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Biomedical subjects

C A Paterson

Publications and source records attributed to C A Paterson.

At least 37 records · Page 2Linked to original sources
PubMed1999

"She looked at me crazy": escalation of conflict through telegraphed emotion.

This exploratory study investigated a feature of adolescent social interaction that can quickly initiate or escalate conflict: telegraphed emotion (a specific emotion conveyed through a subset of its expressive components or through a brief, extreme display of the complete expression; Shields & Maybury, manuscript in preparation). In the form of "the look," telegraphed emotion conveys intensely felt, yet controlled, affect. Using focus groups, teenage females were asked to define the look and to discuss its use, such as who employs it, when it is used, and the consequence of using the look. They reported that the look has different meanings depending on whether it is directed at a friend or a nonfriend, and whether it is used by females or males. With friends, the purpose is to communicate anger, sadness, or disgust; with nonfriends, it indicates disgust or anger, but within a framework of asserting power or status. The look often results in escalation of conflict, especially a verbal or physical fight. These findings are discussed, and the implications for investigating and preventing adolescent interpersonal conflict are explored.

Adolescent↗
PubMed1999

Expression of prostaglandin receptors EP4 and FP in human lens epithelial cells.

PURPOSE: To examine the expression of prostaglandin (PG) receptors EP2, EP4, and FP in a human lens epithelial cell line (HLE-B3) at molecular and pharmacologic levels. METHODS: Reverse transcription-polymerase chain reactions (RT-PCR) were performed with total RNA preparations from HLE-B3 cells using sense and antisense primers for each of the three prostaglandin receptors. The PCR products were hybridized with specific 32P-labeled probes and, for further confirmation, digested with appropriate restriction enzymes. At the pharmacologic level, the expression of EP4 receptors was determined by measuring intracellular cyclic adenosine monophosphate (cAMP) formation in response to PGE2 (EP1, EP2, EP3, and EP4 agonist) and the EP4 receptor-selective antagonist AH23848. The expression of FP receptors in HLE-B3 cells was explored by measuring intracellular [Ca2+]i mobilization. RESULTS: RT-PCR generated DNA products of predicted sizes corresponding to the EP2, EP4, and FP receptors. Hybridization of the PCR products with specific 32P-labeled probes and restriction digestion of the PCR products further confirmed that they were generated from the respective EP2, EP4, and FP mRNAs. The EP receptor agonist PGE2 significantly increased the cAMP level in HLE-B3 cells. The formation of cAMP by PGE2 was concentration-dependently inhibited by the EP4 receptor-selective antagonist AH23848. Stimulation of HLE-B3 cells by the FP receptor agonist fluprostenol increased [Ca2+]i in a time-dependent manner. CONCLUSIONS: The results of the molecular biologic and pharmacologic experiments showed conclusively the presence of EP4 and FP receptor messenger RNAs and proteins, respectively, in HLE-B3 cells.

Actins↗
PubMed1998

Bisulfite genomic sequencing-derived methylation profile of the xist gene throughout early mouse development.

Differential epigenetic modification by methylation of CpG dinucleotides is a candidate mechanism that may identify the alleles of imprinted genes and result in monoallelic expression of either the maternal or the paternal allele. Determination of the allelic methylation status of imprinted genes in the gametes and during early development is constrained by the limiting quantities of genomic DNA available from these early developmental stages. To circumvent this problem we have used bisulfite genomic sequencing to determine the allelic methylation status of the minimal promoter and a 1-kb region within the Xist gene during preimplantation development. We find that the parental Xist alleles are not differentially methylated in these regions. Our findings are discussed in the context of previous conflicting data obtained using methylation-sensitive restriction enzyme digestion followed by PCR amplification to assay for methylation.

Animals↗
PubMed1998

Acute effect of ethanol on lens cation homeostasis.

