bluuebunny/crossref_metadata_2025 huggingface dataset:
Long QT 1 Mutation KCNQ1A344VIncreases Local Anesthetic Sensitivity of the Slowly Activating Delayed Rectifier Potassium Current
Authors: Siebrands Cornelia C. , Binder Stephan , Eckhoff Ulrike , Schmitt Nicole , Friederich Patrick
Published: August 2006

Background

Anesthesia in patients with long QT syndrome (LQTS) is a matter of concern. Congenital LQTS is most frequently caused by mutations in KCNQ1 (Kv7.1), whereas drug-induced LQTS is a consequence of HERG (human ether-a-go-go-related gene) channel inhibition. The aim of this study was to investigate whether the LQT1 mutation A344V in the S6 region of KCNQ1, at a position corresponding to the local anesthetic binding site in HERG, may render drug insensitive KCNQ1 channels into a toxicologically relevant target of these pharmacologic agents. This may suggest that LQTS constitutes not only a nonspecific but also a specific pharmacogenetic risk factor for anesthesia.

Methods

The authors examined electrophysiologic and pharmacologic properties of wild-type and mutant KCNQ1 channels. The effects of bupivacaine, ropivacaine, and mepivacaine were investigated using two-electrode voltage clamp and whole cell patch clamp recordings.

Results

The mutation A344V induced voltage-dependent inactivation in homomeric KCNQ1 channels and shifted the voltage dependence of KCNQ1/KCNE1 channel activation by +30 mV. The mutation furthermore increased the sensitivity of KCNQ1/KCNE1 channels for bupivacaine 22-fold (KCNQ1wt/KCNE1: IC50 = 2,431 +/- 582 microM, n = 20; KCNQ1A344V/KCNE1: IC50 = 110 +/- 9 microM, n = 24). Pharmacologic effects of the mutant channels were dominant when mutant and wild-type channels were coexpressed. Simulation of cardiac action potentials with the Luo-Rudy model yielded a prolongation of the cardiac action potential duration and induction of early afterdepolarizations by the mutation A344V that were aggravated by local anesthetic intoxication.

Conclusions

The results indicate that certain forms of the LQTS may constitute a specific pharmacogenetic risk factor for regional anesthesia.
Anesthetic Preconditioning Enhances Ca2+Handling and Mechanical and Metabolic Function Elicited by Na+–Ca2+Exchange Inhibition in Isolated Hearts
Authors: An Jianzhong , Rhodes Samhita S. , Jiang Ming Tao , Bosnjak Zeljko J. , Tian Ming , Stowe David F.
Published: August 2006

Background

Anesthetic preconditioning (APC) is well known to protect against myocardial ischemia-reperfusion injury. Studies also show the benefit of Na+-Ca2+ exchange inhibition on ischemia-reperfusion injury. The authors tested whether APC plus Na+-Ca2+ exchange inhibitors given just on reperfusion affords additive protection in intact hearts.

Methods

Cytosolic [Ca2+] was measured by fluorescence at the left ventricular wall of guinea pig isolated hearts using indo-1 dye. Sarcoplasmic reticular Ca2+-cycling proteins, i.e., Ca2+ release channel (ryanodine receptor [RyR2]), sarcoplasmic reticular Ca2+-pump adenosine triphosphatase (SERCA2a), and phospholamban were measured by Western blots. Hearts were assigned to seven groups (n = 8 each): (1) time control; (2) ischemia; (3, 4) 10 microM Na+-Ca2+ exchange inhibitor KB-R7943 (KBR) or 1 microM SEA0400 (SEA), given during the first 10 min of reperfusion; (5) APC initiated by sevoflurane (2.2%, 0.41 +/- 0.03 mm) given for 15 min and washed out for 15 min before ischemia-reperfusion; (6, 7) APC plus KBR or SEA.

Results

The authors found that APC reduced the increase in systolic [Ca2+], whereas KBR and SEA both reduced the increase in diastolic [Ca2+] on reperfusion. Each intervention improved recovery of left ventricular function. Moreover, APC plus KBR or SEA afforded better functional recovery than APC, KBR, or SEA alone (P < 0.05). Ischemia-reperfusion-induced degradation of major sarcoplasmic reticular Ca2+-cycling proteins was attenuated by APC, but not by KBR or SEA.

