Biomedical subjects

J A Hughes

Publications and source records attributed to J A Hughes.

At least 19 recordsLinked to original sources

PubMed2007

alpha7 Nicotinic receptor gene delivery into mouse hippocampal neurons leads to functional receptor expression, improved spatial memory-related performance, and tau hyperphosphorylation.

Brain alpha7 nicotinic receptors have become therapeutic targets for Alzheimer's disease (AD) based on their memory-enhancing and neuroprotective actions. This study investigated the feasibility of increasing neuronal alpha7 receptor functions using a gene delivery approach based on neuron-selective recombinant adeno-associated virus (rAAV)-derived vectors. In order to determine whether alpha7 receptor-mediated cytotoxicity was dependent on receptor density, rat alpha7 nicotinic receptors were expressed at high concentrations in GH4C1 cells as measured with nicotine-displaceable [3H]methyllycaconitine (MLA) binding. The potency of GTS-21 (an alpha7 receptor agonist) to induce cell loss was similar in these cells to that seen in pheochromocytoma (PC12) cells expressing nine-times-lower receptor levels, suggesting that cytotoxicity was more dependent on agonist concentration than receptor density. Hippocampal transduction with rat alpha7 nicotinic receptors increased [3H]MLA binding in this region in wild type and alpha7 receptor-knockout (KO) mice without apparent cytotoxicity. No difference was observed in Kd values for MLA binding between endogenous and transgenic receptors. Single cell recordings demonstrated that dentate granule cells that normally have no alpha7 receptor response did so following alpha7 receptor gene delivery in wild type mice. Recovery of alpha7 function was also observed in stratum oriens and stratum radiatum neurons of KO mice following gene delivery. Wild type mice exhibited improved acquisition performance in the Morris water task 1 month after bilateral hippocampal transductions with the rat alpha7 receptor gene compared with green fluorescent protein-transduced controls. However, both groups reached similar training levels and there was no difference in subsequent probe performance. Finally, this gene delivery approach was used to test whether alpha7 receptors affect tau-phosphorylation. Chronic (i.e. 2 month but not 2 week) expression of high levels of alpha7 receptors in hippocampus increased AT8 staining characteristic of hyperphosphorylated tau in that region, indicating that endogenous agonist-mediated receptor activation may be able to modulate this process.

Aconitine↗

PubMed2005

The relationship between enamel softening and erosion caused by soft drinks at a range of temperatures.

OBJECTIVES: Investigations of the erosive potential of soft drinks are usually performed at room or body temperature, but drinks are more frequently served chilled, with ice, or hot. Since the rate of chemical reactions usually increases with temperature, it is predicted that erosion is more severe at high temperatures and reduced at low temperatures. The aim of this study was to investigate the correlation between enamel softening, enamel erosion, and temperature. METHODS: Atomic force microscopy nanoindentation and non-contact optical profilometry were used to assess changes in enamel nanomechanical properties after 5 min and erosive material loss after 30 min exposure to two different non-carbonated soft drinks at 4, 25, 50 and 75 degrees C. RESULTS: For one drink (Robinson's Original Juice Drink), there was a statistically significant difference between nanomechanical properties and erosion depth at all temperatures, with softening and erosion increasing with temperature. For another drink (Ribena ToothKind Juice Drink), there was a slight softening and virtually no material loss, and temperature had no statistically significant impact on erosion. There was a good linear correlation (R2 = 0.94) between nanomechanical properties and material loss. CONCLUSIONS: The difference between the drinks can be explained by their composition. For the erosive drink, material loss increased, and nanohardness decreased, approximately linearly with temperature. The correlation between softening and erosion demonstrated that nanomechanical properties after very short erosion times can be considered a good predictor of bulk material loss after considerably longer erosion times.

Beverages↗

PubMed2004

A methodology for testing the erosive potential of sports drinks.

