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W Smith

Publications and source records attributed to W Smith.

At least 127 records · Page 7Linked to original sources
PubMed1997

The effect of delta-gluconolactone, an oxidised analogue of glucose, on the nonenzymatic glycation of human and rat haemoglobin.

Nonenzymatic glycation of proteins and oxidative stress are considered independent factors important in the development of the complications of diabetes but may be interrelated by the process of autoxidative glycation. This pathway involves monosaccharide autoxidation to a reactive ketoaldehyde analogue and subsequent reaction with protein to form a ketoimine adduct. This study demonstrates that delta-gluconolactone (delta-GL), an oxidised analogue of glucose, is a potent glycating agent in vitro of haemoglobin present in blood samples from insulin-dependent diabetic and non-diabetic human subjects and from spontaneously diabetic, insulin-dependent BB/Edinburgh (BB/E) rats. The percentage glycated haemoglobin after incubation (37 degrees C, 5 h) with delta-GL (25 mmol/l) was significantly (P < 0.002) higher than that observed using an equimolar concentration of glucose. Intravenous administration of delta-GL (1 g/kg) to non-diabetic BB/E rats also significantly increased glycation of haemoglobin (6.0 +/- 0.1% vs 4.9 +/- 0.1%, P < 0.01) whereas intravenous injection of an identical dose of glucose had no significant effect (5.1 +/- 0.1% vs 5.0 +/- 0.2%). These results support the hypothesis that nonenzymatic glycation of proteins involves attachment by both native and oxidised monosaccharides. Further investigation of the interactions between diabetes-associated increases in oxidative stress and glycation on the development and progression of the vascular complications of diabetes is necessary.

Animals↗
PubMed1997

Cytotoxic T-cell clone against rectal carcinoma induced by stimulation of a patient's peripheral blood mononuclear cells with autologous cultured tumor cells.

In an effort to establish cytolytic T lymphocytes (CTLs) against colorectal carcinoma (CRC) by stimulating patients' lymphocytes with autologous tumor cells, we used peripheral blood mononuclear cells (PBMC) from a patient with minimal residual rectal carcinoma following removal of the primary lesion and involved regional lymph nodes as a source to generate CTLs in culture. A CTL line and clone were established from the patient's PBMC following stimulation of PBMC with autologous, cultured tumor cells and interleukin-2. The CTL line and the clone consisted predominantly of CD4+ lymphocytes. The CTL clone expressed two T-cell receptor variable alpha chains (V alpha11 and V alpha22) and one beta chain (Vbeta14). The cytokine secretion pattern of the CTL line was of the Th1-type. Both the CTL line and the clone lysed the autologous rectal carcinoma cells, but not the allogeneic, partially human lymphocyte antigen (HLA)-matched or nonmatched CRC cells, autologous Epstein-Barr virus-transformed B cells, K562 (natural killer target) cells or Daudi (lymphokine-activated killer target) cells. Lysis of autologous tumor cells most likely was HLA class I-restricted. Our unique success in generating CTLs against this tumor type may rest in the inclusion of a patient with minimal residual, rather than advanced, disease.

Amino Acid Sequence↗
PubMed1997

Association of pseudoexfoliation syndrome with increased vascular risk.

PURPOSE: To examine vascular associations with pseudoexfoliation syndrome in view of the wide-spread elastosis now demonstrated in this disorder that affects many tissues, including vessel walls. METHODS: The Blue Mountains Eye Study is a population-based study of eye disease in an area west of Sydney, Australia. Of 4433 eligible persons aged 49 years or older, 3,654 (82.4%) participated. Signs of pseudoexfoliation were graded clinically, after excluding 108 people who had bilateral cataract surgery. RESULTS: Pseudoexfoliation was present in 81 (2.3%) of 3546 participants aged 49 years or older. The prevalence of pseudoexfoliation increased with age and was higher in women and in people with glaucoma. Pseudoexfoliation was statistically significantly associated with a history of angina or hypertension or a combined history of angina, acute myocardial infarction, or stroke. CONCLUSION: Slit-lamp signs of pseudoexfoliation may identify individuals with an increased vascular risk.

