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Biomedical subjects

P J Quinn

Publications and source records attributed to P J Quinn.

At least 19 recordsLinked to original sources
PubMed2004

Is the distribution of alpha-tocopherol in membranes consistent with its putative functions?

Vitamin E acts as an antioxidant and stabilizer of membranes. Other functions of vitamin E unrelated to its effects on membranes are emerging. Vitamin E partitions into the lipid bilayer matrix of membranes. It orients perpendicularly to the plane of the membrane with the hydroxyl group pointing to the lipid-water interface. The vitamin is not randomly distributed in the plane of the membrane but tends to form clusters. These clusters appear to be composed of vitamin E and phosphatidylcholine in a stoichiometry of about one vitamin E per 10 phospholipid molecules. Vitamin E partitions into domains of phosphatidylcholine in model membranes formed from mixtures of phosphatidylcholine and phosphatidylethanolamine irrespective of whether the phosphatidylcholine is in the fluid or gel phase. The creation of domains enriched in vitamin E in membranes is not consistent with an antioxidant function and effects on membrane structure and stability indicate other roles of the vitamin.

Cell Membrane↗
PubMed2003

Salmonella serotypes present on a sample of Irish pig farms.

A survey of the prevalence of Salmonella species infection was conducted on 59 Irish farrow-to-finish pig herds. Faecal samples were collected from the pens of first-stage weaners (growing pigs approximately three to 10 weeks of age), second-stage weaners (approximately 10 to 17 weeks of age) and fatteners, and from the dry sow and farrowing sow houses. The prevalence of infection was estimated to within 5 per cent with a 95 per cent confidence interval. Thirty of the 59 herds were infected, 12 with Salmonella Typhimurium only, eight with Salmonella Derby only and seven with both S Typhimurium and S Derby; serotypes London, Livingstone and Infantis were each isolated from a single herd. Farms in Ireland are assigned to one of three infection categories on the basis of the antibody levels in samples of meat juice taken at slaughter. When a herd was classified as either positive or negative on the basis of the isolation of Salmonella from at least one faecal sample there was no association between the herd's category as determined by meat juice serology and the probability of the isolation of Salmonella from the faecal samples. However, there were differences in prevalence between pigs at different stages of production in herds of different categories. Farrowing sow houses in moderately infected (category 2) herds had significantly lower infection rates (P < or = 0.05) than other herd categories and other stages of production. Pigs from first-stage weaner pens in slightly infected (category 1) herds were more likely to be infected with Salmonella than pigs at any other stage of production or category of herd.

Animals↗
PubMed2003

Direct evidence for recycling of myeloperoxidase-catalyzed phenoxyl radicals of a vitamin E homologue, 2,2,5,7,8-pentamethyl-6-hydroxy chromane, by ascorbate/dihydrolipoate in living HL-60 cells.

Myeloperoxidase (MPO)-catalyzed one-electron oxidation of endogenous phenolic constituents (e.g., antioxidants, hydroxylated metabolites) and exogenous compounds (e.g., drugs, environmental chemicals) generates free radical intermediates: phenoxyl radicals. Reduction of these intermediates by endogenous reductants, i.e. recycling, may enhance their antioxidant potential and/or prevent their potential cytotoxic and genotoxic effects. The goal of this work was to determine whether generation and recycling of MPO-catalyzed phenoxyl radicals of a vitamin E homologue, 2,2,5,7,8-pentamethyl-6-hydroxychromane (PMC), by physiologically relevant intracellular reductants such as ascorbate/lipoate could be demonstrated in intact MPO-rich human leukemia HL-60 cells. A model system was developed to show that MPO/H(2)O(2)-catalyzed PMC phenoxyl radicals (PMC*) could be recycled by ascorbate or ascorbate/dihydrolipoic acid (DHLA) to regenerate the parent compound. Absorbance measurements demonstrated that ascorbate prevents net oxidation of PMC by recycling the phenoxyl radical back to the parent compound. The presence of DHLA in the reaction mixture containing ascorbate extended the recycling reaction through regeneration of ascorbate. DHLA alone was unable to prevent PMC oxidation. These conclusions were confirmed by direct detection of PMC* and ascorbate radicals formed during the time course of the reactions by EPR spectroscopy. Based on results in the model system, PMC* and ascorbate radicals were identified by EPR spectroscopy in ascorbate-loaded HL-60 cells after addition of H(2)O(2) and the inhibitor of catalase, 3-aminotriazole (3-AT). The time course of PMC* and ascorbate radicals was found to follow the same reaction sequence as during their recycling in the model system. Recycling of PMC by ascorbate was also confirmed by HPLC assays in HL-60 cells. Pre-loading of HL-60 cells with lipoic acid regenerated ascorbate and thus increased the efficiency of ascorbate in recycling PMC*. Lipoic acid had no effect on PMC oxidation in the absence of ascorbate. Thus PMC phenoxyl radical does not directly oxidize thiols but can be recycled by dihydrolipoate in the presence of ascorbate. The role of phenoxyl radical recycling in maintaining antioxidant defense and protecting against cytotoxic and genotoxic phenolics is discussed.