This study examined the effect of ethanol on the calcium homeostasis of the bovine lens. After acute exposure of the whole lens to physiologically related ethanol concentration, the calcium content of the lens cortex increased from 0.345 +/- 0.075 to 0.476 +/- 0.047 micromol/g (p < 0.05). In contrast, other cation levels such as sodium, potassium, and magnesium did not change. In the study of the lens calcium transport, ethanol caused an increase in the calcium permeability of the lens lipid membrane by about 12% at 30 mM ethanol. Ethanol did not alter the calcium pump activity at ethanol concentration up to 400 mM. Above 600 mM ethanol, the calcium pump was almost completely inhibited. It has been suggested that moderate to heavy alcohol consumption is a risk factor for cataracts. This study indicates that acute ethanol exposure can cause a loss in the lens calcium homeostasis, which maybe one of the cellular mechanisms to contribute to the cataract development in the alcoholic individual.

Animals↗
PubMed1998

Effects of GLP-1 on gastric emptying, antropyloric motility, and transpyloric flow in response to a nonnutrient liquid.

Glucagon-like polypeptide 1 (GLP-1) may be a major enterogastrone, slowing gastric emptying when released by intestinal nutrients. In six conscious dogs, we studied the effects of GLP-1, on antropyloric motility, gastric emptying, and transpyloric flow after instillation of 500 ml of saline into the stomach. The meal was given and recordings were started 15 min after intravenous bolus and infusion of either saline or three different doses of GLP-1. Intravenous GLP-1 produced a dose-related retardation of gastric emptying associated with a decrease in the number and volume of flow pulses in comparison to saline. This change in transpyloric flow was associated with an inhibition of antropyloric pressure waves, a stimulation of isolated pyloric pressure waves, and an increase in basal pyloric tone induced by intravenous GLP-1 infusion. Our findings show that GLP-1 has a potent dose-dependent inhibitory effect on transpyloric flow and gastric emptying. This effect is temporally associated with inhibition of antral "pumping" and stimulation of pyloric "braking" mechanisms.

Animals↗
PubMed1998

Role of nitric oxide mechanisms in control of pyloric motility and transpyloric flow of liquids in conscious dogs.

The role of nitric oxide (NO) mechanisms in control of pyloric function and transpyloric flow were investigated in six conscious dogs. Antropyloroduodenal motility, transpyloric flow, and gastric emptying were measured 15 min after intravenous injection of 100 ml of either saline, L-arginine (50 mg/kg), L-NNA (5 mg/kg), or L-arginine (50 mg/kg) followed by L-NNA (5 mg/kg). Infusion of L-NNA was associated with retardation of gastric emptying (65 +/- 6%) in the first 30 min, in comparison to the saline (90 +/- 3%) or L-arginine (90 +/- 2%). This effect was prevented by infusion of L-arginine prior to L-NNA, after which 89 +/- 3% of the liquid emptied in 30 min. There was a significant reduction (P < 0.05) in the number and volume of flow pulses, and an increase in pyloric tone (P < 0.05) after L-NNA in comparison to the other three test conditions. There were no differences, however, in the number of antropyloric or isolated pyloric pressure waves under the four conditions. Our findings suggest that NO mechanisms influence gastric emptying and transpyloric flow of nonnutrient liquids by altering the pyloric tone, thus increasing resistance to flow.

Animals↗
PubMed1998

Acute calcification in alkali-injured rabbit cornea treated with synthetic inhibitor of metalloproteinases (SIMP).

