Infrared spectroscopy with multivariate analysis to interrogate endometrial tissue: a novel and objective diagnostic approach.

BACKGROUND: Endometrial cancer is the most common gynaecological malignancy in the United Kingdom. Diagnosis currently involves subjective expert interpretation of highly processed tissue, primarily using microscopy. Previous work has shown that infrared (IR) spectroscopy can be used to distinguish between benign and malignant cells in a variety of tissue types. METHODS: Tissue was obtained from 76 patients undergoing hysterectomy, 36 had endometrial cancer. Slivers of endometrial tissue (tumour and tumour-adjacent tissue if present) were dissected and placed in fixative solution. Before analysis, tissues were thinly sliced, washed, mounted on low-E slides and desiccated; 10 IR spectra were obtained per slice by attenuated total reflection Fourier-transform IR (ATR-FTIR) spectroscopy. Derived data was subjected to principal component analysis followed by linear discriminant analysis. Post-spectroscopy analyses, tissue sections were haematoxylin and eosin-stained to provide histological verification. RESULTS: Using this approach, it is possible to distinguish benign from malignant endometrial tissue, and various subtypes of both. Cluster vector plots of benign (verified post-spectroscopy to be free of identifiable pathology) vs malignant tissue indicate the importance of the lipid and secondary protein structure (Amide I and Amide II) regions of the spectrum. CONCLUSION: These findings point towards the possibility of a simple objective test for endometrial cancer using ATR-FTIR spectroscopy. This would facilitate earlier diagnosis and so reduce the morbidity and mortality associated with this disease.

A randomised controlled trial comparing two different local anaesthetic injection techniques prior to LLETZ.

The aim of this study was to compare two different anaesthetic injection techniques and assess whether one was less painful than the other when used prior to LLETZ. A total of 60 women undergoing LLETZ were randomised into two groups. The control group received local anaesthesia by deep injection into the substance of the cervix. The study group received an equivalent amount of local anaesthetic but this was injected superficially prior to deep injection into the cervix. Pain was assessed using a visual analogue scale. Women in the study arm experienced less pain than controls during injection of local anaesthetic.

Indels and imperfect duplication have driven the evolution of human Complement Receptor 1 (CR1) and CR1-like from their precursor CR1 alpha: importance of functional sets.

This study examines the effects of duplication and insertions-deletions (indels) by comparing human complement receptor 1 (CR1) and human CR1-like (CR1L) with syntenic genes from four other vertebrates (chimpanzee, baboon, rat, and mouse). By phylogenetic analysis, the domains of these genes can be classified into 10 distinct subfamilies (a, b, c, d, e, f, g(-like), h, j, and k), which have been largely conserved throughout vertebrate and invertebrate evolution. In spite of many complex and diverse duplications and indels, the subfamily order of domains (a, j, e, f, b, k, d, g(-like)) has been maintained. The number of domain sets has increased progressively, thereby expanding the functional repertoire.

Genomic analysis reveals a duplication of eight rather than seven short consensus repeats in primate CR1 and CR1L: evidence for an additional set shared between CR1 and CR2.

We report the discovery of previously unrecognised short consensus repeats (SCRs) within human and chimpanzee CR1 and CR1L. Analysis of available genomic, protein and expression databases suggests that these are actually genomic remnants of SCRs previously reported in other complement control proteins (CCPs). Comparison with the nucleotide motifs of the 11 defined subfamilies of SCRs justifies the designation g-like because of the close similarity to the g subfamily found in CR2 and MCP. To date, we have identified five such SCRs in human and chimpanzee CR1, one in human and chimpanzee CR1L, but none in either rat or mouse Crry in keeping with the number of internal duplications of the long homologous repeat (LHR) found in CR1 and CR1L. In fact, at the genomic level, the ancestral LHR must have contained eight SCRs rather than seven as previously thought. Since g-like SCRs are found immediately downstream of d SCRs, we suggest that there must have been a functional dg set which has been retained by CR2 and MCP but which is degenerate in CR1 or CR1L. Interestingly, dg is also present in the CR2 component of mouse CR1. The degeneration of the g SCR must have occurred prior to the formation of primate CR1L and prior to the duplication events which resulted in primate CR1. In this context, the apparent conservation of g-like SCRs may be surprising and may suggest the existence of mechanisms unrelated to protein coding. These results provide examples of the many processes which have contributed to the evolution of the extensive repertoire of CCPs.

Immunogenetic factors in HPV-associated cervical cancer: influence on disease progression.

No HLA allele or specificity was significantly different in frequency between a group of 150 cervical cancer patients from north-west England and controls (corrected P values). HLA-DRB1*1501/DQB1*0602 was non-significantly increased, particularly among patients with HPV16-positive tumours. HLA-B7-positive patients had a significantly poorer clinical outcome than HLA-B7-negative patients. A significant component of the genotypic effect is down-regulation of HLA-B7 expression by the tumour cells.

