Biomedical subjects

S J Robinson

Publications and source records attributed to S J Robinson.

At least 37 records · Page 2Linked to original sources

PubMed1993

Regulated expression of the rat medium chain hydrolase gene in transgenic rape seed.

Medium chain hydrolase (MCH) is an enzyme which regulates the chain length of fatty acid synthesis specifically in the mammary gland of the rat. During lactation, MCH interacts with fatty acid synthase (FAS) to cause premature release of acyl chains, thus providing medium chain fatty acids for synthesis of milk fat. In this study we have investigated the ability of rat MCH to interact with the phylogenetically more distant FAS structure present in plant systems and to cause a perturbation of fatty acid synthesis. In in vitro experiments, addition of purified MCH to rapeseed homogenates was found to cause a significant perturbation of fatty acid synthesis towards medium chain length products. The rat MCH gene was expressed in transgenic oilseed rape using a seed specific rape acyl carrier protein (ACP) promoter and a rape ACP plastid targeting sequence. Western analysis showed MCH protein to be present in transgenic seed and for its expression to be developmentally regulated in concert with storage lipid synthesis. The chimaeric preprotein was correctly processed and immunogold labelling studies confirmed MCH to be localized within plastid organelles. However, fatty acid analysis of oil from MCH-expressing rape seed showed no significant differences to that from control seed.

Animals↗

PubMed1993

The dentinal surface: its influence on dentinal adhesion. Part III.

This final paper in a series of three uses transmission and scanning electron microscopy to compare the category III dentinal adhesive systems--those with shear bond strength values near or exceeding 17 MPa. Contemporary commercial dentinal adhesive systems such as Superbond and Scotchbond Multi-Purpose, chosen for their varied chemical pretreatments, are contrasted; a contemporary commercial system (Prisma Universal Bond 3) with characteristics of category I and II dentinal adhesives is also included for comparison. The shear bond strength values attained with most category III systems are high enough to cause cohesive failure of the dentin during bond strength testing. This result is attributed to a combination of factors that include effective wetting and penetration of the prepared dentinal surface as well as a tendency to leave collagen fibers at the adhesive-dentin interface in an apparently structurally intact state.

Boron Compounds↗

PubMed1993

Adhesives and nonshrinking dental resins of the future.

Much attention has been directed toward producing dentinal adhesives that withstand the forces involved during polymerization shrinkage of composite resins. Studies have shown that an effective dentin-adhesive bond depends on the wetting and penetration characteristics of the dentinal adhesive system and the reactivity of the treated dentinal surface. The structure of the collagen in the demineralized dentinal layer also seems to influence the behavior of the bond. Adhesive systems that do not completely denature the fibrous collagen and leave interwoven banded collagen in the demineralized layer produce superior bond strengths (greater than 20 MPa). Other research efforts seek to develop a nonshrinking high-performance polymer for use as a matrix material for dental composite resins. Spiroorthocarbonates that expand during polymerization have been developed. In combination with a three-component epoxy comonomer, matrix resins have been produced that expand during polymerization and whose mechanical properties, water sorption, solubility, and degree of polymerization are acceptable for dental use. Further developments of nonshrinking composite resins and improved dentinal adhesives will greatly increase the longevity of 21st-century composite resin restorations and should significantly ease clinical placement.

Adhesives↗

PubMed1992

The isolation and functional characterisation of a B. napus acyl carrier protein 5' flanking region involved in the regulation of seed storage lipid synthesis.

Acyl carrier protein (ACP) is a key component of the fatty acid biosynthetic machinery in plants. A 1.4 kb 5' flanking region of a Brassica napus ACP gene (ACP05) was transcriptionally fused to the reporter gene beta-glucuronidase (GUS), and expression of the chimeric gene monitored in transgenic tobacco. GUS activity was found to increase through seed development reaching a maximum value, coincident with the most active phase of storage lipid synthesis that was, on average, 100-fold higher than that observed in leaf. In control plants transformed with CaMV 35S-GUS constructs, GUS activity was similar in leaf and all stages of seed development. Based on average values, the level of GUS expression obtained via the ACP promoter was comparable to that obtained from the CaMV 35S promoter. We therefore conclude that the isolated 5' ACP flanking sequence represents a strong promoter element involved in the developmental regulation of storage lipid synthesis in B. napus seed tissue. Putative regulatory elements in the 5' upstream region of ACP05 were identified by dot matrix analysis and by sequence comparison with the upstream regions from a second seed-expressed rape ACP gene and from an Arabidopsis ACP gene.

