O-9Uncommon subtypes of borderline nuclear changes: cytological features and predictive value

Introduction:  To establish the significance of cytological features which could predict clinically significant endometrial pathology, and therefore guide reporting practice in cervical samples.
Methods:  A retrospective review of SurePath liquid-based cytology (LBC) cervical samples between 2002 and 2006, obtained at screening and colposcopy. These smears contained normal endometrial cells present at inappropriate times of the menstrual cycle, endometrial cells with atypia (borderline change) and with features suspicious / diagnostic of endometrial carcinoma (glandular neoplasia). False negative and false positive cases detected on subsequent histology were also included. The control group comprised negative samples and a few abnormal smears. All smears were randomly assigned and blinded to menopausal status, age, use of oral contraceptive pill and hormone replacement therapy and presence of intrauterine device. Each smear was reviewed for 16 cytologic criteria and a cytological diagnosis was given for each.
Results:  A total of 219 smears were available for review; 137 were negative, out of which 85 contained normal endometrial cells, 41 contained endometrial cells with atypia, 10 contained endometrial cells with features suggestive of adenocarcinoma and 31 contained endometrial cells with features diagnostic of adenocarcinoma. The feature most associated with benign endometrial cells is top hat with central cell condensation. In contrast, the features associated with malignant endometrial cells are smooth nuclear membrane, pale chromatin, small nucleoli and scalloped borders.
Discussion:  The criteria identified in this study do not definitively define a neoplastic process, but appear to be helpful in individual cases. This study emphasises that endometrial changes should be always interpreted with the relevant clinical information, which would otherwise lead to overdiagnosis in premenopausal women.     

O-12The use of control samples in the PAPANICOLAOU technical external quality assessment scheme

Technical external quality assessment (TEQA) in Wales is based on NHSCSP publication 19, which sets out policy and procedures for the scheme. The purpose of EQA is to sustain and improve the quality of patient care by promoting a high standard of performance. Following the introduction of liquid base cytology (LBC) technical limitations, particularly in assessing counterstaining, have been noted. LBC provides the means to address these limitations - As part of a development plan for TEQA in Wales, a control sample procedure was introduced to the scheme. A pooled control sample was composed, containing residual material from six 'matched' negative samples, re-suspended in collection fluid. Aliquots of this sample were distributed for processing and staining, to the 13 laboratories registered with the scheme. The slides produced were assessed at a scheduled TEQA assessment in accordance with the standard criteria. Initial overall scoring for these control samples produced acceptable levels of staining for 12 of the laboratories - one laboratory produced a marginal score. Repeat distributions have shown maintained or improved results. This method provides a prospective quality assessment tool, which counters the emphasis on slide selection and eliminates potential selection bias, whilst introducing consistency and improving comparability across participant laboratories. The method may also prove helpful in identifying technical inconsistencies such as equipment or handling errors that may occur during sample processing prior to or during staining. The control process, which is now used routinely in the Welsh TEQA scheme; is considered complimentary to, and not a replacement for, the selection process established in NHSCSP #19. However, we feel that this development could be considered as a new initiative in the National TEQA scheme. The control process is applicable to all LBC systems in current use.     

O-2Preliminary results of the ARTISTIC study: a randomised trial in screening to improve cytology

