Effects of pilocarpine on salivary flow in patients with Sj?gren's syndrome.

Pilocarpine, a muscarinic-cholinergic agonist drug, has been reported to stimulate salivary flow in patients with salivary gland dysfunction. Previous studies involved heterogeneous groups of patients with salivary gland dysfunction and examined the short-term, single-dose, tablet form of pilocarpine. In this single-blind, placebo-controlled study we examined the long-term effects of pilocarpine administration on patients with definitively diagnosed Sj?gren's syndrome (SS). Nine subjects with SS who received pilocarpine, and nine age- and sex-matched SS control subjects who received a placebo, participated. Baseline predosing sialometric and clinical data were obtained for all subjects. The study group used 2% pilocarpine as a liquid ophthalmic drop preparation, four drops three times per day, for 6-weeks. Identically appearing placebo solution with the same dosing schedule and duration was used for the control subjects. Sialometric and clinical examinations were performed. The results indicated a significant overall increase in both whole unstimulated salivary flow (0.15 +/- 0.03 ml/min in study subjects vs 0.02 +/- 0.001 ml/min in control subjects; p less than 0.001) and parotid stimulated salivary flow (0.14 +/- 0.04 ml/min in study subjects vs 0.009 +/- 0.002 ml/min in control subjects; p less than 0.001) in the pilocarpine group as compared with the placebo group. The results of this study support the use of pilocarpine to increase salivary flow in patients with SS.     

s-IgA and cytokine levels in whole saliva of Sjögren's syndrome patients before and after oral pilocarpine hydrochloride administration: a pilot study

Previous investigations have found elevated levels of s-IgA in the parotid saliva and normal levels in submandibular saliva of patients with Sjögren's syndrome (SS). Fox et al. also found elevated levels of cytokines (i.e., IL-2 and IL-6) in serum, salivary epithelial cells and parotid saliva of patients with SS. The oral administration of pilocarpine hydrochloride stimulates whole and parotid salivary flow. The purpose of this study was to determine the levels of s-IgA and IL-2 and IL-6 in whole saliva before and after administration of pilocarpine hydrochloride in SS subjects. Ten definitively diagnosed SS subjects were enrolled in the study, as were ten controls (C). The mean age was 57.2 years and all subjects were female. Whole unstimulated saliva (WUS) was collected by standard techniques for 5 min, after which the volume and flow rate were determined (mean WUS: SS = 0.047 vs C = 0.480 ml/min). Samples were centrifuged and the immunoglobulin analysis performed on the supernatants by immunoreactivity in a double-sandwich technique as previously described by Rudney et al. Cytokine analysis was performed similarly utilizing commercially available kits from R&;D Systems. The results as analyzed by pairwise t-tests revealed comparable levels of s-IgA in the saliva of the SS patients, as compared to controls at baseline (means±SEM: SS-IgA = 348.1±82.0 vs C-IgA = 284.0±65.1 μg/ml; NS ). Whole salivary flow was significantly increased (328%) in the SS subject group 60 min after the administration of 5 mg pilocarpine hydrochloride (means±SEM: 0.0472±0.017 vs 0.1546±0.054 ml/min; P<0.01). There was no significant change in the concentration of s-IgA in the SS subject group following the pilocarpine dose (means±SEM: SS-IgA = 439.9±121.2 μl/ml; P = NS). There were elevated levels of IL-2 in the saliva of four out of the ten and IL-6 in two out of the ten SS patients, as compared to controls (means±SEM: SS-IL-2 = 127.8±11.4 vs C-IL-2 = 30.8±1.6 pg/ml and SS-IL-6 = 41.4±7.1 vs C-11.6 ± 2.8 pg/ml). There was also a significant decrease in the concentration of IL-2 in the same four out of ten SS subjects following the pilocarpine dose (means±SEM: SS-IL-2 = 32.4±10.3; P<0.01). These preliminary results indicate that s-IgA levels do not change with increased salivary flow following the administration of pilocarpine hydrochloride in patients with Sjögren's syndrome. While cytokines are elevated in the whole saliva of some SS patients, a decrease in IL-2 concentration may occur with increased salivary flow.     

