Analysis of colorectal tumor progression by microdissection and comparative genomic hybridization

This investigation aimed to identify patterns of copy number change in colorectal tumor progression from adenoma to liver metastasis. Fifty-three microdissected sub-regions from 17 cases of colorectal cancer were assigned to one of six histopathologically defined categories: coexisting adenoma, tumor above the muscularis layer, tumor within the muscularis layer, tumor extending through the bowel wall to serosal fat, lymph node metastasis, and liver metastasis. Microdissected samples were treated by a microwave processing step and then used as templates for universal PCR amplification. PCR products were fluorophore labeled and subjected to comparative genomic hybridization. Copy number changes were found in all samples, and every chromosome arm (excluding acrocentric short arms) was affected. More losses than gains were detected, but there were no significant differences between the numbers of changes seen in each category. Each individual sample revealed unique changes, additional to those shared within each case. The most frequently observed gains were of X and 12q. The most common losses were of 8p, 16p, 9p, 15q, 18q, and 10q. Nominally significant associations were observed between metastatic tumor and loss of 12q24.1 or 10p13-14, non-metastatic tumor and loss of 8q24.1, tumor extending to serosal fat and loss of 6q24-25 or gain of 4q11-13, tumor extending to serosal fat and metastatic lesions and loss of 4q32-34 or 22q11-12, and adenoma and loss of 15q24. Loss of 4q32-34 remained highly significant after correction for multiple testing. Adenoma was the only category not to show loss of 17p. These data reveal a genetically heterogeneous picture of tumor progression, with a small number of changes associated with advanced disease.     

Repeated laparoscopic ovarian diathermy is effective in women with anovulatory infertility due to polycystic ovary syndrome

OBJECTIVE: To assess the effectiveness of a second laparoscopic ovarian diathermy (LOD) in women with polycystic ovary syndrome (PCOS). DESIGN: Retrospective study. SETTING: Tertiary infertility clinic. PATIENT(S): Twenty anovulatory infertile women with PCOS who underwent LOD 1-6 years previously. Twelve subjects had previously responded positively to the first LOD, but the anovulatory status recurred (Group I), whereas eight subjects had not responded at all (Group II). INTERVENTION(S): Laparoscopic ovarian diathermy. MAIN OUTCOME MEASURE(S): Rates of ovulation, pregnancy and resumption of menstrual regularity, and biochemical changes. RESULT(S): An overall ovulation rate of 12 out of 20 women (60%) and a pregnancy rate of 10 out of 19 (53%) were achieved after the second LOD. In women (n = 12) who previously responded positively to the first LOD (LOD-sensitive), the ovulation and pregnancy rates were 10 out of 12 (83%) and 8 out of 12 women (67%), respectively, which were significantly (P<.05) higher than 2 out of 8 (25%) and 2 out of 7 (29%) of the previous nonresponders (LOD-resistant). Statistically significant hormonal changes including reduction of luteinizing hormone (LH), testosterone (T), and free androgen index (FAI) after the repeat LOD were only observed in the LOD-sensitive group. CONCLUSION(S): Repeat LOD is highly effective in women who previously responded to the first procedure.     

Immunocytochemical investigation of neurovascular relationships in human tooth pulp

This study sought to explore the anatomical relationships between peptidergic nerves and blood vessels within human primary and permanent teeth. Extracted primary and permanent molars (n = 120) were split longitudinally, placed in Zamboni's fixative and the coronal pulps were processed for indirect immunofluorescence. Ten-micrometre-thick serial frozen pulp sections were triple-labelled using combinations of the following antisera: (1) protein gene-product 9.5 (PGP 9.5), a general neuronal marker; (2) one of the neuropeptides, calcitonin gene-related peptide (CGRP), substance P (SP), vasoactive intestinal polypeptide (VIP) or neuropeptide Y (NPY); and (iii) the lectin Ulex europeus, a label for vascular endothelium. The mid-coronal pulp region was examined, using fluorescence microscopy, to determine the proportion of blood vessels showing a positive innervation (recorded when PGP 9.5-labelled nerves appeared to intersect the vessel wall). In addition, the percentage of these vascular-related nerves expressing each of the above neuropeptides was recorded. Overall, 20% of pulpal blood vessels appeared to have a positive innervation. In the main these were thick-walled arterioles. Capillaries, venules and lymphatics were mostly devoid of an associated innervation. Ninety-two per cent of vascular-related nerves expressed CGRP, 87% expressed SP, 15% expressed VIP and 80% expressed NPY. There were no significant differences in overall innervation or peptide-related innervation between primary and permanent teeth (P < 0.05, ANOVA), indicating that pulpal blood flow is likely to be subject to similar neurological control mechanisms in both dentitions.     

