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Biomedical subjects

T H Lilley

Publications and source records attributed to T H Lilley.

17 recordsLinked to original sources

Visual whiteness ranking of a Vitapan 3D Master shade guide by untrained assessors.

OBJECTIVE: This study was conducted to obtain the ranking order of a 29-tab (including three "bleaching tabs") Vita Shade Guide for perceived "whiteness" by untrained assessors, and to compare this ranking with both the shade guide manufacturer's rank order and a colorimetrically derived rank order. METHODOLOGY: A total of 85 people, not trained in color assessment by shade guides, were asked to rank order the tabs of a Vitapan 3D Master shade guide for perceived "whiteness" under standardized lighting conditions. A whiteness ranking was also obtained colorimetrically using a Minolta CM-2600d spectrophotometer calibrated to give CIE whiteness values. The data were analysed using means, standard deviations, and standard errors. RESULTS: The assessors varied more in their rankings for darker shades than the lighter ones. The order derived by the assessors and the colorimetry order did not match well with the manufacturer's order, especially toward the darker shades. CONCLUSION: For use in studies of whitening products, a new order of the Vitapan 3D Master shade guide tabs has been developed in relation to whiteness.

Adult↗

The measurement of tooth whiteness by image analysis and spectrophotometry: a comparison.

Digital image capturing and analysis techniques have been used to measure the colour of teeth and to compare with spectrophotometric results and visual observations. A non-linear image analysis approach was developed and, for the colour range of human teeth, allows device-dependant digital camera colour data to be quantitatively transformed to Commission Internationale de l'Eclairage (CIE) colorimetric values. With reference to a CIE standard illuminant, two different lighting arrays have been used. For flat and non-translucent white and yellow surfaces, spectrophotometric results showed that this transformation achieves required accuracy. It was found, in all of the present studies, which included measurements on the VITA Lumin Vacuum shade guide and extracted teeth, that spectrophotometry invariably underestimated values of the CIE whiteness index. However, the results from these two types of measurement correlated well. There was also a reasonably good correlation between earlier data obtained by visual assessment and the present data by the two instrumental methods. For extracted teeth, both instrumental methods used in this work did not confirm a whitening effect for 2-min brushing with toothpaste, but did show significant whitening results for bleaching with 15% hydrogen peroxide.

Algorithms↗

Moderation of oral bacterial adhesion on saliva-coated hydroxyapatite by polyaspartate.

AIMS: Synthetic sodium alpha,beta-polyaspartate (PA) has been investigated as a moderator of adhesion and the subsequent biofilm formation by oral bacteria. METHODS AND RESULTS: The inhibition of bacterial adhesion by PA was assessed by (i) a 30-min incubation with Streptococcus sanguis in a microtitre assay with the wells coated with hydroxyapatite (HAP) and (ii) an 18-h challenge with human salivary microflora in a HAP disc assay. In contrast to HAP-coated surfaces, clean polystyrene surfaces in the microtitre assay exhibited no anti-adhesion properties. It has been found that PA significantly and similarly adsorbs onto HAP surfaces in the presence and absence of salivary coating. The HAP disc assay also showed that PA, both in aqueous solutions and in toothpaste, reduced the level of adhered microflora and this effect was enhanced by added propylene oxide-ethylene oxide copolymers. CONCLUSION: The principal finding from this work is the potential role for PA as an inhibitor of dental plaque formation. PA may significantly modify the salivary pellicle. SIGNIFICANCE AND IMPACT OF THE STUDY: This work indicates the use of PA in controlling the development of dental plaque and the formation of bacterial biofilm in general.

Adsorption↗

Multiple interactions between polyphenols and a salivary proline-rich protein repeat result in complexation and precipitation.

Polyphenols (tannins) in the diet not only precipitate oral proteins, producing an astringent sensation, but also interact with dietary proteins and digestive enzymes in the gut, resulting in a variety of antinutritive and toxic effects. Salivary proline-rich proteins (PRPs), which are secreted into the oral cavity, form complexes with and precipitate dietary polyphenols, and thus, they constitute the primary mammalian defense directed against ingested tannins. In order to characterize the interaction, NMR studies were performed which involved titrating a series of polyphenols into a synthetic 19-residue PRP fragment. The results show that the predominant mode of association is a hydrophobic stacking of the polyphenol ring against the pro-S face of proline and that the first proline residue of a Pro-Pro sequence is a particularly favored binding site. Measurement of dissociation constants indicates that the larger and more complex polyphenols interact more strongly with the PRP fragment; the order of binding affinity was determined as procyanidin dimer B-2 > pentagalloylglucose > trigalloylglucose >> proanthocyanidin monomer (-)-epicatechin approximately propyl gallate. Smaller polyphenols can bind with one phenolic ring stacked against each proline residue, whereas larger polyphenols occupy two or three consecutive prolines. The more complex polyphenols interact with the PRP fragment in a multidentate fashion; moreover, they self-associate or stack when bound. Thus, a model is proposed in which multiple polyphenol/polyphenol and polyphenol/PRP interactions act cooperatively to achieve precipitation.