Conclusions

APC plus Na+-Ca2+ exchange inhibition exerts additive protection in part by reducing systolic and diastolic Ca2+ overload, respectively, during ischemia-reperfusion. Less degradation of sarcoplasmic reticular Ca2+-cycling proteins may also contribute to cardiac protection.
Ontogeny of gonadotropin‐releasing hormone (GnRH) neurons in hybrid striped bass Morone sp.: whole‐mount in situ hybridization analysis
Authors: Yashuvi Y. , Klenke U. , Palevitch O. , Abraham E. , Zohar Y. , Gothilf Y.
Published: August 2006

The ontogeny of gonadotropin‐releasing hormone (GnRH) mRNA‐producing neurons was studied in developing hybrid striped bass (white bass Morone chrysops female × striped bass Morone saxatilis male), 1–55 days post‐fertilization (dpf), by whole‐mount in situ hybridization. Neurons that produce salmon (s) GnRH were first detected at 32 h post‐fertilization in the olfactory placodes. These neurons migrated posteriorly during development and reached their final position at the olfactory bulbs‐telencephalon boundary, possibly the terminal nerve ganglion (TNg) by 11 dpf. First signal of chicken (c) GnRH‐II neurons appeared in the midbrain 2 dpf and remained there throughout development. A signal of seabream (sb) GnRH mRNA was first detected at 21 dpf in the preoptic area (POA) and as a bilateral continuum along the ventral telencephalon at 32–55 dpf. The expression of all three forms of GnRH increased throughout development. These results suggest that cGnRH‐II neurons originate in the mid‐brain, and that sGnRH neurons originate in the olfactory placodes and migrate caudally to the TNg. Neurons that express sbGnRH seem to originate at the preoptic area and then to migrate anteriorly along the ventral telencephalon. An olfactory placodal origin of these neurons, however, cannot be ruled out.
Evidence of reproductive isolation among closely related sympatric species of Serrasalmus (Ostariophysii, Characidae) from the Upper Madeira River, Amazon, Bolivia
Authors: Hubert N. , Duponchelle F. , Nuñez J. , Rivera R. , Renno J.‐F.
Published: August 2006

The delimitation of the Serrasalmus species (Characiformes, Serrasalminae) from the Bolivian Amazon (Amazonas, Madeira) was examined using intron length polymorphism assessed by EPIC‐PCR. The six pairs of primers provided 16 polymorphic loci across the species of the region and the allelic diversity ranged between two and 10 alleles per locus. For each locus, the lack of departure from Hardy‐Weinberg expectations in a large number of the populations analysed and the homogenous distribution of linkage disequilibrium between paralogous loci and loci belonging to different intronic systems provided strong lines of evidence that the 16 amplified loci constituted independent neutral markers. Furthermore, allelic diversity was size‐dependent, thereby indicating that insertion‐deletions occurred frequently but randomly in introns, and that intron length polymorphism was a valid marker for investigating the systematics of piranhas. EPIC‐PCR demonstrated that eight of the nine nominal species of piranha of the Upper Madeira were reproductively isolated and identified a new species that differed from its closest morphological and genetic relatives by seven diagnostic or semi‐diagnostic loci. By contrast, no diagnostic or semi‐diagnostic locus was found between S. spilopleura and S. eigenmanni, nor were their allelic frequencies different, thereby questioning the validity of their biological species status, at least in the Upper Madeira. This study, which was one of the first applications of EPIC‐PCR to a large‐scale molecular systematic purpose, demonstrates that it is a rapid, reliable and cost‐effective tool for elucidating issues pertaining to fish systematics.
Perceived Adverse Consequences of Quality Threatening Behaviour in Audit Firms
Authors: Pierce Bernard , Sweeney Breda
Published: August 2006

Research in many countries has consistently shown that audit seniors engage in widespread quality threatening behaviours (QTB). Regarding the consequences of these behaviours, only audit partners’ perceptions of possible consequences have been examined previously. The objective of this study is to examine audit seniors’ perceptions of the consequences of QTB for internal and external groups using semi‐structured interviews with 25 audit seniors in Ireland. Findings suggest that detection of the behaviours is an important factor in determining the consequences for individual auditors, audit firms and the profession but not for the wider business community. In general, interviewees perceived low risk of detection and showed little consideration of the ethical implications of their actions. No communication was perceived to exist between the firms and audit seniors on prohibition of the behaviours or sanctions against those found guilty of engaging in the behaviours. Although the literature highlights the importance of personnel controls in complex environments such as audit firms, these findings suggest that the effectiveness of personnel controls is undermined by a perception that the likelihood of serious consequences is remote and also by weaknesses in the ethical training of auditors. The use of the term dysfunctional to describe QTB is open to question given the absence of a perceived link between the behaviours and consequences for organisational effectiveness found in this study.
Internal Auditors’ Assessment of their Contribution to Financial Statement Audits: The Relation with Audit Committee and Internal Audit Function Characteristics
Authors: Zain Mazlina Mat , Subramaniam Nava , Stewart Jenny
Published: August 2006

