic features of the catecholamine-positive (CA+) cell population that develops in quail neural crest cultures. The number, spatial distribution, and morphology of CA+ and tyrosine hydroxylase-positive (TH+) cells are similar at all ages examined, suggesting that these 2 cell classes are identical. Neither CA+ nor TH+ cell bodies or processes were stained using antisera that recognize the 70 or 160 kDa subunits of chicken neurofilament protein. Other cell bodies and fibers in the cultures (which were CA- and TH-) were stained with these neurofilament antisera. The uptake and storage of 3H-norepinephrine by neural crest cultures containing CA+ cells were inhibited in the presence of desmethylimipramine and by incubation at 0 degrees C, but were unaffected by normetanephrine. Overnight treatment with reserpine eliminated histochemically detectable CA fluorescence from the cultures. Chronic reserpine treatment from day 2 to 7 in vitro prevented the appearance of CA+ cells, while normal numbers of TH+ and somatostatin-like immunoreactive (SLI) cells developed. The number and light-microscopic morphology of the CA+ cells that developed in these cultures were not dramatically altered by either exogenous NGF or 6-hydroxydopamine. Using the method of Grillo et al. (1974), we have demonstrated that the CA+ cells observed in the light microscope corresponded to cells containing abundant cytoplasmic granular vesicles (GV) characteristic of catecholamine storage granules observed in other systems. The GV diameters were quite similar in cells examined after 5, 7, 14, and 21 d in vitro. Most GV were 50-200 nm in diameter and were distributed in a unimodal manner, with the observed modal values in the range of 85-115 nm at the ages examined. The number of GV/micron2 of cytoplasmic area remained quite constant at all ages examined. These data, taken together with other available information, suggest that the CA+ cells that differentiate in our neural crest cultures resemble, in many respects, the small, intensely fluorescent cells found in autonomic ganglia and extra-adrenal chromaffin tissue of many species. At present, we do not know if the CA+ cells that differentiate in our neural crest cultures are a stable endpoint of development or whether they are a developmental intermediate in adrenergic differentiation that is normally observed only transiently during the development of avian sympathetic ganglia in vivo, but that can persist under our tissue culture conditions.

Alterations in components of the pineal melatonin synthetic pathway by acute insulin stress in the rat and Syrian hamster.

Acute insulin stress increased plasma catecholamine levels in both the Syrian hamster and albino rat within 3 h after an intraperitoneal injection of either 5 or 10 units of insulin. In the rat, this stress caused a concurrent increase in pineal serotonin N-acetyltransferase (NAT) activity and melatonin content with no observable change in hydroxyindole-O-methyltransferase (HIOMT) activity. In the hamster, on the other hand, acute insulin stress did not alter pineal NAT activity, but depressed both HIOMT activity and melatonin content up to 3 h after the stress. These results present further evidence that catecholamines do not control hamster pineal melatonin synthesis by the same mechanism as observed in the rat.

The effects of hypoxia on catecholamine dynamics in the rat carotid body.

The catecholamine content of the rat carotid body was assayed using high performance liquid chromatography with electrochemical detection. The concentration of dopamine (DA) was found to predominate over that of norepinephrine (NE) by a small margin (31 pmol/carotid body pair DA; 23 pmol/carotid body pair NE). The turnover rates of carotid body DA and NE were determined from the time-dependent decline in their concentrations following the blockade of synthesis with alpha-methyl-p-tyrosine. Values were obtained (DA t 1/2 = 1.9 h; NE t 1/2 = 2.3 h) which suggested a rapid turnover of carotid body catecholamines. Exposure of rats to conditions of severe hypoxia (5% O2-95% N2) failed to significantly alter either the content or turnover of carotid body catecholamines. By contrast, the concentration of carotid body DOPAC, a reflection of DA utilization, was significantly elevated following hypoxic conditions. Further, in vivo tyrosine hydroxylase activity was assessed by measuring the accumulation of carotid body DOPA after inhibiting L-aromatic amino acid decarboxylase with NSD-1015. Basal tyrosine hydroxylase activity (approximately 14-16 pmol/carotid body pair/h) also was significantly increased by acute hypoxic exposure. These results, in part, suggest that rat carotid body DA may act as a neurotransmitter whose synthesis and release are coupled to stimulus demand.

Cytochemical evidence for the existence of norepinephrine-containing glomus cells in the rat carotid body.

Some investigators have postulated that glomus cells of the rat carotid body contain only dopamine (DA), and that the norepinephrine (NE) measured in the carotid body resides only in sympathetic nerve endings and ganglion cells. To investigate this hypothesis, we employed a pharmacologic drug sequence which depleted all carotid body catecholamines and then selectively restored DA levels while keeping NE levels significantly lowered. Analysis of carotid body catecholamines by high performance liquid chromatography (HPLC) validated this drug regimen. Employing this drug treatment, we examined glomus cells after potassium dichromate cytochemical staining in an effort to distinguish those glomus cell vesicles which still contained appreciable amounts of catecholamine, presumably DA. Glomus cells from rats receiving vehicle or L-dopa (100 mg kg-1 ip) alone had 83 and 97% of their cells stained, respectively. However, glomus cells from reserpinized (5 mg kg-1 ip) animals were largely unstained (89%). Carotid bodies from animals treated with reserpine and then, 24 h later, with L-dopa 90 min prior to sacrifice had about 46% of their glomus cells stained while 54% of the cells were unstained. The results of this last group, coupled with our biochemical data which demonstrated that DA levels were comparable to control values but that NE was 80% depleted, suggest that a significant number of glomus cells did not contain enough catecholamine to react with the dichromate. We believe that these unstained cells may normally contain NE and that glomus cells may be of several types, some containing predominantly DA and others NE.