We have characterised the muscarinic receptor subtypes found in human skin fibroblasts and compared binding levels in cell lines from members of the Alzheimer's disease family with the Swedish amyloid precursor protein (APP) 670/671 mutation. Binding studies with [3H] quinuclidinyl benzilate ([3H]QNB) and the M2/M4 selective antagonist [3H] (+/-)-5,11-dihydro-11-([(2-[(di-propylamino)methyl]-1- piperidinyl]ethyl)amino]carbonyl)-6H-pyrido(2,3-b)(1,4)benzodiazepine-6- one ([3H]AF-DX 384) revealed the presence of a single population of muscarinic receptors on lysed fibroblast membranes. [3H]QNB binding was displaced by a number of selective muscarinic ligands with a rank order of potency: atropine > himbacine > methoctramine > (+/-)-p-fluoro-hexahydro-sila-difenidol hydrochloride > pirenzepine > muscarinic-toxin-3. APP 670/671 mutation carrying cell lines showed 25-35% lower levels of muscarinic receptors labelled with [3H]QNB, [3H]N-methyl scopolamine and [3H]AF-DX 384, compared to controls. This difference was not statistically significant due to large individual variation. It is concluded that muscarinic receptors on adult skin fibroblasts are predominantly of the M2 subtype. Since these cells do not possess M1 and M3 receptor subtypes, they are unlikely to provide a good model for studying muscarinic receptor regulation of APP processing.
Beluga whales have been hunted for food by Native People in the Canadian Arctic since prehistoric time. Here we report the results of analyses of total mercury in samples of liver, kidney, muscle and muktuk from collections over the period 1981-2002. We compare these results with human consumption guidelines and examine temporal and geographic variation. Liver has been analyzed more frequently than other organs and it has been used as the indicator organ. Mercury accumulates in the liver of the whales over time so that the whale ages are usually linked statistically to their levels of mercury in liver. Virtually all the samples of 566 animals analyzed contained mercury in liver at concentrations higher than the Canadian consumption guideline of 0.5 microg g-1 (wet weight) for fish. (There is no regulatory guideline for concentrations in marine mammals in Canada.) Samples from locations in the Mackenzie Delta in the western Canadian Arctic and from Pangnirtung in the eastern Canadian Arctic were obtained more often than from other location and these offered the best chances to determine whether levels have changed over time. Statistical outlier points were removed and the regressions of (ln) mercury in liver on age were used to calculate the level of mercury in whales of age 13.1 years in order to compare age-adjusted levels at different locations. These age-adjusted levels and also the slopes of regressions suggested that levels have increased in the Mackenzie Delta over the sampling period although not in a simple linear fashion. Other locations had fewer collections, generally spread over fewer years. Some of them indicated differences between sampling times but we could not establish whether these differences were simply temporal variation or whether they were segments of a consistent trend. For example, the levels in whales from Arviat were considerably higher in 1999 than in 1984 but we have only two samples. Similarly, samples from Iqaluit in 1994 exceeded considerably those in 1993 and the interval seems too short to reflect any regional temporal trend and more likely represent an extreme case of year-to-year variation. Previous analyses of data from geographically distinct groups had suggested that whales in the western Canadian Arctic had higher levels of mercury than those from the eastern Canadian Arctic. The present analysis suggests that such regional differences have diminished and are no longer statistically significant. No site has indicated significant decreases in more recent samples. The levels of total mercury in the most analyzed organs fell in the order of liver (highest levels), kidney, muscle and muktuk (lowest level). While muktuk had the lowest level of the organs most frequently analyzed, it is the preferred food item from these whales and it still exceeded the consumption guideline in most instances.