Annually in the UK around 250 000 cervical smears show low-grade abnormalities. Alternative management policies following a low-grade smear are cytological surveillance or referral for colposcopy. Their effectiveness and cost-effectiveness, and the potential for human papillomavirus (HPV) testing to triage women to either management, has been debated. Trial of management of borderline and other low-grade abnormal smears (TOMBOLA) (a large RCT) addresses these uncertainties, considering clinical, psychosocial and economic outcomes. 4439 women aged 20–59, resident in Grampian, Tayside or Nottingham with a low-grade smear were randomised to cytological surveillance (six-monthly smears in primary care) or hospital-based colposcopy. At colposcopy, women with visible abnormality were randomised to immediate treatment or biopsy and recall for treatment if necessary. Recruitment HPV status was assessed using PCR techniques. Women were followed for three years to an exit colposcopy. Cumulative incidence of CIN2 or more severe disease (CIN2+) in the colposcopy arm was 7.9% per year, higher than in cytological surveillance (5.8%; OR = 1.43, 95% CI 1.23–1.67). This difference was less marked for CIN3+ (OR = 1.27, 1.04–1.55), suggesting spontaneous regression of some CIN2, and that initial colposcopy can lead to over-treatment. There was little difference in psychosocial outcomes between arms. In comparison of biopsy and recall versus immediate treatment, there was no difference in cumulative incidence of CIN2+ or psychosocial outcomes. There was over-treatment and increased frequency/duration of bleeding with immediate treatment. There was no compelling economic reason to favour any one management method. Testing for HPV does not appear to be effective in triage. Based on these findings, we make management recommendations for women with low-grade smears.     

O‐4CHARACTERISTICS OF FALSE NEGATIVE LBC SAMPLES

Background: Liquid based cytology (LBC) is the method of choice for the processing of samples in the cervical screening programme in England and Wales. Two LBC systems are in use; ThinPrep® and SurePath®. The manufacturers of SurePath® recommend that slides are screened in two separate directions perpendicular to each other (bidirectional screening) during the initial learning period. There is currently no evidence to evaluate whether this method is superior to screening in one direction only (unidirectional). Objectives: To compare the detection rate of high‐grade dyskaryosis in SurePath® preparations between screeners who used a unidirectional method and screeners who used a bidirectional method, to see if the screening method influenced detection rate. Methods: A questionnaire was sent to 23 screening staff at our Cytology Centre in October 2005, enquiring as to the method they used to screen SurePath® slides, as well as the grade of staff and the number of years experience they had screening cervical cytology slides. The staff were divided into two groups based on whether they screened unidirectionally or bidirectionally. A retrospective computerised search of all the slides screened by the staff for the period September 2004 to August 2005 was carried out, and the high‐grade detection rate at primary screening for the groups were calculated. Further analysis was carried out to determine the statistical significance of any difference between the two groups, as well as to account for potential confounding factors. Results: Initial results suggesting a statistically significant difference between the screening methods were later shown to be confounded by experience. Critical analysis of the data and study design reveal that no firm conclusions can be made on the basis of this work. Conclusion: No firm conclusions on screening method can be drawn due to fundamental limitations and confounding factors in this retrospective work‐based study. Recommendations for further studies are made, controlling for confounding factors.     

Synthesis and properties of diastereoisomers of adenosine 5'-(O-1-thiotriphosphate) and adenosine 5'-(O-2-thiotriphosphate).

The chemical synthesis of adenosine 5'-(O-1-thiotriphosphate) (ATPalphaS) and adenosine 5'-(O-2-thiotriphosphate) (ATPbetaS) is described. Both exist as a pair of diastereomers, A and B. The isomers of ATPalphaS can be distinguished on the basis of their different reaction rates with myokinase as well as nucleoside diphosphate kinase. With both enzymes, isomer A reacts fast whereas isomer B reacts considerably more slowly. Phosphorylation of a mixture of isomers of ADPalphaS with pyruvate or acetate kinase yields ATPalphaS, isomer A, whereas the phosphoryl transfer with creatine or arginine kinase yields isomer B. The isomers of ATPbetaS differ in their reactivity with myosin. Isomer A is readily hydrolyzed, whereas isomer B is not. However, isomer B reacts faster with nucleoside diphosphate kinase and ADP than isomer A. Phosphoryl transfer with pyruvate kinase onto ADPbetaS yields ATPbetaS, isomer A, with acetate kinase, isomer B.     

