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D M Kelly

Publications and source records attributed to D M Kelly.

At least 37 records · Page 2Linked to original sources

Temporal relationship of acinar and microvascular changes in caerulein-induced pancreatitis.

A study in rats investigated the temporal relationship between acinar cell changes and alterations in the local microvasculature in oedematous pancreatitis produced by administration of caerulein 5 micrograms kg-1 h-1. Samples were taken from experimental and control animals after 15 min, 30 min, 1 h and 2 h of caerulein infusion. Transmission electron microscopy showed ultrastructural acinar cell changes after 15 min whereas the earliest microvascular changes were seen after 30 min. Ultrastructural alterations in the acinar cells thus preceded local microvascular changes. Microvascular distortion appears to be a consequence and not a cause of pancreatitis in the caerulein model.

Animals↗

Temporal expression of VLA-2 and modulation of its ligand specificity by rat glomerular epithelial cells in vitro.

BACKGROUND: The interaction of glomerular epithelial cells (GEC) with their underlying basement membrane is of critical importance in maintaining normal glomerular function. Little is known regarding their expression and use of extracellular matrix adhesion receptors in normal conditions and during pathogenic states. EXPERIMENTAL DESIGN: To examine the use of such receptors, we have produced monoclonal antibodies that inhibit the function of the rat alpha 2 beta 1 integrin receptor (VLA-2) and the common beta 1 subunit. The monoclonal antibodies have been used to examine the expression and functional use of these receptors by rat glomerular cells cultured in vitro. RESULTS: Rat glomerular visceral epithelial cells are unusual in that, unlike many of the epithelium seen in vivo, these cells do not express VLA-2, an integrin receptor with affinity for laminin and collagen. Our results demonstrate that differentiated GEC, newly isolated from glomeruli, do not use VLA-2 for attachment to collagen and laminin-coated surfaces. However, after 3 days of in vitro growth, approximately 50% of these cells express this receptor and, upon their first in vitro passage, selectively utilize VLA-2 for attachment to collagen but not to laminin-coated surfaces. After long-term maintenance in culture, all GEC express VLA-2, and utilize this receptor for binding to collagen and in their interaction with laminin. In contrast, VLA-2 plays only a partial role in the adherence of mesangial cells to collagen and is not involved in their attachment to laminin-coated surfaces. CONCLUSIONS: These results show that, as GEC become adapted to in vitro growth, they begin to synthesize and use the VLA-2 integrin receptor suggesting a simultaneous downregulation or inactivation of other beta 1 type integrin receptors. This ability to modulate their receptor repertoire may allow GEC to respond to pathologic conditions in vivo.

Animals↗

Pattern discrimination perimetry in patients with glaucoma and ocular hypertension.

We compared the results of the pattern discrimination perimeter to the program 30-2 on the Humphrey Field Analyzer (Humphrey, Inc., San Leandro, California) in 93 consecutive patients with ocular hypertension and glaucoma and 30 control patients. In 20 patients with ocular hypertension, a significantly greater number of glaucomatous defects were noted on pattern discrimination perimetry (ten patients) than on the program 30-2 (two patients) (P less than .05, Wilcoxon signed rank test). The diversity in diagnoses found on pattern discrimination testing was not explained by age, intraocular pressure, refraction, number of glaucoma medicines, race, presence of vascular disease, optic disk status, or pupil size. In contrast, in 73 patients with glaucoma no statistical difference in the severity of diagnoses was noted between perimeters (P greater than .05, Wilcoxon signed rank test). These results suggest the potential value of pattern discrimination perimetry as a visual function test in patients with glaucoma and in defining subsets of patients with ocular hypertension not found with conventional automated perimetry.

Adult↗

Long-term and short-term fluctuation in pattern discrimination perimetry.

We studied threshold fluctuation with the pattern discrimination perimeter in 24 healthy subjects at 56 locations within the central 30 degrees. This perimeter evaluates a subject's ability to discriminate a patterned stimulus measured by a percentage scale. We found an intraindividual fluctuation of 10.52% and an interindividual fluctuation of 8.78%. A statistically increased intraindividual, but not interindividual fluctuation was noted with increasing eccentricity from fixation (P less than .05, Bartlett's test). However, no correlation in fluctuation was noted with advancing age or increasing false-positive errors (P greater than .05, correlation coefficient). Also, no difference in fluctuation between superior or inferior hemifields was observed (P greater than .05, Student's t-test). The average threshold across all subjects was 54.3%, which provided an upper limit of normal, two standard deviations from the mean, of less than 80% for most locations. This study indicates that fluctuation should be considered when interpreting pattern discrimination fields, but that the extent of fluctuation generally allows for an adequate separation between normal and abnormal measurements.

Adult↗

Pulmonary microvasculature in experimental acute haemorrhagic and oedematous pancreatitis.

