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K Kyriacou

Publications and source records attributed to K Kyriacou.

At least 37 records · Page 2Linked to original sources
PubMed1987

Factors affecting the secretion of submandibular salivary kallikrein in cats.

Glandular kallikrein has been assessed in submandibular saliva, homogenates and plasma by the fluorimetric substrate D-Val-Leu-Arg-7-amino-4-trifluoromethylcoumarin (AFC) and histochemically in tissue sections by the 4-methoxy-2-naphthylamide (MNA) analogue. Nerve stimulation was used to produce salivary secretion. Parasympathetic saliva contained low concentrations of kallikrein, independently of any circulating catecholamines from the adrenals. Sympathetic saliva contained very high concentrations of kallikrein; the amounts in individual drops rapidly reached a peak then declined gradually. Adrenergic blocking drugs during mixed parasympathetic and sympathetic stimulation showed that beta-adrenergic effects normally increase the secretion of kallikrein in response to the alpha-adrenergic influence from sympathetic nerve impulses. Small amounts of a glandular kallikrein-like activity are present in the plasma. Effluent blood from the submandibular gland before, during and after stimulation of either nerve gave no indication that submandibular kallikrein passes from the glandular compartment to the blood under conditions of unobstructed salivary flow. Excision of the chorda tympani indicated that parasympathetic nerve impulses are required for the normal resynthesis of submandibular kallikrein. The secretion of salivary kallikrein is essentially an exocrine function but its role in the saliva remains obscure. The results suggest that sudden mobilization of kallikrein may occur at times into the saliva and that a separate population of adrenergic axons, under separate central control, may pass to the striated ducts specially for this purpose.

Animals↗
PubMed1986

Structural and functional studies of the effects of parasympathetic nerve stimulation on rabbit submandibular salivary glands.

The effects were assessed by combining physiological experiments with morphological examination, using light and electron microscopy. Continuous, parasympathetic nerve stimulation, at frequencies varying from 1 to 10 Hz, caused a copious flow of saliva. Both acinar and granular tubule cells showed extensive degranulation, but the effects on tubule cells were the more dramatic. After stimulation, there was a marked loss of acidic mucosubstances from the acinar cells and an almost complete loss of neutral mucosubstances from the granular tubule cells; this was particularly evident with stimulation at higher frequencies (6-10 Hz).

Animals↗
PubMed1985

Use of different derivatives of D-Val-Leu-Arg for studying kallikrein activities in cat submandibular glands and saliva.

Glandular kallikrein shows a special selectivity for D-Val-Leu-Arg-4-methoxy-2-naphthylamide in comparison with other potential oligopeptide substrates and it provides a useful histochemical substrate, although the reaction may not always be specific. However, in cat submandibular saliva, a biochemical assay using the closely related D-Val-Leu-Arg-7-amino-4-trifluoromethylcoumarin (AFC) as substrate, which affords more sensitive detection, showed that soya bean trypsin inhibitor causes no inhibition. This indicates that there are unlikely to be contaminating enzymes competing for the substrate in this body fluid. Support for this observation has been gained by the useful new enzyme overlay membrane technique for fluorescent assessment of reactive bands of enzymes after isoelectric focusing, using membranes of cellulose acetate impregnated with D-Val-Leu-Arg-AFC. Comparison of results after isoelectric focusing of purified cat submandibular kallikrein with samples of cat submandibular saliva confirmed that the substrate is monospecific for kallikrein in saliva of the cat. This knowledge has enabled us to start assessing the dynamics of the secretion of kallikrein by the gland. Testing individual drops of saliva has shown that an amazingly rapid mobilization of kallikrein occurs in high concentrations on sympathetic nerve stimulation. The corresponding oligopeptide-based inhibitor D-Val-Leu-Arg-chloromethyl ketone was found to be strongly inhibitory of the amidase reaction by kallikrein but showed a low specificity for kallikrein. Nevertheless, its effects have been tested in vivo by the intravascular route and it caused an increase in the resting salivary vascular resistance whether administered close-arterially or intravenously. Thus, it would seem that a kallikrein-like protease does influence the background tone in the vessels and the source of this enzyme is thought to be mast cells.

Amino Acid Chloromethyl Ketones↗
PubMed1985

Histochemistry of hydrolytic enzymes in resting submandibular glands of rabbits.