Acyl Carrier Protein↗

PubMed1992

The dentinal surface: its influence on dentinal adhesion. 2.

The first paper in this series presented a categorization of dentinal adhesive systems that was organized roughly according to chronology, chemistry, and shear bond strength values. Category I dentinal adhesives demonstrate the lowest shear bond strength values, approximately 5 to 7 MPa. Category II dentinal adhesives, including experimental ferric and aluminum oxalate pretreatments and two commercial products, demonstrate shear bond strength values between 8 and 14 MPa. This second category of dentinal adhesives is the subject of this paper. An attempt is made to draw a parallel between the penetrative ability of the adhesive (the wettability of the dentinal substrate) and the efficacy of the adhesive bond as determined by shear bond strength values. Category III dentinal adhesives will be discussed in another paper.

Adhesives↗

PubMed1991

The dentinal surface: its influence on dentinal adhesion. Part I.

Three categories of dentinal adhesive are proposed: category I includes adhesives with shear bond strength values between 5 and 7 MPa; category II includes dentinal adhesives with shear bond strengths between 8 and 14 MPa; and category III includes adhesives with shear bond strength values up to 20 MPa. In part I of this article, photomicrographs of the dentinal smear layer and three category I first-generation dentin-adhesive interfaces are presented. The photomicrographs show that the wetting and penetration of the first-generation dentinal adhesives were not adequate to produce high shear bond strengths. When the category I adhesives were tested for shear bond strength, failures occurred at the interface or in the resin adhesive. Future articles will explain wetting and adhesive performance of category II and III adhesives.

Acrylic Resins↗

PubMed1990

Gene encoding the 5.7-kilodalton chlorosome protein of Chloroflexus aurantiacus: regulated message levels and a predicted carboxy-terminal protein extension.

The major light-harvesting pigment of the green filamentous bacterium Chloroflexus aurantiacus is bacteriochlorophyll (Bchl) c, localized in chlorosomes attached to the inner surface of the cytoplasmic membrane. Chlorosomes consist of four polypeptides and associated pigments and lipids. Previous studies of the inducible assembly of the photosynthetic apparatus had indicated that the major chlorosomal polypeptides are present as high-molecular-weight aggregates before the appearance of mature chlorosomes, and a mechanism for posttranslational processing of a polyprotein had been proposed. We have isolated the gene (csmA) encoding the 5.7-kilodalton chlorosomal polypeptide from C. aurantiacus in order to determine whether this protein is synthesized as part of a polyprotein. Analysis of the nucleotide sequence of csmA indicates that the gene is not large enough to encode more than one known chlorosome polypeptide. Transcriptional analysis indicates that csmA is transcribed as a small message whose abundance is regulated in response to oxygen, so that no csmA message is detectable in cells grown aerobically in the dark. Comparison of the sequence predicted by csmA with the peptide sequence of the Bchl c binding protein purified from chlorosomes indicates that this protein is synthesized with a carboxy-terminal extension of 27 amino acids. We discuss possible roles for this carboxy-terminal extension in the assembly of chlorosomes.

Amino Acid Sequence↗

PubMed1988

The effect of dacarbazine on wound healing.

The effects of dacarbazine on wound healing were studied in an animal model. Sixty rats were divided into four groups. Each animal received a standard dorsal midline wound on day 0. Group 1 was injected with normal saline and acted as control; group 2 received 4 mg/kg of dacarbazine daily on days -3, -2, -1 (preoperative); group 3 received dacarbazine on days +1, +2, +3 (perioperatively); and group 4 received dacarbazine on days +5, +6, +7 (postoperatively). Wound bursting strength (WBS) was measured on days +7, +14, and +21. There was a significant decrease in WBS in experimental animals in groups 3 and 4 (perioperative and postoperative administration) compared to control animals, but no significant effect of preoperative dacarbazine on WBS in experimental animals (group 2). It is suggested that these effects are mediated through the late inflammatory phase of wound healing.

