Biomedical subjects

A M Lloyd

Publications and source records attributed to A M Lloyd.

At least 19 recordsLinked to original sources

PubMed2003

The effect of a 2% zinc citrate, 0.3% Triclosan dentifrice on plaque acid production following consumption of a snackfood.

OBJECTIVES: A) To assess plaque lactate production following consumption of three foods (cake, chocolate/caramel bar, sweetened coffee), and B) To measure the effect of a fluoride dentifrice containing 2% zinc citrate and 0.3% Triclosan on plaque lactate and pH drop following consumption of cake. METHODS: A) 10 subjects completed the first study. Plaque samples taken before and at 8,15 and 30 minutes after eating. Samples were analysed for lactate via Capillary Electrophoresis. B) 30 subjects completed the second study. Plaque samples were taken before and after cake and use of test dentifrice or no treatment control. Plaque pH and lactate content were assessed. RESULTS: A) Plaque lactate levels increased after all three foods; peak lactate levels occurred 8 minutes after eating. B) Plaque lactate concentrations after eating cake were 39.2mM for the control treatment and a significantly lower value, 23.6mM, for the test 2% zinc citrate, 0.3% Triclosan dentifrice. After food challenge, pH values were 5.53 for the no treatment group and a significantly higher value of 5.79 for the test dentifrice group. CONCLUSIONS: A toothpaste containing 2% zinc citrate, 0.3% Triclosan can significantly reduce plaque lactate generation and pH drop induced by cake, compared to no treatment control.

Adolescent↗

PubMed2003

The effect of a toothpaste containing 2% zinc citrate/0.3% Triclosan on the glycolysis of plaque bacteria ex vivo after food intake.

OBJECTIVE: To develop an ex vivo plaque pH method to assess the efficacy of a new zinc citrate/Triclosan formulation. METHODS: Study (1) focussed on method development. Study (2) examined the effect of a toothpaste containing 2% zinc citrate/0.3% Triclosan on the pH of plaque after product use and consumption of pizza. Study (3) investigated the effect of the same formulation and a fluoride toothpaste on the pH of plaque without an 'eating occasion'. The pH of plaque samples was measured over 10 minutes in the presence of glucose. RESULTS: The test product significantly reduced the amount of acid produced 30 minutes (p = 0.0035) and 3 hours (p = 0.0018) after brushing (study (2)). In study (3) use of the test product significantly reduced the amount of acid produced 3 hours after brushing (p = 0.0023). No significant benefit was found for the fluoride toothpaste. CONCLUSION: An ex vivo plaque pH method has been developed which can detect changes in acid produced following brushing with different toothpastes. A toothpaste containing 2% zinc citrate/0.3% Triclosan significantly reduced the total acid produced for at least 3 hours after product use. Moreover it has been demonstrated that this effect is detectable even after eating.

Adult↗

PubMed2002

Automated colorimetric screen for apyrase inhibitors.

Apyrases are enzymes that efficiently hydrolyze ATP and ADP and may operate both inside and outside the cell. Although apyrases are important to a variety of cellular mechanisms and uses in industry, there are no available apyrase-specific inhibitors. Colorimetric assays based on the Fiske-Subbarow method for measuring inorganic phosphate are able to detect the release of inorganic phosphate from ATP and other nucleotides. We found that this type of assay could be automated and used to screen for apyrase-inhibiting compounds by assaying for a reduction in released phosphate in the presence of potential inhibitors. The automation of this assay allowed for the successful screening of a commercially available compound library. Several low molecular weight compounds were identified that, when used at micromolar concentrations, effectively inhibited apyrase activity.

Apyrase↗

PubMed2000

Progress in the molecular genetic analysis of trichome initiation and morphogenesis in Arabidopsis.

Arabidopsis trichomes are large unicellular structures that develop on the surface of most shoot-derived organs. In leaves, the number, spacing and shape of trichomes is tightly regulated, and this process has been used as an experimental system to study the control of cell fate and pattern formation. The control of trichome initiation is complex: both the potential of a cell to adopt the trichome cell fate and an intricate signaling pathway determine the pattern of trichome initiation events. Several important new results suggest that trichome initiation and morphogenesis are redundantly regulated by both positive and negative factors. A testable model for the control of trichome initiation is presented.

Arabidopsis↗

PubMed2000

Arabidopsis seed coat development: morphological differentiation of the outer integument.