This paper examines the relation between audit committee characteristics, internal audit function characteristics and internal auditors’ assessment of their contribution to financial statement audits. Using survey data from chief internal auditors of 76 Malaysian publicly‐listed firms, we provide evidence of a positive relationship between internal auditors’ assessment of their contribution to financial statement audits and three audit committee characteristics: the proportion of independent audit committee members, their knowledge and experience of accounting and auditing, and the extent of audit committee review of internal audit programmes, budget and coordination proposals. Further, a positive relationship is found between internal auditors’ evaluation of their contribution to the financial statement audit and internal audit function characteristics including size, prior experience of staff in auditing, time availability and the closeness of the function’s relationship with the external auditor. The results indicate that more effective audit committees and well‐resourced internal audit units tend to be positively associated with the internal auditors’ assessment of their contribution to the external audit.
Small interfering RNA targeting epidermal growth factor receptor enhances chemosensitivity to cisplatin, 5‐fluorouracil and docetaxel in head and neck squamous cell carcinoma
Authors: Nozawa Hiroshi , Tadakuma Takushi , Ono Takeshi , Sato Masaki , Hiroi Sadayuki , Masumoto Kazuma , Sato Yasunori
Published: August 2006

Overexpression of epidermal growth factor receptor (EGFR) has been found in various epithelial malignancies, including head and neck squamous cell carcinoma (HNSCC), and is associated with increased tumor growth, metastasis, resistance to chemotherapeutic agents and poor prognosis. As such, EGFR is a potential target for antitumor therapy and several EGFR inhibitors have been investigated in preclinical or clinical settings. In the present study, we used small interfering RNA (siRNA) to downregulate EGFR expression while evaluating the effect of EGFR siRNA on cell proliferation, and the combined effects with cisplatin, 5‐fluorouracil (5‐FU) and docetaxel in HNSCC. Furthermore, HNSCC xenografts were treated with EGFR siRNA alone or in combination with cisplatin, and tumor growth was examined. EGFR expression, proliferation, angiogenesis and apoptosis index were evaluated by immunohistochemistry. The results showed that EGFR siRNA efficiently downregulated EGFR expression and inhibited cell growth of HNSCC. Treatment with EGFR siRNA in combination with cisplatin, 5‐FU and docetaxel enhanced chemosensitivity with a significant increase in apoptosis. EGFR siRNA delivered by atelocollagen enhanced the antitumor effect of cisplatin in the HNSCC xenograft model. These cumulative results suggest that EGFR siRNA combined with cisplatin, 5‐FU and docetaxel may be a feasible strategy to enhance the effects of chemotherapy in patients with HNSCC. (Cancer Sci 2006; 97: 1115–1124)
Role of phosphatidylinositol‐3 kinase/Akt pathway in bladder cancer cell apoptosis induced by tumor necrosis factor‐related apoptosis‐inducing ligand
Authors: Oka Natsuo , Tanimoto Shuji , Taue Ryuichi , Nakatsuji Hiroyoshi , Kishimoto Tomoteru , Izaki Hirofumi , Fukumori Tomoharu , Takahashi Masayuki , Nishitani Masaaki , Kanayama Hiro‐omi
Published: August 2006

TRAIL/Apo2L is a pro‐apoptotic cytokine that is capable of inducing apoptosis in a wide variety of cancer cells but not in normal cells. Among various molecular strategies by which cancer cells evade apoptosis, PI3K/Akt signaling represents a dominant survival pathway. In this report, we investigated the role of PI3K/Akt pathway in TRAIL‐induced apoptotic death in human bladder cancer cells. We observed that RT4 cells had very low level of constitutively active Akt and were sensitive to TRAIL, whereas UM‐UC‐3 and T24 cells had higher levels of constitutively active Akt and were resistant to TRAIL. Downregulation of constitutively active Akt by PI3K inhibitors, wortmannin and LY294002, reversed cellular resistance to TRAIL. However, transfecting constitutively active Akt into RT4 cells increased Akt activity and inhibited TRAIL‐induced apoptosis. These results suggest that elevated Akt activity protects UM‐UC‐3 and T24 cells from TRAIL‐induced apoptosis, and the PI3K/Akt signaling might inhibit apoptotic signals. Thus, the modulation of Akt activity by combining pharmacological drugs or genetic alterations of the Akt expression could induce cellular responsiveness to TRAIL and PI3K/Akt signaling pathway could serve as a novel target for therapeutic intervention in bladder cancer. (Cancer Sci 2006; 97: 1093–1098)
Changes in tropical forest vegetation along an altitudinal gradient in the Udzungwa Mountains National Park, Tanzania
Authors: Lovett Jon C. , Marshall Andrew R. , Carr Jeff
Published: August 2006

