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J Dillon

Publications and source records attributed to J Dillon.

At least 19 recordsLinked to original sources
PubMed2006

Molecular characterization of the metabotropic glutamate receptor family in Caenorhabditis elegans.

mGluRs (metabotropic glutamate receptors) are G-protein-coupled receptors that play an important neuromodulatory role in the brain. Glutamatergic transmission itself plays a fundamental role in the simple nervous system of the model organism Caenorhabditis elegans, but little is known about the contribution made by mGluR signalling. The sequenced genome of C. elegans predicts three distinct genes, mgl-1, mgl-2 and mgl-3 (designated Y4C6A.2). We have used in silico and cDNA analyses to investigate the genes encoding mgls. Our results indicate that mgl genes constitute a gene family made up of three distinct subclasses of receptor. Our transcript analysis highlights potential for complex gene regulation with respect to both expression and splicing. Further, we identify that the predicted proteins encoded by mgls harbour structural motifs that are likely to regulate function. Taken together, this molecular characterization provides a platform to further investigate mGluR function in the model organism C. elegans.

Amino Acid Sequence↗
PubMed2004

The photochemical oxidation of A2E results in the formation of a 5,8,5',8'-bis-furanoid oxide.

With age and certain diseases, the retinal pigment epithelium (RPE) accumulates a complex mixture of fluorophores generally called lipofuscin. This mixture has been implicated in those diseases via several possible mechanisms including photochemically mediated damage and detergent effects. Recently, a component of that mixture has been identified, a bis-retinoid pyridinium compound called A2E. Furthermore, the photolysis of this compound has been shown to lead to a series of photooxidation products each differing by 16 amu; with the addition of up to a total of nine oxygen atoms to A2E. It has been suggested that these products consist of a series of epoxides forming along the acyclic side chains. The present study reinvestigates this hypothesis and demonstrates that the bis-oxygenated product is in fact a relatively stable 5-8 5'-8'-bis-furanoid oxide. This is in agreement with the oxidation products observed for a homologue of A2E, beta-carotene. In addition, it was found that the bis-oxygenated photoproduct of A2E contained a mixture of symmetrical and unsymmetrical oxidized products where the second oxygen was present in the cyclohexenyl ring structure in the unsymmetrical product. Most importantly, both of these oxidation products of A2E were detected in lipofuscin extracts from human RPE cells, suggesting that these processes occur in vivo.

Furans↗
PubMed2004

Determining calmodulin binding to metabotropic glutamate receptors with distinct protein-interaction methods.

mGluRs (metabotropic glutamate receptors) are G-protein-coupled receptors that modulate synaptic transmission. The eight mammalian mGluRs form three groups based on sequence and functional similarities: group I (1 and 5), group II (2 and 3) and group III (4, 6-8) mGluRs. In the present study, we used a Y2H (yeast two hybrid) screen to identify proteins that interact with the C-terminal intracellular tail of mGluR3. Prominent among the candidate receptor interacting proteins was calmodulin, a Ca(2+) sensor known to bind identifiable sequences in group I and III mGluRs. The Y2H method was used to investigate calmodulin binding to mGluRs but failed to confirm the documented interaction with group III mGluRs. Furthermore, subsequent biochemical analysis showed that calmodulin does not interact with group II mGluRs. This illustrates that certain Ca(2+)-dependent interactions are not recapitulated in yeast. Moreover, it highlights the necessity for supporting biochemical data to substantiate interactions identified with Y2H methods.

Amino Acid Sequence↗
PubMed2003

Fluoxetine improves functional work capacity in women with premenstrual dysphoric disorder.

Interference with social and occupational functioning is a key criterion for premenstrual dysphoric disorder (PMDD) and distinguishes it from the less severe premenstrual syndrome (PMS). We conducted a post hoc analysis of the results of a previously reported study evaluating the efficacy of fluoxetine in the management of PMDD, to determine the extent to which women with PMDD perceived impairment in their functional work capacity during the luteal phase of their menstrual cycle. The effects of two doses of fluoxetine vs placebo in alleviating PMDD symptoms and restoring normal work capacity during this period were assessed. We measured baseline follicular vs luteal phase presence of 8 patient-rated functional work capacity-related symptoms on the Premenstrual Tension Scale-Self Rated in 320 women who met diagnostic criteria for late luteal phase dysphoric disorder, now known as PMDD. Women were then randomized to double-blind treatment with either fluoxetine 20 mg/d, fluoxetine 60 mg/d, or placebo daily for 6 menstrual cycles. All 8 work capacity-related symptoms were more likely to be present in the baseline luteal phase than in the baseline follicular phase. A statistically significant improvement from baseline to the average treatment score for the work capacity subscale was detected for both fluoxetine groups compared to the placebo group. This beneficial response to fluoxetine was evident by the first cycle of treatment. Our results demonstrate that fluoxetine at a relatively low dose of 20 mg/d quickly reduced symptoms that negatively affect work capacity and was well tolerated.