OBJECTIVES: The aim of the study was to develop and test a methodology in situ, which simulated the consumption of sports drinks. A secondary aim was to assess the acceptability of the method to sedentary participants. METHODS: To select the sports drink for the study in situ, five commercially available sports drinks were examined for erosive potential in vitro. The study in situ was a single centre, 2-period, 2-treatment crossover study to compare the erosive effect of a commercially available sports drink (Test), with that of mineral water (Control), over 10 day periods on 10 healthy volunteers. Subjects wore upper removable appliances containing two human enamel specimens from 9 a.m. to 5 p.m. The regimen of intake of the drinks was 350 ml in 10, 5-min rest, 650 ml in 25, 5-min rest, 500 ml in 10 and 5-min rest. Measurements of enamel loss were made on samples after 5 and 10 days by profilometry. RESULTS: The in situ study showed a statistically significant difference in erosive potential between the test and control beverages. No specimen exposed to the control beverage displayed appreciable erosion. Erosion occurred with the test drink, but to a variable degree between subjects. The subjects unanimously found the drinking regimen unpleasant. CONCLUSIONS: The sports drink caused significantly more erosion in situ than water and as seen in other studies, there was marked variation in susceptibility to erosion between subjects. The new drinking regimen was designed to simulate pre, during and post-exercise intake. Although all the sedentary subjects participating in this study reported that they found the volume of fluids consumed over a short period of time excessive it is unlikely that this would prove problematic in the exercise environment.

Adult↗

PubMed2004

Modification of soft drinks with xanthan gum to minimise erosion: a study in situ.

OBJECTIVE: To compare the erosive effect of a new blackcurrant/calcium drink with xanthan gum, on enamel in situ, with a low erosive product, a conventional fruit beverage and water. DESIGN: The study was single centre, randomised, single blind, 4 treatment crossover design. SETTING: This study, conducted in 2000, employed volunteers working at the Bristol Dental Hospital, UK. SUBJECTS, MATERIALS AND METHODS: 16 subjects (>or=18 years) wore two enamel samples in a removable acrylic appliance. The drinks tested were (A) New blackcurrant/calcium/gum drink (test product), (B) Original blackcurrant/calcium drink, (C) Conventional blackcurrant drink (positive control) and (D) Water (negative control), for 15 day study periods. Drinking was supervised, with 250 ml imbibed four times/day between 9.00am-5.00pm. Profile measurements of specimens were made at baseline, 5, 10 and 15 days. MAIN OUTCOME MEASURES: Paired t-tests compared erosion by surfometry with selected pairs of formulations. RESULTS: Of 16 screened subjects (3 male, 13 female), mean age 34.2 years, 1 subject failed to complete the study. A caused significantly less enamel loss than C, with no statistically significant differences from B at any time points measured. Of 43 treatment emergent adverse events, none were considered related to the study formulations. CONCLUSIONS: A retained low erosive properties similar to B, with additional benefits of taste flexibility and beverage stability.

Adolescent↗

PubMed2004

Cutaneous haemangioma: prevalence and sonographic characteristics of associated hepatic haemangioma.

AIMS: To determine the prevalence of hepatic haemangiomas (HH) occurring with multiple or large cutaneous haemangiomas (CH), and compare the ultrasound (US) characteristics of HH with morphology of concurrent CH. MATERIALS AND METHODS: Retrospective and prospective sonographic and clinical analysis was undertaken on 69 patients referred for US to exclude silent HH. This comprised 62 patients referred over 6 years with six or more CH or one large CH (#10878;5 cm) and seven patients referred over 1 year with three to five small CH. RESULTS: HH occurred in 45% miliary CH and 40% mixed small and large CH, but also in 12-14% of patients with one large or three or more small CH. None had HH progression. There was association between HH sonographic characteristics and CH number and morphology. CONCLUSION: Clinically silent HH occurred in nearly one in four of infants presenting overall with six or more small CH or large CH. Previous studies have documented HH mortality between 18 and 80%, although this was not shown in our study. Therefore, if clinical suspicion exists, US is indicated to enable early steroid treatment if significant HH are present.

Child↗

PubMed2004

The protective effect of fluoride treatments against enamel erosion in vitro.