Biocatalytic conversion of unnatural substrates by recombinant almond R-HNL isoenzyme 5

Hydroxynitrile lyases (HNLs, EC 4.1.2.10, EC 4.1.2.11, EC 4.1.2.37, EC 4.1.2.39) enantioselectively catalyse the reversible addition of HCN to ketones or aldehydes, thereby forming chiral cyanohydrins, which is of special interest for industrial bio-conversions. We cloned the gene for the HNL isoenzyme 5 (PaHNL5) of the almond tree (Prunus amygdalus) and overexpressed it in the methylotrophic yeast Pichia pastoris. This opened new ways for the synthesis of (R)-cyanohydrins. The characterisation of PaHNL5 revealed high activity for the natural substrate and high enantioselectivity. For further improvement of enzyme properties such as higher activity for the conversion of unnatural substrates, a high throughput cultivation and screening system has been created, which allows the employment of P. pastoris as production host for high throughput cultivation and screening of thousands of enzyme variants. The synthesis and cleavage of 2-chlorobenzaldehyde cyanohydrin were used for the demonstration of enzyme activity of recombinant PaHNL5 with a non-natural substrate and for the development of a high throughput screening procedure.     

Identifying important areas for butterfly conservation in Italy

The current study combines the use of niche modelling with a site prioritization method to identify important areas for butterfly conservation in Italy. A novel machine learning method (bagging predictors) was used to predict the distribution of 232 species of butterflies across the Italian Peninsula. The results of the models were used to identify high-value sites with a multispecies prioritization method called zonation. In order to identify important areas for species of conservation concern, we incorporated a species weighting scheme to zonation analyses. We also used the results of the zonation analyses to identify a series of management landscapes on the basis of the similarity in species composition among sites. The basic zonation showed that most important areas for butterfly conservation are located in the Alps, the Appennine, the Apulia region and in the island of Sardinia. The inclusion of a species weighting scheme in the zonation analyses revealed the importance of two new areas located in Southern Italy and emphasized the importance of the Alps for species of conservation concern. The landscape identification procedure selected a series of landscapes, which provide protection to a full range of species ranging from the Alps to Mediterranean areas. Our study shows that the areas selected in our analyses should be given high priority in future conservation plans and monitoring schemes.     

O-8Detection of human telomerase gene (TERC) amplification in cervical neoplasia: a retrospective study of 50 patients with normal smears or mild or moderate dyskaryosis

Circulation E of the North West non gynaecological cytology EQA scheme was split into two with one half receiving conventional glass slides and the other half receiving digital images supplied by the SlidePath company over the web. More of the participants eligible participated in the slide half (43/65) than the digital half (17/41).
Similar results were obtained for seven out of ten of the scoring cases and for both the educational cases. However for one case eight of the digital participants could not obtain the images over the web. In the remaining three scoring cases the digital participants were less good at obtaining the correct answer compared to the slide based participants. The use of this virtual microscopy has shortened the circulation length considerably. The digital participants complained about the speed the digital slides appeared on their computers and whilst most agreed that this is the way forward they are still uncomfortable with their performance being marked on their digital submissions. Delegates will have the opportunity of viewing still images taken for each case from this circulation.     

Retrospective review of 65 atrioesophageal fistulas post atrial fibrillation ablation

Background

Although a rare complication of catheter based ablation for atrial fibrillation (AF), atrioesophageal fistula (AEF) is a serious and fatal event [[1], [2], [3], [4], [5]]. Most reports of AEF are single cases or small case series.

Genetic evidence for a microtubule-destabilizing effect of conventional kinesin and analysis of its consequences for the control of nuclear distribution in Aspergillus nidulans

Conventional kinesin is a microtubule-dependent motor protein believed to be involved in a variety of intracellular transport processes. In filamentous fungi, conventional kinesin has been implicated in different processes, such as vesicle migration, polarized growth, nuclear distribution, mitochondrial movement and vacuole formation. To gain further insights into the functions of this kinesin motor, we identified and characterized the conventional kinesin gene, kinA, of the established model organism Aspergillus nidulans. Disruption of the gene leads to a reduced growth rate and a nuclear positioning defect, resulting in nuclear cluster formation. These clusters are mobile and display a dynamic behaviour. The mutant phenotypes are pronounced at 37 degrees C, but rescued at 25 degrees C. The hyphal growth rate at 25 degrees C was even higher than that of the wild type at the same temperature. In addition, kinesin-deficient strains were less sensitive to the microtubule destabilizing drug benomyl, and disruption of conventional kinesin suppressed the cold sensitivity of an alpha-tubulin mutation (tubA4). These results suggest that conventional kinesin of A. nidulans plays a role in cytoskeletal dynamics, by destabilizing microtubules. This new role of conventional kinesin in microtubule stability could explain the various phenotypes observed in different fungi.     