Aged↗
PubMed1997

Serum beta carotene, alpha tocopherol, and age-related maculopathy: the Blue Mountains Eye Study.

PURPOSE: To assess associations between serum beta carotene, alpha tocopherol, and age-related maculopathy. METHODS: We studied 156 subjects with age-related maculopathy matched for age, sex, and month of blood collection to 156 control subjects without age-related maculopathy. Subjects were identified from the Blue Mountains Eye Study: those with late age-related macular degeneration and early age-related maculopathy using examination and grading of retinal photographs, and control subjects without age-related maculopathy randomly sampled from the study population. RESULT: Neither serum alpha tocopherol nor beta carotene was significantly associated with age-related maculopathy. CONCLUSION: These findings provide no evidence of a protective association between serum alpha tocopherol or beta carotene and age-related maculopathy.

Australia↗
PubMed1997

Identification of pore-forming subunit of P-type calcium channels: an antisense study on rat cerebellar Purkinje cells in culture.

Treatment of cerebellar neurones in culture with an antisense oligonucleotide (ODN) against alpha1A, reduced the whole-cell P-type calcium channel current relative to mismatch ODN treated controls (p < 0.001). Therefore, AgaIVA (50 nM) reduced whole-cell calcium current in mismatch and antisense treated cells by 70 +/- 4 and 19 +/- 3%, respectively.

Animals↗
PubMed1997

Is there an association between migraine headache and open-angle glaucoma? Findings from the Blue Mountains Eye Study.

OBJECTIVE: To determine whether an association exists between migraine headache history and open-angle glaucoma (OAG). DESIGN: Population-based, cross-sectional study. PARTICIPANTS: Subjects were 3654 people aged 49 or older; 82% of permanent residents from an area west of Sydney participated. INTERVENTION: All participants underwent an interview and a detailed eye examination, including automated perimetry and stereo optic disc photography. MAIN OUTCOME MEASURES: Open-angle glaucoma was diagnosed in subjects with matching typical glaucomatous visual field defects and pathologic optic disc cupping, independent of intraocular pressure level. The diagnosis of migraine history (typical or nontypical) was based on participant responses to specific questions, consistent with International Headache Society criteria. RESULTS: Open-angle glaucoma prevalence increased exponentially with age, with rates of 0.4%, 1.3%, 4.7%, and 11.4% among persons aged less than 60 years, between 60 and 69 years, between 70 and 79 years, and 80 years or older, respectively. The frequency of reporting a past history of typical migraine headache declined with increasing age, with rates of 23.1%, 16.2%, 12.8%, and 10.4% for corresponding age groups. For all age groups combined, there was no significant association between typical migraine headache and OAG (odds ratio [OR], 1.3; 95% confidence interval [CI], 0.8-2.2), after multivariate adjustment. However, after stratifying into 10-year age groups, increased odds for OAG were found for people giving a history of typical migraine headache and aged 70-79 years (OR, 2.5; 95% CI 1.2-5.2), after adjusting for variables found associated with glaucoma. This association was marginally stronger for high-pressure OAG cases (OR, 2.7; 95% CI, 1.1-5.6). CONCLUSIONS: These data suggest the possibility of an association between history of typical migraine headache and OAG, which could be modified by age.

Aged↗
PubMed1997

Prevalence and associations of epiretinal membranes. The Blue Mountains Eye Study, Australia.