Punch-biopsy of minor salivary glands in the diagnosis of Sjögren's syndrome

Abstract— The aim of the present investigation has been to examine whether the histopathologic changes in the minor salivary glands in patients with Sjögren's syndrome are of the same nature as the changes in the major salivary glands. Punch-biopsies were taken from the lower labial mucosa in 14 patients; 12 of these had a verified Sjögren's syndrome. In 10 patients changes in accordance with those of a benign lymphoepithelial lesion were present in the minor salivary glands. The technique was found valuable in the diagnosis of general disorders with salivary gland involvement.     

Salivary gland function and changes in patients with oral lichen planus

Abstract – Saliva analysis, sialography and histopathologic examination of labial salivary glands were performed on patients with oral lichen planus. Diseases connected with salivary gland function were also recorded. Saliva analysis regarding secrection rate, pH and buffer capacity in unstimulated and stimulated saliva was permormed on 39 patients. 87% of the patients exhibired a low or very low unstimulated secretion rate, the mean value being 0.14 ml/min. The rate of stimulated saliva, pH and buffer capicity did not deviate from normal reference values. Sialographic examination was performed on 18 patients, corresponding to 36 major salivary glands. Radiologic changes were seen in 89% patients. Histopathologic examination was performed on 15 patients. Lymphocytic infiltration, acinar atrophy, fibrosis, fatty degeneration or ducral changes were observed in the minor glands of all patients. Different degrees of acinar atrophy were present in 93% of the patients. Lymphorytic infiltration was seen in 12 patients (80%) of whom three exhibited focal accumulation as in Sjögren's syndrome. Since decreased salivary secretion and symptoms of joint diseases and keratoconjunctivitis sicca were frequently present, over a third of the patients showed clinical signs comparable to those of Sjögren's syndrome. A high frequency of gastrointestinal and endocrine diseases was also recorded, which suggests that a general exo and endocrine influence may be present in patients with oral lichen planus.     

The level of cariogenic microlorganisms in patients with Sjögren's syndrome

Sixteen patients with caries-inacthe Sjögren's syndrome with low parotid salivary flow rates (≤ 0.25 mL/min) and 18 caries-inactive control subjects with higher salivary flow rates were compared. Mutans streptococci (MS) and lactobacilll (LB) counts were measured by means of Dentocult® SM strip mutans and LB assays. The group with Sjögren's syndrome displayed higher counts of MS (P = 0.014) and LB (p = 0.003) when compared with controls. The results of this study indicate that patients with caries-inactive Sjögren's syndrome and low salivary flow may have higher colonization of cariogenic mlcro-organisms than healthy individuals.     

Analysis of the concentration and output of whole salivary constituents in patients with Sjögren's syndrome

In Sjögren's syndrome, salivary glands are affected, resulting in a diminished salivary flow. In the present study, the protein composition, sialic acid content and the amounts of calcium and phosphate of stimulated whole saliva from 43 patients with Sjögren's syndrome, were compared with those of control saliva samples from 17 healthy subjects. The absolute concentrations of albumin, cystatin C. cystatin S. total IgA and total protein, but not amylase, were increased significantly in both primary and secondary Sjögren's syndrome. The output/min of total protein, albumin, amylase, and IgA was, however, decreased in Sjögren patients. These results suggest that the diminished output of salivary defence factors, rather than their absolute concentrations, may be related to the oral health problems seen in Sjögren's syndrome patients.     

Minor salivary gland immunohistology in the diagnosis of primary Sjögren's syndrome

Background:  Focal lymphocytic infiltrates of minor salivary glands are considered target-organ related signs of Sjögren's syndrome. The percentages of plasma cells expressing IgA, IgG and IgM in minor salivary gland biopsies have also been suggested as useful in establishing a diagnosis of Sjögren's syndrome, and this study aimed at evaluating this method.
Methods:  All biopsies from patients under investigation for Sjögren's syndrome ( n  = 210) at our department during 4 years were analyzed for IgA, IgG and IgM producing cells by immunohistochemistry, and related to Sjögren classification parameters.
Results:  A focus score ≥1 was observed in 67/210 patients and the frequency of IgA producing cells was <70% in 42/210 patients. Sufficient clinical data for classification of disease were available for 57/210 patients. Patients were classified as having primary Sjögren's syndrome (pSS) ( n  = 9), secondary Sjögren's syndrome (sSS) ( n  = 12) or non-Sjögren's syndrome (non-SS) ( n  = 36). IgA expressing cells were significantly decreased ( P  < 0.01) and IgG expressing cells significantly increased ( P  < 0.02) in patients with pSS compared to non-SS. Also, increased numbers of salivary gland IgG producing plasma cells correlated with increased IgG serum levels ( P  < 0.001). However, there was no significant difference between sSS and non-SS with regard to IgA, IgG or IgM expressing cells in the glands.
Conclusions:  Our results support previous reports indicating the relevance of quantitative evaluation of Ig isotype expression in plasma cells in the clinical investigation of Sjögren's syndrome and further indicate a difference in plasma cell populations between pSS and sSS.     