Amino Acid Sequence↗

Tannin interactions with a full-length human salivary proline-rich protein display a stronger affinity than with single proline-rich repeats.

The protein IB5 has been purified from human parotid saliva. This protein contains several repeats of a short proline-rich sequence. Dissociation constants have been measured at several discrete binding sites using 1H-NMR for the hydrolysable tannins (polyphenols) beta-1,3,6-tri-O-galloyl-D-glucopyranose, beta-1,2,4,6-tetra-O-galloyl-D-glucopyranose and beta-1,2,3,4,6-penta-O-galloyl-D-glucopyranose and the condensed proanthocyanidin (--)-epicatechin. The dissociation constants for trigalloyl glucose and pentagalloyl glucose were 15 X 10(-5) and 1.7 X 10(-5) M, respectively, which are 115 and 1660 times stronger than those previously measured under the same conditions for a single repeat of a mouse salivary proline-rich protein. The increase in affinity is ascribed to intramolecular secondary interactions, which are strengthened by the rigidity of the interacting molecules.

Amino Acid Sequence↗

Study of the interaction between salivary proline-rich proteins and a polyphenol by 1H-NMR spectroscopy.

The interaction between salivary proline-rich proteins and plant polyphenols (tannins) in the oral cavity and their subsequent precipitation influences the taste, texture and nutritional value of food; it is thought to be responsible for the astringency of many foods and beverages. To investigate the interaction, two-dimensional 1H-NMR studies have been carried out on the binding of a representative polyphenol, pentagalloyl glucose, to two synthetic peptides (19 and 22 residues in length) that are typical of the repeat sequence of mouse salivary proline-rich protein MP5. Intermolecular nuclear Overhauser effects and chemical shift changes show that the main binding sites on the peptides are proline residues together with the preceding amide bond and amino acid. The interaction is principally a hydrophobic association between a galloyl ring and the pyrrolidine ring face containing the C alpha proton, but secondary hydrogen-bonding effects help to stabilise the complex. Very similar interactions are seen for both peptides. The conformation of the peptides remains extended on binding. The chemical shift changes seen for many of the peptide protons can be fitted to a simple binding curve with dissociation constant of around 40 mM, but some protons show evidence of cooperative binding involving several galloyl groups. Higher concentrations of pentagalloyl glucose lead to a reduced off-rate from the complex and eventual precipitation which implies that precipitation is caused by a kinetic competition between aggregation and dissociation of the complex.

Amino Acid Sequence↗

Polyphenols, astringency and proline-rich proteins.

Recent, NMR and precipitation, studies of molecular recognition of proline-rich proteins and peptides by plant polyphenols are described and rationalized. The action of polysaccharides and caseins in the moderation of the astringent response, which is engendered by polyphenols present in foodstuffs and beverages, is described. The possible influence of plant cell wall glycoproteins on the process of lignification is discussed in the light of the observed affinity of phenolic substrates for prolyl residues in protein structures.

Amino Acid Sequence↗

E. coli penicillin acylase isolation by selective release, aqueous two-phase partitioning and ultrafiltration.

A procedure has been developed for the purification of penicillin acylase from E. coli fermentation broths. The method described is based primarily on selective release from biomass, charge-directed partitioning in an aqueous two-phase system, and the use of ultrafiltration membranes to recycle the modified polyethylene glycol required and to further purify the enzyme. Each step has been discussed and comparisons have been made with other approaches, where possible. Suggestions have been made for optimisation in process engineering. The approach developed may be applicable to other beta-lactam antibiotic acylases or more generally to some of the other E. coli periplasmic proteins.

Biological Assay↗

Structure of N-acetyl-L-leucyl-L-prolinamide monohydrate.

C13H23N3O3.H2O, Mr = 287.36, orthorhombic, P2(1)2(1)2(1), a = 6.3863 (8), b = 14.346 (2), c = 17.247 (2) A, V = 1580.1 (6) A3, Z = 4, Dx = 1.208 g cm-3, lambda(Cu K alpha) = 1.54178 A, mu = 7.04 cm-1, F(000) = 624, room temperature, final R = 0.037 for 1693 reflections with I greater than or equal to 2.5 sigma (I) and 257 variables. The peptide linkage is in a trans conformation. The pyrrolidine ring adopts a puckered C gamma-endo conformation and the leucine side chain is in a twisted [g-(tg-)] conformation. The crystal packing is stabilized by a three-dimensional network of hydrogen bonds which involve all the donor groups. The water molecule plays an important part in the crystal packing, being engaged in three hydrogen bonds with different N-acetylamide molecules.