The distribution of several hydrolytic enzymes was investigated in rabbit submandibular glands at both the light and electron microscopical levels. Glands were fixed by either immersion or perfusion fixation with a variety of fixatives containing 1-2% glutaraldehyde and 2-4% formaldehyde in 0.1 M cacodylate buffer at pH 7.2. Light microscopically, the acinar cells showed some staining for ATPase, acid phosphatase and nonspecific esterases but showed weak staining for thiamine pyrophosphatase. Acid phosphatase staining occurred most strongly in granular tubule cells. Staining for esteroproteases was confined to the periluminal rims of intercalary and striated ducts. Alkaline phosphatase was very sensitive to glutaraldehyde and was confined to myoepithelial cells. Electron microscopical observations revealed the presence of acid phosphatase reaction product in lysosomes, immature granules and in GERL-like structures, the last being much more conspicuous in the granular tubule cells. ATPase reaction product was localized to the basal and luminal plasma membranes and lumina of both acinar and granular tubule cells. The Golgi complex of these two types of cells exhibited only moderate amounts of reaction product for thiamine pyrophosphatase. Alkaline phosphatase activity, on the other hand, was exclusively localized to myoepithelial cells in their plasma membranes and sometimes in the nuclear envelope.

Acid Phosphatase↗
PubMed1982

Kallikrein-like activity in salivary glands using a new tripeptide substrate, including preliminary secretory studies and observations on mast cells.

The tripeptide substrate D-val-leu-arg-4-methoxy-2-naphthylamine gave a precise localization of reaction product in cryostat sections of aldehyde-fixed salivary glands from a number of species, with Fast Blue B as the capture reagent. In submandibular glands, there was strong staining of the granules in granular tubules of rats and hamsters and somewhat less in mice. Submandibular striated ducts showed variable periluminal staining in a finer granular form; it was abundant in guinea-pigs, strong in cats but somewhat less pronounced in dogs. Parotid glands contained less reactivity with none detectable in hamsters and guinea-pigs. In the rabbit, neither gland showed any reaction. Mast cells were densely stained in glands from cats and dogs; they were less reactive in rats and unstained in the other species. The closely related 7-amino-4-trifluoromethylcoumarin derivative of the tripeptide has been found highly satisfactory for assessing activity in submandibular saliva from cats. Preliminary functional studies indicate that an extensive rapid secretion of enzyme occurs into saliva on sympathetic stimulation, with a corresponding depletion of reactive material from the striated ducts in tissue sections. Far less mobilization of enzyme occurs into saliva on parasympathetic stimulation with no obvious change in the histochemical reaction of striated ducts. The possible significance of these findings in cats is discussed. Extensive qualitative and quantitative studies are required to evaluate enzyme and substrate specificities in each species. Nevertheless, derivatives of D-val-leu-arg offer great promise for the functional testing of kallikrein-like reactivity both by histochemical means on cells and biochemically in their secretions.

Animals↗
PubMed1980

Localisation of esteroproteases in 'resting' salivary glands from different species and the effects of the organophosphorus inhibitor E600.

The distribution of reaction sites for esteroproteases has been assessed in 'resting' salivary glands from rats, guinea-pigs, cats, dogs and rabbits using the new substrate N-acetyl-L-methionine alpha-naphthyl ester (Lexow et al. 1979). Two main types of activity were observed in the parenchyma. 1) Condensed staining was present on the luminal sides of certain ducts in some, but not all, glands, and this type of staining was always most conspicuous in the submandibular glands of each species. It is likely that this periluminal esteroprotease activity arises from secretory enzymes that will pass into the saliva. Their greater presence in submandibular glands may have association with specialised licking activities. 2) Variable diffuse cytoplasmic staining was present in certain acinar cells from some, but not all, glands. This was most pronounced in the mucous cells of zygomatic glands from dogs. It is considered that the enzymes demonstrated in acinar cells are involved in processing secretory products rather than in being secretory themselves. The possibility that this includes 'signal peptidase' is discussed. Ductal activity in rats, cats and guinea-pigs had some relationship with sites of tryptophan staining but not in rabbits or dogs. Any relationship with acinar staining was less evident. Mast cells, showing strong esteroprotease staining, were present in variable numbers in the different glands. Preliminary attempts to qualify the enzymes being demonstrated were made by using the inhibitor E600. This substance inhibited all activity in acinar cells from each species, and in rabbits and dogs it also inhibited the ductal activity as well. However, in rats and guinea-pigs it had little or no effect on the ductal activity and in cats it had only a small inhibitory effect on the ductal activity. E600 had no obvious inhibitory effects on mast cell activity.

Animals↗
PubMedDate not supplied

Ultrastructural pathology of the heart in patients with beta-thalassaemia major.

Patients with beta-thalassaemia major frequently suffer from hypersiderosis which leads to hemochromatosis of major organs such as the heart and liver. Little information exists about the ultrastructural pathology of the human heart in beta-thalassaemia patients. Five Cypriot patients with elevated blood ferritin and intractable heart failure were investigated. Cardiac biopsies from these patients were studied by light and electron microscopy, as well as by X-ray microanalysis. Ultrastructural examination revealed the presence of disrupted myocytes showing loss of myofibers, dense nuclei, and a variable number of pleomorphic electron dense granules. These cytoplasmic granules or siderosomes consisted of iron-containing particles as confirmed by X-ray microanalysis. It is likely that the ultrastructural changes observed in myocytes of patients with beta-thalassaemia are largely due to iron deposition.

