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Cystic fibrosis screening by sweat analysis: a critical review of techniques.

Currently employed methods for the assay of electrolytes in sweat, relevant to the diagnosis of cystic fibrosis, are critically evaluated with particular reference to the measurement of electrical conductivity. Attention is drawn to the factors responsible for potentially large errors. On the basis of experimentation, supported by laboratory practice, recommendations are made for the reduction of these errors to clinically acceptable levels. Data concerning sodium and potassium content, electrical conductivity and osmolality of undiluted sweat collections are compared. The feasibility and comparative advantages of measuring sweat osmolality with a vapour pressure osmometer are discussed.

Child

The significance of drug analysis of sweat in respect to rapid screening for drug abuse.

Morphine and methamphetamine, which are excreted in the sweat, are detected by the use of routine serological and physicochemical techniques for urinary examinations. Screening for drug abuse can be done with the same accuracy of that of urine. Rapid excretion of the drug via kidney (within one day) is followed by a slow but steady excretion of the sweat gland. Methamphetamine given orally in a dose of 10 mg is excreted in the sweat at a constant rate (1.4 microgram/ml). No significant difference of the amount excreted by both systems is found. Alveolar lining seems to prevent the elimination of the volatile methamphetamine via respiration. Not only narcotics and stimulants, but also many alkaloids and barbituarates are excreted in the sweat and detected quantitatively by the same principles. The toxicological analysis of the sweat promises a new scope of forensic investigation.

Forensic Medicine

Comparative cytokeratin analysis of sweat gland ducts and eccrine poromas.

Human eccrine sweat gland ducts and benign and malignant eccrine poromas were studied for the expression of various cytokeratins (CK) and vimentin by applying immunoperoxidase and immunofluorescence microscopy to frozen or paraffin-embedded sections, and using two-dimensional gel electrophoresis and immunoblotting. In acrosyringia and dermal eccrine ducts, the luminal cells exhibited intense staining for CKs 1/10/11 and 19. The periluminal cell layers of acrosyringia contained CKs 1/10/11, while CK 5 was absent. In contrast, the basal cell layer of dermal ducts was only positive with the antibody against CK 5, i.e. a pattern resembling that seen in epidermal basal cells. CK 9 was detected only in keratinocytes peripherally surrounding acrosyringia. In benign poromas, gel electrophoresis revealed that CKs 5 and 14 were predominant, with CKs 6, 16 and 17 being minor components. At the immunohistochemical level CKs 1/10/11 and 19 could be further detected with varying frequency in scattered or clustered cells and/or duct-like structures. Occasionally, CK 9-positive cells were observed. Malignant poromas displayed a similar overall gel-electrophoretic pattern. Their immunohistochemical staining patterns were also similar to (albeit rather more variable than) those seen in benign poromas. Our results show that, with respect to their CK expression pattern, the majority of poroma cells resemble the basal cells of both the dermal ducts and the epidermis, while only minor and variable subpopulations acquire features present in ductal/acrosyringial luminal cells that would be indicative of poral differentiation. Thus, the matrix cells of poromas seem to be most closely related to basal cells located at the transition between the glandular epidermal ridge and dermal eccrine duct, being in no way analogous to the cells of the adult acrosyringium above the basal cell level.

Adenoma, Sweat Gland

A simple and disposable sweat collector.

Apart from in cystic fibrosis, where sweat analysis provides valuable diagnostic information, sweat yields remain an overlooked biological fluid. Technical problems (dilution, condensation, contamination, evaporation, etc.) linked to currently available collection procedures are of concern and thwart their use. To overcome some of these technical difficulties, an original sweat-collection technique is described. A collection capsule is created inside a flexible, adhesive and disposable anchoring membrane pasted onto the skin. A fluid-tight window is positioned in the upper part of the pocket and gives access to its content. Through the collection window, complete emptying of the sweat collector can be achieved repeatedly by suction using a vacutainer tube inserted in a tube holder equipped with a long dull needle. With prior addition of a suitable marker, fractional samplings can also be performed using a precision micropipette. This collecting method allows for kinetic studies on sweat rate and sweat content. The limited bias-inducing manipulations linked to the described technique, coupled with the ease of performing kinetic studies on sweat volume and content, make this original tool a reliable and accurate sweat-collection technique.