Antioxidants↗
PubMed2002

A study of greyhounds with tonsillar enlargement and a history of poor racing performance.

Fifteen greyhounds with tonsillar enlargement were subjected to detailed investigation. Affected greyhounds exhibited coughing, poor racing performance and tonsillar lymphoid hyperplasia over a period of months. Each of the 15 affected animals had evidence of respiratory tract disease. Twelve had non-specific respiratory tract disease, two had pneumonia and one had pulmonary infiltration with eosinophils (PIE). Histopathological examination of the tonsils from affected dogs revealed that greyhounds with tonsillar enlargement are more likely to have tonsillar lymphoid hyperplasia than tonsillitis. As a result, lymphoid hyperplasia would be a suitable term to describe this tonsillar condition. Respiratory tract diseases, rather than tonsillar hyperplasia, was the more likely cause of the poor racing performance of affected dogs. The aetiological relationship, if any, between respiratory disease and tonsillar enlargement is unclear from this study and requires further investigation.

Animals↗
PubMed2001

Direct detection of a microlens in the Milky Way.

The nature of dark matter remains mysterious, with luminous material accounting for at most approximately 25 per cent of the baryons in the Universe. We accordingly undertook a survey looking for the microlensing of stars in the Large Magellanic Cloud (LMC) to determine the fraction of Galactic dark matter contained in massive compact halo objects (MACHOs). The presence of the dark matter would be revealed by gravitational lensing of the light from an LMC star as the foreground dark matter moves across the line of sight. The duration of the lensing event is the key observable parameter, but gives non-unique solutions when attempting to estimate the mass, distance and transverse velocity of the lens. The survey results to date indicate that between 8 and 50 per cent of the baryonic mass of the Galactic halo is in the form of MACHOs (ref. 3), but removing the degeneracy by identifying a lensing object would tighten the constraints on the mass in MACHOs. Here we report a direct image of a microlens, revealing it to be a nearby low-mass star in the disk of the Milky Way. This is consistent with the expected frequency of nearby stars acting as lenses, and demonstrates a direct determination of a lens mass from a microlensing event. Complete solutions such as this for halo microlensing events will probe directly the nature of the MACHOs.

Journal Article↗
PubMed2001

Nitric oxide-dependent pro-oxidant and pro-apoptotic effect of metallothioneins in HL-60 cells challenged with cupric nitrilotriacetate.