Fluoride is known to reduce enamel solubility during the caries process. The aim of this investigation was to determine whether fluoride preparations affect erosion attributed to citric acid and citric acid-based soft drinks. Flat enamel specimens embedded in epoxy resin were prepared from caries free, human third molar teeth extracted from patients aged 18-35 years. Specimens were polished to have a profile within a tolerance of +/-0.3 microm measured on a profilometer. Specimens were taped to leave a 2 mm window of exposed enamel. Groups of specimens were exposed to citric acid and soft drinks with and without the addition of sodium fluoride or exposed to the same solutions after pre-treatment with fluoride products. Enamel loss was measured by profilometer after 10, 20 and 30 min of acid exposure. The different acidic solutions varied significantly in the amount of erosion produced both with and without the addition of fluoride. In addition, the different fluoride products differed significantly in the protective effect afforded. Both fluoride application methods reduced in mean terms, enamel erosion at all time points and by all acidic solutions. The majority of differences were <25% and as the study was powered to show differences as significant at or above this level few reached statistical significance. Fluoride applied to enamel either in acidic solutions or as a pre-treatment, reduces enamel erosion; however, the actual clinical benefit appears low. More studies are required, including investigations in situ.

Adolescent↗

PubMed2003

Evaluation of spinal ultrasound in spinal dysraphism.

AIMS: The aims of this study were to evaluate the role of spinal ultrasound in detecting occult spinal dysraphism (OSD) in neonates and infants, and to determine the degree of agreement between ultrasound and magnetic resonance imaging (MRI) findings. MATERIALS AND METHODS: Eighty-five consecutive infants had spinal ultrasound over 31 months. Of these, 15 patients (age 1 day-7 months, mean 40 days; nine male) had follow-up MRI. Ultrasound and MRI findings were correlated retrospectively. RESULTS: Six out of 15 (40%) ultrasound examinations showed full agreement with MRI, seven of 15 (47%) had partial agreement, and two of 15 (13%) had no agreement. In the present series ultrasound failed to visualize: four of four dorsal dermal sinuses, three of four fatty filum terminales, one of one terminal lipoma, two of four partial sacral agenesis, three of four hydromyelia and one of 10 low-lying cords. CONCLUSION: Agreement between ultrasound and MRI was good, particularly for the detection of low-lying cord (90%). Therefore we recommend ultrasound as a first-line screening test for OSD. If ultrasound is abnormal, equivocal or technically limited, MRI is advised for full assessment.

Female↗

PubMed2003

Development of low erosive carbonated fruit drinks. 1. Evaluation of two experimental orange drinks in vitro and in situ.

OBJECTIVES: To determine the in vitro erosive potential and in situ erosive effect of two new formulation low calorie carbonated orange drinks with that of two conventional diet products and water. METHODS: In the in vitro study, six specimens of deciduous and permanent enamel were randomly allocated to each of the five products and a '4h' protocol employed. In the in situ study, 15 healthy volunteers participated in a single centre, single blind, 5-phase crossover study, conducted according to Good Clinical Practice, and employing a validated model. RESULTS: The in vitro erosive potential of the experimental formulations was less than that of the comparators at all time points. Conversely, the observed erosive potential of both experimental formulations was greater than that of the control. Consistent statistically significant differences were found in relation to permanent enamel only. Unfortunately, the in situ study did not produce results entirely consistent with those of the in vitro study. Notably, a generally progressive loss of enamel was observed in specimens exposed to the control. CONCLUSIONS: The data from the in vitro study show the experimental formulations to have low comparative erosivity. However, the methodologies in vitro and in situ somewhat unusually do not correlate in ranking the erosivity of drinks. The results of this study should therefore be viewed with caution, further research being clearly warranted.

Adolescent↗

PubMed2003

Development of low erosive carbonated fruit drinks 2. Evaluation of an experimental carbonated blackcurrant drink compared to a conventional carbonated drink.

