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Biomedical subjects

E Odell

Publications and source records attributed to E Odell.

15 recordsLinked to original sources

A minimally invasive immunocytochemical approach to early detection of oral squamous cell carcinoma and dysplasia.

Squamous dysplasia of the oral cavity indicates increased risk of progression to squamous cell carcinoma (SCC). An important advance would be the development of a minimally invasive assay for identification of oral SCC and dysplasia. We have investigated the suitability in this context of immunostaining oral smears for minichromosome maintainance proteins (MCMs), sensitive and specific biomarkers of cell cycle entry. Immunohistochemical examination of 66 oral tissue samples showed a greater frequency of Mcm-2 expression in surface layers of moderate/severe dysplasia and SCC compared to benign keratosis/mild dysplasia. Immunocytochemistry for Mcm-2/Mcm-5 was performed on 101 oral smears. Conventional smears included 23 from normal mucosa, benign proliferative disease and mild dysplasia, all of which were MCM negative. Of 52 conventional smears of SCC tissue samples, 18 were inadequate. However, MCM-positive cells were present in 33/34 adequate samples. Of 26 liquid-based cytology smears, 19 out of 20 smears from SCC were adequate and all were MCM positive. Six smears from benign lesions were adequate and MCM negative. We conclude that MCMs are promising markers for early detection of oral SCC and dysplasia, particularly in a liquid-based cytology platform. Detection of MCMs would be amenable to automation and potentially applicable in the developing world. Further studies are now warranted.

Biomarkers, Tumor↗

Brachyury, a crucial regulator of notochordal development, is a novel biomarker for chordomas.

Chordomas are malignant tumours that occur along the spine and are thought to derive from notochordal remnants. There is significant morphological variability between and within chordomas, with some showing prominent areas of chondroid differentiation. Our microarray data from a broad range of connective tissue neoplasms indicate that, at the transcriptional level, chordomas resemble cartilaginous neoplasms. Here we show that chordomas express many genes known to be involved in cartilage development, but they also uniquely express genes distinguishing them from chondroid neoplasms. The brachyury transcription factor, known to be involved in notochordal development, is only expressed by chordomas. Using a polyclonal antibody, we show that brachyury is expressed in the embryonic notochord and in all 53 chordomas analysed, labelling both chondroid and chordoid areas of these tumours. In contrast, the protein was not detected in over 300 neoplasms, including 163 chondroid tumours. Brachyury was not detected in the nucleus pulposus, arguing against the hypothesis that this tissue derives directly from the notochord. These data provide compelling evidence that chordomas derive from notochord and demonstrate that brachyury is a specific marker for the notochord and notochord-derived tumours.

Biomarkers, Tumor↗

Role of microchimerism in the pathogenesis of oral lichen planus.

OBJECTIVE: Microchimerism of persistent fetal cells has been implicated in some cell-mediated autoimmune diseases. This study examines the hypothesis that fetal microchimerism plays a role in the pathogenesis of lichen planus (LP) affecting the oral cavity. STUDY DESIGN: Mucosal biopsies of 12 women with oral LP (OLP) were tested for the presence of both male cells and male DNA originating from prior pregnancies or prior blood transfusions. Six male patients with OLP served as a control group. Biopsies were analyzed for the presence of Y-chromosome-positive cells by fluorescence in situ hybridization (FISH) with X- and Y-specific DNA probes. To confirm the FISH findings, we used fluorescent polymerase chain reaction (PCR) to identify Y-chromosome sequences in DNA extracted from mucosal lesions. RESULTS: Using FISH technology, all the 18 biopsy samples showed good hybridization results. In females, Y-chromosome-specific signals were not detected by FISH at any site of the lesions. PCR amplification demonstrated a single peak at the locus specific for the X-chromosome. CONCLUSION: Male DNA microchimerism was not present in any of the investigated lesions, suggesting that microchimerism because of persisting male fetal cells is unlikely to play a major role in the pathogenesis of OLP.

Adult↗

Mechanical evaluation of mandibular defects reconstructed using osteogenic protein-1 (rhOP-1) in a sheep model: a critical analysis.