This study describes changes in woody vegetation in the Mwanihana forest, Udzungwa Mountains National Park, Tanzania, over an altitude range of 470–1700 m. Two methods, fixed‐ and variable‐area plots, are compared to elucidate altitudinal variation in tropical forest structure, diversity and community composition. Six 25 m × 100 m fixed area plots recorded a total of 2143 woody stems of ≥3 cm d.b.h. from 204 species. The 78 variable‐area plots recorded the nearest twenty trees of ≥20 cm d.b.h. to an objectively chosen point, giving a total of 1560 stems in 9.1 ha from 156 species. A linear trend of increasing stem density with altitude was seen for variable‐area plots. Species diversity is highest at high elevations. There was no clear zonation of elevational vegetation types. Restricted range taxa occur at all altitudes sampled. The study also revealed some methodological considerations. Bias in sample size and plot area can be tested by employing two sampling methods. Of the two methods used, fixed area plots are preferred as variable area plots are impractical in tangled understorey. Plot size must be controlled for in order to make reliable observations of diversity. Sampling along a continuous or near‐continuous altitudinal gradient with sufficient replication is also important.
Dynamics of male residence and female oestrus during a breeding season of blue monkeys in the Kakamega Forest, Kenya
Authors: Mugatha S. M. , Ogutu J. O. , Cords M. , Maitima J. M.
Published: August 2006

Blue monkeys (Cercopithecus mitis stuhlmanni) are seasonal breeders with a dynamic mating system in which typical one‐male social units are regularly disrupted and replaced by multi‐male ones. The number of males in the group is correlated with the number of oestrous females. We used observations of male presence and female oestrus on individual days during a 6‐month period to assess whether the presence of multiple males in a group stimulates female oestrus or whether oestrous females attract multiple males to the group. We confirmed prior observations with our finding that the number of males in a group was significantly correlated (rs = 0.435, P < 0.0001) with the number of oestrous females across 126 observation days. A transition matrix did not show an obvious relationship between day‐to‐day changes in the numbers of oestrous females and males. However, cross‐correlation analysis provided stronger support for the idea that the number of oestrous females attracts males to a group than for the idea that the influx of strange males stimulates oestrus in female blue monkeys. Autocorrelation analysis showed that while female oestrus appeared to show a high degree of synchrony, as expected in a seasonal breeder, there was no evidence that the number of males accompanying a group of females influenced the likelihood of other males joining or leaving the group. Overall, our results confirm that female oestrous behaviour stimulates changes in male residence patterns. However, other observations suggest that changes in male residence may also stimulate female oestrus in some circumstances.
Amphibian abundance and diversity in Meru National Park, Kenya
Authors: Victor Wasonga D. , Bekele Afework , Lötters Stefan , Balakrishnan Mundanthra
Published: August 2006

The diversity and abundance of amphibians were investigated in Meru National Park, Kenya, using transect sampling, drift‐fence and pitfall trapping and opportunistic collecting. A total of 430 individuals under seven genera (Amietophrynus, Hemisus, Hyperolius, Phrynobatrachus, Phrynomantis, Ptychadena, Xenopus) comprising eleven species were sampled in three different habitats (apart from this, two additional species are known from Meru National Park): Acacia‐wooded grassland; Combretum‐wooded grassland; Acacia–Commiphora bushland. The sex ratio for almost all species was balanced (chi‐square, χ2; P > 0.5) and was not affected by habitat type (ANOVA: F = 8.3026, P = 0.6914). Shannon–Weaver Index (2.227) and Simpson's Index (8.244) were relatively high, and most of the eleven species sampled appeared to have a relatively even distribution (Shannon's Evenness Index, E = 0.927). However, Hemisus marmoratus and Phrynomantis bifasciatus were exclusively recorded in Acacia‐wooded grassland and in low abundances. There was a positive linear relationship of body weight against snout–vent length for two randomly selected anurans (Hyperolius glandicolor, Phrynobatrachus natalensis) among all three vegetation communities.
O‐3 A STUDY TO DETERMINE WHETHER THERE IS SIGNIFICANT CELL LOSS ON THE DISCARDED THINPREP BROOM WHEN USED FOR CERVICAL CYTOLOGY
Authors: Umana A. , Kubba A. A. , Dunsmore H. , Kokhan A. R. , Herbert A.
Published: August 2006