Embryonic stem cells: advances toward potential therapeutic use

Established lines of human pluripotent stem cells provide a convenient tool for investigating cell differentiation in a way that is pertinent to human embryonic development, providing insights into the causes of birth defects and diseases such as cancer that involve aberrant cell proliferation and differentiation. In principle, human pluripotent stem cells, including embryonic stem and embryonic germ cells, are capable of differentiating into all of the cell types that are present in the adult human. They therefore have the potential to provide a source of tissues for replacement in diseases in which native cell types are inactivated or destroyed. Intense media and public interest has surrounded the announcement of human pluripotent stem cells derivation, focusing on the ethical implications of embryo-related work and on the prospects of an unlimited source of tissues for transplantation-based treatments. Recent studies have focused on identifying method for culture of these cells and inducing their differentiation into specific cell types.     

A multidisciplinary online educational database

BACKGROUND: Before the introduction of the Trent online educational database there was no central resource that potential purchasers could use to access information on clinically relevant courses and programmes of professional development. The online database enables cancer centres, cancer units, and education and training confederations to identify current provision of cancer education and training courses in Trent. CONCLUSION: The electronic online evaluation identified that the majority of users felt the information they found was relevant and that they would recommend the database to colleagues. They also felt that it was easy to find the information they required and the information was presented in a useful and appropriate format.     

Development of a plasma-polymerized surface suitable for the transplantation of keratinocyte-melanocyte cocultures for patients with vitiligo

The purpose of this study was to develop a convenient methodology for the coculture of autologous melanocytes and keratinocytes for grafting of patients with vitiligo. While grafting of pure melanocytes may achieve repigmentation, the inclusion of keratinocytes ensures rapid reepithelialization. Previously we have used confluent sheets of keratinocytes (with melanocytes present) to transfer cells. However, we found that as the keratinocyte density increased, melanocyte number and function were downregulated. Accordingly in this study we explored combinations of three culture surfaces and three media, seeking to achieve subconfluent culture of primary keratinocytes with a reasonable density of melanocytes, using cells immediately after isolation from skin. For this in vitro study, the surfaces studied were uncoated glass coverslips, and glass coverslips coated with collagen I or a nitrogen-containing plasma polymer. The results show that both the substrate surface and the medium composition influence the proliferation and survival of melanocytes. Keratinocytes and melanocytes could be successfully cocultured on a chemically defined plasma polymer substrate using a serum-free medium.     

Plasma-polymerized surfaces for culture of human keratinocytes and transfer of cells to an in vitro wound-bed model

The aim of this study was to develop plasma-polymerized surfaces suitable for the attachment and culture of human keratinocytes and that would allow their subsequent transfer to a wound-bed model. Keratinocyte attachment has been assessed on a carrier polymer, either untreated or treated with a hydrocarbon plasma polymer, collagen I, or carboxylic-acid-containing plasma copolymers. Cell attachment was poor on the "bare" carrier polymer and hydrocarbon plasma polymer (PP) surfaces. Cell attachment was good and comparable on collagen I-coated carrier polymer and carrier polymer plasma coated with carboxylic acid functionalities. After 24 h of cell culture, surfaces were inverted so that cells were adjacent to a de-epidermalized dermis (DED) for 4 days. After 4 days in contact with DED, the surfaces were removed and the level of residual cells and cells transferred to DED were assessed using a cell viability assay. Cell transfer from the collagen I-coated surface was on the order of 90%. Transfer from the carrier polymer surface and the hydrocarbon-coated surface was poor while cells cultured on acid-containing surfaces showed high levels of transfer. Cell transfer was greatest from those surfaces containing the highest level of acid functionality (ca. 21%). Cell transfer was not significantly affected by the choice of carrier polymer material although some sample-to-sample variation was seen. To determine that plasma-polymerized surfaces could be used clinically, selected samples were sterilized with ethylene oxide. Subsequent analysis and cell culture indicated that the surface chemistry and cell-transfer capability of these plasma-polymerized surfaces were unaffected by the sterilization procedure. Plasma-polymerized carboxylic-acid-containing surfaces show great promise in the field of wound healing, encouraging keratinocyte attachment and permitting keratinocyte transfer to a wound bed.