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Biomedical subjects

C Robinson

Publications and source records attributed to C Robinson.

At least 19 recordsLinked to original sources
PubMed1992

Protein targeting across the three membranes of the Euglena chloroplast envelope.

A system has been developed for the import in vitro of precursor proteins into Euglena chloroplasts, which have three envelope membranes. Preparation of functional chloroplasts with intact envelope membranes has been optimized. Import of the precursor (50 kDa) for the tetrapyrrole biosynthesis enzyme porphobilinogen deaminase (PBGD), and processing to the mature size (40 kDa), occurred at 25 degrees C in the light and the presence of ATP, with an estimated efficiency of 62%. Pretreatment of the chloroplasts with proteases abolished this import, suggesting the involvement of specific protein receptors. The presequence of PBGD was found to be cleaved by Escherichia coli leader peptidase to an intermediate form (46 kDa). A construct in which the first 30 residues of the presequence (presumed to be the region removed by leader peptidase) had been deleted was no longer imported. Neither prePBGD nor the truncated precursor were imported into pea chloroplasts, although both bound to the pea chloroplast envelope. Conversely, a chimeric construct, in which the mature PBGD protein was fused downstream of the transit peptide for pea ferredoxin-NADP reductase, was efficiently imported into pea chloroplasts and processed to the mature size. However, this was not imported into Euglena chloroplasts, although again it bound to them. These results provide preliminary evidence for the possibility of two functional domains within the Euglena PBGD presequence. The implications of these findings with respect to the evolution of Euglena chloroplasts are discussed.

Amino Acid Sequence↗
PubMed1992

Carboxyl-terminal processing of the D1 protein and photoactivation of water-splitting in photosystem II. Partial purification and characterization of the processing enzyme from Scenedesmus obliquus and Pisum sativum.

The D1 polypeptide of photosystem II (PSII) is synthesized as a precursor that is processed by cleavage at the carboxyl terminus during assembly of the active PSII complex. A mutant of the green alga Scenedesmus obliquus, LF-1, inactive in water-splitting, lacks the D1 processing activity but assembles otherwise normal PSII complexes containing the precursor D1 molecule. We have isolated and partially purified a soluble protease from sonicated thylakoids of both wild-type S. obliquus and Pisum sativum which will process the precursor D1 molecule in PSII-enriched membranes from the LF-1 mutant to the mature size. After processing (but not before), photoactivation of these PSII membranes in the presence of manganese restores water-splitting to levels seen after photoactivation of PSII membranes from dark-grown, wild-type, cells. The protease is unable to process D1 in intact thylakoids from the LF-1 mutant but processes D1 if present during sonication of the thylakoids, indicating that processing of the carboxyl-terminal extension of D1 occurs in the lumen of the thylakoid. The processing protease from both S. obliquus and P. sativum is a single subunit enzyme of native molecular mass 33-35 kDa. Processing rate is optimal at pH 6.5. Processing in vitro is evident within 5 min and is markedly inhibited by millimolar concentrations of divalent cations (Cu, Zn greater than Mn greater than Ca, Mg) but not by any known inhibitors of the major classes of proteases. The protease is inactive against the precursors of other thylakoidal proteins and is thus distinct from the thylakoidal amino-terminal processing enzyme involved in the removal of transit peptides from cytoplasmically-synthesised proteins imported into the thylakoid lumen.

Algal Proteins↗
PubMed1992

Proton gradient-driven import of the 16 kDa oxygen-evolving complex protein as the full precursor protein by isolated thylakoids.

The biogenesis of the lumenal 16 kDa protein of the photosynthetic oxygen-evolving complex was analysed using an assay for the import of proteins by isolated thylakoids. The precursor protein is imported with high efficiency in the light in both the presence and absence of stromal extract. Import is almost completely blocked in the dark or if the uncoupler nigericin is present in the light. The data indicate that transport across the thylakoid membrane is driven by a proton motive force in which the proton gradient is the dominant component, and that the full precursor protein can be transported across the thylakoid membrane without prior cleavage by the stromal processing peptidase.