PURPOSE: The purpose of the study is to determine the prevalence and associations of epiretinal membranes in a defined older Australian population and to assess their influence on visual acuity. METHODS: Three thousand six hundred fifty-four persons 49 years of age or older, representing 88% of permanent residents from an area west of Sydney, underwent a detailed eye examination, including stereo retinal photography. Epiretinal membranes were diagnosed clinically and from photographic grading. RESULTS: Signs of epiretinal membranes were found in 243 participants (7%; 95% confidence interval [CI], 6.1, 7.6), bilateral in 31%. The prevalence was 1.9% in persons younger than 60 years of age, 7.2% in persons 60 to 69 years of age, 11.6% in persons 70 to 79 years of age, and 9.3% in persons 80 years of age and older, with slightly higher rates in women. Two stages were identified: an early form without retinal folds, termed "cellophane macular reflex" present in 4.8%, and a later stage with retinal folds, termed "preretinal macular fibrosis" (PMF), found in 2.2% of the population. Preretinal macular fibrosis, but not cellophane macular reflex, had a small, significant effect on visual acuity. Preretinal macular fibrosis was significantly associated with diabetes, after age-gender adjustment, in subjects without signs of diabetic retinopathy (odds ratio, 3.2; 95% CI, 1.4, 7.2). Preretinal macular fibrosis also was associated with increased fasting plasma glucose (odds ratio, 1.2; 95% CI, 1.1, 1.3). Epiretinal membranes were found in 16.8% of persons who had undergone cataract surgery in one or both eyes (including PMF in 3.7%), in 16.1% of retinal vein occlusion cases (PMF in 12.5%), both significantly higher rates than in subjects without these conditions (P < 0.0001), and in 11% of persons with diabetic retinopathy (PMF in 3.6%), not significantly higher (P = 0.17). CONCLUSIONS: This study has documented the frequency and mild effect on vision of epiretinal membranes in an older population. Diabetes was associated significantly with idiopathic cases, whereas well-known associations with past cataract surgery and retinal disease were confirmed.

Adult↗
PubMed1997

Open-angle glaucoma and diabetes: the Blue Mountains eye study, Australia.

PURPOSE: The authors explore the relationship between diabetes and open-angle glaucoma in a defined older Australian population. METHODS: Three thousand six hundred fifty-four people 49 to 96 years of age, living west of Sydney, underwent a detailed eye examination. This included automated perimetry, stereo optic disc photographs, and applanation tonometry; in addition, fasting plasma glucose levels were ascertained. Glaucoma was diagnosed if matching visual field and optic disc cupping were present, without reference to intraocular pressure (IOP) level. Ocular hypertension (OH) was diagnosed if IOP in either eye was > or = 22 mm and glaucomatous disc and visual field changes were absent. RESULTS: Glaucoma prevalence was increased in people with diabetes, diagnosed from history or elevated fasting plasma glucose level (5.5%), compared with those without diabetes (2.8%; age-gender adjusted odds ratio [OR] 2.12, 95% confidence intervals [CI] 1.18-3.79). Ocular hypertension was also more common in people with diabetes (6.7%), compared with those without diabetes (3.5%; OR 1.86, CI 1.09-3.20). Diabetes was present in 13.0% of people with glaucoma, compared with 6.9% of those without glaucoma. This increase was highest for previously diagnosed glaucoma cases (16.7%; OR 2.82, CI 1.35-5.87). However, in 67% of such cases, glaucoma was diagnosed before the diabetes. For those not receiving glaucoma treatment, IOP was consistently slightly higher in people with diabetes, with the age-gender adjusted mean IOP 0.6 mm higher. CONCLUSIONS: The significant and consistent association between diabetes and glaucoma found in our study, which appeared independent of the effect of diabetes on IOP, suggests that there is a real association between these two diseases.

Aged↗
PubMed1997

The expression of voltage-dependent calcium channel beta subunits in human cerebellum.