Introduction:  The aims of this study were to find out whether cells could be lost on the discarded ThinPrep (TP) broom. A similar study with SurePath (SP), whose methodology recommends leaving the broom in the vial, showed that cells were lost if the broom was discarded and that the amount was operator dependent.1

Methods:  Three experienced colposcopists (C1, C2 and C3) collected TP samples from 306 women. The broom was rinsed thoroughly in a vial (V1) and then rinsed and retained in a second vial (V2). Slides from each case were randomised as A or B to conceal whether they were from V1 or V2, double‐screened and a cytological diagnosis and cell count recorded for each one. Cells were counted in 10 high‐power fields (HPF) and the cellularity classified as follows: high (HC) > 20 per HPF, intermediate (IC) 10–20 per HPF and low (LC) <10 in order to quantitate the cells potentially discarded in V2.

Results:  The percentages with high‐grade and low‐grade abnormal cytology decreased with the degree of cellularity of the slide. Abnormal cytology was found in 96/133 (72.2%) of HC, 85/131 (64.9%) of IC and 15/42 (35.7%) of LC in V1 and in 5/8 (62.5%) of HC, 35/54 (64.8%) of IC and 86/244 (35.2%) of LC in V2. The cellularity of V2 was HC or IC in 62/306 (20.3%) and varied between the colposcopists: C1 39/109 (35.8%); C2 20/103 (19.4%); C3 3/94 (3.2%). C1 would have discarded the most cells but had the lowest percentage with LC in V1: 9/109 (8.3%) while the average was 42/306 (13/7%). Abnormal cytology in V2 was also found in V1 in 113/126 (89.1%).

Discussion:  Abnormal cells would have been discarded with the TP broom and the amount varied between the colposcopists. This finding could have an affect on adequacy of the sample used for routine diagnosis.

Reference:  1. Bigras G, Rieder MA, Lambercy J‐M et al. Keeping collecting device in liquid medium is mandatory to ensure optimized liquid‐based cervical cytology sampling. J Lower Genital Tract Dis 2003;7:168–74.
O‐5 UNIDIRECTIONAL VERSUS BIDIRECTIONAL SCREENING OF SUREPATH® LIQUID BASED CYTOLOGY SLIDES –PITFALLS & PERILS OF STUDY DESIGN
Authors: Holbrook M. , Roebuck J. , Rana D. N. , Marshall J. , Roberts S. , Desai M.
Published: August 2006

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.
O‐8 FALSE NEGATIVE BRONCHIAL CYTOLOGY
Authors: Feeley L. , Royston D.
Published: August 2006

Introduction and Aims:  The sensitivity of bronchial cytology (brushings, washings) is in the region of 70–80%. Therefore a significant number of false negatives occur. The main aims of this study were to: (i) To identify negative bronchial washings and brushings performed in 2004 with malignant follow‐up and (ii) To ascertain the cause of the false negative result where possible. 56 patients with negative bronchial cytology and subsequent malignant follow‐up on a pulmonary specimen were identified. These cases formed the basis of the study.

Results:  In our series peripheral tumour location and specimen inadequacy accounted for the majority of false negative results. Malignant cells were missed in a minority of cases (5 of 56). An explanation for failure of diagnosis in 4 of the 5 cases was paucity of malignant cells. In one case of small cell carcinoma, malignant cells were abundant, but were not recognized.

Recommendations:  What can be done to reduce the false negative rate?

(1) In cases where radiology has indicated that the tumour is peripherally located patients could proceed directly to fine needle aspiration.

(2) Improved communication between clinicians and pathologists is required in relation to: (a) specimen adequacy and (b) degree of clinical suspicion.
O‐9 AUDIT ON BK VIRUS DETECTION IN RENAL TRANSPLANT RECIPIENTS
Authors: Bhatt N. , McDermott N. , Young M.
Published: August 2006

Introduction:  BK virus nephropathy is an emerging cause of renal transplant failure accounting for allograft loss in 45–50% of recipients between 2–60 months post‐transplantation. The Renal Transplant Unit in Royal Free Hospital has devised a local surveillance programme for screening renal transplant patients at weekly intervals by urinary cytology (UC) and electron microscopy (EM), and confirmation of positive results by plasma PCR and allograft biopsy.

Objective:  (i) To monitor the implementation of local guidelines; (ii) To compare UC with EM and (iii) To identify areas for improvement.

Methods:  Decoy cell and EM positive cases were retrieved from the WinPath database for new renal transplant recipients (n = 55) during 1st November 2004 to 31st October 2005 in Royal Free Hospital. Plasma PCR was retrieved for positive cases.