Biological Transport, Active↗
PubMed1992

Immunohistochemical investigation of epidermal growth factor receptor expression during periods of accelerated rat incisor eruption.

The effect of accelerated odontogenesis of the rat mandibular incisor on the expression of receptors for epidermal growth factor (EGF) was examined using specific monoclonal antibodies to the receptor molecule. Acceleration of odontogenesis was achieved by regular trimming of the tooth crown. At normal eruption rates the major area of cross-reactivity was over the secretory ameloblasts. Some labelling of the papilla and preameloblasts was evident. When proliferation was increased the major area of effect was at the preodontoblast/odontoblast boundary where there was a marked increase in labelling, initially at the proximal end of the cell adjacent to the basal lamina. The ameloblasts did not show such a dramatic increase in receptor numbers. Increase in labelling was also evident in the remainder of the papilla. The results suggest that an increase in proliferation with normal morphogenesis is associated with an overall increase in the numbers of EGF receptors, particularly in a population of cells immediately before and after elongation and differentiation of odontoblasts.

Ameloblasts↗
PubMed1992

The effect of periodontal disease on sulphated glycosylaminoglycan distribution in the sheep periodontium.

The glycosylaminoglycan (GAG) distribution in the soft periodontal tissues of the sheep was investigated topographically in healthy tissue and the changes associated with periodontal disease determined and correlated with increasing disease severity. Total sulphated GAG content was determined spectrophotometrically and the proportions of individual GAGS measured after separation by cellulose acetate electrophoresis. In healthy tissue, total sulphated GAG distribution was found to mirror that described previously for collagen, being highest in the gum-pad region and adjacent to alveolar bone. This changed markedly in relation to periodontal disease. Early disease-related changes in GAGs were first detected adjacent to alveolar bone. In severely diseases tissue, an overall increase in sulphated GAG, particularly in the gum-pad region, was present. Increasing disease severity was accompanied by a decrease in dermatan sulphate and a concomitant increase in the proportion of chondroitin-4-sulphate. These changes may be due to the combined effects of chronic inflammation and alterations to the mechanical loading of the tissue owing to loss of attachment.

Animals↗
PubMed1992

The thickness of the sound and periodontally diseased human cementum.

Cementum of teeth previously analysed for fluoride was re-examined in order to determine whether or not periodontal disease had affected the thickness of the tissue. In the periodontally diseased teeth the cementum was thinner than normal in the middle region of the root. The apical cementum, however, was significantly thicker in periodontally diseased than in the sound teeth of subjects over 60 yr old. This may account for the higher total fluoride levels previously reported in the apical cementum of these same teeth.

Adult↗
PubMed1992

Excessive production of reactive oxygen metabolites by inflamed colon: analysis by chemiluminescence probe.

Reactive oxygen metabolites (ROMs) are involved in inflammatory diseases and are postulated to contribute to tissue injury in colitis. To determine whether excessive ROMs are generated by inflamed colonic mucosa and to identify possible sources and type of ROMs, mucosal ROMs were estimated in rats and humans using a chemiluminescence probe. Colitis was induced in rats by intracolonic injection of acetic acid or intraperitoneal injection of mitomycin C. Intact, inflamed colon in rats produced more ultraweak chemiluminescence than normal colon. Inflamed mucosal scrapings from both rat models produced significantly more luminol-enhanced chemiluminescence. Addition of catalase, an H2O2 scavenger, or azide, a myeloperoxidase inhibitor, into the media significantly decreased chemiluminescence from inflamed mucosal scrapings. Indomethacin, an antioxidant cyclo-oxygenase inhibitor, also decreased chemiluminescence, but MK-866, a 5-lipoxygenase inhibitor, had no effect. Colonic biopsy specimens obtained during colonoscopy from patients with ulcerative colitis also produced more catalase-inhibitable chemiluminescence than normal colonic mucosa. These data indicate that excessive ROMs are produced by inflamed colonic mucosa in both humans and rats, which may contribute to tissue injury.

Adult↗
PubMed1992

Thermal analysis of commercial Gutta-percha using differential scanning calorimeter and dynamic mechanical thermal analysis.