The beta subunits of voltage-dependent calcium channels, exert marked regulatory effects on the biophysical and pharmacological properties of this diverse group of ion channels. However, little is known about the comparative neuronal expression of the four classes of beta genes in the CNS. In the current investigation we have closely mapped the distribution of beta1, beta2, beta3 and beta4 subunits in the human cerebellum by both in situ messenger RNA hybridization and protein immunohistochemistry. To our knowledge, these studies represent the first experiments in any species in which the detailed localization of each beta protein has been comparatively mapped in a neuroanatomically-based investigation. The data indicate that all four classes of beta subunits are found in the cerebellum and suggest that in certain neuronal populations they may each be expressed within the same cell. Novel immunohistochemical results further exemplify that the beta voltage-dependent calcium channel subunits are regionally distributed in a highly specific manner and studies of Purkinje cells indicate that this may occur at the subcellular level. Preliminary indication of the subunit composition of certain native voltage-dependent calcium channels is suggested by the observation that the distribution of the beta3 subunit in the cerebellar cortex is identical to that of alpha(1E). Our cumulative data are consistent with the emerging view that different native alpha1/beta subunit associations occur in the CNS.

Aged↗
PubMed1997

The development of Ca2+ channel responses and their coupling to exocytosis in cultured cerebellar granule cells.

Using single-cell imaging, we investigated developmental changes in the modulation of KCl-evoked Ca2+ entry by various voltage-dependent Ca2+ channels and the coupling of these channels to exocytosis in cultured cerebellar granule neurons. A component of the KCl-evoked Ca2+ elevation sensitive to nifedipine and localized at cell somata, decreases with culture age. A component blocked by 200 nM omega-Agatoxin-IVA increases with age and whilst localized primarily at the cell somata, also becomes evident at the neurites. The change in activity between nifedipine-sensitive Ca2+ channels and omega-Agatoxin-IVA-sensitive Ca2+ channels occurs at 13 days in vitro at cell somata. A component of Ca2+ entry insensitive to nifedipine and 200 nM omega-Agatoxin-IVA is localized primarily at the neurites and is apparent at all ages. Single-cell imaging of exocytosis using FM1-43 destaining indicates that the residual, but not the nifedipine- or omega-Agatoxin-IVA-sensitive components of Ca2+ entry, modulates exocytosis. However cells cultured for 20-26 days develop a component of Ca2+ entry at the neurites which is sensitive to 200 nM omega-Agatoxin-IVA and omega-Conotoxin-MVIIC and which partially controls release. Immunolocalization studies reveal that binding sites for omega-Conotoxin-GVIA are present throughout development, even though this toxin does not inhibit KCl-evoked [Ca2+]c elevations or exocytosis. 300 nM omega-Agatoxin-IVA labels both somata and, at later developmental stages, neurites, consistent with the functional studies.

Animals↗
PubMed1997

Immunohistochemical and in situ mRNA hybridisation techniques to determine the distribution of ion channels in human brain: a study of neuronal voltage-dependent calcium channels.

The molecular, structural and functional characterisation of ion channels in the CNS forms an area of intense investigation in current brain research. For strategic and logistical reasons, rodents have historically been the species of choice for these studies. The examination of human CNS tissues generally presents the investigator with specific challenges that are often less problematic in animal studies, e.g. post-mortem delay/agonal status, and thus both the experimental design and techniques must be manipulated accordingly. Since much pharmaceutical interest is currently focused on neuronal ion channels, the examination of their expression in human brain material is of particular importance. We describe here the details of methods that we have developed and used successfully in the study of the expression of voltage-dependent calcium channels (VDCCs) in human CNS tissues. Presynaptic neuronal VDCCs control neurotransmitter release and are important new drug targets. They are composed of three subunits, alpha 1, beta and alpha 2/delta and multiple gene classes of each protein have been identified. Little is known, however, about the distribution of neuronal VDCCs in the human central nervous system, although initial studies have been performed in rat and rabbit.

Autoradiography↗
PubMed1997

Preparation and purification of antibodies specific to human neuronal voltage-dependent calcium channel subunits.