PURPOSE: The aim of this study was to evaluate the ultrastructure of acute calcification observed in alkali-injured rabbit corneas treated with synthetic inhibitor of metalloproteinases (SIMP). METHODS: A 12-mm alkali burn was made on the right corneas of 16 rabbits. Eight eyes were treated with topical phosphate-buffered SIMP, and eight control eyes were given the buffer only. Six corneas (five in the SIMP group and one in the control group) developed chalky white opacification in the central area of the cornea within 6-30 days. The corneas were excised for the following examinations: (a) light microscopy, (b) energy-dispersive x-ray analysis, (c) electron microscopy, and (d) high-angle x-ray diffraction. RESULTS: The results confirmed that there was stromal calcification in these corneas, as shown by positive van Kossa staining, needle-shaped calcium crystal aggregates, and increased calcium in the opaque areas of the corneas. Proteoglycans in the corneal stroma of both the SIMP group and the control group significantly decreased after alkali burns. Very few proteoglycan filaments could be identified in the calcified areas of the stroma. X-ray diffraction reflections with a Bragg spacing of 3.44 +/- 0.04 A were observed from all the calcified corneas treated with SIMP. The intermolecular spacings of collagen fibrils in the calcified stroma (1.24 +/- 0.34 nm) were significantly lower than those in the noncalcified stroma (1.64 +/- 0.04 nm) and the control corneas (1.64 +/- 0.02 nm). The results indicate that the collagen molecules within the fibrils of the calcified stroma are packed more closely than normal. CONCLUSION: The comparison of the x-ray diffraction patterns from the calcified corneas and chicken bone suggests that the calcium deposits in these corneas are in a form similar to those found in bone. The higher rate and more extensive calcification in the SIMP-treated corneas may result from the inhibitory action of SIMP on debris processing, but the mechanism is unknown.

Acute Disease↗
PubMed1997

Detection of EP2, EP4, and FP receptors in human ciliary epithelial and ciliary muscle cells.

We have examined the expression of three prostaglandin (PG) receptors, EP2, EP4, and FP, in a nonpigmented ciliary epithelial cell line (ODMCl-2) and in human ciliary muscle (HCM) cells. Total RNA preparations from either ODMCl-2 or HCM cells were subjected to reverse transcription-polymerase chain reaction (RT-PCR) with sense and antisense primers for each of the three PG receptors. The RT-PCR generated DNA products of predicted sizes corresponding to the EP2, EP4, and FP receptors in both ODMCl-2 and HCM cells. PCR products corresponding to each receptor were hybridized with specific 32P-labeled probes and, for further confirmation, digested with appropriate restriction enzymes. Pharmacological studies with the EP2 receptor-selective agonist butaprost resulted in a significant increase in the cyclic AMP level in ODMCl-2 cells. The stimulation of cyclic AMP in ODMCl-2 cells by PGE2 and 11-deoxy PGE1, the respective EP1/EP2/EP3/EP4 and EP2/EP3/EP4 receptor agonists, was concentration-dependently inhibited by the EP4 receptor-selective antagonist AH23848. These results conclusively demonstrate the presence of both mRNA and protein for EP2, EP4, and FP receptors in ODMCl-2 and HCM cells.

Alprostadil↗
PubMed1997

Calcium permeability in large unilamellar vesicles prepared from bovine lens cortical lipids.

The loss of calcium homeostasis in the lens is thought to play an important role in cataract formation. Although both lens Ca(2+)-ATPase and membrane lipid permeability are essential to calcium homeostasis, membrane permeability has been studied less extensively. In the present study, the calcium permeability of large unilamellar vesicles (LUVs) prepared from bovine lens cortical lipids has been characterized. When a calcium gradient had been established by the addition of CaCl2 to the external medium, calcium influx through the membrane was monitored by calcium concentration sensitive changes in Fura-2 fluorescence as a function of time. The calcium permeability coefficient, P, was 4.46 x 10(-13) cm s-1 at 37 degrees C. This value was about 4 fold higher than that for LUVs prepared from egg phosphatidylcholine and many times higher than that for LUVs prepared from sphingomyelin. These results provide a basis for future studies of factors that influence the permeability of lens cell membranes to calcium.

Animals↗
PubMed1997

Generation second messengers by prostanoids in the iris-sphincter and ciliary muscles of cows, cats and humans.