Amino Acid Sequence↗

The prediction of water activities in multicomponent systems.

The molecular bases of the equilibrium water activity concept are discussed and illustrated using some simplified examples. It is shown, using a relatively refined treatment, that at the moment we are not in a position to predict water activities, even for simple systems. A method is described which allows the prediction of the water activity of multicomponent systems from the properties of solutions containing one and two solutes. The method can also be used to predict solubilities in systems containing more than one solute, and this is illustrated with an example.

Chemical Phenomena↗

Cryoprotectant toxicity and cryoprotectant toxicity reduction: in search of molecular mechanisms.

Cryoprotectant toxicity is a fundamental obstacle to the full potential of artificial cryoprotection, yet it remains in general a poorly understood phenomenon. Unfortunately, most relevant biochemical studies to date have not met the basic criteria required for demonstrating mechanisms of toxicity. A model biochemical study of cryoprotectant toxicity was that of Baxter and Lathe, which demonstrated that alteration of a specific enzyme (fructose diphosphatase, or FDPase) was the cause of impaired glycolysis after treatment with and removal of dimethyl sulfoxide (D). FDPase alteration by D was reported to be preventable by the simultaneous presence of amides. This protection could be due to a "counteracting solute" effect similar to that employed by nature, but we find no meaningful correlation between the general protein stabilizing or destabilizing tendency of the cryoprotectant medium and its toxicity. Baxter and Lathe postulated that the effect of D arises from hydrogen bonding between D and the epsilon amino groups of surface lysine residues on FDPase, and it was found that molecules which resembled this group could block the alteration induced by D, presumably by competing with lysine residues for association with D. However, we find that the interaction between D and lysine in the presence of water is actually thermochemically repulsive, and that the presence of formamide does not affect the interaction between D and lysine, implying no useful complex formation between formamide and D. We were also unable to demonstrate that the blocking compounds consistently reduce toxicity when added to D rather than substituting for D, contrary to predictions based on complex formation between blocking compounds and D. In summary, it seems that present concepts of cryoprotectant toxicity are in need of serious revision.

Amides↗

Traditional herbal medicines--the role of polyphenols.

The polyphenol composition of some traditional herbal remedies is reviewed. Polyphenols probably act in such remedies by virtue of their astringent action and current views on the molecular basis of this property are outlined and discussed.

Chemical Phenomena↗

Natural astringency in foodstuffs--a molecular interpretation.

The structures of plant polyphenols (vegetable tannins) are briefly reviewed. Their interactions with proteins, polysaccharides, and the alkaloid caffeine are discussed at the molecular level, and these fundamental properties are related to the quality of astringency that polyphenols possess. The various ways in which astringency may be modified and ultimately lost are outlined in relation to the aging of red wines, the formation of nonbiological hazes in beers and lagers, and the ripening of fruit.

Astringents↗

Ferritin as an iron-storage protein: mechanisms of iron uptake.

The major physiological role of the ubiquitous protein ferritin is to store iron, which it does as a mineral core within a protein shell. Using evidence from a variety of sources we suggest that this versatile protein responds in its mechanism of iron uptake to changes in environmental conditions.

Ferritins↗

Lipid phase transitions in model biomembranes. The effect of ions on phosphatidylcholine bilayers.

Differential scanning calorimetry has been used to study the endothermic phase behaviour of some model biomembranes (i.e. phosphatidylcholine-water systes) in the presence of a wide range of alkaline, alkaline earth and heavy metal salts. Studies and comparisons were made of both cation and anion effects. Shifts occur in the temperatures of both the pre-transition and main transition endotherms. The observed shifts are smaller than those which have been reported for charged lipids, and no evidence has been found for the formation of specific complexes. Electron microscopic studies on freez-fractured dispersions of phosphatidylcholine-water-salt systems show that with some salts the typical rippled surface observed with L-alpha-dimyristoyl phosphatidylcholine, when in the gel state, is replaced by a smooth surface.

Anions↗

Synergy of binary poly(oxypropylene-oxyethylene) copolymers in reducing retention of Streptococcus sanguis to hydroxyapatite.

Binary poly(oxypropylene-oxyethylene) block copolymer systems have been investigated as a non-bactericidal approach to reduce the retention of an oral bacterium, Streptococcus sanguis. Using a previously validated hydroxyapatite-coated microtitre model to simulate the tooth, the performance of copolymer pairs was measured experimentally. A synergy index and an efficacy index were defined to describe the reduction of bacterial retention by the binary systems that comprise the copolymer pairs. Relationships between the synergy and efficacy indices and their associated compositions are given. The results obtained have been rationalised using a previously developed theoretical approach in conjunction with the binding energetics of species to surfaces.

Bacterial Adhesion↗