Intracellular safeguarding functions of metallothioneins (MTs) include sequestering transition and heavy metals, scavenging free radicals and protecting against electrophiles. We report that MT protection against Cu-induced cytotoxicity can be reversed and pro-oxidant and pro-apoptotic effects can be induced in HL-60 cells exposed to NO. We demonstrate that in ZnCl(2)-pretreated HL-60 cells loaded with copper nitrilotriacetate (Cu-NTA), exposure to an NO donor, S-nitroso-N-acetyl penicillamine, resulted in S-nitrosylation and oxidation of MT cysteines. This disruption of MT Cu-binding thiolate clusters caused loosening and release of redox-active Cu, enhanced redox-cycling activity of Cu and increased peroxidation of major classes of membrane phospholipids. We also found that Cu-induced oxidative stress in ZnCl(2)-pretreated/Cu-NTA-loaded HL-60 cells was accompanied by apoptosis documented by characteristic changes of nuclear morphology, internucleosomal DNA cleavage, externalization of phosphatidylserine, release of cytochrome c from mitochondria into cytosol and activation of caspase-3. We conclude that in Cu-challenged cells, NO can reverse the protective role of MTs and convert them into pro-oxidant, pro-apoptotic implements.

Annexin A5↗
PubMed2001

Cholesterol favors phase separation of sphingomyelin.

The phase behavior of mixed lipid dispersions representing the inner leaflet of the cell membrane has been characterized by X-ray diffraction. Aqueous dispersions of phosphatidylethanolamine:phosphatidylserine (4:1 mole/mole) have a heterogeneous structure comprising an inverted hexagonal phase H(II) and a lamellar phase. Both phases coexist in the temperature range 20-45 degrees C. The fluid-to-gel mid-transition temperature of the lamellar phase assigned to phosphatidylserine is decreased from 27 to 24 degrees C in the presence of calcium. Addition of sphingomyelin to phosphatidylethanolamine/phosphatidylserine prevents phase separation of the hexagonal H(II) phase of phosphatidylethanolamine but the ternary mixture phase separates into two lamellar phases of periodcity 6.2 and 5.6 nm, respectively. The 6.2-nm periodicity is assigned to the gel phase enriched in sphingomyelin of molecular species comprising predominantly long saturated hydrocarbon chains because it undergoes a gel-to-fluid phase transition above 40 degrees C. The coexisting fluid phase we assign to phosphatidylethanolamine and phosphatidylserine and low melting point molecular species of sphingomyelin which suppresses the tendency of phosphatidylethanolamine to phase-separate into hexagonal H(II) structure. There is evidence for considerable hysteresis in the separation of lamellar fluid and gel phases during cooling. The addition of cholesterol prevents phase separation of the gel phase of high melting point sphingomyelin in mixtures with phosphatidylserine and phosphatidylethanolamine. In the quaternary mixture the lamellar fluid phase, however, is phase separated into two lamellar phases of periodicities of 6.3 and 5.6 nm (20 degrees C), respectively. The lamellar phase of periodicity 5.6 nm is assigned to a phase enriched in aminoglycerophospholipids and the periodicity 6.3 nm to a liquid-ordered phase formed from cholesterol and high melting point molecular species of sphingomyelin characterized previously by ESR. Substituting 7-dehydrocholesterol for cholesterol did not result in evidence for lamellar phase separation in the mixture within the temperature range 20-40 degrees C. The specificity of cholesterol in creation of liquid-ordered lamellar phase is inferred.

Animals↗
PubMed2001

Kinetic phase behavior of distearoylphosphatidylethanolamine dispersed in glycerol.

Phase behavior of distearoylphosphatidylethanolamine dispersed in excess glycerol has been examined by differential scanning calorimetry. Transformation from lamellar-gel to lamellar crystalline phase was found to take place at temperatures near 74.9 degrees C upon cooling and near 76.3 degrees C during heating scans. The transition can also be observed under isothermal conditions at temperature in this range. The kinetics of the transformation from lamellar-gel to lamellar-crystal phase was analyzed by the well-known Avrami equation. The apparent Avrami exponents were found to be approximately 1.6. The effective dimensionality of the growth pattern can then be set as 1, after taking into account the contribution of nucleation at the examination temperatures. The activation energy of the phase transition was estimated as approximately 255 kJ mol(-1). The data are discussed in terms of development of successful cryoprotective strategies using glycerol.

