Age-related percutaneous penetration part 2: effect of age on dermatopharmacokinetics and overview of transdermal products.

Transdermal drug delivery allows for a constant rate of drug administration and prolonged action, which can be beneficial to elderly patients who are often polymedicated. Several studies have compared dermatopharmacokinetics in the young and elderly with conflicting results. Despite the potential limitations of age-related changes in skin factors and cutaneous metabolism, marketed transdermal products generally do not report age-related differences in pharmacokinetics. This overview discusses the current data, summarizes marketed product findings and highlights the importance of further studies to evaluate age-related dermatopharmacokinetics.

Age-related percutaneous penetration part 1: skin factors.

Changes in the skin that occur in the elderly may put them at increased risk for altered percutaneous penetration from pharmacotherapy along with potential adverse effects. Skin factors that may have a role in age-related percutaneous penetration include blood flow, pH, skin thickness, hair and pore density, and the content and structure of proteins, glycosaminoglycans (GAGs), water, and lipids. Each factor is examined as a function of increasing age along with its potential impact on percutaneous penetration. Additionally, topical drugs that successfully overcome the barrier function of the skin can still fall victim to cutaneous metabolism, thereby producing metabolites that may have increased or decreased activity. This overview discusses the current data and highlights the importance of further studies to evaluate the impact of skin factors in age-related percutaneous penetration.

Adenovirus-mediated transduction of intestinal cells in vivo.

The intestinal tract has many features that make it an attractive target for therapeutic gene transfer. In this study, replication-defective adenoviral vectors were used to explore parameters that may be important in administering gene therapy vectors to the intestine. After surgically accessing the intestine, an E1-, E3-deleted adenoviral vector encoding beta-galactosidase (beta-Gal) was directly injected into various regions of the small and large intestine of rats and rabbits. Significant transduction of the tissue was observed and histochemical staining was used to identify enterocytes as the primary targets of gene transfer. Expression of beta-Gal did not differ substantially when the virus was administered to the duodenum, ileum, or colon. When the vector was directly administered to segments of the distal ileum containing a Peyer's patch, transgene expression was approximately 10-fold higher than in segments lacking a Peyer's patch. In the Peyer's patches, a high level of expression was localized to epithelial cells, potentially M cells, overlying the lymphoid follicle domes. Transduction of these cells could have application in DNA-mediated oral vaccination. Administration of an adenoviral vector encoding a secreted alkaline phosphatase to the lumen resulted in expression and secretion of this gene product into the circulation. This finding demonstrates the potential of enterocytes to serve as heterotopic sites for the synthesis of heterologous gene products that would be secreted into the lumen of the intestinal tract or into the bloodstream.

Prolonged activation of inhibitory somatostatin receptors increases adenylate cyclase activity in wild-type and Gs alpha-deficient (cyc-) S49 mouse lymphoma cells.

Many cells develop enhanced adenylate cyclase activity after prolonged exposure to drugs that acutely inhibit the enzyme and it has been suggested that this adaptation may be due to an increase in Gs alpha. We have treated wild-type and Gs alpha-deficient cyc- S49 mouse lymphoma cells with a stable analogue (SMS 201-995) of the inhibitory agonist somatostatin. After incubation with SMS for 24 h, the forskolin-stimulated cAMP synthetic rate in intact cyc- cells was increased by 76%, similar to the increase found in the wild-type cells. Forskolin-stimulated adenylate cyclase activity in the presence of Mn2+ was also increased in membranes prepared from SMS-treated cyc- cells; however, guanine nucleotide-mediated inhibition of adenylate cyclase activity was not changed despite a small decrease in inhibitory Gi alpha subunits detected by immunoblotting. Pretreatment of cyc- cells with pertussis toxin prevented SMS from inducing the enhancement of forskolin-stimulated cAMP accumulation in intact cells. After chronic incubation of cyc- cells with SMS, exposure to N-ethylmaleimide, which abolished receptor-mediated inhibition of cAMP accumulation, did not attenuate the enhanced rate of forskolin-stimulated cAMP synthesis compared to N-ethylmaleimide-treated controls. These results with cyc- cells demonstrate that an adaptive increase in adenylate cyclase activity induced by chronic treatment with an inhibitory drug can occur in the absence of expression of Gs alpha.

Occupational sensitivity to Tenebrio molitor Linnaeus (yellow mealworm).

