Decreased salivary matrix metalloproteinase-8 reflecting a defensive potential in juvenile parotitis.

OBJECTIVE: Matrix metalloproteinases MMP-2 and MMP-9 have been associated with juvenile parotitis. However, the role of MMP-8 has not been addressed previously. This work focuses on salivary MMP-8 and -9 levels in juvenile parotitis. METHODS: During a five-year period at Helsinki University Hospital, a tertiary care hospital, 41 patients aged 17 or under, were identified as having parotitis; from 36 of these patients, saliva samples were collected for MMP-8 IFMA (time-resolved immunofluorometric assay) analyses. Control saliva samples were collected from 34 age- and gender-matched children admitted for an elective surgery who had no history of parotitis. For comparison, salivary levels of MMP-9, tissue inhibitor of matrix metalloproteinase (TIMP-1), MMP-8/TIMP-1 ratio, human neutrophil elastase (HNE), and myeloperoxidase (MPO) were analyzed by ELISA. Additionally, salivary MMP-8 levels were compared to historical saliva samples from 18 adult gingivitis patients as well as to 10 healthy adult controls. RESULTS: The median (25%, 75% percentile) MMP-8 concentration in saliva of parotitis patients was significantly lower than MMP-8 concentration in saliva of their controls [50.4ng/ml (37.5, 72.9) vs. 148.5ng/ml (101.2, 178.5) p<0.0001] and lower than in patients with gingivitis [347.9ng/ml (242.6, 383.2) p<0.0001] or healthy adult controls [257.2ng/ml (164.9, 320.7) p<0.0001]. The MMP-8/TIMP-1 ratio was lower than in controls [0.13 (0.05-0.02) vs. 0.3 (0.17-0.46) p<0.0001]. The median MMP-9 concentration in saliva of parotitis patients was significantly higher than in controls [143.9ng/m (68.8-189.0) vs. 34.9ng/ml (16.3-87.6) p<0.0001]. Neither HNE, MPO, nor TIMP-1 alone separated the patients from the control groups. CONCLUSIONS: MMP-9 was up-regulated in juvenile parotitis saliva, suggesting that MMP-9 may play a destructive role in juvenile parotitis, as others have suggested. The present novel findings reveal a decreased salivary MMP-8 concentration, suggesting that MMP-8 may reflect in juvenile parotitis down-regulated or anti-inflammatory immune characteristics.

Anti-inflammatory action of cholecystokinin and melatonin in the rat parotid gland.

OBJECTIVE: To define the influence of cholecystokinin and melatonin on the inflammatory response of the lipopolysaccharide-exposed rat parotid gland. MATERIALS AND METHODS: Bacterial lipopolysaccharide was infused retrogradely into the parotid duct. The degree of inflammation three hours postadministration was estimated from the activity of myeloperoxidase, reflecting glandular neutrophil infiltration. RESULTS: The myeloperoxidase activity of the lipopolysaccharide-exposed gland was 10-fold greater than that of the contralateral gland. Combined with sulphated cholecystokinin-8 (10 or 25 µg kg(-1) , given twice intraperitoneally) or melatonin (10 or 25 mg kg(-1) x 2) the lipopolysaccharide-induced response was elevated 4.6- and 3.5-folds at the most. The cholecystokinin-A receptor antagonist lorglumide reduced the inhibitory effect of cholecystokinin-8, while the melatonin 2-preferring receptor antagonist luzindole had no effect on the melatonin-induced inhibition. Unselective nitric oxide-synthase inhibition abolished the increase in myeloperoxidase activity, whereas inhibition of inducible or neuronal nitric oxide-synthase (of non-nervous origin) halved the inflammatory response. CONCLUSION: Some hormones may contribute to anti-inflammatory action in salivary glands in physiological conditions. They are potential pharmacological tools for treating gland inflammation. The inflammation, as judged from the myeloperoxidase activity, was entirely dependent on nitric oxide-synthase activity, indicating that the hormones directly or indirectly reduced the generation of nitric oxide.

Association between the occurrence of matrix metalloproteinases 2 and 9 in parotid saliva with the degree of parotid gland damage in juvenile recurrent parotitis.