A morphological description of the differentiation of the outer integument of the Arabidopsis thaliana seed is presented. The period covered starts at about the octant embryo stage, extends to the mature seed, and concludes beyond that at the initial stages of seed imbibition. During this period the two-cell-layered outer integument goes through a dramatic differentiation process. The outer cell layer secretes mucilage in a ring between the plasma membrane and the outer cell wall at the corners of the cell. This secretion forces the cytoplasm into a columnar shape in the center of the cell. Before and during this process, starch granules are produced, initially at the center of the outer wall and later within the column. Late in differentiation, the starch granules are degraded as the cell produces a highly reinforced wall surrounding the columnar protoplast and at the radial walls between adjacent cells. This results in a cell containing large amounts of mucilage surrounding and completely outside of a highly reinforced columella. The mucilage and outer wall then dehydrate to leave the columella and radial walls visible as the epidermal plateau and reticulations visible on the mature seed. The inner cell layer of the outer integument also produces and degrades starch granules concomitantly with the outer layer but produces no mucilage. In the mature dry seed the collapsed outer wall remains connected to the top of the columella and the radial walls, but these connections are rapidly broken as the mucilage fully hydrates.

Arabidopsis↗

PubMed2000

GL3 encodes a bHLH protein that regulates trichome development in arabidopsis through interaction with GL1 and TTG1.

Arabidopsis trichome development and differentiation is a well-studied model for plant cell-fate determination and morphogenesis. Mutations in TRANSPARENT TESTA GLABRA1 (TTG1) result in several pleiotropic defects including an almost complete lack of trichomes. The complex phenotype caused by ttg1 mutations is suppressed by ectopic expression of the maize anthocyanin regulator R. Here it is demonstrated that the Arabidopsis trichome development locus GLABRA3 (GL3) encodes an R homolog. GL3 and GLABRA1 (GL1) interact when overexpressed together in plants. Yeast two-hybrid assays indicate that GL3 participates in physical interactions with GL1, TTG1, and itself, but that GL1 and TTG1 do not interact. These data suggest a reiterated combinatorial model for the differential regulation of such diverse developmental pathways as trichome cell-fate determination, root hair spacing, and anthocyanin secondary metabolism.

Amino Acid Sequence↗

PubMed1999

Antisense expression of the CK2 alpha-subunit gene in Arabidopsis. Effects on light-regulated gene expression and plant growth.

The protein kinase CK2 (formerly casein kinase II) is thought to be involved in light-regulated gene expression in plants because of its ability to phosphorylate transcription factors that bind to the promoter regions of light-regulated genes in vitro. To address this possibility in vivo and to learn more about the potential physiological roles of CK2 in plants, we transformed Arabidopsis with an antisense construct of the CK2 alpha-subunit gene and investigated both morphological and molecular phenotypes. Antisense transformants had a smaller adult leaf size and showed increased expression of chs in darkness and of cab and rbcS after red-light treatment. The latter molecular phenotype implied that CK2 might serve as one of several negative and quantitative effectors in light-regulated gene expression. The possible mechanism of CK2 action and its involvement in the phytochrome signal transduction pathway are discussed.

Arabidopsis↗

PubMed1998

Comparing treatment intensities of tactile-thermal application.

The purpose of this study was to investigate the relationships of four intensities of tactile-thermal application (TTA) to changes in duration of stage transition (DST) and performance on a newly designed scale of penetration and aspiration by groups of patients made dysphagic by stroke. Patients were randomly assigned to receive 150, 300, 450, or 600 trials of TTA during each of 2 weeks. Data on the time required to provide such treatment, the actual number of trials clinicians were able to provide, and on the influence of the four intensities are provided. No single intensity emerged as the most therapeutic. It is suggested that subsequent studies with larger groups include intensities between 300 and 550.

Aged↗

PubMed1996

Enhancement of somatic intrachromosomal homologous recombination in Arabidopsis by the HO endonuclease.

The HO endonuclease promotes gene conversion between mating-type alleles in yeast by a DNA double-strand break at the site of conversion (the MAT-Y/Z site). As a first step toward understanding the molecular basis of homologous recombination in higher plants, we demonstrate that expression of HO in Arabidopsis enhances intrachromosomal recombination between inverted repeats of two defective beta-glucuronidase (gus) genes (GUS- test construct). One of these genes has the Y/Z site. The two genes share 2.5 kb of DNA sequence homology around the HO cut site. Somatic recombination between the two repeats was determined by using a histochemical assay of GUS activity. The frequency of Gus+ sectors in leaves of F1 plants from a cross between parents homozygous for the GUS- test construct and HO, respectively, was 10-fold higher than in F1 plants from a cross between the same plant containing the GUS- test construct and a wild-type parent. Polymerase chain reaction analysis showed restoration of the 5' end of the GUS gene in recombinant sectors. The induction of intrachromosomal gene conversion in Arabidopsis by HO reveals the general utility of site-specific DNA endonucleases in producing targeted homologous recombination in plant genomes.