Adolescent

A modified ion-selective electrode method for measurement of chloride in sweat.

A modified method of analysis of sweat chloride concentration with an ion-selective electrode is presented. The original method of sweat chloride analysis proposed by the Orion Research Corporation (Cambridge, Massachusetts 02139) is inadequate because it produces erratic and misleading results. The modified method was compared with the reference quantitative method of Gibson and Cooke. In the modified method, individual electrode pads are cut and placed in the electrodes rather than using the pads supplied by the company; pilocarpine nitrate (2,000 mg/l) is used in place of pilocarpine HCl (640 mg/l); sodium bicarbonate as the weak electrolyte is used instead of K2SO4. A 10-minute period for sweat accumulation is employed rather than a zero-time collection as in the original Orion method. The modification has been studied for reproducibility in individuals, reproducibility between right and left arm in individuals; it has been compared extensively with the quantitative method of Gibson and Cooke, both in normal individuals and in patients with cystic fibrosis. There is excellent agreement between the modified method and the quantitative reference method. There appears to be a slight bias toward higher concentrations of chloride from the right arm compared with the left arm, but this difference is not medically significant.

Adolescent

Techniques for microdrop analysis of fluids (sweat, saliva, urine) with an energy-dispersive X-ray spectrometer on a scanning electron microscope.

Volumes of 10(-10) liter of sweat, saliva, and urine were analyzed with an energy-dispersive spectrometer (EDS) X-ray analyzer on a standard scanning electron microscope (SEM). Results of the EDS analysis of diluted and undiluted samples were verified by comparison with results of analyses of the same sample prepared by conventional quantitative methods. Except for Cl in concentrated urines and Mg in low concentrations, good agreement was found between the methods. The analysis provides quantitation of most elements of biological interest present in concentrations of about 1 mM, while it demonstrates good linearity (r greater than 0.99) throughout a wide range of commonly encountered biological concentrations. Reproducibility of the analysis is on the order of 2% and the minimal determinable concentration is generally between 0.5 and 1.0 mM for S, P, K, and Ca and slightly more than 1.0 mM for Mg.

Body Fluids

Cytokeratin expression in mucinous sweat gland carcinomas: an immunohistochemical analysis of four cases.

Four mucinous sweat gland carcinomas were examined for the distribution of cytokeratin (CK) polypeptides using immunohistochemical techniques on paraffin-embedded sections. All the tumour specimens reacted with monoclonal antibodies to CK 7, CK 8, CK 18 and CK 19. Antibodies to CK 1, CK 1/2/10/14, CK 1/5/10/11, CK 13, CK 14 and CK 20 did not stain any of the carcinomas. The results add additional support to the notion that mucinous sweat gland carcinoma represents a tumour histogenetically related to the eccrine secretory coil. Furthermore, the absence of CK 20 might significantly contribute to the differentiation of this tumour from cutaneous metastases from gastrointestinal carcinomas.

Adenocarcinoma, Mucinous

Analysis of the proteins in sweat and urine by agarose-gel isotachophoresis.

A sensitive and practical method is described for the analysis of the proteins contained in human sweat and urine which does not require pre-concentration of the sample. Technical details are provided of the agarose-gel isotachophoresis and the proteinograms of normal and pathological urine samples, as well as proteinograms of human sweat. The method can also be applied as an electro-concentration system in a field-strength gradient. By means of this electro-concentration system, and in combination with immunodiffusion against monospecific antisera, a detection limit of albumin of 50 ng/ml has been obtained.

Albumins

Autonomic dysfunction in the Landry-Guillain-Barre syndrome.