We have examined the generation of second messengers after stimulation of feline, bovine, human iris-sphincter and ciliary muscles by selected prostaglandins (PGs). The tissues, labeled or unlabeled with 3H-myo-inositol, were stimulated by a range of concentrations of 16,16-dimethyl PGE2, 11-deoxy PGE1, 17-phenyl trinor PGE2 and PGF2alpha. In both tissues of all three species, 16,16-dimethyl PGE2 and 11-deoxy PGE1 stimulated the formation of cyclic AMP. Butaprost, an EP2 receptor agonist, which was tested only in feline ciliary muscle, generated cyclic AMP. In the feline iris-sphincter and in bovine and feline ciliary muscles, 17-phenyl trinor PGE2, an EP1 receptor agonist, significantly increased inositol phosphate turnover. The FP receptor agonist, PGF2alpha stimulated inositol phosphate turnover in the bovine, feline, and human iris-sphincter muscles and in human ciliary muscles. Feline and bovine ciliary muscles did not respond to PGF2alpha. These results suggest that EP1 receptors are present in feline iris-sphincter muscle and in bovine and feline ciliary muscles. The EP2 receptors exist in both tissue. These results also suggest the presence FP receptors in bovine, feline, and human iris-sphincter and in human ciliary muscles. Bovine and feline ciliary muscles do not appear to express FP receptors.

Animals↗
PubMed1997

Calcium ATPase activity and membrane structure in clear and cataractous human lenses.

PURPOSE: Maintenance of calcium homeostasis is imperative for the clarity of the lens. Ca(2+)-ATPase is essential for the removal of cytosolic calcium, either across the plasma membrane or through intracellular organelles such as the endoplasmic reticulum. In this study, membranes prepared from clear lens epithelium were compared to membranes prepared from cataractous lens epithelium. METHODS: Human lens membranes were prepared by a protocol utilizing homogenization and centrifugation. Ca(2+)-ATPase activity was measured biochemically using Gamma-32P labeled ATP. Lipid order was measured using infrared and Raman spectroscopy. RESULTS: Ca(2+)-ATPase activity was similar in membranes prepared from cataractous lenses that were classified as nuclear subcapsular, nuclear and brunescent cataracts. Ca(2+)-ATPase activity was approximately 50% less in membranes prepared from cataractous lenses in comparison to clear lenses. Because clear lenses from Indian donors was unavailable, clear human lenses were used as a qualitative control for the measurement for Ca(2+)-ATPase activity. Lipid order was measured in lens fibers from cataractous and clear lenses from the United States donors. Lipid order increased from 55% in the hydrocarbon chains from clear lens fibers to 84% in cataractous lens fibers. CONCLUSIONS: These findings support the hypothesis that membranes are deranged in cataractous tissue, which should lead to altered levels of calcium.

Adult↗
PubMed1997

Effect of hybrid peptides of cecropin A and melittin in an experimental model of bacterial keratitis.

Synthetic peptides, ranging from 12 to 18 residues, containing partial sequences from natural cecropin A and melittin were tested for activity in an experimental pseudomonas keratitis model in rabbits. In separate experiments, two Pseudomonas aeruginosa strains: (a) a clinical isolated strain, and (b) an American Type Culture Collection (ATCC) strain, were inoculated into the stroma of one cornea of each rabbit. Peptides were topically applied at 0.1% in phosphate-buffered saline (PBS) and compared with PBS alone and 0.3% gentamicin eye drops. Clinical evaluation, based on the McDonald-Shadduck scale, was performed during a > 48-h period after the bacterial inoculation. The peptide-treated animals showed significantly lower (p < 0.05) inflammatory signs and lower anterior-segment bacterial damage compared with PBS-treated animals, after the first 6 h. The antiinflammatory/antimicrobial activity was non significantly differnt (p > 0.05) from that in animals treated with gentamicin. We conclude that peptides keeping the sequence KWKLFKK from cecropin A and at least the sequence VLKVL from melittin show promise as novel agents in topical ocular therapy of bacterial keratitis.

Administration, Topical↗
PubMed1996

Lipid-protein interactions in human and bovine lens membranes by Fourier transform Raman and infrared spectroscopies.