Animals↗

PubMed1987

Effects of exogenous guanosine on chromatophore differentiation in the axolotl.

Guanosine is shown to dramatically alter the pigment phenotype of axolotls by suppressing melanization and enhancing the biosynthesis and deposition of purine-derived pigments. Phenotypic changes caused by guanosine are manifested by altered chromatophore differentiation patterns such that few black pigment cells (melanophores) differentiate (and those that do are punctate and necrotic in appearance), whereas the development of yellow (xanthophore) and reflecting (iridophore) pigment cells is enhanced. Mechanisms for changes in chromatophore differentiation, and thus pattern formation, are discussed, including the possibility that pigment cells may undergo transdifferentiation in vivo.

Aging↗

PubMed1986

The pigmentary system of developing axolotls. III. An analysis of the albino phenotype.

The albino mutant in the Mexican axolotl (Ambystoma mexicanum) is analysed with respect to the differentiation of pigment cells. Pigment cells were observed with the transmission electron microscope in order to determine any unusual structural characteristics and to determine what happens to each of the cell types as development proceeds. Chemical analyses of pteridine pigments were also carried out, and the pattern of pteridines in albino animals was found to be more complex than, and quantitatively enhanced (at all developmental stages examined) over, the pattern observed in comparable wild-type axolotls. The golden colour of albino axolotls is due primarily to sepiapterin (a yellow pteridine) and secondarily to riboflavin (and other flavins). Coincident with enhanced levels of yellow pigments, xanthophore pigment organelles (pterinosomes) in albino skin reach a mature state earlier than they do in wild-type axolotl skin. This morphology is conserved throughout development in albino animals whereas it is gradually lost in the wild type. Unpigmented melanophores from albino axolotls are illustrated for the first time, and in larval albino axolotls the morphology of these cells is shown to be very similar to xanthophore morphology. In older animals xanthophores are easily distinguished from unpigmented melanophores. Iridophores seem to appear in albino skin at an earlier stage than they have been observed in wild-type skin. Morphologically, wild-type and albino iridophores are identical.

Albinism↗

PubMed1986

The pigmentary system of developing axolotls. IV. An analysis of the axanthic phenotype.

The axanthic mutant in the Mexican axolotl (Ambystoma mexicanum) was analysed with respect to the differentiation of pigment cells. Transmission electron micrographs revealed the presence of melanophores and cells that are described as unpigmented xanthophores in axanthic skin. Iridophores apparently failed to differentiate in axanthic axolotls (a pattern similar to that observed in melanoid axolotls). Chromatographic analyses of skin extracts confirmed that there are no pteridines (xanthophore pigments) in axanthic skin, suggesting that the axanthic gene may affect pteridine biosynthesis at some point early in the biosynthetic pathway. Why iridophores fail to differentiate in these animals is not known, but this, too, may be related to an inability to synthesize pigments properly. Xanthophore and iridophore pigments both presumably derive from purine precursors. Finally, all axanthic animals were found to be infected by a virus. Electron microscopic results demonstrated the presence of numerous macrophages in the dermis of the skin, occupying positions typical of pigment cells. The virus was localized primarily in macrophages, but was also observed in pigment cells. The virus is, as yet, uncharacterized but is thought to contribute to the low survivability of axanthic adults.

Ambystoma↗

PubMed1984

The pigmentary system of developing axolotls. I. A biochemical and structural analysis of chromatophores in wild-type axolotls.