Calorimetry, Differential Scanning↗
PubMed2001

Quantitative analysis of phospholipid peroxidation and antioxidant protection in live human epidermal keratinocytes.

To characterize oxidative stress in phospholipids of normal human epidermal keratinocytes we metabolically labeled their membrane phospholipids with a natural oxidation-sensitive fluorescent fatty acid, cis-parinaric acid, and exposed the cells to two different sources of oxidants--a lipid-soluble azo-initiator of peroxyl radicals, 2,2'-azobis(2,4-dimethyl-valeronitrile), AMVN, and a superoxide generator, xanthine oxidase/xanthine. We demonstrated that both oxidants induced pronounced oxidation of four major classes of cis-parinaric acid-labeled phospholipids-phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and phosphatidylinositol-in normal human epidermal keratinocytes that was not detectable as any significant change of their phospholipid composition. Vitamin E was effective in protecting the cells against phospholipid peroxidation. Since viability of normal human epidermal keratinocytes was not changed either by labeling or exposure to oxidants the labeling protocol and oxidative stress employed are compatible with the quantitative analysis of phospholipid peroxidation in viable cells.

Adult↗
PubMed2001

The effect of breastfeeding on child development at 5 years: a cohort study.

OBJECTIVE: It is uncertain to what degree the relationship between breastfeeding and later cognitive development is a true biological effect, or is confounded by psychosocial factors. The study aim was to further investigate this relationship and the effect of duration of breast feeding on cognitive development. METHODS: A total of 3880 children were followed from birth. Breastfeeding duration was measured by questionnaire at 6 months of age and a Peabody Picture Vocabulary Test Revised (PPVT-R) was administered at 5 years. PPVT-R scores were adjusted for the effects of a large array of biological and psychosocial confounders. The relationship between breastfeeding and the mean PPVT-R scores were examined using analysis of variance and multiple linear regression. RESULTS: A strong positive relationship was demonstrated between breastfeeding and the PPVT-R scores with increasing scores with increased duration of breastfeeding. After adjusting for a wide range of biological and social factors, the adjusted mean for those breastfed for 6 months or more was 8.2 points higher for females and 5.8 points for males when compared to those never breastfed. CONCLUSION: These findings suggest a significant benefit to child development is conferred by breastfeeding and is related independently to longer periods of breastfeeding.

Adult↗
PubMed2000

The distribution of alpha-tocopherol in mixed aqueous dispersions of phosphatidylcholine and phosphatidylethanolamine.

The effect of alpha-tocopherol on the structure and phase behaviour of mixed aqueous dispersions of phosphatidylcholine and phosphatidylethanolamine has been examined by synchrotron X-ray diffraction. Equimolar mixtures of dioleoylphosphatidylethanolamine:dioleoylphosphatidylcholine and dimyristoylphosphatidylcholine:dioleoylphosphatidylethanolamine did not show evidence of phase separation of an inverted hexagonal structure typical of alpha-tocopherol and phosphatidylethanolamine from lamellar phase. Mixed dispersions of dioleoyl derivatives of phosphatidylethanolamine:phosphatidylcholine (3:1) form a typical miscible gel phase at low temperatures but which phase separates into lamellar liquid-crystal and inverted hexagonal phases at temperatures greater than 65 degrees C. The presence of 1, 2 or 5 mol% alpha-tocopherol caused a decrease in the temperature at which the inverted hexagonal phase appears. Phase separation of non-lamellar phase from lamellar gel phase can be detected in the presence of 7.5 and 10 mol% alpha-tocopherol, indicating a limited capacity of the phosphatidylcholine to incorporate alpha-tocopherol into the lamellar domain. A partial phase diagram of the ternary mixture has been constructed from the X-ray scattering data. It was concluded that there is no preferential interaction of alpha-tocopherol with phosphatidylethanolamine in mixed aqueous dispersions containing phosphatidylcholines.

Lipid Bilayers↗
PubMed2000

The structure and thermotropic phase behaviour of dipalmitoylphosphatidylcholine codispersed with a branched-chain phosphatidylcholine.