Chemical alarm signals in fish are passively released into the water from ruptured epidermal cells, and induce instant fright responses in conspecifics. Fish also display alarm responses to injured heterospecific skin, as well as to scent of piscivorous predators that have ingested prey. A conspicuous alertness to visual disturbances has also been observed in fish following long-term exposure to extracts of filtered, homogenized skin, but the chemical inducers of such vigilance are actually unknown. We tested if a continual exposure to water-soluble alarm signals, from either conspecifics or heterospecifics, affects alertness of fish. Based on previous experience, it was assumed that alertness could be detected following visual disturbances. Naïve crucian carp were initially exposed to the aqueous extracts of centrifuged skin homogenates, from either conspecifics, or from one out of four heterospecific species (tench, perch, Arctic charr, and brown trout). Darting movements, inter-individual distances, and vertical distribution were used to measure behavioral fright responses released by the test stimuli. After seven weeks of continual exposure to the same extracts, behavioral observations were repeated during visual disturbance. Compared with fish that were long-term exposed to skin extracts of tench or charr, crucian carp exposed to extracts of conspecifics, or to extracts of trout or perch, displayed lower inter-individual distances before being visually disturbed. However, no apparent fright responses were observed following such disturbances, and fish that had been continually exposed to conspecific chemical alarm signals displayed feeding behavior. Our results revealed that fish under assumed continual stress, induced by long-term presence of water-soluble alarm cues, only moderately changed their behavioral pattern. This further demonstrates that the aqueous part of extracts from homogenized skin does not contain any causative agents for inducing any conspicuous alertness.
Psoriatic arthritis is a chronic systemic disease in which patients develop persistent inflammation of the skin and joints, leading to disability and joint damage. The extracellular component hyaluronan (HA) plays an important role in regulatory processes such as inflammation, wound healing and tumour progression. At any site of inflammation HA can be depolymerized to low-molecular weight fragments, which, in turn, induce an array of inflammatory mediators that can lead to chronic inflammation. This study describes the serum concentration and dermal distribution of HA, its receptor CD44 and the metalloproteinases 3 and 9 in skin biopsies from patients with different types of psoriatic arthritis. Fifty-one patients with psoriatic arthritis were included in the study and classified as oligo- or poly-arthritic PsA with and without treatment. Biopsies were obtained from both involved and non-involved skin and compared with biopsies from healthy individuals. Serum HA was analysed for estimation of the total turnover of HA. The main findings were an overall redistribution of HA in both involved and non-involved psoriatic skin and an epidermal imbalance between HA and CD44. The structurally and functionally important basement membrane zone was found to be disintegrated and devoid of HA irrespective of the type of articular involvement, treatment or skin affection.
Little data exist on vitamin D deficiency related with intake, especially for the Canadian population. The purpose of this study was to develop and evaluate a food frequency questionnaire (FFQ) with 37 items for rapid assessment of vitamin D intake in healthy young adults of diverse ancestry. We recruited 107 subjects in Southern Ontario during the late winter of 2007 who completed an FFQ twice (FFQ-1 and FFQ-2, repeated for reproducibility assessment) and a 7-day food diary (for validation). Serum 25-hydroxyvitamin D (25(OH)D), the major biomarker of vitamin D nutritional status, and skin melanin were determined. The FFQ results were highly correlated with 7-day diary results and with serum 25(OH)D concentrations (r = 0.529, P
Cholesterol 1,2,3 9 (TM) is being promoted as a non-invasive way to measure cholesterol that has accumulated in a person's skin. The test received a medical device licence from Health Canada in January 2001. It was approved by the U.S. Food and Drug Administration (FDA) in June 2002. This test is not intended to be used as a screening tool for coronary artery disease in the general population. Evidence from non-randomized, non-blinded clinical trials suggests a correlation between higher skin cholesterol levels and the presence of severe coronary arterial lesions. At this point, technical improvements and more robust evidence are required to determine the significance of this technology in clinical practice.