Gutta-percha is the most widespread root canal filling material and knowledge of its chemical and thermomechanical properties is of a great interest to its use in clinical dentistry. In the present investigation, the results from differential scanning calorimeter were compared with those obtained by dynamic mechanical thermal analysis. A significant correlation was established between these results and the chemical composition of six different commercial samples of gutta-percha.

Calorimetry, Differential Scanning↗
PubMed1992

Effect of moisture and aging on gutta-percha.

The purpose of this article was to study the plasticizing effect of controlled levels of different humidities on native and commercial gutta-percha samples. It was expected that the results might provide the practicing endodontist with invaluable insight into the mechanical properties of commercial gutta-percha cones (i.e. retrievability, malleability, flexibility, torsional, and bending ability). The "conditioning" of the gutta-percha samples depended greatly on the percentage of compatible and incompatible substances. Mixtures of native gutta-percha with compatible substances (e.g. wax) showed greater elongation, tensile strength, and number of rotations but lower deformation angle than mixtures of native gutta-percha with incompatible substances (e.g. salts, metal oxides). The observed lower values for tensile strength and torsional strain and higher value for elongation and number of rotations for wet samples compared with the dry ones were attributed to the plasticizing effect on gutta-percha of the insertions of water molecules in the polymer chains.

Drug Storage↗
PubMed1992

Impact of a shielded safety syringe on needlestick injuries among healthcare workers.

OBJECTIVES: Evaluate the impact of a shielded 3 cc safety syringe on needlestick injuries among healthcare workers. DESIGN: Surveillance study. SETTING: Three medical centers. RESULTS: The total number of needlesticks from all sources rose from 134 during the baseline period to 140 during the study phase. However, the overall rate of needlesticks involving 3 cc syringes decreased from 14/100,000 inventory units to 2/100,000, and the frequency declined substantially at each of the participating medical centers. CONCLUSIONS: These results suggest that shielded syringes and related technology may offer potential for reducing the risk of needlestick injuries among hospital and other healthcare employees. This study should be viewed as a preliminary effort to assess the potential of shielded-syringe technology for reducing the risk of needlestick injuries among healthcare workers. The results are encouraging, but more clinical experience with this new device and longer follow up are essential to provide the data necessary for a definitive assessment of efficacy.

Humans↗
PubMed1992

Perceptions of pediatricians' helpfulness: a national study of mothers of young disabled children.

A national sample of 503 mothers of young children (up to six years of age) with developmental disabilities was surveyed to determine the extent to which their family pediatricians are helpful in providing information and support. The respondents ranged from lower-middle to upper class, with the mean SES reflective of middle socio-economic status. Their children had a variety of disabilities that qualified them to receive early intervention services. The majority of the mothers found their pediatricians helpful about issues related directly to their child's medical condition. However, few found them helpful with issues relating to the effect of the disability on their child's development, or to their own personal or family needs.

Child↗
PubMed1992

Enamel ultrastructure in pigmented hypomaturation amelogenesis imperfecta.

Hypomaturation amelogenesis imperfecta (AI) is a hereditary condition of enamel that is presumed to result from defects during the maturation stage of enamel development. This study characterized the enamel ultrastructure and enamel crystallite morphology, as well as the distribution of organic material in enamel affected with pigmented hypomaturation AI. Enamel exhibiting autosomal recessive pigmented hypomaturation AI was sectioned or fractured and examined using light microscopy, scanning electron microscopy and transmission electron microscopy. Enamel samples were treated with 30% NaOCl or 8 M urea to remove organic components and determine the effect of deproteinization on crystallite morphology. These were compared with untreated normal enamel samples. The enamel crystallites in hypomaturation AI exhibited considerable variability in size and morphology. Examination of deproteinized tissue indicated that the AI crystallites had a thick coating, presumably of organic or partially mineralized material, which was not visible in normal enamel. The results of this investigation provide further evidence that hypomaturation AI is associated with the retention of organic material that is most probably enamel protein. Enamel protein retention is likely to be involved in the inhibition of normal crystallite growth resulting in the morphological crystallite abnormalities associated with this disorder.