A biochemical and transmission electron microscopic description of the wild-type pigment phenotype in developing Mexican axolotls (Ambystoma mexicanum) is presented. There are three pigment cell types found in adult axolotl skin - melanophores, xanthophores and iridophores. Both pigments and pigment cells undergo specific developmental changes in axolotls. Melanophores are the predominant pigment cell type throughout development; xanthophores occur secondarily and in fewer numbers than melanophores; iridophores do not appear until well into the larval stage and remain thereafter as the least frequently encountered pigment cell type. Ultrastructural differences in xanthophore organelle (pterinosome) structure at different developmental stages correlate with changes in the pattern of pteridine biosynthesis. Sepiapterin, a yellow pteridine, is present in larval axolotl skin but not in adults. Riboflavin (also yellow) is present in minimal quantities in larval skin and large quantities in adult axolotl skin. Pterinosomes undergo a morphological "reversion" at some point prior to or shortly after axolotls attain sexual maturity. Correlated with the neotenic state of the axolotl, certain larval pigmentary features are retained throughout development. Notably, the pigment cells remain scattered in the dermis such that no two pigment cell bodies overlap, although cell processes may overlap. This study forms the basis for comparison of the wild type pigment phenotype to the three mutant phenotypes-melanoid, axanthic and albino-found in the axolotl.

Ambystoma↗

PubMed1984

The pigmentary system of developing axolotls. II. An analysis of the melanoid phenotype.

The melanoid mutant in the Mexican axolotl (Ambystoma mexicanum) is analysed with respect to the differentiation of pigment cells. Pigment cells were observed with the transmission electron microscope in order to determine any unusual structural characteristics and to determine what happens to each of the cell types as development proceeds. Chemical analysis of pteridine pigments was also carried out, and changes in pteridine biosynthesis were found to correlate well with changes in xanthophore morphology and number. In melanoid axolotls, as development proceeds, melanophore numbers increase, xanthophores decrease, and iridophores fail to differentiate at all. This is considered to result from: (a) conversion of xanthophores (that are present in young larvae) to melanophores; (b) the gradual programming of the majority of chromatoblasts to become, exclusively, melanophores, and (c) the failure of some chromatoblasts (possibly iridoblasts) to differentiate altogether. The ultrastructural and chemical evidence presented in this study is compared to similar data for wild-type axolotls, and a mechanism regarding how the melanoid gene might act is suggested.

Ambystoma↗

PubMed1982

Photosynthetic Electron Transfer in Preparations of the Cyanobacterium Spirulina platensis.

Electron transfer activity in intact trichomes of Spirulina platensis (Nordst.) Geitl. can be observed with either CO(2) or methylviologen as the Hill acceptor. Ferricyanide cannot penetrate the intact trichomes, but photoreduction of this oxidant can be observed when mediated by lipophilic oxidants such as p-phenylenediamine or 2,5-dimethyl-p-benzoquinone. The insensitivity of these reactions to dibromothymoquinone indicates that they are due largely to the activity of photosystem II. Direct photoreduction of ferricyanide can be observed in spheroplasts of Spirulina, indicating that such preparations have altered permeability properties when compared with intact trichomes. Preparation of these spheroplasts, which are osmotically fragile, requires that intact trichomes be washed with KCl and EDTA to induce lysozyme sensitivity and thereby allow digestion of the cell wall. The KCl/EDTA washing procedure used for spheroplast preparation alters the permeability of Spirulina trichomes, as evidenced by the ability of these preparations to photoreduce ferricyanide. This photoreduction reaction is insensitive to dibromothymoquinone, and is stimulated by high concentrations of divalent cations. During assays, the reaction is inhibited by the inclusion of polyethyleneglycol as an osmotic protectant. Photoreduction of methylviologen and NADP(+) is also observed in the washed trichomes, along with an endogenously catalyzed photoreduction of O(2) to H(2)O(2). Photophosphorylation cannot be observed in the washed preparations, but cyclic photophosphorylation with phenazinemethosulfate is observed after mild sonication. These results indicate that KCl/EDTA-washed trichomes of S. platensis retain the full range of energy transducing capacities associated with thylakoid membranes of the intact trichomes; the washing procedure facilitates spheroplast formation and alters, but does not abolish, permeability barriers in these preparations.

Journal Article↗

PubMed1981