OBJECTIVE: The study was aimed to investigate whether the occurrence of matrix metalloproteinases (MMP) 2 and 9 in parotid saliva of juvenile recurrent parotitis (JRP) patients is associated with the degree of glandular involvement. STUDY DESIGN: Thirty-three JRP patients were included. Involvement of parotid gland was assessed by sialography. Parotid saliva was assayed for MMP-2 and MMP-9 by zymography. Medical charts were examined for number of recurrences, disease laterality, and time of follow-up. Logistic regression analysis between occurrence of either MMP, the clinical parameters, and sialographic staging was conducted. RESULTS: None of the clinical parameters under analysis were found to be associated with degree of sialographic involvement. Statistical associations were found between presence of MMP-9 and MMP-2 in parotid saliva and sialographic stage (P = .017; odds ratio [OR] 6.5, 95% confidence interval [CI] 1.4-30.4; and P = .009; OR 6.1; 95% CI 1.6-23.7; respectively). CONCLUSIONS: Occurrence frequency of MMP-2 and MMP-9 in parotid saliva from affected glands of JRP patients was associated with degree of gland damage.

Molecular alterations of parotid saliva in infantile chronic recurrent parotitis.

Infantile chronic recurrent parotitis (ICRP) is an insidious disease whose etiopathogenesis remains an enigma. Alterations in the physical appearance of parotid saliva from ICRP patients have been frequently reported. However, sialochemical studies in regard to ICRP are very rare. The aim of this study was to determine whether saliva of ICRP patients presents major physicochemical and biochemical alterations compared with saliva from paired healthy controls. Parotid, whole, and submandibular/sublingual saliva was collected at an asymptomatic stage from 33 ICRP patients (5-16 y old, both sexes) and from 33 sex- and age-matched healthy controls. Saliva was analyzed for protein concentration, mode of protein diffusion on cellulose membranes, unidimensional sodium dodecylsulfate (SDS)-polyacrylamide gel electrophoresis protein profiles and zymographic profiles of metalloproteinase 2 (MMP-2) and metalloproteinase 9 (MMP-9). Parotid saliva of ICRP patients showed an increased protein concentration, altered mode of protein diffusion, a higher frequency of polypeptide bands of 43, 37, 33, 29, 26, 16, and 10 kD, higher asymmetry in the polypeptide profiles of both contralateral parotid saliva, and an increase in the frequency of MMP-2 and MMP-9. Parotid saliva of patients with ICRP is molecularly altered with respect to normal saliva. Some of the molecular differences could be related to the etiopathogenesis of the disease.

Indicadores de actividad amilasa en la enfermedad parotídea. Parotiditis crónica recurrente infantil / Amylase activity markers for parotid disease: infant recurrent chronic parotiditis

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[Fluorometric method of determining prolylendopeptidase activity in human erythrocytes in normal and pathologic conditions]. / Fluorometricheskii metod opredeleniia aktivnosti proliléndopeptidaz v éritrotsitakh liudei v norme i pri patologicheskikh sostoianiiakh.

A highly sensitive fluorometric method for determination of prolylendopeptidase (PE) activity in human erythrocyte hemolysates in the presence of hemoglobin has been developed. The method is based on measurement of fluorescence of 4-methyl-7-aminocoumarine released in the course of enzymatic reaction from the substrate Z-glycyl-proline-4-methylcoumarine-7-amide. A correlation was introduced for the quenching of fluorescence by hemoglobin. The method is suitable for the determination of PE activity in human erythrocyte hemolysates in various pathological states. The dependence of PE activity on the incubation time, protein and substrate concentrations were studied using the 1,200-fold purified preparations of prolylendopeptidase II. The values of PE activity in erythrocyte hemolysates of healthy donors and in those of patients with odontogenic phlegmons of maxillary-facial area were virtually identical. PE activity in erythrocyte hemolysates of stored blood was 5 times lower than that in the cell hydrolysates of fresh blood. The PE activity was not observed in blood serum of fresh and stored blood of healthy persons and of patients with acute inflammatory processes of maxillary-facial area, as well as in blood serum of patients with hepatitis and glomerulopephritis.

[Glandular kallikrein in chronic recurrent parotitis]. / Glanduläres Kallikrein bei chronisch-rezidivierender Parotitis.

Salivary kallikrein was measured in the parotid saliva of 11 patients suffering from chronic recurrent parotitis and 15 healthy control persons. While in the control group kallikrein levels from 1-4 U/l could be registrated, the results obtained from patients were significantly elevated and varied between 23,45 U/l and 89,41 U/l. Release of kallikrein from destroyed duct cells is suggested to be the origin of the elevated salivary levels of this enzyme in chronic recurrent parotitis. A leakage of glandular kallikrein into the interstitial fluid and thereby release of kallidin from kininogen is discussed as a possible mechanism in the development of this disease.

Amylase as an additional marker of salivary gland neoplasms. An immunoperoxidase study.