Tests of autonomic function have been performed on 7 patients with the Landry-Guillain-Barre syndrome and 5 control subjects. These included a sweat test, and measurement of both postural hypotension and baroreflex sensitivity. Sweating was definitely abnormal in 5 patients. Postural hypotension was present in 4 patients. The baroreflex sensitivity was significantly reduced (p less than 0.01) in the patients when compared with the control group. The results demonstrate that both the sympathetic and parasympathetic nervous systems may be affected in the Landry-Guillain-Barre syndrome.

Adult

Women with climacteric symptoms: a target group for prevention of rapid bone loss and osteoporosis.

The relations of vasomotor symptoms to the rate of bone loss and to the response of forearm bone mineral density (BMD) to hormone replacement therapy (HRT) were analyzed in a 2-year non-randomized study. Forty peri/postmenopausal women who were given HRT for climacteric symptoms were compared with untreated control women, individually matched for age and length of time since the last menstrual period. The women who received HRT gained, on average, about 2% in BMD, while the control women lost about 6% (mean group difference 8%; 95% confidence interval (CI) 5.7-10.2). Adjustment for potential confounders did not change the results. Sweating frequency was inversely correlated with serum estradiol levels (p = 0.05). Among untreated women the rate of bone loss was higher in those who had frequent sweating initially than in those with less frequent sweating (9% vs. 4%, mean difference 4.3%; 95% CI 0.7-7.8, p = 0.023). Among women who received HRT, those who had the highest frequency of sweating initially, compared with those with a lower frequency, showed a greater gain in bone density (mean difference 4%; 95% CI 1.2-6.8, p = 0.007). In multivariate analysis adjusting for covariates, sweating frequency remained an independent determinant of change in bone density in women both with and without HRT. When sweating frequency and serum estradiol levels were compared in a multivariate analysis, only sweating frequency showed an independent association with rate of bone loss. The findings indicate that women with severe climacteric symptoms may have an excessive rate of bone loss and should therefore be considered as a special target group for prevention of osteoporosis.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged

[Electrophysiological analysis of the functions of the sweat gland fibers in various diseases].

The authors relate the data on vegetative sweating fibers studied by the method of evoked skin sympathetic potentials in patients with diabetes mellitus, Charcot-Marie atrophy, spinal amyotrophy, Guillain-Barre syndrome, essential hyperhidrosis, and in those with Raynaud's phenomenon and thyrotoxicosis. A remarkable decrease of conduction by sweating fibers was revealed in patients with diabetes mellitus, Charcot-Marie atrophy and Raynaud's phenomenon; a moderate decrease was shown by the patients with Guillain-Barre syndrome and by those with spinal amyotrophy. The conclusion is made about the high information content of the method.

Adult

Roles of Ca and cAMP on C1 channel activity in cystic fibrosis sweat clear cells as studied by microsuperfusion and cell volume analysis.

In an attempt to study regulation of C1 channels in intact sweat secretory coils in cystic fibrosis and controls in the least invasive manner, isolated secretory coils were superfused with various drugs and K-efflux was determined as an indirect measure of C1 movement. C1 channel activity was also determined from the drug-induced cell volume increase in gramicidin (GC)-treated dissociated eccrine clear cells. We observed that while MCh-induced K-efflux from the CF secretory coils was entirely normal, K-efflux in the presence of isoproterenol (ISO), forskolin (FK), or IBMX was absent in CF, suggesting that these agents failed to stimulate C1 movement. C1 channel activity of dissociated CF clear cells, as studied by cell volume analysis, was entirely normal when stimulated by Ca-elevating agents but was defective when stimulated by cAMP-elevating agents. TPA (phorbol ester) does not appear to stimulate C1 channel activity nor does it modify the effect of other agents. The following observations from the present and previous studies are not necessarily consistent with the traditional thesis that the observed C1 movement is due to cAMP: CT-cAMP had no effect on cell swelling or on K-efflux; ISO is more potent in accumulating tissue cAMP than IBMX yet the latter is more potent in stimulating K-efflux; IBMX increases cytoplasmic [Ca] yet is unable to stimulate K-efflux in CF; K-efflux stimulated by cAMP-elevating agents was inhibited by removal of Ca from the bath; and, cell swelling of GC-treated cells in response to cAMP elevating agents was inhibited by removal of Ca. The inability of IBMX to stimulate C1 channels in the face of elevated cytoplasmic [Ca] and cAMP in CF cells deserves further scrutiny.