Loss of HLA class I expression in prostate cancer: implications for immunotherapy.

OBJECTIVES: There is currently no reliable predictor of the metastatic potential of apparently localized prostate cancer in an individual patient or satisfactory treatment for patients with advanced disease. One of the factors that may influence tumor progression is the cellular arm of the immune response, and central to this is the human leukocyte antigen (HLA) system, which acts to restrict T-cell recognition of potential tumor antigens. It has been reported in some cancers that down regulation of HLA class I expression by the tumor cells is associated with poor prognosis. In this report, HLA class I and II expression have been investigated in both benign and malignant prostate disease, first to define the extent of altered HLA expression and second to assess whether HLA expression may be related to disease progression. METHODS: HLA expression was assessed by immunohistochemistry utilizing a set of monoclonal antibodies that recognize both monomorphic determinants and the commoner HLA class I allelic products. RESULTS: In contrast to the normal HLA class I expression of the benign tissue, complete loss of HLA class I expression occurred in 34% of primary prostate cancers and 80% of lymph node metastases. When individual allelic expression was assessed, the minimum estimate of down regulation was 85% in the primary prostate cancers and 100% of the metastases. CONCLUSIONS: This investigation has demonstrated a higher rate of HLA class I loss than has been reported in other tumors and would suggest that the immune system may have an important role in the progression of prostate cancer, as well as having implications for the design and success of immunotherapy regimens in advanced disease.

Frequency of down-regulation of individual HLA-A and -B alleles in cervical carcinomas in relation to TAP-1 expression.

The development of cervical carcinoma is strongly associated with specific types of human papillomaviruses (HPVs). A role for cellular immunity in cervical disease is supported by the increased occurrence of HPV-associated lesions in immunosuppressed individuals. Upon viral infection or malignant transformation, ensuing alterations in gene expression result in the generation of novel sets of peptides which can form complexes with specific HLA class I heavy chains and beta 2-microglobulin. These are then expressed at the cell surface as potential targets for specific T cells. In this study of 100 carcinomas HLA-A and -B class I expression by the tumour cells was down-regulated at one or more alleles in at least 73% of cervical carcinomas. Interference with the transporter associated with antigen presentation (TAP), which translocates cytosolic peptides from endogenously synthesised proteins (e.g. viral) into the lumen of the endoplasmic reticulum was found in 38% of the HLA class I down-regulated tumours. Loss of expression for common HLA class I alleles ranged from 36% to 71%, and such changes might be expected to influence specific immunogenic peptide presentation and consequent immune recognition. These results underline the importance of single as well as multiple allelic loss in cervical neoplasia and have important implications for attempts to intervene immunologically in cervical cancer.

The association of an HPV16 oncogene variant with HLA-B7 has implications for vaccine design in cervical cancer.

HLA-restricted cytotoxic T-lymphocyte (CTL) recognition of human papillomavirus (HPV) oncogene products may be important in the control of the HPV infections associated with the development of cervical cancer. We have identified, in HLA-B7 individuals, a consistent variation in the HPV16 E6 oncoprotein sequence, which alters an HLA-B7 peptide binding epitope in a way likely to influence immune recognition by CTLs. These results illustrate a biologically relevant mechanism for escape from immune surveillance of HPV16 in HLA-B7 individuals. Thus, both HLA type and HPV16 strain variation need to be considered in the screening of at-risk individuals and for the rational design of anti-HPV vaccines.

Loss of transporter protein, encoded by the TAP-1 gene, is highly correlated with loss of HLA expression in cervical carcinomas.

Malignant tumor cells can escape CD8+ cytotoxic T cell killing by downregulating class I major histocompatibility complex (MHC) expression. Stable class I MHC surface expression requires loading of the heavy chain/light chain dimer with antigenic peptide, which is delivered to class I MHC molecules in the endoplasmic reticulum by the presumed peptide transporter, encoded by the transporter associated with antigen presentation (TAP) 1 and 2 genes. We have investigated whether loss of class I MHC expression frequently observed in different cancers could result from interference with TAP function. A polyclonal antiserum, raised against a bacterial glutathione S-transferase/human TAP-1 fusion protein, was used for the immunohistochemical analysis of TAP-1 expression in 76 cervical carcinomas. Results showed loss of TAP-1 expression in neoplastic cells in 37 out of 76 carcinomas. Immunohistochemical double staining procedures in combination with HLA-specific antibodies revealed congruent loss at the single cell level of TAP-1 and HLA-A/B expression in 28 out of 37 carcinomas. The remaining samples expressed HLA(-A) in the absence of TAP-1 (n = 6) or showed loss of HLA(-A/B) while TAP-1 was expressed (n = 3). These data strongly indicate that inhibition of peptide transport by downregulation of TAP-1 is a potential strategy of malignant cells to evade immune surveillance.

A retrospective analysis of 94 patients with CIN and false negative cervical smears taken at colposcopy.