OBJECTIVE: Previous studies demonstrated that a non-carbonated blackcurrant juice drink with added calcium produced little erosion of enamel in vitro and in situ by comparison with other low pH fruit drinks. The aim of this study is to evaluate the erosive effects in situ of an experimental formulation carbonated drink with added calcium, compared to a conventional carbonated drink over a 20 day period using a similar design as in previous studies. METHOD: The study was a single centre, single blind, randomised placebo controlled three cell crossover design involving 15 volunteers. The test drinks were an experimental carbonated ready-to-drink (RTD) blackcurrant drink, a conventional carbonated RTD orange drink and water. Two enamel samples were retained in situ, in the midline of the hard palate on an upper removable acrylic appliance. Drinks were 250 ml volumes consumed four times per day during 20 working days. Measurements of enamel loss were made on enamel samples after days 2, 5, 10, 15 and 20 by profilometry. RESULTS: The experimental carbonated blackcurrant drink caused significantly less enamel loss than the conventional carbonated orange drink at all time points measured, but was not significantly different from water at days 2, 5 and 20. CONCLUSIONS: This study has shown that it is possible to modify carbonated soft drinks in a manner similarly shown with non-carbonated soft drinks, to minimise dental erosion.

Adult↗

PubMed2002

Plasmid delivery in the rat brain.

Neurodegenerative diseases as a class do not have effective pharmacotherapies. This is due in part to a poor understanding of the pathologies of the disease processes, and the lack of effective medications. Gene delivery is an attractive possibility for treating these diseases. For the paradigm to be effective, efficient, safe and versatile vectors are required. In this study we evaluated three plasmid delivery systems for transgene expression in the rat hippocampus. Two of these systems were designed to have enhanced intracellular biodegradability. It was hypothesized that this system would be less toxic and could increase the free (non-vector) associated plasmids within the cell, leading to increased transgene activity. Polyethylenimine (PEI) and r-AAV-2 (recombinant adeno associated virus-2) were used as positive, non-viral and viral controls respectively, in the in vivo experiments. The results from the studies indicate there is a distinct difference between the various vectors in terms of total cells transfected, type of cell transfected, and toxicity. Non-viral systems were effective at transfecting both neurons and glia cells within the hippocampus, while the r-AAV-2 transfected mainly neurons. In summary, plasmid-mediated systems are effective for transgene expression within the brain and deserve further study.

Animals↗

PubMed2002

Dentine hypersensitivity: the effects of brushing toothpaste on etched and unetched dentine in vitro.

Although many people have exposed dentine, only a percentage exhibit symptoms of dentine hypersensitivity. This has been ascribed to opening and closing of the dentinal tubules by for example, smear layer changes or tubular occlusion. The aims of this study were to examine the surface morphological changes of etched and unetched dentine in vitro, attributed to the effects of toothbrushing with and without toothpastes designed for the alleviation of dentine hypersensitivity. A total of 96 etched and 96 unetched human dentine specimens were brushed with various toothpastes and water for 1, 2, 5 or 10-min periods in a toothbrushing machine and subsequently examined under scanning electron microscopy for surface changes. Analyses of brushed etched specimens demonstrated that time and treatment were significant variables (P < 0.05) for tubule occlusion. Further, the interaction between time and treatment was significant (P < 0.05). The artificial silica based paste was significantly better for all time intervals at occluding the dentine tubules. All toothpastes investigated caused dynamic changes to the smear layer of the unetched dentine, opening tubules. However, the artificial silica based paste resulted in occluded rather than patent tubules. All of the pastes evaluated had the capacity to remove the smear layer but some could then occlude tubules through the contained abrasives.

Acid Etching, Dental↗

PubMed2001

Membrane fluidity effects of estratrienes.

Estrogens have demonstrable neuroprotective effects. This fact has lead to the proposed use of estrogens for the prevention and/or treatment of Alzheimer's disease. The exact protective mechanism estrogens provide is not fully understood. In this report, a potential non-genomic mechanism for estratrienes involving alterations in membrane fluidity was studied. Steroids, such as estrogen, are known to be membrane-active and can alter the lipid packing. In this study we used fluorescent methodologies to address the effect of naturally occurring steroids (17alpha and 17beta-estradiol, testosterone, and progesterone) and new estratriene analogs on membrane fluidity using liposomes and HT-22 hippocampal cells. The study's results indicate steroids, based on the estratriene nucleus, can modulate lipid packing as evidenced by (1) decreased membrane fusion events and (2) decreased membrane fluidity. The effects on the membrane were both time and concentration dependent. It was also demonstrated through rational design estratriene analogs can be synthesized with enhanced membrane effects. Finally, in a glutamate-induced toxicity HT-22 model, we also demonstrated cellular protection with the estratriene-based molecules and analogs. The data suggest the plethora of cellular actions of estrogens may relate to or be influenced by membrane effects of the steroid.