Adolescent↗
PubMed2001

Synthesis and purification of 3-hydroxykynurenine-O-beta-glucoside, a primate lens ultraviolet filter, and its application in a two-step assay for beta-glucosidase activity.

3-Hydroxykynurenine-3-O-beta-glucoside (3-HKG) functions in the primate lens as a filter of 295- 400-nm light, thereby protecting the retina from damaging UV radiation. Although extensive studies have been conducted to determine the functional role of 3-HKG in the primate lens, an efficient method for its synthesis and purification has yet to be developed. Several procedures have been reported for the synthesis of 3-HKG; however, these procedures either result in low yields or require numerous sequential reactions and purification steps. In this study, we report a two-step synthesis of 3-HKG with a one-step purification and a two- to eightfold increase in yield over previously reported methods. Additionally, an assay was developed to confirm the presence of a beta-glycosidic linkage in the purified reaction product and we propose a method by which 3-HKG can be used as a general probe of beta-glucosidase activity. The assay consists of adding glucose oxidase to the 3-HKG/glucosidase solution and then allowing the hydrogen peroxide, generated from the interaction of glucose with glucose oxidase, to oxidize 3-hydroxykynurenine to xanthomattin (XAN) and 4,6-dihydroxyquinolinequinone carboxylic acid (DHQCA). Both XAN and DHQCA absorb strongly between 400 and 500 nm and the color change of the solution can be seen by eye. In addition, XAN fluoresces in the visible region with lambda(max) = 527 nm.

Animals↗
PubMed2001

Community participation in road safety: barriers and enablers.

The objectives of the paper were to identify barriers to community participation in road safety activities and to recommend strategies to reduce those barriers. Information was obtained from: a review of relevant literature, from the authors' extensive community-based experiences, from undertaking community-based research, and from interviews with key stakeholders in road safety in Western Australia. Ten significant barriers to community participation were identified. These are classified into two groups: personnel and planning issues. The former includes: reasons why people are often reluctant to become involved in projects in their communities, a lack of leadership, and a lack of skills. The latter includes: inappropriate program foci, inappropriate program evaluation, lack of resources, and a lack of sustainability. It was concluded that the barriers to community involvement in road safety initiatives are similar in many parts of Australia and overseas and apply to a wide range of health promotion interventions besides road safety. Awareness of the barriers and knowledge of their solutions can ensure that appropriate steps are taken to maximise the likelihood of community participation.

Accidents, Traffic↗
PubMed2001

Deletion of the cell-division inhibitor MinC results in lysis of Neisseria gonorrhoeae.

The minCDE genes involved in division site selection in Neisseria gonorrhoeae were identified using raw data from the N. gonorrhoeae genome project and are part of a cluster of 27 genes. When gonococcal min genes were heterologously expressed as a cluster in Escherichia coli, minicells and filaments were produced, indicating that gonococcal min genes disrupted cell division in other genera. The insertional inactivation of the minC gene of N. gonorrhoeae CH811 resulted in a strain (CSRC1) with decreased viability and grossly abnormal cell division as observed by phase-contrast and electron microscopy analysis. Western blot analysis of N. gonorrhoeae CSRC1 confirmed that MinC(Ng) was not produced. Complementation of CSRC1 by integrating a minC-6xHis tag fusion at the proAB locus by homologous recombination restored viability and 1.9 times wild-type levels of MinC(Ng) expression. This slight increase of expression caused a small percentage of the complemented cells to divide aberrantly. This suggested that the 6xHis tag has partially affected the stability of MinC, or that the chromosomal position of minC is critical to its regulation. Comparison of MinC proteins from different bacteria showed a homologous region corresponding to residues 135-230 with five conserved amino acids. Overexpression of MinC(Ng) in wild-type E. coli cells induced filamentation and an E. coli minC mutant was successfully complemented with minC(Ng). Therefore, the evidence indicates that MinC from N. gonorrhoeae acts as a cell-division inhibitor and that its role is essential in maintaining proper division in cocci.