Results:  Up to eight samples were sent for UC at random intervals from 47 patients, and up to 7 were sent for EM from 42 patients. Eleven were UC‐positive and one was EM‐positive. PCR was not requested in UC‐positive cases and 3 out of 10 were positive retrospectively. The EM‐positive case was PCR‐positive but UC‐negative.

Discussion:  Urine Cytology is more appropriate and cost effective for screening than Electron Microscopy. During the period covered by this audit, samples were sent in an inconsistent fashion after allograft dysfunction rather than for screening. A screening protocol has been agreed and a re‐audit is planned.
O‐10 THE ACCURACY OF CERVICAL CYTOLOGY: A COMPARISON BETWEEN THE THINPREP IMAGING SYSTEM AND CONVENTIONAL METHODS
Authors: Davey E , Irwig L , Macaskill P , Chan S , D'Assuncao J , Richards A , Farnsworth A
Published: August 2006

Douglass Hanly Moir is a large Australian laboratory which has recently introduced the ThinPrep Imaging System (TPI) for reading ThinPrep slides, which is still performed using a split‐sample technique. The Imager is a computerized system which identifies 22 fields for the cytologist to review using automated light microscopy. We compared the accuracy of TPI and conventional cytology (CC) during normal laboratory operation. The ThinPrep sample was prepared after taking a conventional Pap smear. TPI and CC reading was done without knowledge of the result of the other reading. The final cytology report issued to the referring doctor reflected the more severe of these two results. Histology results for all cases in which TPI and CC cytology results showed more than minimal disagreement were sought from the NSW Pap Test Register. Of 55 164 split sample pairs, 3.1% of CC of slides and 1.8% of TPI slides were unsatisfactory. There were 1758 women for whom there was more than minimal discrepancy between TPI and CC cytology results. TPI gave the more severe result in 1193 of the 1758 cases. In cases where only one of each pair of discrepant cytology results was CIN1 or higher grade, TPI detected 133 cases of high‐grade histology among 380 biopsies (35%), whereas CC detected 62 cases among 210 biopsies (29.5%). A repeat analysis based on reading of histology by one pathologist blinded to initial Pap smear result showed a similar result. Reading times were measured over 5 months for both TPI and CC for twenty cytologists who read both types of smears. On average, they read 13.3 TPI slides per hour and 6.1 CC slides per hour. This study provides evidence that cervical cytology read using the TPI detects more histological high‐grade disease than does CC. Further evidence shows that reading times are significantly reduced for cytologists using the TPI.
O‐12 BLAND DYSKARYOSIS: A NEW PITFALL IN THINPREP® LIQUID BASED CYTOLOGY
Authors: Lynch M. A. , Rana D. N. , Desai M. , Denton K.
Published: August 2006

Liquid based cytology (LBC) has improved cell visualization and preservation in cervical cytology. There has been a reduction in inadequate rate and some data to suggest an increase in sensitivity for dyskaryosis. Training for LBC has focused on differences in distribution of abnormal cells, but in most cases the morphological appearance of the dyskaryotic cells themselves is similar to that seen in conventional cytology. We are describing a new presentation of dyskaryosis which may be a cause of false negative cytology. We have referred to this as ‘Bland dyskaryosis’ because cells appear deceptively bland on low power examination, and can be misinterpreted as metaplastic or endocervical cells. Bland dyskaryosis cells are seen in groups. The architecture of the group is very disorganized, and adjacent cells show variation in size. Cells have a high nuclear/cytoplasmic ratio and smooth nuclear membranes. Chromatin is finely granular and evenly distributed. This is an unusual presentation of high‐grade dyskaryosis and we feel that there is a learning curve in laboratories converting to liquid based cytology. The spectrum of appearances of squamous dyskaryosis needs to be delineated to allow further increases in sensitivity for dyskaryosis.
P‐10 TO LYSE OR NOT TO LYSE?
Authors: McQueen F. , Fiddler J.
Published: August 2006

Introduction:  A glacial acetic acid wash performed retrospectively or prospectively on visibly bloodstained cervical ThinPrep® specimens can reduce the unsatisfactory rate and increase the number of diagnostic epithelial cells. This study was undertaken to determine which specimens are most likely to benefit from a prospective glacial acetic acid wash.

Methods:  Bloodstained ThinPrep® specimens selected for routine lysing prior to processing were macroscopically assessed and scored based on the level of blood present (+ to +++). Both unlysed and lysed slides were prepared from each specimen and microscopically examined.