Effect of 0.1% pilocarpine mouthwash on xerostomia: double‐blind,randomised controlled trial

The objective of this study was to evaluate the effect of 0.1% pilocarpine mouthwash in xerostomic patients. Sixty volunteers were randomly allocated to two groups. The experimental group used 0.1% pilocarpine solution, and the control group used 0.9% saline. The short‐ and long‐term effects of pilocarpine were investigated by measuring the severity of oral dryness, minor salivary flow rates and unstimulated whole salivary flow rate at predetermined times. The severity of oral dryness was decreased in both groups at 0, 30 and 60 min after mouthwashing, with no significant difference between the groups. Buccal and labial secretions were increased in both groups, but only the experimental group exhibited increased palatal secretion. Labial and palatal secretions, but not buccal secretion, differed between the groups. The unstimulated whole salivary flow rate was increased in the experimental group and differed from that in the control group. After 4 weeks, the severity of oral dryness was decreased in both groups and did not differ between them. The oral dryness at night or on awakening significantly decreased in both groups, with no significant difference between them, but the oral dryness at other times of the day and the difficulty in swallowing foods were not significantly changed in both groups. Minor salivary and unstimulated whole salivary flow rates did not increase in both groups. Until 1 h after mouthwashing, 0.1% pilocarpine mouthwash increased minor salivary and unstimulated whole salivary secretions, but was not superior compared with 0.9% saline at relieving subjective oral dryness.     

Natural history of HIV-associated salivary gland disease.

To describe the natural history of HIV-associated salivary gland disease, which is characterized by enlarged major salivary glands and/or xerostomia in HIV-infected persons, we assessed 22 patients at an initial and follow-up examinations (median span of examinations, 15 months). Sixteen patients (73%) had bilateral parotid gland enlargement, 17 had symptoms of dry mouth, and 11 had both conditions. Parotid gland enlargement remained unchanged in 10 patients, it progressed in 2, and it regressed in 4 during treatment with zidovudine or steroids. Those patients with parotid gland enlargement had a significantly lower mean stimulated parotid flow rate (0.27 ml/min/per gland) than a control group of HIV+ persons without salivary gland disease (0.48 ml/min/per gland) (p less than 0.05), whereas the mean unstimulated whole salivary flow rates did not did not differ significantly between the two groups. The mean salivary flow rate of the study group did not change during the observation period. When HIV-associated salivary gland disease was diagnosed, 5 patients (23%) had AIDS, and at follow-up 10 (46%) had AIDS. Seven of these had Kaposi's sarcoma. The mean peripheral blood CD4 cell count was 280 and 225 per mm3 at the initial and follow-up examinations, respectively. The corresponding CD8 counts were 1138 and 900. The pathogenesis of HIV-associated salivary gland disease may include hyperplasia of intra-parotid lymphoid tissue. Because HIV-associated salivary gland disease can clinically resemble Sj?gren's syndrome, the differential diagnosis of bilateral parotid enlargement should include HIV infection.     

Cytokines in Sjögren's syndrome

Cytokines play a central role in the regulation of immunity and are often found to be deregulated in autoimmune diseases. Sjögren's syndrome is a chronic autoimmune disease characterized by inflammation and loss of secretory function of the salivary and lachrymal glands. This review highlights the current knowledge of the expression and the function of pro- and anti-inflammatory cytokines both locally and systemically in Sjögren's syndrome patients. In the salivary glands, saliva and serum of these patients, many pro-inflammatory cytokines are upregulated. Concomitantly, most anti-inflammatory cytokines are not detectable or are expressed at low levels. Besides a role in inflammation, cytokines are also thought to be involved in salivary gland dysfunction by directly interfering with the epithelial cells in the glands. Future research on the role of novel cytokines in Sjögren's syndrome in combination with a better understanding of the effect of cytokines on exocrine dysfunction will aide the identification of the best therapeutic targets for Sjögren's syndrome.     