Bacterial Proteins↗
PubMed2001

The efficacy of fluoxetine in improving physical symptoms associated with premenstrual dysphoric disorder.

OBJECTIVE: To determine the effectiveness of fluoxetine in alleviating physical symptoms of premenstrual dysphoric disorder. DESIGN: Randomised, double-blind, placebo controlled, parallel study. SETTING: Canadian University based outpatient clinics. Participants Four hundred and five subjects, of whom 320 with prospectively determined premenstrual dysphoric disorder were randomised. METHODS: Randomised women were assigned to fluoxetine 20 or 60 mg/day or placebo. Common physical symptoms associated with premenstrual dysphoric disorder including breast tenderness, bloating, and headache were evaluated by visual analog scales and the self-rated and observer premenstrual tension syndrome scales. OUTCOME MEASURES: Luteal phase change from mean baseline scores to mean treatment scores for all scales. RESULTS: Fluoxetine treatment was statistically superior to placebo, with no significant differences between the two fluoxetine dosages in their effects on physical symptoms. CONCLUSION: Daily fluoxetine treatment is superior to placebo in improving the most common physical symptoms associated with premenstrual dysphoric disorder.

Adult↗
PubMed2001

Photochemically modified alpha-crystallin: a model system for aging in the primate lens.

The purpose of this study was to quantitatively study the changes that occur upon irradiation of 3-hydroxykynurenine (3-HK) in the presence of alpha-crystallin under conditions similar to those in the lens. The samples were prepared in 10 mM phosphate buffer at pH 7.4, bubbled with O2 or Ar and irradiated with 300-400 nm light. The amount of light absorbed by the samples (Iabs) was measured using azobenzene as an actinometer. Modifications to alpha-crystallin were monitored by ultraviolet-visible and fluorescence spectroscopy. Aerobic samples had increased absorption around 320 nm and above 400 nm while the 3-HK maximum at 368 nm decreased. The isolated modified protein showed that there was increased absorption throughout the spectrum. Changes in the anaerobic samples were similar to those of the aerobic but occurred more slowly. As irradiation time increased fluorescence emission of the isolated protein red shifted and quantum yields of fluorescence (phi f) were calculated at different irradiation time intervals by comparison to 3-HK. By comparing OD320/OD365 for the model system to values from primate lenses, Iabs can be correlated with age and transmission of the sample in the blue region of the spectrum and thus allows lenticular aging to be quantitated.

Aging↗
PubMed2001

The nitrite/collagen reaction: non-enzymatic nitration as a model system for age-related damage.

The effects of age seen in long-lived connective tissue proteins are thought to be the result of post-translational modifications by reactive molecules. One such molecule is the nitrite ion. Human nitrite exposure results predominately from endogenous production of nitric oxide as well as inhalation of cigarette smoke and ingestion of cured meats. Although nitrite reactions with various proteins have been studied previously with regard to carcinogenesis, the specific reaction with collagen and its role in age-related damage has never been examined. We describe the reaction of nitrite with type I collagen at neutral pH and body temperature. The incubation of collagen with nitrite results in an increase in cross-linking, the accumulation of a yellow chromophore, and a depletion of tyrosine residues. Similar changes also are found in aged human collagen. In addition, 3-nitro-tyrosine, which has recently been used as a marker for peroxynitrite mediated damage, is produced from this reaction. Thus, we propose non-enzymatic nitration as an in vitro model system for human collagen age-related damage.

Aging↗
PubMed2001

Acceptability and patterns of endometrial bleeding in estradiol-based HRT regimens: a comparative study of cyclical sequential combinations of trimegestone or norethisterone acetate.