Neuronal voltage-dependent calcium channels (VDCCs) each comprising of alpha 1, alpha 2 delta, and beta subunits, are one mechanism by which excitable cells regulate the flux of calcium ions across the cell membrane following depolarisation Studies have shown the expression of several alpha 1 and beta subtypes within neuronal tissue. The comparative distribution of these in normal human brain is largely unknown. The aim of this work is to prepare antibodies directed specifically to selected subunits of human neuronal VDCCs for use in biochemical and mapping studies of calcium channel subtypes in the brain. Previous studies have defined DNA sequences specific for each subunit Comparison of these sequences allows the selection of unique amino acid sequences for use as immunogens which are prepared as glutathione-S-transferase (GST) fusion proteins in E. coli. Polyclonal antibodies raised against these fusion proteins are purified by Protein A chromatography, followed by immunoaffinity chromatography and extensive adsorptions using the appropriate fusion protein-GST Sepharose 4B columns. The resultant antibodies are analysed for specificity against the fusion proteins by ELISA, and by immunofluorescence and Western immunoblot analysis of recombinant HEK293 cells stably transfected with cDNAs encoding alpha 1, alpha 2 delta and beta subunits.

Animals↗
PubMed1997

In vivo and in vitro evidence of altered nitric oxide metabolism in the spontaneously diabetic, insulin-dependent BB/Edinburgh rat.

1. Altered vasoreactivity may contribute significantly to the pathogenesis of diabetic vascular complications. This study investigated the effect of (a) insulin-related diabetes, and (b) chronic in vivo administration of N(omega)-nitro-L-arginine ester (L-NAME), a nitric oxide (NO) synthase inhibitor, on mean arterial pressure and in vitro vascular reactivity to noradrenaline in mesenteric arterial bed preparations from spontaneously diabetic, insulin-dependent and treated BB rats, the best animal model of insulin-dependent mellitus (IDDM) currently available. Four groups of animals from the Edinburgh colony (BB/E) of spontaneous diabetic BB rats were studied: age-matched (mean +/- s.e. mean = 156 +/- 2d) non-diabetic (glycated haemoglobin = 3.8 +/- 0.1%) and insulin-treated diabetic (glycated haemoglobin = 6.2 +/- 0.5%; duration of diabetes = 56 +/- 4 d) groups were either L-NAME treated (oral dose = 27 +/- 1 mg kg-1 d-1; duration of treatment from 30 until 153 days of age) or untreated. Although our diabetic BB/E rats do not achieve overall normoglycaemia, individual adjustment of the daily insulin dose administered to every diabetic rat achieves better glycaemic control than previous groups studying altered vascular reactivity and endothelial dysfunction in this animal model of diabetes. 2. Mean arterial pressure (measured directly via indwelling carotid arterial cannulae) was not significantly different between non-diabetic (116 +/- 3 mmHg; n = 10) and diabetic (122 +/- 2 mmHg; n = 12) BB/E rats. L-NAME treatment significantly (P < 0.001) increased mean arterial pressure in both groups (165 +/- 6 mmHg; n = 9 and 142 +/- 4 mmHg; n = 6 respectively) but the degree of hypertension observed in L-NAME-treated diabetic rats was significantly (P < 0.01) attenuated compared to non-diabetic rats treated with L-NAME. 3. Mesenteric arterial bed preparations were cannulated under anesthesia, excised and intralumenally perfused ex vivo with noradrenaline (0.2-20 microM). Basal perfusion pressures were not significantly different in mesentery preparations from non-diabetic (27.0 +/- 2.6 mmHg) and diabetic (27.1 +/- 3.2 mmHg) BB/E rats. There was no significant difference in maximal response above basal perfusion pressure (MAX) or pEC50, defined as the negative log of the agonist concentration required to give 50% of the maximal response above basal perfusion pressure, to noradrenaline in untreated non-diabetic (166 +/- 7 mmHg and 5.74 +/- 0.05 respectively) and diabetic (170 +/- 11 mmHg and 5.59 +/- 0.05) BB/E rats. 4. In vivo treatment of non-diabetic and diabetic BB/E rats with L-NAME had no significant effect on basal perfusion pressure (25.9 +/- 4.3 mmHg and 28.5 +/- 3.9 mmHg respectively). L-NAME treatment in vivo increased (P < 0.001) MAX to noradrenaline of non-diabetic rats (224 +/- 8 mmHg) but did not affect the value for diabetic rats (178 +/- 14 mmHg). L-NAME treatment did not alter after the pEC50 values in either group (5.71 +/- 0.05 and 5.65 +/- 0.05). 5. Consistent with previous studies using vascular preparations from spontaneously diabetic BB rats, mesentery preparations from diabetic BB/E rats (n = 12) exhibited a significantly reduced vasodilator response to acetylcholine (F value = 4.4, P < 0.05) across the concentration range studied compared to non-diabetic BB/E rats (n = 12) although there was no significant difference in maximal relaxation (diabetic 53.1 +/- 4.3% vs non-diabetic 55.7 +/- 5.5%) or pEC50, (diabetic 6.92 +/- 0.25 vs non-diabetic 7.49 +/- 0.22). There was no significant (F value = 0.8, P > 0.1) difference in the response to GTN between preparations from non-diabetic and diabetic rats (maximal relaxation: 49.6 +/- 3.7% vs 48.5 +/- 4.3%; pEC50: 7.84 +/- 0.12 vs 7.89 +/- 0.22 respectively). 6. In conclusion, vascular responsiveness to noradrenaline is not impaired in spontaneously diabetic BB/E rats with significantly better glycaemic control than those used in previous studies. However, following chronic L-NAME treatment, diabetic BB/E rats exhibit attenuated hypertension and an absence of enhanced vascular responsiveness to noradrenaline in vitro compared to similarly treated non-diabetic rats. These results, together with the significantly impaired endothelium-dependent vasodilatation and unchanged endothelium-independent vasodilatation in vitro of preparations from diabetic BB/E rats, are consistent with the hypothesis that functional changes in the synthesis and metabolism of NO (rather than altered vascular responsiveness to NO) occur in diabetes. Our results indicate that good glycaemic control alone is insufficient to prevent these abnormalities in NO availability and further studies to characterize the origin of these changes are necessary.