Results:  Fifty‐eight specimens (32 scored +, 12 ++ and 14+++) were assessed. Three unlysed slides prepared from the ++ specimens and 12 from +++ specimens were evaluated as unsatisfactory due to excessive blood and inadequate numbers of squamous cells. In contrast, only one of the unlysed slides from the 32 + specimens was considered to be unsatisfactory. Almost all the lysed slides were satisfactory and generally more cellular than the unlysed slides. Abnormal cells were found in four cases (both unlysed and lysed paired slides).

Discussion:  Although the acetic acid wash increases cellularity of bloodstained ThinPrep® specimens, prospectively lysing all bloodstained specimens is an unnecessary procedure. Lysing only very heavily bloodstained specimens prior to processing will reduce laboratory workload, costs and the possibility of specimen mix up. Occasionally a retrospective wash may be required but screening staff should be aware that although blood may be present on an unlysed ThinPrep® slide, a lysed slide should not be requested unless there are insufficient numbers of squamous cells present.
P‐11 THE IMPACT OF INTRODUCING LIQUID BASED CYTOLOGY INTO A ROUTINE SCREENING LABORATORY
Authors: Gregory L. , Dudding N. , Smith J. H. F.
Published: August 2006

With the exception of information from the liquid based cytology (LBC) pilot site there has been limited data to date on the impact of the introduction of SurePath™ LBC in the NHSCSP. We will present data to show the impact on a medium sized laboratory (42 000 requests per annum) over the first phases of rollout. Data from before, during and post conversion, a period of 28 months, shows the following trends: (1) A significant fall in the inadequate rate (2) A slight decrease in the borderline / mild dyskaryosis rate (3) A small increase in the high‐grade squamous dyskaryosis reporting rate (4) An increase in both the number and specificity of glandular lesions reported (5) A largely unchanged positive predictive value for high‐grade abnormalities (6) A significant increase in laboratory productivity, unrelated to inadequate rate but through increased speed of screening. Although our experience is not directly comparable to the experience of the LBC pilot, our observations may well reflect that we were fortunate to be trained by staff from one of the LBC sites and thereby benefited directly from their experience.
P‐19 THE LABORATORY AND CLINICAL ADVANTAGES OF PROMPT CERVICAL SMEAR HISTOLOGY MISMATCH AUDIT
Authors: Payne F. B. , McKean M. E. , Lorimer M. Y. , Adam K. A. , Smart L. M.
Published: August 2006

ABC guidelines require correlation of cervical biopsy and smear results for calculation of positive predictive values. This need only involve retrospective review of cytology with histology considered the ‘gold standard’. Because NHSCSP recommend longer follow up for high than low grade CIN, it is important that high grade CIN is not underdiagnosed. We undertook prompt mismatch review, including both smear and biopsy review to fulfil an improved audit function and facilitate appropriate patient management.

Method:  All colposcopy referral smears were entered into an Access database and correlated with the subsequent colposcopic biopsy. All non‐correlating smears were reviewed independently and a consensus result reached. When the consensus remained high grade the histology was reviewed similarly and, if upgraded, a supplementary report was issued.

Results:  Over 6 months, 578 women referred to colposcopy with abnormal cytology had biopsies. In 491 (83%) cases, the referral smear and biopsy correlated within one grade. Of 51 smear (9%) ‘undercalls’ relative to the biopsy, 29 (5%) were upgraded to correlate on review. Of 36 smears (6%) classed as ‘overcalls’, 7 were true cytological overcalls and downgraded. 10 biopsies were upgraded, 5 from low to high grade CIN. 19 (3%) high grade smears remained non‐correlating. Correlation increased to 91% after smear and histology review.

Conclusion:  This system of mismatch correlation enables clinicians to receive amended results early enough for modification of treatment and follow up based on an accurate CIN grade. Tabulated results highlight non‐correlating cases where the biopsy may be non‐representative and identify cases for clinico‐pathological conferences. Multi‐observer review of true cytology undercalls and overcalls has an educational role and improved accuracy of CIN grading increases PPVs. Although labour intensive, this approach offers significant advantages and audit criteria may be refined to focus on relevant issues.
P‐24 MUCINOUS ASCITES ‐ AN APPROACH TO CYTOLOGICAL CLASSIFICATION
Authors: Lynch M. A. , Rana D. N.
Published: August 2006