Alzheimer Disease↗

PubMed2001

Transgene delivery with a cationic lipid in the presence of amyloid beta (betaAP) peptide.

The ability of a cationic lipid to deliver plasmid DNA (pDNA) in presence of the neurotoxic fragment of amyloid beta-peptide was evaluated. Pre-treatment of cells with betaAP (25-35) peptide resulted in a modest increase in transgene expression. When betaAP (25-35) peptide was mixed with the pDNA/liposome complex and used, the complexes lost their ability to transfect. However, the reverse sequenced betaAP (35-25) peptide demonstrated no significant differences in transgene expression in pre-treated cells, and in cells where betaAP (35-25) peptide was mixed with pDNA/liposome complexes and transfected. The amount of pDNA delivered to the cells was decreased in presence of betaAP (25-35) as measured with flow cytometry using fluorescently labeled liposomes. The decreased endocytosis may be due to their rod-like structure formation as demonstrated by electron microscopy and atomic force microscopy (AFM). These results demonstrate that betaAP (25-35) peptide may interfere with gene delivery with cationic systems.

Amyloid beta-Peptides↗

PubMed2001

Pharmacokinetics of plasmid DNA in the rat.

PURPOSE: The pharmacokinetics of plasmid DNA after IV bolus administration in the rat by following supercoiled (SC), open circular (OC), and linear (L) pDNA forms of the plasmid. METHODS: SC, OC, and L pDNA were injected at 2,500, 500, 333, and 250 microg doses. The concentrations in the bloodstream of OC and L pDNA were monitored. RESULTS: SC pDNA was detectable in the bloodstream only after a 2,500 microg dose, and had a clearance of 390(+/-50) ml/min and Vd of 81(+/-8) ml. The pharmacokinetics of OC pDNA exhibited non-linear characteristics with clearance ranging from 8.3(+/-0.8) to 1.3(+/-0.2) ml/min and a Vd of 39(+/-19) ml. L pDNA was cleared at 7.6(+/-2.3) ml/min and had a Vd of 37(+/-17) ml. AUC analysis revealed that 60(+/-10) % of the SC was converted to the OC form, and nearly complete conversion of the OC pDNA to L pDNA. Clearance of SC pDNA was decreased after liposome complexation to 87(+/-30) ml/min. However the clearance of OC and L pDNA was increased relative to naked pDNA at an equivalent dose to 37(+/-9) ml/min and 95(+/-37) ml/min respectively. CONCLUSIONS: SC pDNA is rapidly metabolized and cleared from the circulation. OC pDNA displays non-linear pharmacokinetics. Linear pDNA exhibits first order kinetics. Liposome complexation protects the SC topoform, but the complexes are more rapidly cleared than the naked pDNA.

Animals↗

PubMed2001

The effect of pH on the erosion of dentine and enamel by dietary acids in vitro.

The reported incidence of tooth erosion caused by acidic soft drinks has been increasingly documented. Citric and phosphoric acids are the two main dietary acids present in these soft drinks. Many variables need to be determined in order to assess risk factors for dental erosion caused by beverage consumption including pH, titratable acidity, pKa, buffering capacity, hence the aim of these in vitro investigations. Methodologies included profiling flat enamel and dentine samples (< +/- 0.3 microm profile) from unerupted human third molars. Groups of five specimens were placed in acidic solutions adjusted with alkali over the available pH range; citric, phosphoric and hydrochloric acid were adjusted with sodium hydroxide and citric acid with trisodium citrate. Tissue loss was calculated by profilometry. Results showed that under these conditions citric acid caused far more erosion over the pH range employed than phosphoric acid for both tissue types. Citric acid compared with hydrochloric acid highlighted dissolution and chelation effects. Phosphoric acid caused minimal erosion over pH 3 for enamel and pH 4 for dentine. These factors could be considered in order to reduce the erosivity of acidic soft drinks.

Adult↗

PubMed2000

Quantifiable analysis of human immunoglobulin heavy chain class-switch recombination to all isotypes.