Tenebrio molitor is an abundant stored-grain pest in the northern United States. We evaluated an individual with work-related symptoms of rhinoconjunctivitis on exposure to this insect. Prick skin tests with extracts prepared from the larval, pupal, and adult-life stages were positive for the patient and for another individual with allergy to a closely related species of beetle, Alphitobius diaperinus. Specific IgE antibodies to the extracts were demonstrated by RAST. RAST inhibition demonstrated immunologic cross-reactivity between the life stages of T. molitor and also between T. molitor and A. diaperinus, as well as slight cross-reactivity with blowfly. The proteins in the extracts of each life stage were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. More than 15 protein bands were detected in each of the extracts, although the patterns of separation were different for each life stage. After immunoblotting and autoradiography, six different IgE-binding proteins were identified in the larval extract, five in the pupal extract, and seven in the adult extract, with similar IgE-binding patterns noted for the larval and adult extracts. We conclude that this patient developed IgE-mediated sensitivity to T. molitor antigens as the result of occupational exposure. This study confirms the fact that beetles of the Tenebrionid family are potentially significant allergens for workers exposed to grains or grain products.

Allergens involved in occupational asthma caused by baby's breath (Gypsophila paniculata).

Although the floral industry deals with many potential allergens, few examples of occupational asthma exist in this industry. A 22-year-old florist experienced symptoms of rhinitis, conjunctivitis, and asthma on exposure to baby's breath. To determine the contribution of baby's breath to the patient's symptoms, an extract of baby's breath was prepared. Prick skin tests with a 1:10(-5) wt/vol concentration of the extract produced an immediate response, whereas nonexposed atopic and normal control subjects did not react. The patient's asthmatic response to baby's breath was confirmed by bronchial challenge that caused an immediate fall in FEV1 of 26.2% from baseline after inhalation of 88 breath units of the extract. With a direct RAST, the patient's serum bound 38 times the amount of IgE bound by the negative control. IgE binding in the RAST was inhibited by the baby's breath extract but not by unrelated inhibitors (ragweed and tree pollens). Immunoblotting demonstrated IgE binding to 13 protein bands in the extract with molecular weights ranging from 11.5 to 68 kd. Serum from a patient previously reported to have sensitivity to baby's breath recognized five protein bands. Three proteins with molecular weights of 27, 31, and 37 kd were recognized by both patients' sera. We conclude that this patient developed IgE-mediated sensitivity to multiple allergens in baby's breath. This study confirms the importance of this plant as a potential cause of occupational asthma in the floral industry.

Occupational sensitivity to Alphitobius diaperinus (Panzer) (lesser mealworm).

Alphitobius diaperinus is an important beetle in the grain and poultry industries. We evaluated three individuals with work-related symptoms of asthma, rhinitis, conjunctivitis, urticaria, and angioedema on exposure to the insect. Prick skin tests with extracts prepared from the larval, pupal, and adult life stages were positive in all three patients. Specific IgE antibodies to these extracts were demonstrated by RAST or radioimmunoassay. RAST and radioimmunoassay inhibition confirmed the specificity of IgE binding and further demonstrated immunologic cross-reactivity between the three life stages. Peripheral blood leukocytes from two of the individuals demonstrated significant histamine release when they were compared with cells from nonexposed atopic and normal control subjects. The proteins in the extracts of each life stage were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. More than 30 protein bands were detected in each of the extracts; however, the patterns of separation were different for each life stage. After immunoblotting and autoradiography, IgE-binding proteins were recognized by sera from all three individuals in the larval extract at 90 kilodaltons (kd), in the pupal extract at 90, 64, and 38 kd, and in the adult extract at 84 kd. Additionally, several other proteins were identified as being allergenic in some of the patients. We conclude that these three patients developed IgE-mediated sensitivity to A. diaperinus antigens as the result of occupational exposure. To our knowledge, this is the first description of sensitivity to this grain beetle.

Soybean flour asthma: detection of allergens by immunoblotting.

A 43-year-old woman developed asthma 6 years after beginning work in a food-processing plant in which soybean flour was used as a protein extender. Symptoms of sneezing, coughing, and wheezing would begin within minutes of exposure to soybean flour and resolve 2 hours after exposure ceased. Skin tests were positive to a soy extract prepared from the flour. Airway hyperreactivity was confirmed by a positive bronchial challenge to methacholine. Bronchial challenge with soybean flour produced an immediate increase in specific airway resistance from 5.0 to 22.7 L. cm of H2O/L/sec. There was no response to challenge with lactose. The patient's allergic response to soy-flour extract was further characterized by several immunologic methods. IgE binding to soy-flour protein by direct RAST was 5.98 times that of a normal control serum. The soy-flour extract was separated by dodecyl sulfate-polyacrylamide gel electrophoresis. Twenty-four protein bands were detected in the crude soy-flour extract. After immunoblotting and subsequent autoradiography, nine proteins with molecular weights ranging from 54,500 to 14,875 were found. Cross-reactivity studies with other legumes demonstrated apparent immunologic identity between a component in green pea extract and a soybean protein with a molecular weight of 17,000. The clinical significance of this cross-reactivity is not known. We conclude that in this case of occupational asthma to soybean flour, multiple allergens were involved. Immunoblotting may be useful in identifying the allergens involved in occupational asthma.