Osteoinductive bone morphogenetic proteins (BMPs) have been used extensively in experimental and clinical orthopaedic research. It is a natural progression for these growth regulators to be tested in the craniofacial region. The aim of this investigation was to analyse the mechanical properties of the sheep mandibles reconstructed using recombinant human osteogenic protein type 1 (rhOP-1). A unilateral 35 mm osteoperiosteal continuity defect was created at the parasymphyseal region of the mandible in six adult sheep. The animals were sacrificed 3 months after surgery and mechanical properties of the regenerated bone at the operated sides (OS) were compared to the corresponding bone at the non-operated side (NOS). The regenerated tissue at the OS were then submitted for histological and histomorphometric analysis. Although all the animals achieved complete bony union, a wide range of mechanical properties was found. The rhOP-1-induced bone achieved a mean of 36% of the strength of the bone at the NOS (P < 0.05). The mean value of the stiffness of the OS was 24% of the NOS (P < 0.05). While half of the samples of the OS had 'weak' mechanical properties (9-25% strength compared to NOS) and a low stiffness (6-18%), the rest showed relatively higher strength (47-63%) and were stiffer (35-47%). Unlike the NOS, the operated sides failed under tensile stresses and cracks initiated at the superior border of the mandible. The wide mechanical variations suggest that further basic bone biology research is needed to provide better understanding of the cellular and molecular events which take place during the process of osteoinduction.

Activin Receptors, Type I↗

Histological assessment of bioengineered new bone in repairing osteoperiosteal mandibular defects in sheep using recombinant human bone morphogenetic protein-7.

UNLABELLED: Numerous experimental studies have been published about osteoinductive bone morphogenetic proteins (BMPs). However, to our knowledge there has been no detailed histological study of a mandibular defect in a large mammal, reconstructed using BMPs. We describe here the histological features of rhBMP-7-induced bone in mandibular defects in sheep. METHODS: A 35 mm osteoperiosteal defect was created at the parasymphyseal region of the mandible in six adult sheep. The continuity of the mandible was maintained using a bony plate, and rhBMP-7 was applied on a type I collagen carrier. Bone labels were injected at selected time intervals during the follow-up period. The animals were killed after 3 months and bone samples were examined histologically, histomorphometrically, and by fluorescence microscopy. RESULTS AND CONCLUSIONS: We found a mixture of woven and lamellar bone that contained many cells with large nuclei. This had not reorganised to form cortical bone and the rhBMP-7-induced bone was more porous than the native bone. The newly-formed bone restored both endosteal and periosteal layers. rhBMP-7-induced bone was biocompatible and induced no ossification of soft tissue or abnormal growth of nearby vital structures. The mineral apposition rate was 1.98 microm/day (range 0.62-5.63 microm/day), a value close to that reported in humans. This suggests that BMPs have a limited effect in accelerating the rate of mineralisation, but promote the pre-mineralisation processes, and perhaps the formation of woven bone.

Animals↗

3-D optical coherence tomography of the laryngeal mucosa.

Laryngeal carcinoma is one of the commonest primary head and neck malignancy and the need for early identification is very important for successful treatment. Outpatient fibreoptic examination of the larynx is unreliable in differentiating benign, pre-malignant and malignant lesions, and therefore surgeons have to rely on biopsies for a definitive diagnosis. This is an invasive procedure requiring general anaesthesia and may have a detrimental effect on the patient's voice. Conventional imaging modalities (ultrasound, computed tomography and magnetic resonance imaging) have a limited resolution and hence cannot give sufficient information on the extent or nature of laryngeal lesions. The aim of our study is to investigate the feasibility of optical coherence tomography (OCT) in imaging the normal larynx, to lay the foundations for an investigation of its ability to differentiate between benign and malignant disease. Ten tissue specimens from normal larynges were imaged with an 850 nm OCT system that was capable of providing both B-scan (longitudinal or cross-section) images as well as C-scan (en-face or images at constant depth). The en-face OCT mode allowed us to reconstruct 3-D OCT images of the tissue examined. Imaged specimens were processed with standard histopathological techniques and sectioned in the plane of the B-scan OCT images. Haematoxylin-eosin stained specimens were compared with the OCT images thus collected. Preliminary results showed good correlation between OCT images and histology sections in normal tissue.

Humans↗

Effects of local anesthesia on substance P and CGRP content of the human dental pulp.

The aim of the study was to determine immunoreactive Substance P (iSP) and immunoreactive calcitonin gene-related peptide (iCGRP) content in the human dental pulp and whether local anesthesia has an effect on the neuropeptide content. Dental pulps were obtained from patients, who underwent surgical extraction of all 4 impacted wisdom teeth under general isoflurane gas anesthesia. There was a very high interindividual variation in tissue content, with small variation in levels found in teeth from the same patient. Pulps obtained from lower teeth without local anesthesia contained an average of 131 +/- 62 fmol/mg protein of iCGRP and 15 +/- 9 fmol/mg iSP (n = 10). With additional mandibular block anesthesia the values were 194 +/- 71 fmol/mg iCGRP (statistically significant, p = 0.0356, Mann-Whitney-rank-sum-test) and 12 +/- 3.6 fmol/mg iSP. The results suggest that local anesthesia attenuates neuropeptide release in the human dental pulp during surgical extraction.