Innervation pattern and Ca2+ signalling in labial salivary glands of healthy individuals and patients with primary Sjögren's syndrome (pSS)

Abstract: We have characterised the innervation pattern and intracellular Ca²⁺-signalling in labial salivary glands (LSG) of 16 patients with primary Sjögren's syndrome (pSS) and 27 healthy controls. Numerous immunoreactive nerve fibers (IRF) containing vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase activating peptide (PACAP) were found around acini, ducts and blood vessels. Substance P (SP)-, neuropeptide Y-, tyrosine hydroxylase- and nitric oxide synthase-IRF were mainly surrounding ducts and blood vessels. The majority of pSS patients had inflamed LSG and the presence of focal lymphocytic infiltrates (FI) were more frequent and pronounced as compared with healthy controls. In areas with normal or diffusely inflamed LSG tissue, pSS patients demonstrated the same distribution of IRF as healthy controls with similar histology. However, IRF were absent in central areas of FI both in pSS and age-matched healthy controls. Although all pSS patients had hyposalivation, stimulation with acetylcholine, norepinephrine, phenylephrine, isoproterenol, VIP, PACAP, SP, adenosine 5'-triphosphate and uridine 5'-triphosphate induced the same increase in the intracellular free Ca²⁺ concentration in LSG acini from both pSS patients and healthy controls, indicating the presence of functional receptor systems in vitro .     

Management of dry mouth in Sjögren's syndrome

The management of dry mouth is essential for patients with Sjögren's syndrome. The symptomatic treatment has included using air humidifiers, rinsing the mouth with water or mouthwash, the application of a salivary substitute and administration of secretagogues. There are three secretagogues suitable for the alleviation of dry mouth in Sjögren's syndrome patients in Japan; cevimeline hydrochloride hydrate (cevimeline), pilocarpine hydrochloride, and anetholtrithione. A relationship between the effect of cevimeline on saliva secretion and the degree of salivary gland destruction evaluated by sialography and histopathological findings in the labial minor salivary glands has been reported. These diagnostic approaches could provide useful prognostic information on the efficacy of cevimeline in Sjögren's syndrome patients. Concomitantly, a bite guard was suggested as an effective lubricating device because it maintains the lubricants in the proper location. In addition, the management of the complications of dry mouth, such as tooth caries, periodontitis and oral candidiasis, which all lead to a reduction in the QOL, is also important. Both the prevention and treatment of erythematous candidiasis is especially important in the management of Sjögren's syndrome.     

Histologic lesion in labial salivary glands of patients with systemic lupus erythematosus

In an attempt to evaluate the presence or incidence of Sj?gren's syndrome in patients with systemic lupus erythematosus (SLE), 72 randomly selected patients with SLE, regardless of sicca manifestations, underwent labial salivary gland biopsy. Thirty-seven patients (51%) had normal labial salivary gland tissue. Seventeen patients (24%) had a mild perivascular infiltration of 15 to 20 lymphocytes per focus. In the remaining 18 patients (25%) heavy infiltration of more than 50 lymphocytes per focus was observed. From the latter group five specimens showed perivascular distribution of the infiltrates. About 40% of patients with any infiltration had keratoconjunctivitis sicca. Patients with severe lymphocytic infiltration in minor salivary gland tissue rarely had kidney involvement and often had lymphadenopathy and circulating rheumatoid factor, cryoglobulins, and antibodies to SS-A (Ro) and SS-B (La) antigens. Vasculitis was not found more frequently in any group of patients. Our results suggest that perivascular lymphocytic infiltrates in the labial salivary glands of patients with SLE may be the initial histologic lesion of Sj?gren's syndrome.     

The effect of parotid salivary flow rate on the levels of salivary antimicrobial proteins in patients with Sj?gren's syndrome.