Animals↗
PubMed1997

Knowledge and beliefs about common eye diseases.

PURPOSE: To ascertain the level of knowledge of common causes of blindness in an adult Australian population and to relate this to use of eye care services. METHODS: A population-based study of common eye diseases in an urban population aged 49 years or older was conducted. The questions were concerned with the awareness and knowledge of and the ability to describe three common eye diseases, namely cataract, glaucoma and age-related macular degeneration (AMD). RESULTS: Awareness of cataract (98%) and glaucoma (93%) were high in this population, but awareness of AMD was low (20%). Among people who were aware of the target eye disease, only 29% showed some knowledge of glaucoma, 26% showed some knowledge of AMD and 20% showed some knowledge of cataract; this was also low in people who had previous eye treatment, such as cataract surgery. Knowledge was related to education level, occupational prestige and knowledge of other eye diseases. After excluding people with a previous eye disease diagnosis, those people who were aware and had some knowledge of eye disease accessed eyecare services more frequently. CONCLUSIONS: Knowledge of common eye diseases is generally lacking. Age-related macular degeneration is the leading cause of blindness in Australia, yet only 20% of the present study population had heard of it. As there are often no early symptoms for glaucoma, community awareness of this disease and the need for screening of people at risk may allow timely diagnosis and more effective therapy before advanced visual field loss has occurred. An informed public is more likely to present earlier with visual symptoms before irreversible visual loss has occurred and is more likely to comply better with recommended therapy.

Adult↗