Anesthesia, Dental↗

Chronic sinusitis.

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Aspergillosis↗

Zinc as a growth factor for Aspergillus sp. and the antifungal effects of root canal sealants.

Noninvasive aspergillosis of the maxillary sinus is frequently associated with extrusion of root canal cements into the sinus, and it has been suggested that zinc oxide-based materials might promote infection because zinc is a growth factor for Aspergillus sp. To investigate this possibility pure zinc oxide-eugenol cement and five other cements were tested for antifungal activity against five strains of Aspergillus fumigatus, one strain of A. niger, and one strain of A. flavus by zone inhibition and by spectrophotometric measurement of growth rates at various zinc concentrations. All cements had antifungal activity. Eugenol-containing materials had the most activity, but this activity was reduced with mixing and setting. No growth enhancement attributable to zinc was detected by either method. To determine whether contaminated materials might introduce spores into the sinus, 63 samples taken from one clinic were cultured. Aspergillus sp. was isolated from two.

Acrylic Resins↗

Interaction of human myeloperoxidase with nitrite.

EPR (electron paramagnetic resonance) and optical spectroscopy show that human neutrophil myeloperoxidase is converted from ferric high-spin to low-spin by the addition of nitrite. The Soret peak shifts from 429 to 447 nm and new peaks appear in the visible region at 573 and 627 nm; the EPR g-values change from 6.84, 5.02, 1.95 to 2.55, 2.31, 1.82. Small differences are seen in the EPR (but, not optical) spectra of myeloperoxidase isoenzyme I compared to isoenzymes II and III. The reaction with nitrite is detectable by EPR in intact neutrophils and is thus a possible in vivo monitor of NO/nitrite production by these cells.

Chlorides↗

Prediction of domain organisation and secondary structure of thyroid peroxidase, a human autoantigen involved in destructive thyroiditis.

Organ specific autoimmune diseases are relatively common immunological disorders in man which include thyroid autoimmune disease, insulin-dependent diabetes mellitus and myasthenia gravis. The target autoantigens in some of these diseases have recently been characterised. In thyroid autoimmune disease this includes the key enzyme, thyroid peroxidase (TPO), which is involved in the generation of thyroid hormone. Structural knowledge about autoantigens such as thyroid peroxidase will allow a greater understanding of the interaction between autoantigens and the aberrant immune response, and facilitate the development of strategies for antigen-specific therapeutic manipulation. We report here a prediction of the secondary structure of thyroid peroxidase, together with the results of circular dichroic spectroscopy of a homologous purified enzyme. A combination of 3 secondary structure prediction programs has been used, following multiple sequence alignment, and TPO has been found to consist mainly of alpha-helical conformation, with little beta-sheet present. This structure prediction, together with knowledge of the exon-intron boundaries allows a model for the domain organisation of the TPO molecule to be proposed.

Amino Acid Sequence↗

Thyroid microsomal/thyroid peroxidase autoantibodies show discrete patterns of cross-reactivity to myeloperoxidase, lactoperoxidase and horseradish peroxidase.

The recent cloning of the thyroid peroxidase (TPO) has shown that it is identical to the thyroid microsomal antigen (TMA), a potent antigen involved in autoimmune thyroid disease (ATD), which shares significant sequence homology with myeloperoxidase. The present study shows that autoantibodies (aAb) to the TMA/TPO antigen cross-react with human leucocyte myeloperoxidase, bovine lactoperoxidase and horseradish peroxidase. Cross-reactivity to myeloperoxidase was only apparent by ELISA using reduced and alkylated antigen preparations or by immunoblotting following denaturation with SDS. Sequential absorption of sera on SDS-denatured thyroid microsomes immobilized on Sepharose-4B followed by absorption on native microsomes removed all aAb specificities to TMA/TPO and the three peroxidase preparations, giving compelling evidence on the genuine cross-reactive nature of these aAbs. Sera from different patients contain different qualitative and quantitative specificities of aAb to the TMA/TPO antigen, confirming the polyclonal nature of this autoimmune response.

Absorption↗