Synthesis of 5-deoxy-d-xylo-hexose and 5-deoxy-l-arabino-hexose,and their conversion into adenine nucleosides

Anti-Markovnikov hydration of the olefinic bond of 5,6-dideoxy-1,2-O-isopropylidene-3-O-p-tolylsulfonyl-α- d-xylo-hex-5-enofuranose (4) and methyl 5,6-dideoxy-2,3-di-O-p-tolylsulfonyl-α-l-arabino-hex-5-enofuranoside (11) by the addition of iodine trifluoroacetate, followed by hydrogenation in the presence of a Raney nickel catalyst in ethanol containing triethylamine, afforded 5-deoxy-1,2-O-ísopropylidene-3-O-p-tolylsulfonyl-α-d-xylo-hexofuranose (6) and methyl 5-deoxy-2,3-di-O-p-tolylsulfonyl-α-d-arabino-hexofuranoside (14), respectively. 5-deoxy-d-xylo-hexose and 5-deoxy-l-arabino-hexose were prepared from 6 and 14, respectively, by photolytic O-detosylation and acid hydrolysis. Syntheses of 9-(5-deoxy-β-d-xylo-hexofuranosyl)-adenine and 9-(5-deoxy-α-l-arabino-hexofuranosyl)adenine are also described. Application of the sodium naphthalene procedure, for O-detosylation, to 11 is reported in connection with an alternative synthetic route to methyl 5-deoxy-α-l-arabino- hexofuranoside.     

The gene conversion hypothesis of MHC evolution: a review

 Gene conversion is often invoked to explain the evolution of sequence patterns observed in major histocompatibility complex (MHC) genes and their alleles. This is the gene conversion hypothesis of MHC sequence evolution. These observations and their interpretation probably belong in a larger theoretical framework, namely the evolution of systems of resistance to rapidly evolving pathogens. This review looks critically at the evidence in favor of the gene conversion hypothesis in this context. We conclude that the case for the existence of an adaptive mechanism in the MHC favoring gene conversion mutations is not proven.     

A review of thermal-chemical conversion of lignocellulosic biomass in China

Biomass, a renewable, sustainable and carbon dioxide neutral resource, has received widespread attention in the energy market as an alternative to fossil fuels. Thermal-chemical conversion of biomass to produce biofuels is a promising technology with many commercial applications. This paper reviewed the state-of-the-art research and development of thermal-chemical conversion of biomass in China with a special focus on gasification, pyrolysis, and catalytic transformation technologies. The advantages and disadvantages, potential of future applications, and challenges related to these technologies are discussed. Conclusively, these transformation technologies for the second-generation biofuels with using non-edible lignocellulosic biomass as feedstocks show prosperous perspective for commercial applications in near future.     

Dream intensity profile as an indicator of the hysterical tendencies to dissociation and conversion.

The present study explored the clinical utility of the Dream Intensity Scale (DIS) by evaluating the degree to which the components of dream intensity can serve as indicators of the tendencies to hysterical dissociation and conversion, taking neuroticism and psychological boundaries into consideration. The discriminant models constituted by the DIS scales alone differentiated participants who manifested clinically significant dissociative or conversion symptoms from the remaining sample, with the accuracy rate ranging between 67.2% and 71.5%. The discriminative power of the DIS was so impressive that the inclusion in the models of neuroticism, psychological boundaries, and dissociative or conversion levels—the key factors associated with hysteria—raised the correct classification rate by less than 9%. The relationships between neuroticism, boundary thinness, dissociative features, epileptic-like symptoms, and subjective dream intensity were discussed. (PsycINFO Database Record (c) 2010 APA, all rights reserved)