Arabidopsis↗

PubMed1995

A glutathione S-transferase involved in vacuolar transfer encoded by the maize gene Bronze-2.

Glutathione S-transferases (GSTs) are enzymes that detoxify heterocyclic compounds (xenobiotics) by covalently linking glutathione to the substrate, forming a glutathione S-conjugate. A glutathione pump in the vacuolar membrane of barley actively sequesters herbicide-glutathione S-conjugates; glutathionation allows recognition and entry of the conjugates into vacuoles. The protein encoded by the Bronze-2 gene in maize performs the last genetically defined step in anthocyanin biosynthesis, resulting in the deposition of red and purple pigments in the vacuoles of maize tissues. We show here that Bz2 encodes a GST with activity in maize, transformed Arabidopsis thaliana plants and Escherichia coli. We demonstrate that anthocyanins extracted from maize protoplasts expressing BZ2 are conjugated with glutathione, and that vanadate, a known inhibitor of the glutathione pump in plant vacuolar membranes, inhibits the accumulation of anthocyanins in the vacuole. These results provide a biochemical function for BZ2, and suggest a common mechanism for the ability of plants to sequester structurally similar but functionally diverse molecules in the vacuole.

Amino Acid Sequence↗

PubMed1994

Epidermal cell fate determination in Arabidopsis: patterns defined by a steroid-inducible regulator.

The Arabidopsis mutant ttg lacks both trichomes (epidermal hairs) and anthocyanin pigments. Trichomes and anthocyanins are restored by the constitutive expression of the maize transcriptional regulator (R). The expression of an R-glucocorticoid receptor chimeric protein results in a steroid hormone-dependent, conditional allele of R that functions in whole Arabidopsis plants. The response of the chimeric protein to pulses of hormone was used to define the pattern and timing of trichome formation on the developing leaf epidermis. Each adaxial epidermal leaf cell appears to have an equal probability of differentiating into a trichome; there is a temporal zone of decision for trichome cell fate that proceeds as a wave from the tip to the base of developing leaves.

Alleles↗

PubMed1994

The TTG gene is required to specify epidermal cell fate and cell patterning in the Arabidopsis root.

The control of cell fate was investigated in the root epidermis of Arabidopsis thaliana. Two distinct types of differentiated epidermal cells are normally present: root-hair-bearing cells and hairless cells. In wild-type Arabidopsis roots, epidermal cell fate was found to be correlated with cell position, with root-hair cells located over radial walls between cortical cells, and with hairless cells located directly over cortical cells. This normal positional relationship was absent in ttg (transparent testa glabrous) mutants (lacking trichomes, anthocyanins, and seed coat mucilage); epidermal cells in all positions differentiate into root-hair cells. The opposite condition was generated in roots of transgenic Arabidopsis expressing the maize R (R-Lc) gene product (a putative TTG homologue) under the control of a strong promoter (CaMV35S), which produced hairless epidermal cells in all positions. In both the ttg and R-expressing roots, epidermal cell differentiation was affected at an early stage, prior to the onset of cell elongation or root-hair formation. The ttg mutations were also associated with abnormalities in the morphology and organization of cells within and surrounding the root apical meristem. The results indicate that alterations in TTG activity cause developing epidermal cells to misinterpret their position and differentiate into inappropriate cell types. This suggests that, in wild-type roots, TTG provides, or responds to, positional signals to cause differentiating epidermal cells that lie over cortical cells to adopt a hairless cell fate.

Arabidopsis↗

PubMed1994

Functional expression of the yeast FLP/FRT site-specific recombination system in Nicotiana tabacum.

The FLP/FRT site-specific recombination system of Saccharomyces cerevisiae was expressed in stably transformed tobacco plants. The FLP protein efficiently catalyzes recombination between two directly repeated FLP recombination target (FRT) sites, deleting the sequence between them. In the constructs tested here, this deletion places the CaMV 35S promoter adjacent to a hygromycin resistance marker; transcriptional activation of the marker allows direct selection of recombination events. After crossing plants containing an integrated FLP expression construct with plants containing a FLP substrate, F1 seedlings can be selected directly for hygromycin resistance, indicating that recombination occurs at, or very early after zygote formation. Molecular analysis confirmed the expected recombination product.

Agrobacterium tumefaciens↗

PubMed1993

The HAT4 gene of Arabidopsis encodes a developmental regulator.