OBJECTIVE: The purpose of this study was to examine the effect of salivary flow rate on the levels of antimicrobial salivary proteins in 24 patients with Sj?gren's syndrome and 22 age- and race-matched healthy control subjects. METHOD AND MATERIALS: Parameters examined included stimulated salivary flow rate, total salivary protein, lactoferrin, lysozyme, amylase, and secretory immunoglobulin A. RESULTS: The mean total salivary protein and the mean salivary amylase were significantly greater in patients than in controls. However, no significant difference was observed in the mean stimulated salivary flow rates or the levels of lactoferrin, lysozyme, or secretory immunoglobulin A of patients and controls. To examine the effect of salivary flow rate on the levels of salivary antimicrobial protein, the levels of these proteins in patients with salivary flow rate of < or = 0.3 mL/min per gland were compared to those in healthy controls with salivary flow rate > or = 0.4 mL/min per gland. Analyses showed the levels of lactoferrin to be significantly higher among patients than among controls. CONCLUSION: The levels of salivary amylase and lactoferrin may be influenced by the levels of salivary output in patients with Sj?gren's syndrome. The relationship between salivary flow rate and the levels of amylase and lactoferrin is not clear at the present time.     

Salivary gland involvement in autoimmune thyroiditis, with special reference to the degree of association with Sj?gren's syndrome.

From a total of 63 patients with autoimmune thyroiditis, 19 cases were further investigated to determine the degree of concomitant morphologic and functional salivary gland changes. For comparison, 21 of a total of 28 cases of primary Sj?gren's syndrome were also examined. Of the 19 cases of autoimmune thyroiditis, 11 showed various degrees of salivary gland involvement on the basis of an analysis of lower lip salivary gland biopsy specimens, scintigraphy of the parotid, and unstimulated whole sialometry. Six of these cases fulfilled the criteria of primary Sj?gren's syndrome. A remarkably high proportion of dark-staining acini was observed in the lower lip biopsy specimens of our patients with thyroiditis (8 of 19, 42%) and less among our patients with primary Sj?gren's syndrome (5 of 21, 24%). We conclude that significant involvement of salivary glands may occur in cases of autoimmune thyroiditis, which indicates that common mechanisms may frequently be operative in the development of thyroid and salivary gland immune disease.     

The prevalence of xerostomia and salivary gland hypofunction in a cohort of HIV-positive and at-risk women

The association of xerostomia and salivary gland hypofunction with HIV infection has been established for men but not for women. We investigated the prevalence of these conditions in a national cohort (n = 733) of HIV-positive and at-risk HIV-negative women. Participants in this prospective cross-sectional study were recruited from the Women's Interagency HIV Study (WIHS) at five outpatient USA clinics. Xerostomia was assessed based on "yes" responses to a dry-mouth questionnaire. Samples of unstimulated whole and chewing-stimulated whole saliva were collected under standardized conditions. The major salivary glands were also evaluated clinically. The prevalence of dry-mouth complaint, the absence of saliva upon palpation, and zero unstimulated whole saliva (flow rate = 0 mL/min) were significantly (p = 0.001) higher in HIV-positive women. Adjusted odds of zero unstimulated whole saliva were significantly (p = 0.02) higher in HIV-positive women vs. HIV-negative women (OR = 2.86; 95% CI, 1.23 to 6.63). Significant (p = 0.03) univariate association was found between zero unstimulated whole saliva and CD4 counts. Adjusted odds of zero unstimulated whole saliva were significantly (p = 0.02) higher for HIV-positive women with CD4 < 200 compared with those with CD4 > 500 (OR = 2.61; 95% CI, 1.17 to 5.85). Chewing-stimulated flow rates were not significantly different between seropositive and seronegative women. The prevalence of xerostomia and salivary gland hypofunction appears to be significantly higher in HIV-positive women relative to a comparable group of at-risk seronegative women. Immunosuppression levels measured by CD4 cell counts are significantly associated with xerostomia and salivary gland hypofunction in a population of HIV-positive women.     

Anti-salivary antibodies in primary Sjögren's syndrome

Earlier studies have described an antibody that recognized salivary ductal epithelium in sera from 15–50% of patients with primary Sjögren's syndrome; however, the specific salivary antigen in those studies was not identified. The present study further investigated this unknown salivary antigen. Twenty-nine of 31 patients (94%) with primary Sjögren's syndrome demonstrated IgG antinuclear antibodies that bound to an epithelial cell line with ductal characteristics derived from a human salivary gland. Seventy-seven percent of these patients had serum antibodies that bound to ductal cells of normal human parotid tissue after formalin fixation. Western blots of cell extracts, immunofluorescence, and adsorption studies indicated that SS-A/Ro and SS-B/La were the antigens recognized in the salivary cell line. The pattern of fluorescence seen when anti-SS-B/La bound to normal parotid tissue was identical to the fluorescence pattern of the anti-salivary ductal antibodies described in earlier literature.     