The pulmonary microvasculature was examined in two experimental models of acute pancreatitis by scanning electron microscopy of microvascular corrosion casts. Haemorrhagic pancreatitis was induced in eight male Sprague-Dawley rats using an intraductal injection of 5 per cent sodium taurocholate. Oedematous pancreatitis was induced in seven male Sprague-Dawley rats using an intravenous infusion of supramaximal doses of caerulein (5 micrograms/kg per hour). The pulmonary vessels were cast using a polymer resin and the cast studied by scanning electron microscopy at 3 and 12 h in those with haemorrhagic and at 1 and 4 h in those with oedematous pancreatitis. Vascular abnormalities were present in both models at the initial study time with abruptly terminating vessels being more prominent in the caerulein model. At the later times, however, the abnormalities in the sodium taurocholate model were much more severe, with a substantial loss of vascular density, tortuosity and abrupt terminations of those vessels present. Microvascular abnormalities may be responsible for some of the pulmonary changes seen in oedematous and haemorrhagic pancreatitis.

Animals↗

Antigen processing and presentation by glomerular visceral epithelium in vitro.

Although macrophages are considered the prototype of antigen presenting cells (APC), recent studies have emphasized the potential role of several parenchymal and mesenchymal cells in this process. We have studied the capacity of cultured glomerular visceral epithelial cells (GEC) to act as effective APC and compared this capacity with that demonstrated by peritoneal macrophages. Affinity-purified and in vitro propagated rat GEC were exposed to hen egg lysozyme, keyhole limpet hemocyanin, and cationic ferritin. As effector cells, we used antigen-specific T cell hybridomas; the level of antigen presentation was assessed by determining the level of interleukin 2 (IL-2) present in tissue culture supernatants. Cytokine-treated GEC were capable of processing and presenting all antigens in a dose-dependent manner. Crucial for antigen presentation were intracellular processing of antigen and the presence of Ia on the cell surface. Our findings indicate that GEC can act as effective APC, and further suggest that this capacity may be relevant to cell-mediated immune injury at the level of the glomerular capillaries in vivo.

Animals↗

Enhancement of experimental actinomycosis in mice by Eikenella corrodens.

The infectivity of Actinomyces israelii in a susceptible-weanling-mouse was increased by the presence of Eikenella corrodens in the inoculum. A minimal infecting dose of 1.7 X 10(7) CFU of A. israelii was required to establish chronic lesions after an intraperitoneal injection. When E. corrodens (3.8 X 10(7) CFU) was included in the inoculum, chronic lesions were established with a dose of 8.5 X 10(4) CFU of A. israelii. E. corrodens alone did not produce persistent lesions. Viable E. corrodens could be recovered from chronic mixed actinomycotic lesions in numbers that often equaled or exceeded the populations of A. israelii in the lesions. The duration of acute actinomycotic infections caused by A. viscosus was temporarily extended by the presence of E. corrodens. The cellular inflammatory response and overall morphology of mixed experimental lesions containing A. israelii and E. corrodens did not appear to be significantly different from those of pure-culture lesions containing A. israelii alone. E. corrodens cells could not be readily discerned in stained histological sections of mixed experimental lesions.

Actinomycosis↗

The intramembrane topography of the mannitol-specific enzyme II of the Escherichia coli phosphotransferase system.

The D-mannitol-specific Enzyme II of the phosphoenolpyruvate-dependent phosphotransferase system of Escherichia coli is an integral cytoplasmic membrane protein responsible for concomitant transport and phosphorylation of this hexitol. We have investigated the intramembrane topography of this enzyme/permease using proteases, membrane-impermeable reagents, and antibodies against the purified protein. The results of these experiments suggest that this protein spans the membrane in a single orientation with a sizeable proportion of its mass extending into the cytoplasm, but with little of the polypeptide exposed at the outside surface of the membrane. Such an orientation is consistent with the reception and transport roles of the mannitol Enzyme II in E. coli.

Cell Membrane↗

Substrate and phospholipid specificity of the purified mannitol permease of Escherichia coli.

D-Mannitol is transported and phosphorylated by a specific enzyme II of the phosphotransferase system of Escherichia coli. This protein was purified previously in detergent solution and has been partially characterized. As one approach in understanding the structure and mechanism of this enzyme/permease, we have tested a number of sugar alcohols and their derivatives as substrates and/or inhibitors of this protein. Our results show that the mannitol permease is highly, but not absolutely, specific for D-mannitol. Compounds accepted by the enzyme include those with substitutions in the C-2(= C-5) position of the carbon backbone of the natural substrate as well as D-mannonic acid, one heptitol and one pentitol. All of these compounds were both inhibitors and substrates for the mannitol permease except for D-mannoheptitol, which was an inhibitor but was not phosphorylated by the enzyme. No compound examined, however, exhibited an affinity for the enzyme as high as that for its natural substrate. We have also investigated the phospholipid requirements of the mannitol permease using phospholipids purified from E coli. The purified protein was significantly activated by phosphatidylethanolamine, but little activation was observed with phosphatidylglycerol or cardiolipin. These observations partially delineate requirements for interaction of sugar alcohols and phospholipids with the mannitol permease. They suggest approaches for the design of specific active site probes for the protein, and strategies for stabilizing the enzyme's activity in vitro.