The structure and thermotropic phase behaviour of a fully hydrated binary mixture of dipalmitoylphosphatidylcholine and a branched-chain phosphatidylcholine, 1, 2-di(4-dodecyl-palmitoyl)-sn-glycero-3-phosphocholine, were examined using differential scanning calorimetry, synchrotron X-ray diffraction and freeze-fracture electron microscopy. The branched-chain lipid forms a nonlamellar phase when dispersed alone in aqueous medium. Mixed aqueous dispersions of the two phospholipids containing less than 33 mol% of the branched-chain lipid form lamellar phases over the whole temperature range were studied (4 degrees C to 60 degrees C). When present in proportions greater than 33 mol% it induces a hexagonal phase in mixed aqueous dispersions with dipalmitoylphosphatidylcholine at temperatures above the fluid phase transition. At temperatures below 35 degrees C a hexagonal phase coexists with a gel bilayer phase. The lamellar<-->nonlamellar transition can be explained satisfactorily on the basis of the shape of the molecule expressed in terms of headgroup and chain cross-sectional areas. At temperatures below 35 degrees C macroscopic phase separation of two gel phases takes place. Freeze-fracture electron microscopy revealed that one gel phase consists of bilayers with a highly regular, periodic superstructure (macro-ripples) whereas the other phase forms flat, planar bilayers. The macro-ripple phase appears to represent a relaxation structure required to adapt to the packing constraints imposed by the incorporation of the branched-chain lipid into the dipalmitoylphosphatidylcholine host bilayer. The data suggest that structural changes that take place on cooling the mixed dispersion below the lamellar<-->nonlamellar phase transition temperature cannot be adequately described using the molecular form concept. Instead it is necessary to take into account the detailed molecular form of the guest lipid as well as its physical properties.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed2000

The effect of dimethyl sulphoxide on the structure and phase behaviour of palmitoleoylphosphatidylethanolamine.

The thermotropic phase behaviour and structure of a nonbilayer-forming lipid, 1-palmitoyl-2-oleoyl-phosphatidylethanolamine, dispersed in water and in aqueous solutions of up to 50 wt% dimethyl sulphoxide (DMSO) have been characterised using synchrotron X-ray diffraction methods. It was found that the presence of DMSO in the solvent induced an increase in the temperature of lamellar-gel to lamellar-liquid-crystal phase transition and a decrease in the temperature of the lamellar-liquid-crystal to inverted-hexagonal phase transition of the phospholipid. The presence of DMSO also caused a decrease in the X-ray repeat spacings of all the phases studied. Electron density profiles of the phospholipid dispersed in water and 50 wt% DMSO in the bilayer gel state were calculated. The presence of 50 wt% DMSO caused the apparent disappearance of the solvent layer separating phospholipid bilayers in the gel state. The results suggest that DMSO contributes to the bilayer electron density profile and that the amphiphilic solvent molecules partition into the interfacial region.

Dimethyl Sulfoxide↗
PubMed2000

Phospholipid signaling in apoptosis: peroxidation and externalization of phosphatidylserine.

The role of phospholipids in apoptosis signaling and the relationship between oxidation of phosphatidylserine and its redistribution in the plasma membrane were studied. A novel method for detection of site-specific phospholipid peroxidation based on the use of cis-parinaric acid as a reporter molecule metabolically integrated into membrane phospholipids in living cells was employed. When several tissue culture cell lines and different exogenous oxidants were used, the relationship between the oxidation of phosphatidylserine and apoptosis has been revealed. The plasma membrane was the preferred site of phosphatidylserine oxidation in cells. It was shown that selective oxidation of phosphatidylserine precedes its translocation from the inside to the outside surface of the plasma membrane during apoptosis. A model is proposed in which cytochrome c released from mitochondria by oxidative stress binds to phosphatidylserine located at the cytoplasmic surface of the plasma membrane and induces its oxidation. Interaction of peroxidized phosphatidylserine with aminophospholipid translocase causes inhibition of the enzyme relevant to phosphatidylserine externalization.

