Josie A. Beeley

Salivary proteins in rheumatoid arthritis and Sjögren's syndrome: one-dimensional and two-dimensional electrophoretic studies.

Parotid saliva from patients with rheumatoid arthritis and Sjögrens syndrome contains elevated levels of multiple anionic proteins of pI ˜3.75—4.75 and Mr ˜32 000. Further studies on these components involving narrow range pH 3.5—5.0 immobilized pH gradients (IPGs) and two‐dimensional (2D) electrophoresis with narrow‐ or broad‐range IPGs in the first dimension have confirmed their association with these disorders. Immunoblotting showed that these proteins include multiple forms of tissue kallikrein. Treatment with neuraminidase results in removal of these anionic substances from the pH 3.75—4.75 region of gels, thereby indicating that heterogeneity arises from differences in sialation of the carbohydrate residues. The results of treatment with endo‐β‐N‐acetylglucosaminidase (Endo F) or peptide N‐glycosidase (PNGase F) and comparison of the results with studies on human urinary kallikrein suggest that proteins other than kallikrein may also comigrate in the anionic region of gels and that deglycosylation of kallikrien is incomplete in the experimental conditions used, probably because of inadequate denaturation. The paper also reviews the limitations of current criteria used in the investigation of salivary gland function associated with connective tissue disorders and the diagnosis of these. It assesses the potential of electrophoresis in forming the basis of new diagnostic techniques and furthering the understanding of the nature of these diseases. The findings presented in this paper could make a key contribution to this.

Effect of extracellular polysaccharides on diffusion of NaF and [14C]-sucrose in human dental plaque and in sediments of the bacterium Streptococcus sanguis 804 (NCTC 10904)

It has been postulated that extracellular polysaccharides form a barrier to diffusion in dental plaque. Diffusion coefficients, D, were measured for NaF and [14C]-sucrose in glucan-free and glucan-containing sediments of Strep. sanguis 804 at 37 degrees C. There was a tendency for NaF and [14C]-sucrose to diffuse faster as the carbohydrate concentration in the sediments increased. NaF diffused only 38 per cent more slowly in cell-free glucan sediment than in water, suggesting that glucan per se does not form a barrier to diffusion. The diffusion coefficient for NaF was positively correlated with carbohydrate concentration in individual plaque samples from 15 subjects and incubation of 3 plaque samples with sucrose resulted in both an increase in carbohydrate concentration in the plaque and an increase in D for NaF. Thus the presence of extracellular polysaccharides in plaque leads to slightly faster rates of diffusion. Nevertheless, the total time for diffusion through plaque may be increased if the presence of extracellular polysaccharides results in thicker layers of plaque.

Clinical applications of electrophoresis of human salivary proteins

Human salivary proteins have been studied by electrophoresis in denaturing and non-denaturing polyacrylamide gel electrophoresis (PAGE) as well as by isoelectric focusing (IEF) and two-dimensional procedures, and the clinical applications of this have been reviewed. Whilst non-denaturing PAGE is useful in studying polymorphisms, sodium dodecylsulphate PAGE appears to be otherwise preferable. Immobilized pH gradients containing carrier ampholytes (CAs) give better resolution than CA-based IEF and overcome the problems of cathode drift and loss of basic material. Proline-rich proteins stain poorly with conventional procedures and special techniques are necessary. In clinical studies, findings must be viewed over and above the large number of polymorphisms which occur normally. Studies relating salivary protein and peptide profiles to dental caries susceptibility are encouraging. Specific protein abnormalities have been associated with connective tissue disorders and could form the basis of new non-invasive diagnostic procedures. Protein differences associated with cystic fibrosis and diabetes mellitus, however, merit reinvestigation with the new procedures now available. Detection of HIV antigens in saliva is a new area of research. In the light of new techniques available and new information which has arisen from DNA studies, future prospects for the clinical applications of electrophoresis of saliva look good.