In other systems, proteins have been shown to alter the molecular structures of lipids in the cell membrane bilayer. We wished to determine if proteins altered the structure of lens lipids. The structure of lipid hydrocarbon chains in urea purified human lens membrane vesicles containing intrinsic, hydrophobically bound proteins was compared to the structure of lipids in vesicles without protein. Fourier transform Raman spectroscopy was used to characterize lipid and protein structure. To study lipid interactions with extrinsic, surface bound proteins, the lipid structure was compared in bovine lipid vesicles with and without alpha-crystallin bound to the surface of the membrane. Lipid structure was studied using Fourier transform infrared spectroscopy. No change in lipid structure was detected even at protein/lipid weight ratios of two to one. Human lens intrinsic proteins contained a high amount of a helical structure (60%), but did not alter hydrocarbon chain interactions.

Adolescent↗
PubMed1996

Comparative studies on prostanoid receptors in human non-pigmented ciliary epithelial and mouse fibroblast cell lines.

To examine the expression of functional prostanoid receptors in the human non-pigmented ciliary epithelial (ODMC1-2) and mouse fibroblast cell lines (NIH 3T3) we have measured the generation of the second messengers, cyclic AMP, inositol phosphates and the mobilization of intracellular calcium ([Ca2+]i) following stimulation by prostaglandin receptor agonists. The amount of cyclic AMP generated was measured by a protein binding method. Radiolabeled inositol phosphates were separated using ion exchange columns and quantified by counting the radioactivity. For intracellular calcium measurements, Fura 2-AM loaded cells were stimulated by PG receptor agonists and the calcium activated fluorescence was measured in a spectrofluorometer. In the ODMC1-2 cell line, the formation of cyclic AMP was stimulated by prostaglandin E2, butaprost and 11-deoxy-prostaglandin E1. The stimulation of cyclic AMP production by prostaglandin E2 was partially inhibited by the EP4 receptor antagonist AH23848. Prostaglandin E2 and 11-deoxy-prostaglandin E1 stimulated the formation of cyclic AMP in NIH 3T3 cells. In ODMC1-2 cells, total inositol phosphate turnover was not increased by 17-phenyl-trinor-prostaglandin F2 alpha, 17-phenyl-trinor-prostaglandin E2 or 11-deoxy-prostaglandin E1. In contrast, all these agonists, with the exception of 11-deoxy-prostaglandin E1, significantly increased total inositol phosphates in NIH 3T3 cells. In the NIH 3T3 cell line, only prostaglandin F2 alpha and 17-phenyl-trinor-prostaglandin F2 alpha increased [Ca2+]i in a dose-dependent manner; in ODMC1-2 cells, neither these agonists nor 17-phenyl-trinor-prostaglandin E2 increased [Ca2+]i. The present studies suggest that in ODMC1-2 cells, EP2 and EP4 receptors but neither EP1 nor FP receptors are expressed; these studies also imply, NIH 3T3 cells express EP2 and FP receptors, while EP1 receptors appear to be absent in this cell line.

3T3 Cells↗
PubMed1995

ATPase activities of rabbit and bovine lens epithelial microsomes: a continuous fluorimetric assay study.

Purine nucleoside phosphorylase (EC 2.4.2.1) catalyzes the irreversible phosphorolysis of 7-methylguanosine (m7Guo), a fluorescent guanosine analogue. Using purine nucleoside phosphorylase and m7Guo, a continuous fluorimetric assay for microsomal ATPases from rabbit and bovine lens is described. The decrease in m7Guo fluorescence intensity at 380 nm, which represents the hydrolysis of ATP, is linear with time up to exhausting all m7Guo. The rate of the fluorescence decrease depends on the sample protein concentration. In the presence of ATPase inhibitors, ion-specific ATPase activities in the lens were determined from the difference of the fluorescence decay rates. Using the fluorimetric assay thapsigargin-sensitive Ca-ATPase in the bovine lens epithelium has been characterized. The fluorimetric assay provides a number of advantages over previous membrane ATPase assays.

Adenosine Triphosphate↗