Pilocarpine for the treatment of salivary glands' impairment caused by radioiodine therapy for thyroid cancer

OBJECTIVES: To study the effect of single-dose pilocarpine hydrochloride 5 mg on the whole unstimulated and stimulated salivary flow in patients suffering from thyroid cancer treated with radioiodine therapy, and to investigate the changes in vital signs during the treatment. SUBJECTS AND METHODS: Five such patients were referred with complaints of dry mouth, rampant caries, and pain in the parotid gland region or history of chronic recurrent suppurative sialodenitis. A single dose of 5 mg pilocarpine hydrochloride was administered to each patient and blood pressure, heart rate, body temperature and salivary secretion rate were monitored at 1, 2 and 3 h. RESULTS: A significant elevation of unstimulated and stimulated saliva flow rate was observed in four patients without significant alteration of the monitored vital signs. CONCLUSIONS: Treatment with pilocarpine hydrochloride may be beneficial in the case of impaired salivary function in patients treated with radioiodine.     

Analysis of residual saliva and minor salivary gland secretions

Residual saliva and minor salivary gland secretions are important for the maintenance of oral mucosal wetness. Salivary proteins and glycoproteins are the major components of the oral mucosal film, which functions as a moisture retainer and a protective barrier. Here, the correlations between the amounts of residual saliva and minor salivary gland secretions and their protein concentrations were investigated in 30 normal healthy individuals. The thickness of the mucosal film was measured at six mucosal surfaces and minor salivary gland secretion rate was measured at two mucosal surfaces. The thickness of residual saliva was determined by placing filter-paper strips against the mucosa at each site for 5 s and then measuring the volume electronically with a Periotron 8000 micro-moisture meter. The unstimulated rate of minor salivary gland secretion was measured for 30 s by the same method. Unstimulated whole salivary flow rate was measured with the spitting method. The total protein concentration of all salivary samples was measured by bicinchoninic acid assay. Before the experiment, the intra-/inter-examiner reliability of the method using the Periotron and the filter-paper strips was investigated. With a range of 0.4256-0.8846, the intraclass correlation coefficient, measured within and between examiners, was indicative of good reliability. The oral mucosal site with the thinnest coat of residual saliva was the anterior hard palate. Mucosal wetness on the hard palate and buccal mucosa showed significant positive correlations with the unstimulated whole salivary flow rate and significant negative correlations with the total protein concentration of residual saliva. Mucosal wetness on the upper and lower labial mucosa also showed significant negative correlations with the total protein concentration of residual saliva. Mucosal wetness on the soft palate was correlated with the minor salivary gland secretion rate (r=0.477, P<0.01). Among the minor salivary glands, the secretion rate of soft palate glands in females showed a significant correlation with the unstimulated whole salivary flow rate (r=0.563, P<0.05) and a significant negative correlation with its total protein concentration (r=-0.525, P<0.05). These data suggested that oral mucosal wetness and minor salivary gland secretions could be influenced by various factors differently according to mucosal sites.     

Study of saliva secretion and the salivary fluoride concentration of the human minor labial glands by a new method

Unstimulated and stimulated flow rate from minor lower labial glands and the fluoride concentration of resting whole and labial saliva were measured over 15 min using a novel method avoiding eversion of the lips. Resting salivary flow rate was measured as 1.09 ± 0.44 μl/min/cm² and stimulated flow rate as 3.13 ± 1.05 μl/min/cm². Secretion rates were significantly (p<0.001) increased during periods of continuous speaking. The increase in secretion elicited by labial movements and speaking may result from mechanical stimulation and/or activity of myoepithelial cells. Fluoride concentrations in resting whole saliva and in unstimulated minor labial gland saliva were 0.066 ± 10.048 and 0.181 ± 0.073 parts/10⁶, respectively. The secretory capacity of the minor labial glands and the high F concentration in their secretions suggests a significant contribution to the F content of whole saliva. Our non-invasive method permits collection from the minor labial glands of a volume large enough for chemical analysis. It should prove useful for studying the effects of different secretory stimuli.