The presence of amylase in normal and neoplastic salivary gland tissue was investigated by immunoperoxidase techniques. Apart from normal and inflamed parotid glands, different kinds of tumours were studied with regard to amylase: acinic cell tumours, adenocarcinomas, adenoidcystic carcinomas, salivary duct carcinomas, mucoepidermoid tumours and squamous cell carcinomas. Amylase could be seen in acinic cell tumours, but not in other neoplasms. The results were discussed with respect to the diagnostic implications.

Relation of age to isoenzyme pattern and total activity of amylase in serum.

Pancreatic and salivary isoenzymes of amylase were determined in serum from 70 subjects. Thin-layer gel/isoelectric focusing was used to separate the isoenzymes. Because other studies (J. Lab. Clin. Med. 90: 141-151, 1977) show that the major isoamylases have isoelectric points between 5.8 and 7.2, we focused the sera on polyacrylamide gel plates with a pH gradient from 5.5 to 8.5. The separated amylase fractions were made visible by direct incubation with a commercially available dye-starch polymer. Isoelectric focusing proved to be convenient, precise, and reproducible, and it can be used as a routine analysis to detect even slight changes in serum amylase distributions. We found that the isoamylase distribution is age dependent, whereas total amylase activity shows no correlation with age.

[Effect of the kallikrein-like enzyme salivain on the course of experimental acute parotitis]. / Vliianie kallikreinopodobnogo fermenta salivaina na techenie ostrogo éksperimental'nogo parotita.

Mechanical trauma of rat parotid gland led to an increase in the size of the gland (due to edema) with simultaneous activation of unspecific proteases, which were estimated at pH 7.6 using casein as a substrate. Administration of kallikrein-like salivary enzyme salivain into non-traumatized parotid gland caused also the same alterations. Complex application of two factors (trauma and salivain) resulted in distinct increase of the gland edema and the proteolytic activity. Administration of trasilol (inhibitor of proteases) decreased the pathogenic effects of salivain. After administration of salivain the alkaline phosphatase activity was shown to decrease in the parotid salivary gland.

[Significance of the determination of multiple forms of urinary amylases for the diagnosis of exocrine pancreas diseases]. / Zur Bedeutung der Bestimmung multipler Formen der Amylasen des Harns für die Diagnostik von Erkrankungen des oxokrinen Pankreas.

The multiple amylases of the urine were examined in healthy persons and in patients with acute and chronic pancreatitis as well as with parotitis using the polyacryl amide gel disk electrophoresis. A separation into about 7 amylase-containing fractions was achieved which could be coordinated to the pancreatic juice and to saliva. In acute pancreatitis the pancreas amylases increased, in chronic pancreatitis, on the other hand, they decreased. In parotitis the salivary amylases increased. For diagnostic purposes the formation of quotients from certain fractions was useful. The examination of the multiple amylases of the urine according to the method mentioned leads to an improved clinico-chemical diagnostics of diseases of the exocrine pancreas and gives the possibility of their course control.

A cause of hyperamylasemia associated with chronic liver disease.

The cause of hyperamylasemia associated with chronic liver disease is unclear. In an attempt to identify the tissue of origin of hyperamylasemia in 3 patients with chornic active hepatitis their serum was isoelectrically focused. The isoamylase patterns obtained were compared to those of pancreatic and salivary amylase. The apparent salivary gland origin of the excessive blood amylase in the patients studied was substantiated by radiological demonstration of parotid sialoectasia in one patient and histological evidence of sialoadenitis in another. Further evidence was the coincident isoelectric points of the predominant isoamylase in the sera of the liver disease patients and of patients with parotid inflammatory disease. Hyperamylasemia associated with chronic liver disease may be of salivary gland origin and as such forms part of the spectrum of extrahepatic manifestations of chronic active hepatitis.

Associations between polymorphic variety and disease susceptibility in two New Guinea populations.

During the Australian/British IBP studies on KarKar Island and at Lufa in the Eastern Highlands, Papua New Guinea, information was collected on the epidemiology and genetic constitution of the same subjects. Advantage of this special situation has been taken to determine whether any associations exist between the genetic markers and the disease states. Those found and which appear real include Rhesus D(u) with proteinuria; MN with splenomegaly and hepatomegaly; Ss with parotid enlargement; acid phosphatase, glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogense and haemoglobin J- Tongariki with presence of malarial parasites; phosphoglucomutase with proteinuria and parotid enlargement; haptoglobin with proteinuria and with splenomegaly and hepatomegaly. These associations are discussed in terms of the probabilities of their arising from heterogeneity in population structure, linkage disequilibrium and pleiotropy.