Adult↗
PubMedDate not supplied

Correlation between morphology, immunohistochemistry and molecular pathology in hereditary and sporadic breast cancer cases.

Breast cancer still represents a serious health problem and is currently the most frequent malignancy in the female population in developed countries. In Cyprus, there are 300 new cases annually. In the present study, histology, electron microscopy, immunohistochemistry, and Western blot analysis were used to investigate 100 cases of invasive breast carcinoma. In addition, mutation analysis for the BRCA1 gene was carried out in patient DNA from 26 families with multiple cases of breast/ovarian cancers. Of note are the results of molecular biology which show that there are no germline truncating mutations in the BRCA1 gene in these 26 Cypriot breast cancer families. Furthermore, Western blot analysis revealed the presence of multiple BRCA1 bands in homogenates of tumor and normal tissues, and immunoelectron microscopy showed the presence of nuclear staining for BRCA1 antibodies.

Adult↗
PubMedDate not supplied

Morphological methods in the diagnosis of mitochondrial encephalomyopathies: the role of electron microscopy.

Mitochondrial encephalomyopathies (MEs) encompass a heterogeneous group of disorders that frequently present a diagnostic challenge to clinicians. Historically, MEs were diagnosed by finding ragged red fibers in the muscle biopsy and confirmatory evidence was provided by the presence of numerical and/or ultrastructural abnormalities in mitochondria. In most centers diagnosis involves clinical evaluation and the morphological, histochemical, and biochemical investigation of a skeletal muscle biopsy. However, with the availability of mitochondrial DNA analysis, the necessity and role of morphological methods and, in particular, electron microscopy has been questioned. The aim of this study was to delineate the role of electron microscopy in the diagnosis of MEs.

Adolescent↗
PubMedDate not supplied

Thin glomerular basement membranes in patients with hematuria and minimal change disease.

A detailed morphometric analysis of glomerular basement membrane (GBM) thickness was carried out on biopsies from 16 patients exhibiting normal histology and unremarkable immunofluorescence. Eleven of these patients presented with proteinuria, 8 in the nephrotic syndrome range, while 5 had hematuria as well. The remaining 5 patients presented with hematuria only. Eight patients had an initial diagnosis of minimal change disease, 4 were diagnosed as thin-membrane nephropathy, 2 had Alport syndrome, and the remaining 2 had hypertensive nephropathy. Quantitative morphometric analysis of GBM identified 3 subsets of patients. The first subset consisted of 6 patients: 5 adults, with an average GBM width of 361 +/- 34 nm, and 1 child. The second subset included 8 patients with thin GBMs and a mean thickness of 253 +/- 15 nm. The last subset comprised 2 patients with Alport syndrome showing marked variability in GBM thickness. This study has confirmed the presence of thin GBMs in hematurics, but has also revealed GBM thinning in 50% of patients with a diagnosis of minimal change disease.

Adolescent↗
PubMedDate not supplied

Ultrastructural diagnosis of mitochondrial encephalomyopathies revisited.

Mitochondrial encephalomyopathies (MEs) are a heterogeneous group of multisystem disorders with extreme variability in clinical phenotype. Due to their complex nature, accurate diagnosis requires a coordinated approach, based on clinical and various laboratory data. Despite the introduction of biochemical assay of mitochondrial enzymes and the availability of mtDNA mutation analysis, the diagnosis of MEs still relies heavily on morphological methods. The latter include histology, histochemistry, and electron microscopy. A comparative study was undertaken to define the contemporary role of electron microscopy in the morphological diagnosis of MEs. Muscle biopsies from 20 patients with MEs, 9 children and 11 adults, were evaluated by histology, enzyme histochemistry, and electron microscopy. The results clearly demonstrate that electron microscopy is of importance in providing essential diagnostic information in pediatric patients, but is of lesser value in the diagnosis of adult cases, where it provides only supplementary information.

Adult↗
PubMedDate not supplied

Crystalloid structures in retroperitoneal paragangliomas: a light and electron microscopic study.

Samples from three clinically functional retroperitoneal paragangliomas were studied by light and electron microscopy. The tumors exhibited a Zellballen pattern histologically, and ultrastructurally all three neoplasms consisted of cells containing catecholamine granules. Prominent cytoplasmic crystalloids were present in all cases. The crystalloids were identified in routine histologic sections, demonstrated eosinophilia, and stained with periodic acid-Schiff, Giemsa, phloxine-tartrazine, and azan stains. Ultrastructurally the crystalloids were osmiophilic, often appeared as slender needles, were membrane bound, and demonstrated a periodicity of 9 nm. The crystalloids, unlike the catecholamine granules, were negative for catecholamine fluorescence. X-ray microanalysis, however, revealed the selective presence of chromium in both catecholamine granules and crystalloids.

Adult↗