Somatic recombinational events, including the immunoglobulin heavy chain class-switch, are a normal feature of B-cell maturation. To enable comprehensive and sensitive class-switch analysis in ex vivo human B cells, we have developed multiple digestion-circularization PCR (DC-PCR) techniques for quantifiable detection of switching to all immunoglobulin isotypes. This technology was validated by extensive sequencing of PCR products, tests with control non-lymphoid cells and B-cell lines of known isotypic specificities, and by demonstrating DC-PCR selectivity in a model system. With tonsillar B-cell DNA, switching to gamma 3, gamma 1, alpha1, gamma 2, gamma 4 and alpha2 isotypes was reproducibly detectable among different individuals. Levels of epsilon switching were relatively low and usually required higher total amounts of template DNAs for detection. Quantitation of alpha1 class switching in a panel of human tonsillar whole B cells was performed by the internal-competitor approach, and showed a pattern consistent with previous studies on IgA+ tonsillar cells. We demonstrate that these assays can rapidly show germline status or specific switch rearrangements in B lymphoid cell lines.

B-Lymphocytes↗

PubMed2000

Adeno-associated virus mediated gene transfer into primary rat brain neuronal and glial cultures: enhancement with the pH-sensitive surfactant dodecyl 2-(1'-imidazolyl) propionate.

This study evaluated the effects of a novel, pH-sensitive surfactant, dodecyl 2-(1'-imidazolyl) propionate (DIP), on cationic lipid mediated transfection in primary rat brain neuronal and glial cultures. The cationic lipid complex DOTAP/DOPE (1, 2-dioleoyl-3-trimethylammonium propionate and dioleoyl phosphatidylethanolamine, respectively) was added over a range of concentrations (0-120 microg/ml) with DNA concentration kept constant (1.6 microg/ml). The neuron-specific enolase (NSE) and cytomegalovirus (CMV) promoters were found to drive green fluorescent protein (GFP) expression in neuron-enriched and glial cultures, respectively, using adeno-associated virus (AAV) derived constructs. NSE-driven GFP expression was not observed in glial cultures. Addition of DOTAP/DOPE increased transfection efficiency over a wide range of lipid concentrations (5-50 microg/ml) keeping DNA concentration constant (1.6 microg/ml). Addition of DIP to the lipid/DNA complex increased maximum transfection efficiencies in glial and neuronal cultures 2-3-fold. Transfection efficiencies were at their maximum with a similar total lipid concentration (50 microg/ml) in both cell-types in the presence of DIP. Neuronal cultures were more sensitive than glia to the toxic actions of DOTAP/DOPE, with or without DIP. These results indicate that AAV-mediated gene-transfer to neurons and glia can be facilitated by addition of a pH-sensitive surfactant to cationic liposome/DNA complexes and that endosomal escape could be a limiting factor in transgene expression.

Animals↗

PubMed2000

Effects of pH and concentration of citric, malic and lactic acids on enamel, in vitro.

OBJECTIVE: Dental erosion associated with soft drink consumption probably results from the contained dietary acids in the formulations. The pH value of any formulation is an important variable in acid erosion but not necessarily the only important factor. The aim of this study was to measure enamel erosion by citric, malic and lactic acids at pH values and acid concentrations representative of a range found in soft drink formulations and to determine the effect of adding calcium to citric acid. METHODS: Flat ground enamel samples were prepared from unerupted human third molar teeth. Groups of five specimens were placed in citric, malic and lactic acid solutions of different pH and acid concentration for three by 10 min exposures at 35 degrees C. Enamel loss was measured by profilometry. Enamel specimens were also exposed to citric acid solutions containing calcium at different pH values and at the same pH with different concentrations of calcium. RESULTS: Numerical data and contour plots for each acid showed a similar pattern for increasing erosion with decreasing pH and increasing acid concentration and vice versa for decreasing erosion. Increasing the concentration of calcium in a fixed pH citric acid solution resulted in decreased erosion. This effect was most marked at higher pH. CONCLUSIONS: This study has shown that under highly controlled conditions the erosion of enamel by solutions of dietary acids is influenced by the interplay of pH, acid concentration and presence of calcium. These variables and in particular the concentration of calcium could be manipulated to produce soft drinks with reduced erosivity to enamel.

Beverages↗