Pink Teeth Appearing as a Post-Mortem Phenomenon

Biochemical investigations into the nature of the post-mortem occurrence of pink teeth are described. Spectral studies and iso-electric focusing indicated the presence of haemoglobin or other haem derivatives. The presence of carbon monoxide was also observed. It is suggested this unstable pink colouration results from the formation of a complex, possibly a haem-carbon monoxide complex, which differs from the more stable carbon-monoxy-haemoglobin.

Genetic analysis of human salivary ?-amylase isozymes by isoelectric focusing

The isozymes of human salivary α-amylase have been separated by thin-layer gel isoelectric focusing in a pH 4–8 gradient followed by a starch-iodine zymogram procedure. The normal isozyme pattern (N) consisted of five major isozymes together with a number of minor ones. In addition, seven variant isozyme phenotypes were also observed, which had a combined frequency of 11.7% in a random population of 368 individuals. Analysis of familial data is indicative of the inheritance of autosomal codominant alleles.

An improved procedure for isoelectric focusing of human salivary proteins

Abstract Isoelectric focusing in thin layers of polyacrylamide gel is a powerful technique for the separation of human salivary proteins. The method described involves prior de-salting and concentration, followed by isoelectric focusing, in a pH gradient of 4–8, of sample volumes up to 0.1 ml. With whole saliva, 30–40 protein bands were clearly evident on staining with Coomassie Brilliant Blue; and α-amylase zymogram procedure revealed 9 or 10 isozymes. Within 8 individuals, a number of reproducible protein variants were apparent.

A European Core Curriculum in Cariology: the knowledge base.

This paper is part of a series of papers towards a European Core Curriculum in Cariology for undergraduate dental students. The European Core Curriculum in Cariology is the outcome of a joint workshop of the European Organization for Caries Research (ORCA) together with the Association for Dental Education in Europe (ADEE), which was held in Berlin from 27 to 30 June 2010. This paper presents a closer look at the knowledge base as presented in the European Core Curriculum in Cariology. It comprises not only traditional basic sciences, such as anatomy and histology, but also emerging sciences such as molecular biology and nanotechnology and also fields such as behavioural sciences and research methodology. The different supporting competences are elaborated and explained. The problems of implementing a curriculum that truly integrates this foundation knowledge into the clinical teaching are discussed.

Adapting to changes in molecular biosciences and technologies

Dental education, like any other educational programme in a research-intensive university environment, must be research led or at least research informed. In this context, as the research and knowledge base of dentistry lies in the biological and physical sciences, dental education must be led by advances in research in both these areas. There is no doubt that biotechnology and nanotechnology have, over the past 25 years, led research in both these areas. It is therefore logical to assume that this has also impacted on dental education. The aim of this paper is twofold; on one hand to examine the effects of biotechnology and nanotechnology and their implications for dental education and on the other to make recommendations for future developments in dental education led by research in biotechnology and nanotechnology. It is now generally accepted that dental education should be socially and culturally relevant and directed to the community it serves. In other words, there can be no universal approach and each dental school or indeed curriculum must apply the outcomes in their own social, cultural and community settings.

The Production and Evaluation of an Antiserum for the Detection of Human Saliva

An antiserum raised in rabbits against human whole saliva cross-reacted with human serum and semen. Affinity chromatography of the antiserum on columns of immobilised human serum and seminal protein absorbed out these cross-reacting antibodies leaving a major antibody which reacted with α-amylase in saliva and pancreatic juice. The absorbed antiserum was evaluated as a reagent for the specific detection of human saliva in stains of forensic interest ; in a blind trial consisting of 87 samples, 32 positives, 1 false negative and no false positives were obtained. The antiserum appears to be highly specific for human saliva and does not cross-react with that of eight species of popular domestic animals.

Immunological studies on human salivary albumin

Abstract The presence of a plasma albumin-like component in human mixed saliva was confirmed. Immunochemical studies, however, suggest that salivary albumin appears to be non-antigenic in the rabbit. The possibility of differences between plasma and salivary albumins is-discussed.