Animals↗
PubMed2000

Ripple phases induced by alpha-tocopherol in saturated diacylphosphatidylcholines.

The effect of alpha-tocopherol on the structure and phase behavior of dilauroyl-, dimyristoyl-, dipalmitoyl-, and distearoyl-phosphatidylcholines was examined using X-ray diffraction and freeze-fracture electron microscopic methods. A ripple phase was observed in all of the mixtures at temperatures well below the pretransition temperature of the corresponding pure phospholipid. Freeze-fracture studies indicated that with proportion of alpha-tocopherol less than 5 mol% a ripple phase with large periodicity (50-150 nm) predominated and with about 10 mol% alpha-tocopherol a ripple phase of periodicity about 16 nm was formed. With more than 10 mol% alpha-tocopherol planar bilayers tended to be formed. Partial phase diagrams of mixed aqueous dispersions of saturated phosphatidylcholines and alpha-tocopherol over temperature ranges about the gel to liquid-crystal phase boundary have been constructed. Alpha-tocopherol-enriched domains form ripple phases that coexist with regions of lamellar gel phase of the pure phospholipid in mixtures containing less than 10 mol% alpha-tocopherol. The presence of increasing amounts of alpha-tocopherol in the phospholipid causes an increase in the proportion of ripple phase at the expense of pure phospholipid bilayer indicating that the alpha-tocopherol-enriched domains might possess a defined stoichiometry of the two constituents.

Dimyristoylphosphatidylcholine↗
PubMed2000

Hydration of DMPC and DPPC at 4 degrees C produces a novel subgel phase with convex-concave bilayer curvatures.

Hydration of dimyristoyl- and dipalmitoylphosphatidylcholines at 4 degrees C results in the formation of a characteristic subgel phase designated Pcc. Examination of the phase by freeze-fracture electron microscopy shows convex-concave deformations of the planar bilayer which are of two types. A smaller type with a radius of curvature of about 20 nm predominates in DMPC, and a larger type with about 70 nm radii of curvatures dominates in DPPC. The Pcc phase can also be formed in samples hydrated at temperatures above the main phase transition if the dispersion is frozen slowly and subsequently incubated at 4 degrees C for several days. The subgel Pcc phase was distinguished from the subgel Lc phase by the temperature of transition, packing of the acyl chains on the basis of wide-angle X-ray diffraction, and 2H-NMR spectra characteristic of a 'solid-ordered' phase. Vibrational spectra of the carbonyl and phosphate regions are consistent with a partially reduced hydration state. The origin of the convex-concave bilayer deformation is believed to result from constraints imposed by limiting hydration of the headgroup and a frustration arising from the spontaneous curvature of both monolayers.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed2000

Elimination of volatile chemicals in disinfectant evaluation procedures by freeze drying.

Standard methods for the removal or inactivation of disinfectants from reaction mixtures are often unsatisfactory when dealing with high concentrations of particulate organic matter. A method employing freeze drying was developed to eliminate residual disinfectant activity from reaction mixtures. Following the addition of volatile chemicals to sterilized cattle slurry, samples were removed from the mixture, freeze dried, and the dried material was reconstituted with diluent containing Escherichia coli as the test micro-organism to determine if the volatile chemicals had been removed. Twelve chemicals and one commercial disinfectant (Virkon S) were evaluated at different concentrations. The chemicals selected were acetone, ammonium hydroxide, carbon tetrachloride, chloroform, dichloromethane, ethyl alcohol, formalin, hydrogen peroxide, isopropyl alcohol, peracetic acid, sodium hypochlorite and xylene. Eight chemicals, acetone, ammonium hydroxide, carbon tetrachloride, chloroform, dichloromethane, ethyl alcohol, isopropyl alcohol and xylene were removed by freeze drying. The remaining four chemicals and the commercial disinfectant, which were not removed by freeze drying, could not be evaluated by this method.

Animals↗