The HAT4 gene from the plant Arabidopsis thaliana encodes a homeo domain protein that contains a leucine zipper motif. Homeo domain-leucine zipper (HD-Zip) proteins have not been found in animal systems, suggesting that HAT4 may define a new family of transcription factors that regulate higher plant development. To explore this possibility, functional studies of HAT4 were carried out in yeast and in transgenic plants. Point mutants of HAT4 isolated in yeast define functionally critical residues within the HD-Zip domain, many of which correspond to highly conserved positions in known homeo domains and leucine zippers. Transgenic plants bearing constructs that alter HAT4 expression exhibit a series of interesting developmental phenotypes, including changes in morphology and developmental rate. Thus, the HAT4 gene of Arabidopsis encodes an HD-Zip protein that functions as a novel developmental regulator.

Amino Acid Sequence↗

PubMed1992

Arabidopsis and Nicotiana anthocyanin production activated by maize regulators R and C1.

Anthocyanin pathway-specific transcriptional activators R and C1 from the monocot maize were expressed in two dicots, Arabidopsis thaliana and Nicotiana tabacum. Expression of R caused augmented anthocyanin pigmentation in both plant species and augmented trichome (hair) production in Arabidopsis. Alone, C1 had no effect. Hybrid transgenic Arabidopsis expressing both C1 and R produced anthocyanins in root, petal, and stamen tissues that normally never express anthrocyanins. When R was expressed in the transparent testa glabrous (without anthocyanins and trichomes) mutant of Arabidopsis, the deficiency was complemented and both anthocyanins and trichomes were restored.

Anthocyanins↗

PubMed1991

A steroid-inducible gene expression system for plant cells.

Promoters that allow the selective induction of gene expression in vivo constitute an important methodology in eukaryotic organisms such as yeast and the fruit fly, but to date no such system has been described for higher plants. Given the fact that mammalian steroid receptors can function as hormone-dependent inducers of gene expression in heterologous systems, the feasibility of using mammalian steroid hormones as selective inducers of plant gene expression was investigated. Here it is shown that the glucocorticoid receptor expressed in plant cells is capable of activating a test gene linked to glucocorticoid response elements, providing the transfected plant cells are treated with glucocorticoid hormone. Nanomolar concentrations of glucocorticoids are sufficient to induce gene expression more than 150-fold, without causing detectable alterations in the physiology of the cultured plant cells. These findings indicate that glucocorticoid induction of steroid-responsive promoters should provide a general method for regulating gene expression in plant cells and imply that such a system might ultimately function in whole plants such as Arabidopsis thaliana.

Base Sequence↗

PubMed1988

Antiplaque effects and mode of action of a combination of zinc citrate and a nonionic antimicrobial agent.

The effect upon plaque growth of adding a nonionic antimicrobial agent, triclosan, to a dentifrice containing zinc citrate was established in short-term in vivo studies. Plaque regrowth was inhibited by brushing with dentifrices which contained either zinc citrate or triclosan. When both were combined in the same dentifrice, the inhibition of overnight plaque regrowth was significantly greater. In two 4-day non-brushing studies, the dentifrices containing both zinc citrate and triclosan were applied either undiluted by the use of a cap splint or as 23% suspensions in water. Both methods resulted in significant reductions in plaque accumulation, with the greater activity being observed for the undiluted application of the dentifrice. Analysis of results of the overnight plaque studies for individual teeth revealed that the two agents had a complementary inhibitory action on plaque regrowth, zinc citrate being more effective on existing plaque whereas triclosan inhibited plaque formation on clean surfaces. The dentifrice containing both agents was effective against both existing plaque and new plaque formation. It is concluded that the addition of triclosan to a dentifrice containing zinc citrate improves its antiplaque potential.

Adolescent↗

PubMed1986

Tomato golden mosaic virus A component DNA replicates autonomously in transgenic plants.

Phenotypically normal petunia plants carrying chromosomal inserts of either the tomato golden mosaic virus (TGMV) A or the B component DNA, as single or tandem inserts, were obtained using an Agrobacterium tumefaciens Ti plasmid-based transformation system. Southern hybridization analysis revealed that the tandem, direct-repeat A plants contained free single and double stranded A component DNAs. No free B component DNA was detected in plants carrying tandem repeats of the B component. Progeny of self-fertilized plants appeared normal. In contrast, one-quarter of the progeny from tandem A by tandem B plant crosses showed chlorotic lesions on their leaves similar to virus symptoms. The significance of these results and the use of this method for the study of virus functions involved in TGMV replication and symptom production are discussed.

DNA Replication↗