OBJECTIVE: This randomized, double-blind, multicenter study was planned to compare the efficacy and tolerance of a novel oral regimen containing estradiol (2.0 mg) sequentially combined with trimegestone, at a daily dose of either 0.25 mg or 0.5 mg, with a standard hormone replacement therapy containing estradiol and norethisterone acetate (E2 + NETA) in the treatment of climacteric symptoms. METHODS: The study was conducted over 13 cycles, each of 28 days, and involved 487 subjects, of whom 349 completed the study. RESULTS: All three treatments were equally effective in alleviating hot flushes and showed a progressive and significant reduction in the value of the Kupperman index. The treatments diminished equally effectively urogenital signs and symptoms. All treatments were well tolerated and the incidences of adverse events associated with each treatment were similar across the treatment groups. The duration of expected withdrawal bleeding was shorter in the estradiol + trimegestone 0.5 mg group than in the estradiol + trimegestone 0.25 mg or E2 + NETA group. CONCLUSION: All treatments were effective and well tolerated, providing significant relief from climacteric symptoms. Treatment with estradiol + trimegestone 0.5 mg provided the most favorable bleeding pattern.

Argentina↗
PubMed2000

The Nitrite/alpha crystallin reaction: a possible mechanism in lens matrix damage.

Recent work has explored the potential deleterious role that nitric oxide (NO) and its derivatives may have in human disease. The many by-products of NO include nitrite ion, which accumulates in the anterior chamber during ocular inflammation and can be derived from cigarette smoking. Cigarette smoking has been strongly linked to nuclear cataract formation, although the mechanism remains unknown. We have previously observed that nitrite reactions with the matrix proteins elastin and collagen produce damaging effects that mimic those observed in age- and smoking-related illnesses. In the present study we report on the reaction of nitrite with alpha crystallin, the major lens matrix protein. Incubations at neutral pH and body temperature of nitrite with alpha crystallin resulted in protein modifications indicative of oxidative damage and similar to changes seen in the aging lens as well as cataracts. These include increased fluorescence, yellowing and protein cross-linking. L-kynurenine, a tryptophan derivative, was identified as a reaction product. L-kynurenine was also formed from the reaction of nitrite with free tryptophan. Thus, this non-enzymatic nitration of alpha crystallin provides a novel mechanism by which lens proteins may be damaged in vivo. Since human exposure to nitrite is increased by cigarette smoking, this reaction could provide an explanation for the association between nuclear cataracts and smoking.

Aging↗
PubMed2000

Triple therapy for Helicobacter pylori eradication: a comparison of pantoprazole once versus twice daily.

BACKGROUND: Proton pump inhibitor-based triple therapy is recommended as treatment for Helicobacter pylori eradication. The proton pump inhibitor may be given once or twice daily. However, little information is available on how these two treatment strategies compare. METHODS: H. pylori-positive patients (two positive test results) with endoscopy-proven healed duodenal ulcer or non-ulcer dyspesia were randomly allocated to 1 week of double-blind treatment with pantoprazole 40 mg once or twice daily, plus clarithromycin 250 mg and metronidazole 400 mg twice daily. Eradication was defined as a negative 13C-urea breath test (13C-UBT) and histology, 4-5 weeks post-treatment. The follow-up phase comprised 12 months off therapy, with 13C-UBT at 6 and 12 months. RESULTS: Two hundred and four patients received treatment: pantoprazole once daily (x1), n=104; twice daily (x2), n=100. Eradication rates were 84% in both the pantoprazole x1 and pantoprazole x2 groups by modified intention-to-treat analysis and 89% and 87%, respectively, by per protocol analysis. Metronidazole resistance was found in 44% of pre-treatment cultures of H. pylori. Eradication rates were similar in susceptible (72%) and resistant (75%) strains. During follow-up, recrudescence of infection occurred in 3/118 patients. CONCLUSION: When using pantoprazole plus clarithromycin and metronidazole, the proton pump inhibitor can be used once daily without loss of efficacy.

2-Pyridinylmethylsulfinylbenzimidazoles↗
PubMed2000

Transmission spectra of light to the mammalian retina.

A simple method has been developed to determine the optical properties of the anterior segment of the intact eye. This consists of a probe that is inserted into the posterior sclera and detects light passing through the anterior segment. The probe is connected to a charge-coupled device spectrophotometer via a fiber optic bundle. It was determined that the young rat anterior segment transmits light down to 300 nm, whereas calf and rabbit eyes transmit no UVB and only part of the UVA to the posterior segment. The absorbing species in these animals is most likely NAD(P)H, which has an absorption maximum at approximately 345 nm and is associated with zeta-crystallin. A young primate anterior segment transmits almost no UV with a steep increase in transmission at CA 400 nm. Because some experiments employed a light tube that is used to illuminate the retina during vitrectomies, this method can be used to determine the transmission spectra of the anterior segment of humans in vivo.