Escherichia coli↗

Persistence of associated gram-negative bacteria in experimental actinomycotic lesions in mice.

Mixed actinomycotic infections were established in a susceptible weanling mouse model by using combinations of Actinomyces israelii and Eikenella corrodens or A. israelii and Actinobacillus actinomycetemcomitans. Acute lesions caused by either of the gram-negative organisms alone were resolved within a few weeks; however, these organisms persisted up to 3 months in chronic lesions in combination with A. israelii.

Actinobacillus↗

Insulin clearance by perfused rat lung.

The clearance of insulin has been demonstrated in the perfused in situ rat lung. Porcine insulin added to the perfusion medium at 50 microunits/l was cleared at 6.3 U/h in lungs from fed animals, 6.1 U/h in fasted animals and 15.7 U/h in diabetic rat lungs. At a higher concentration of insulin (500 microunits/l) increased clearance was observed in lungs from both fed, fasted and diabetic animals, 33.0, 36.0 and 56.5 U/h respectively. It is suggested that the clearance of insulin by the lung is dependent on metabolic state of the animal.

Animal Nutritional Physiological Phenomena↗

The importance of 5-hydroxytryptamine for the induction of harmine tremor and its antagonism by dopaminergic agonists assessed by lesions of the midbrain raphe nuclei.

The brain lesion technique was used to destroy the ascending 5-hydroxytryptamine (5-HT) system at its cell bodies in the dorsal and medial raphe nuclei in order to assess the importance of 5-HT for the induction of harmine tremor and its antagonism by the dopaminergic agonists, L-DOPA, apomorphine and d-amphetamine. Lesions of the medial or dorsal raphe nucleus reduced the intensity of harmine tremor. The remaining tremor was generally resistant to further reduction by the dopaminergic agonists. 5-hydroxytryptophan was shown to enhance tremor: this effect was reduced both by the raphe lesions and by treatment with L-DOPA. The data are discussed in terms of the possible relationship between 5-HT and dopamine.

Alkaloids↗

Nomifensine: a potent dopaminergic agonist of antiparkinson potential.

Nomifensine was shown to be a potent stereotypic agent in rat. Its effect was resistant to a-methylparatyrosine pretreatment but was abolished by combined reserpine/alpha-methylparatyrosine and by haloperidol. Electrolytic lesions placed in dopamine-containing areas of the extra-pyramidal, mesolimbic and amygdaloid systems indicated an effect in all areas, but the globus pallidus and substantianigra were shown to be most important for its action. Also the effect of nomifensine was reduced by lesions of the medial and/or dorsal raphé nuclei. A circling behaviour was recorded following nomifensine administration to animals with unilateral electrolytic lesions of the substantia nigra or asymmetric lesions of the medial raphé nucleus. These effects were resistant to alpha-methylparatyrosine and inhibited by haloperidol. Nomifensine reduced the intensity of harmine-induced tremor. The M2-metabolic of nomifensine mimicked the effects of the parent compound on peripheral administration bu the onset of action was more rapid and the duration shorter. The M2-metabolite was active on intrastriatal injection to induce stereotyped/hyperactive behaviour and contralateral asymmetries. In all experimental situations nomifensine was compared with apomorphine and d-amphetamine (dopamine and L-Dopa where appropriate). Nomifensine/metabolite was shown to be a potent dopaminergic agonist with an action mainly dependent upon functioning of the extrapyramidal system and partly independent of presynaptic mechanisms.

Animals↗

The importance of extrapyramidal function for the induction and antagonism of harmine tremor.

The brain lesion technique was used to investigate the role of the paleostriatum and the nigro-neostriatum in harmine-induced tremor and in its antagonism by dopaminergic agonists, apomorphine, 1-dopa, piribedil, d- and l-amphetamine. Bilateral lesions of the caudate--putamen or substantia nigra failed to modify the intensity of tremor or its antagonism by dopaminergic agonists. Bilateral lesions of the globus pallidus markedly reduced the intensity of tremor and the results indicated that such lesions were also able to reduce the effectiveness of dopaminergic agonists as tremor antagonists. The data suggest that the integrity of the paleostriatum is more important than that of the neostriatum for the mediation of harmine tremor and its antagonism by dopaminergic agonists. The results are discussed in relation to the proposed clinical relationship between paleostriatal dopamine dysfunction and tremor mechanisms.

Alkaloids↗