1-Methyl-3-isobutylxanthine

Changes in cholinergic sweat gland activation in diabetic neuropathy identified by computerised sweatspot analysis.

Peripheral small-fibre denervation has been reported to result in decreased activation of eccrine sweat glands to muscarinic cholinergic agents. Using computerised image-analysis of pilocarpine-activated sweatspot prints of a 4 cm2 area of the dorsum of the foot in 79 randomly selected diabetic patients we have identified a group of neuropathic patients (18%) with decreased sweatspot activation (less than 20/cm2), and a smaller group (6%) of younger patients with less marked neuropathy who had increased activation (greater than 132/cm2), probably resulting from denervation supersensitivity. The associations between sweatspot density and other conventional tests of peripheral nerve function were weak. The prevalence of abnormal sweatspot density, 24%, was similar to that of other tests, except thermal thresholds at the feet (35-37%), which were not correlated with sweatspot activation, suggesting that diabetic neuropathy has differing effects on afferent and efferent small fibres. The method is rapid and reproducible (median coefficient of variation 14%) and its ability to identify patients with increased, as well as decreased, peripheral nerve function may be of value in the characterisation and longitudinal follow-up of small-fibre abnormalities in diabetes.

Adult

Monoclonal antibody analysis of keratin expression in carcinomas of sweat glands.

Characteristics of keratins of five carcinomas of sweat gland origin were immunohistochemically investigated with several antikeratin monoclonal antibodies with differing specificities. Specimens were obtained from two cases of mucinous carcinoma of the skin, two cases of classic type of eccrine adenocarcinoma, and a case of eccrine porocarcinoma. The tumor cells of mucinous carcinoma expressed only simple epithelial keratins. In a case of eccrine adenocarcinoma, simple epithelial keratin 19 was diffusely expressed. The expression of the other simple epithelial keratins was confined to the luminal cells, whereas the remaining tumor cells further expressed stratified epithelial keratins. Eccrine porocarcinoma and a second case of eccrine adenocarcinoma did not express simple epithelial keratins, although stratified epithelial keratins were diffusely expressed. These data suggest that carcinomas of sweat glands express various combinations of simple and stratified epithelial keratins. Development of additional data along these lines may help to further define their classification.

Adenocarcinoma

Christ-Siemens-Touraine syndrome--a clinical and genetic analysis of a large Brazilian kindred: II. Affected males.

We describe 13 males with Christ-Siemens-Touraine syndrome from one family. History and examination were supplemented by three sweat tests and dermatolglyphic analysis. Some of the patients had two uncommon findings (onychodystrophy and excessive lacrimation), and five had an "incomplete" form of the syndrome. Four signs (distal phalanges of fingers and toes radially and tibially deviated, respectively; facial hypochromic spots; large occipitofrontal circumference) seem to be described for the first time. The segregation proportion in the sibships with at least an affected male was found to be normal (1:1) in 44 series of data (43 from the literature), where a high ascertainment bias was present (155 affected and 68 normal males).

Adolescent

Christ-Siemens-Touraine syndrome--a clinical and genetic analysis of a large Brazilian kindred: I. Affected females.

A total of 27 women of a Brazilian kindred are described as having one or more signs of the Christ-Siemens-Touraine syndrome. The history and physical examination were supplemented by four sweat tests and dermatolglyphic analysis. It is suggested that this syndrome has two forms -- a major form (in males) and a minor one (in females). Two signs verified in some of our patients (mosaic patchy distribution of body hair and radial deviation of distal phalanges of index fingers) seem to be here described for the first time. A review of the literature shows a corrected sex ratio between 1 M: 1.21 F and 1 M: 2.38 F among affecteds. Since the manifestation rate of the gene among carriers was estimated at about 0.70, the actual sex ratio is expected to be not lower than 1 M: 1.40 F. Contrary to a general opinion, affected females outnumber affected males.

Adult