Amelogenesis Imperfecta↗
PubMed1992

A flexible and rapid pH cycling procedure for investigations into the remineralisation and demineralisation behaviour of human enamel.

A rapid and flexible model system has been developed to study human enamel behaviour under conditions of oscillating pH similar to those experienced in the mouth. The model uses realistic time intervals and operates at in vivo temperatures (35-37 degrees C). Mineral loss or gain is measured quantitatively on a volume basis. Results indicate that for single 1-day periods with three 20-min exposures to acid, 1% v/v of enamel mineral was lost. These results, together with the histological appearance of the tissue, are in close agreement with previous studies of enamel caries.

Calcium↗
PubMed1992

Influence of occlusion on the fluoride distribution in rat molar cementum.

This study was undertaken to examine the influence of occlusion on the fluoride distribution in cementum following an experiment in which the occlusion in rats was locally altered by extracting the upper left molar. These and control rats with normal occlusion were given water containing 0 or 100 ppm fluoride for 12 weeks. The fluoride distributions in cementum from both first lower molars of the same animal were compared. The fluoride concentrations had increased throughout the tissue as a result of increased fluoride administration, irrespective of any changes in occlusion. They were, as usual, generally highest at or near the cementum surface and decrease towards the interior of the tissue. Where there had been a change in occlusion, the thickness of cementum was less than that of the contralateral tooth, but, despite this, the fluoride profiles in contralateral teeth were similar in both experimental and control rats. In the experimental rats, on the other hand, the total fluoride tended to be lower, and the mean fluoride tended to be higher in left molars without antagonists. These findings were never seen in the control rats. It was concluded that the alteration in occlusion influenced the fluoride distribution in the cementum through its effect on the rate of cementum formation.

Animals↗
PubMed1992

Determination of volumes of fluid in the mouth by fluoride dilution.

A dilution technique, with fluoride as a marker, has been developed to measure the volume of saliva and other fluids in the mouth. Immediately after swallowing, a small amount of fluoride solution is placed in the mouth, mixed with the oral fluids, and expelled from the mouth. The fluoride concentration of the expelled oral fluid is measured and the volume of fluid in the mouth at the time of spitting calculated from the fluoride dilution. Mean values of 0.75 +/- (SD) 0.28 ml for males and 0.72 +/- (SD) 0.16 ml for females were recorded. The accuracy of the volume determination is about +/- 0.10 ml in vivo. This technique has provided a convenient and rapid means of determining saliva volumes. It could also be used to determine rates of secretion and may prove useful in the clinic for assessing a patient's salivary competence.

Adult↗
PubMed1992

The role of albumin in developing rodent dental enamel: a possible explanation for white spot hypoplasia.

The uptake of serum albumin by maturation-stage rodent enamel and the resulting effects on the growth of enamel crystallites were investigated in vitro. Albumin uptake was demonstrated by means of gel electrophoresis and confirmed by Western blotting with use of monoclonal antibodies. Measurement of crystal size was carried out by direct TEM measurement of enamel crystallite outlines after incubations in metastable solutions of calcium phosphate. The ability of endogenous enamel enzymes to degrade albumin was investigated by substrate-specific zymography. The results showed that albumin could be taken up by maturation-stage enamel and produce inhibition of crystallite growth. There was no detectable proteolytic activity in the enamel against albumin substrate, which suggests that albumin entering enamel by extravasation in vivo may produce incomplete tissue maturation, resulting in a white, opaque appearance on eruption.

Animals↗
PubMed1992

Improving documentation of aggressive behavior in nursing home residents.

1. Although 66% of the 1.3 million elderly nursing home residents in the US exhibit aggressive tendencies, research indicates that caregivers underdocument aggressive incidents. As a result, the extent of aggression is unknown. 2. This study found that LPNs reported the largest percentage (41%) of the 51 documented aggressive incidents, which reportedly occurred most frequently in the bathroom (49% of the time) between 5 AM and noon (86% of the time). Most aggressive residents (55.1%) never had visitors. 3. This study concludes that nursing practice would benefit from a consistent definition of aggression, an easy-to-use clinical instrument to collect data on aggression, and caregiver education programs on assessing aggression.

Aged↗