Animals↗
PubMed2000

Heat and concentration effects on the small heat shock protein, alpha-crystallin.

alpha-Crystallin, a major protein of the mammalian lens, plays a vital role in maintaining the structural stability and transparency of the lens. It performs this function through chaperone-like activity; it has recently been reported that heating alpha-crystallin enhances this ability. The present studies, using both time-resolved and steady-state fluorescence methods, were carried out to compare the conformational changes that result from heating with those that result from increasing protein concentration (up to 70 mg/mL). The relative fluorescence quantum yield from tryptophan (Trp) present in alpha-crystallin increases and then decreases with a concomitant shift of the emission maximum to longer wavelengths when either heating times or protein concentrations are increased. The time profile of fluorescence decay was resolved into three components with lifetimes of ca 0.5, 3 and 7 ns and emission maxima of ca 340, 342 and 350 nm, respectively. With longer heating time or increasing concentrations the contribution from the longer-lived component increases at the expense of the shorter-lived species. These data indicate that with heating or at higher concentrations the internal Trp residues move to the surface of the protein giving a more hydrophobic exterior and possibly explain the reported increased chaperone activity upon heating. As a result of the concentration studies, alpha-crystallin may be more efficient in its chaperone activity in vivo than has been determined by in vitro experiments.

Animals↗
PubMed2000

Age-related changes in the absorption characteristics of the primate lens.

PURPOSE: To quantitate aging of the primate lens by changes in the absorption characteristics that are related to the yellowing of lens protein. METHODS: The lenses of lower primates and humans were sectioned anterior to posterior every 0.25 mm, and the UV-visible spectrum of each section was measured to determine the cumulative spectra along the visual axis. The ratio of the absorbance at 320 nm (formed with aging) to the absorbance at 365 nm (present in the young lens) was correlated with the age of the lens. RESULTS: In the young primate UV-B is transmitted to the retina, and UV-A is transmitted to the nucleus of the lens. By puberty, changes in the absorption characteristics of the lens that are associated with the yellowing of lens protein prevented most of the UV-B from reaching the retina and by the eighth decade, the transmittances at 320 and 365 nm to the nucleus of the lens were approximately 40% and 79%, respectively. A linear relationship between the ratio of absorbance at 320 to 365 nm and age was found for both lower primates and humans to the age of 80 years. This is surprising, because the maximum life span of the lower primate is approximately 35 years, whereas humans may live 100 years. CONCLUSIONS: These data suggest that the observed spectral changes associated with the yellowing of the lens are the result of a chronological process, such as chemical or photochemical modifications, not biological aging.

Absorption↗
PubMed2000

An action spectrum for UV-B radiation and the rat lens.

PURPOSE: To determine an action spectrum for UV-B radiation and the rat lens and to show the effect of the atmosphere and the cornea on the action spectrum. METHODS: One eye of young female rats was exposed to 5-nm bandwidths of UV-B radiation (290, 295, 300, 305, 310, and 315 nm). Light scattering of exposed and nonexposed lenses was measured 1 week after irradiation. A quadratic polynomial was fit to the dose-response curve for each wave band. The dose at each wave band that produced a level of light scattering greater than 95% of the nonexposed lenses was defined as the maximum acceptable dose (MAD). Transmittance of the rat cornea was measured with a fiberoptic spectrophotometer. The times to be exposed to the MAD in Stockholm (59.3 degrees N) and La Palma (28 degrees N) were compared. RESULTS: Significant light scattering was detected after UV-B at 295, 300, 305, 310, and 315 nm. The lens was most sensitive to UV-B at 300 nm. Correcting for corneal transmittance showed that the rat lens is at least as sensitive to UV radiation at 295 nm as at 300 nm. The times to be exposed to the MAD at each wave band were greater in Stockholm than in La Palma, and in both locations the theoretical time to be exposed to the MAD was least at 305 nm. CONCLUSIONS: After correcting for corneal transmittance, the biological sensitivity of the rat lens to UV-B is at least as great at 295 nm as at 300 nm. After correcting for transmittance by the atmosphere, UV-B at 305 nm is the most likely wave band to injure the rat lens in both Stockholm and La Palma.

Animals↗