This report investigates the reasons for false negative cervical cytology in 94 out of 630 patients (15%) in whom cervical intraepithelial neoplasia (CIN) was diagnosed on colposcopically directed biopsy. Cervical smears were taken immediately before biopsy and the cases with false negative cytology were compared with those whose cytology was abnormal. Patients with false negative cytology were more likely to have been younger (P < 0.01), to have had fewer pregnancies (P < 0.001), to have had a less severe grade of dyskaryosis on their referral smear (P < 0.001), to have had no endocervical cells on the smear (P < 0.05), to have had a less severe grade of CIN on biopsy (P < 0.001), to have had no punctation visible at colposcopy (P < 0.01), and to have had no mosaic pattern seen at colposcopy (P < 0.05). We found no effect attributable to the patient's menstrual history, the interval between referral smear and colposcopy clinic visit, the smear taker or the type of spatula used to take the smear.

Cultured fetal tectal tissue grafted to the midbrain of newborn rats: morphology of grafts and innervation by host retinal and cortical axons.

It has been shown previously that tectal tissue obtained from young embryos can be successfully transplanted to the neonatal rat brain. In the present study, tecta from E15 rat embryos were maintained as free-floating explants for 3-14 days in vitro (DIV) before being transplanted to the midbrain of newborn rats. We wished to determine how explant culture affected (i) graft survival, (ii) the subsequent morphological and histochemical development of tectal grafts and (iii) the specificity with which host retinal and cortical axons grew into and innervated the graft neuropil. Grafts were examined 6-40 weeks posttransplantation. Host retinal input was assessed by injecting the host eyes with either [3H]proline, horseradish peroxidase (HRP) or wheat-germ agglutinin conjugated HRP. The host cortical projection was examined using anterograde degeneration techniques. Frozen tissue sections were also stained for Nissl, neurofibrils or reacted for acetylcholinesterase (AChE). All 3 DIV and 7 DIV explants survived transplantation and many grew considerably in size within the host brain. 14 DIV grafts were smaller and were found in only 50% of host brains. The cellular organization, fibre architecture and pattern of AChE staining in cultured grafts was similar to that found in non-cultured tectal transplants. Furthermore, host retina and cortex projected into the grafts in a manner similar to their innervation of non-cultured tectal tissue.(ABSTRACT TRUNCATED AT 250 WORDS)

The treatment of bimaxillary protrusion. A cephalometric consideration of changes in the inter-incisal angle and soft tissue profile.

Thirty bimaxillary protrusion cases, who had four first premolars extracted were compared with thirty Class I malocclusion cases who, likewise, had similar extractions and treatment. The changes, both in incisor relationships and the concurrent soft tissue profiles are documented. During treatment, the inter-incisal angle was increased by 20 degrees and relapsed by 4 degrees post-treatment. The protuberance of the soft tissue profile was significantly reduced by 4 degrees concurrently (Holdaway Angle). It is concluded that worthwhile treatment objectives can be achieved in these cases.

Bimaxillary protrusion in the Caucasian: a cephalometric study of the morphological features.

The skeletal morphology of bimaxillary dental protrusion has been investigated in a comparative cephalometric study. Because this is considered a subset of Class I malocclusions, the null hypothesis is that there should be no significant skeletal differences between this group and a Class I control group. There were 30 Caucasians in each group with no bias for age and sex differences between them. Eighteen radiographic landmarks were identified from which 33 skeletal, dental and soft tissue parameters were computed. The bimaxillary group had an average interincisal angle of 115 degrees versus the controls 135 degrees, and showed the following morphological features which persisted over a 5-year growth period: A shorter posterior cranial base. A longer and more prognathic maxilla. Similar mandibular dimensions and prognathism. A mild Class II skeletal pattern. A smaller upper and posterior face height. Diverging facial planes. A procumbent soft tissue profile with a low lip line. These findings indicate that there is a distinctive difference between the underlying skeletal patterns found in the two groups.

The holographic storage of study models.

The storage and recall of dental study models pose major problems for orthodontists. This problem may be solved by taking holographic films of study models. A method is described for their fabrication and some preliminary measurements are reported. The image produced by this method has a measurable three-dimensionality which can be produced at a very reasonable cost.

Effects of glycopyrrolate and cimetidine on gastric volume and acidity in patients awaiting surgery.

Glycopyrrolate or cimetidine was administered before operation to patients undergoing elective surgery. After the induction of anaesthesia, the stomach contents were retrieved and the volume and pH measured. Neither drug diminished the volume of gastric contents compared with control. Glycopyrrolate produced little diminution in hydrogen ion concentration. Cimetidine caused a marked increase in pH with a mean [h+] 3.2 x 10(-3) g litre-1 compared with 1.4 x 10(-2) g litre-1 in the controls and 1.1 x 10(-2) g litre-1 in the glycopyrrolate group. Seventy-seven per cent of the patients receiving cimetidine had a pH greater than 2.5.