BACKGROUND: Dietary xylitol has been shown to increase the amounts of newly synthesized collagen, and to decrease fluorescence of the collagenase-soluble fraction in the skin of both healthy and diabetic rats. As in diabetic rats, a decreased rate of collagen synthesis and increased collagen fluorescence has also been detected in the skin of aged rats. We hypothesize that dietary xylitol supplementation may protect against these changes during aging. OBJECTIVE: The purpose of the present study was to investigate whether a long-term dietary supplementation can protect against the decrease in the amounts of newly synthesized collagen, and against the increase in fluorescence in the collagenase-soluble fraction in the skin of aged rats. METHODS:Twenty-four male Sprague-Dawley rats were used in the study. After weaning, the rats were divided into 2 groups of 12 animals. The rats in the control group were fed a basal RM1 diet, while the rats in the experimental group were fed the same diet supplemented with 10% xylitol. After 20 months, the rats were killed and pieces of skin from their dorsal areas were excised. The thickness of the samples was measured with a micrometer screw gauge. The collagen contents of rat skin were measured as hydroxyproline, and glycosylation as fluorescent intensity of collagen. Statistical significances of the differences between the groups were determined using the unpaired t test. RESULTS: No general side effects were detected in the rats during the experimental period. The skin of the xylitol-fed rats was a little thicker than that of the control rats. The hydroxyproline content of the acid-soluble fraction was significantly greater in the xylitol group as compared to the controls. However, there were no significant differences in the hydroxyproline content of the collagenase-soluble fraction between the groups. The fluorescence of the collagenase-soluble fraction was significantly smaller in the xylitol-fed aged rats than in the aged rats fed the basal diet. CONCLUSIONS: The results of this study indicate that xylitol caused an increase in the amount of newly synthesized collagen and a decrease in collagen fluorescence in the skin of aged rats.
Microdialysis, a new bioanalytical sampling technique enables measurement of substances in the extracellular space. This initial study investigates the technique's usefulness in the field of percutaneous absorption of solvents, using ethanol as test substance. Microdialysis probes are equipped at the tip with a semi-permeable polycarbonate membrane which permits passive diffusion of substances. Ethanol does not damage the membrane. In vitro recovery for ethanol is good. Probes were inserted via a guide into the skin of the ventral forearm in 7 volunteers. 99.5% ethanol was applied to the skin in excess in a glass reservoir. The probe was perfused at a flow of 1 microliter/min. 50 microliters samples were analysed by gas chromatography. Absorption of ethanol was demonstrated in all subjects. Values from the 9 probes inserted ranged from 10 micrograms/ml to 800 micrograms/ml. The variation may be explained by inter-test or inter-individual variability in ethanol absorption. Individual metabolic capacity may be of importance. The method opens new possibilities in the investigation of skin barrier function in man.
The development of cancer involves epithelial-stromal interactions. Alterations in the synthesis and deposition of type I and III collagens are related to the tumor morphology. Skin carcinogenesis in experimental animals provides a reliable model for the development of neoplasia. Ultraviolet (UV) irradiation is the main etiological factor for epidermal dysplasia and malignant tumors in man, but also for dermal degeneration. Non-neoplastic dermal changes and skin tumors induced by ultraviolet irradiation and 7,12-dimethylbenz(a)anthracene were investigated in various mouse strains with different susceptibilities to tumor formation. UVB irradiation resulted in an increased immunoreactivity of collagens in the dermis, in comparison with 7,12-dimethylbenz(a)anthracene. Increased synthesis and deposition of type I and III collagens were found in the stroma adjacent to benign alterations. In well-differentiated squamous cell carcinomas, a similar induction of collagen synthesis and deposition was observed. The destruction of fibrillary structures was more pronounced during the decrease of differentiation from moderately to poorly differentiated squamous cell carcinomas. Anaplastic carcinomas with spindle cell morphology displayed a delicate meshwork of reticular fibers and collagen III, and abnormal expression of mRNA for collagens in some malignant cells with epithelial characteristics. The underlying stroma reacts to the development of epithelial tumors in a reproducible way, which is related to the carcinogenic agent involved.