Pseudomyxoma peritonei (PMP) is the name given to the clinical syndrome and the heterogeneous group of conditions characterised by mucinous ascites and mucinous implants. Patients with PMP are treated with a combination of cytoreductive surgery, peritonectomy and hyperthermic intraperitoneal chemotherapy (HIPEC). A dedicated PMP outpatient clinic refers samples of mucinous ascites to our laboratory. Peritoneal fluid samples in disseminated peritoneal adenomucinosis (DPAM) contain abundant extracellular mucin together with epithelium showing mild to moderate (low grade) cytological atypia. DPAM is associated with the presence of an appendiceal mucinous adenoma. Peritoneal mucinous carcinomatosis (PMCA) is associated with the presence of an appendiceal or intestinal mucinous adenocarcinoma and mucinous ascites containing mucinous epithelium with the cytological features of a carcinoma. PMCA –I (intermediate) cases are those with peritoneal lesions demonstrating predominantly features of DPAM but with foci of well differentiated mucinous adenocarcinoma. Cases classified as PMCA‐D (discordant) are characterised by an atypical adenoma of the appendix and associated peritoneal mucinous adenocarcinoma. There is a recognised statistically significant difference in survival in cases reported as DPAM, cases with intermediate features and PMCA. Therefore we advocate reporting cases of mucinous ascites in conjunction with histology according to the following diagnostic categories: (1) Mucin without the presence of any epithelial cells (2) Disseminated peritoneal adenomucinosis (DPAM) (3) PMCA‐I/D (4) Peritoneal mucinous carcinomatosis (PMCA). Cases histologically diagnosed initially as DPAM may after a period of time undergo dedifferentiation and re‐present as PMCA. It is important that the cytopathologist recognise and classify the features of the cells present within the peritoneal fluid. It is essential that the histological features are correlated with the cytological diagnosis and that any differences in morphology are documented for accurate survival prediction and appropriate clinical management. We present the clinicopathological and cytological features in the cases of PMP which we have encountered in our department and describe our cytological approach to classification.
P‐8 THE LOCAL IMMUNE RESPONSE TO LOW‐GRADE CIN – ALTERATIONS IN T‐CELL AND MACROPHAGE SUBSETS
Authors: Ahmed S. M. , Reid W. M. N. , Poulter L. W.
Published: August 2006

Introduction:  Despite the cervix having the largest concentration of lymphocytes and macrophages in the lower genital tract, there is limited knowledge of these cell types in CIN. This study was undertaken to compare T‐cell and macrophage populations in women with and without low‐grade CIN.

Methods:  Control group: 10 women with a normal recent cervical smear result, mean age 30 years. Study group: 10 women with low‐grade CIN, mean age 33.2 years. Colposcopically directed cervical biopsies were taken from control women and from areas of CIN in the study group, and studied using immunoperoxidase and immunofluorescence staining.

Results:  T‐cell numbers remained constant. In CIN, there were increased proportions of recruited (CD8+CD5+), activated (CD8+HLA‐DR+) and cytolytic (CD8+TIA‐1+) CD8 T‐cells. There were more activated CD4+HLA‐DR+ T‐cells in CIN. Memory T‐cell (CD8+CD45RO+ and CD4+CD45RO+) proportions were reduced. Total macrophage numbers (CD68+) remained constant, but activated macrophages (CD68+HLA‐DR+) rose. Phagocytic (RFD7+) macrophage proportions decreased with a concomitant increase in suppresser macrophages (RFD1+RFD7+).

Discussion:  Despite cellular numbers remaining unchanged, there were significant changes in T‐cell and macrophage populations in women with low‐grade CIN. There is recruitment of activated and cytolytic CD8 T‐cells. The swing to a predominantly suppressive macrophage phenotype may predispose to CIN. Carcinoma of the cervix occurs more frequently in women who are immunosuppressed. Our data suggests that local and systemic immune mechanisms may be relevant in the response to HPV‐induced neoplasia.
DEFINING DYSKARYOSIS – THE BSCC CLASSIFICATION IN 2006
Authors: Denton K.
Published: August 2006

The terminology used for reporting cervical samples in the UK is the BSCC classification, which has evolved over many years. In 2002 the BSCC held a consensus conference to review the BSCC classification, with the intention of providing clearer results for women, improving concordance with other terminologies and facilitating consistency with new scientific developments and technologies. The consensus conference was well attended and robust. In the intervening years there have also been other further advances on morphometry, data on outcomes, data from EQA and other sources. Liquid Based cytology (LBC) has been implemented by the NHS CSP. All of these developments have impacted on the proposed classification, which will be presented in full. The term ‘dyskaryosis’ will be retained and several of the current reporting categories will be relatively unchanged, though additional information on LBC will be provided.

The major proposed changes are:

(1) A move to a single category of ‘High Grade Dyskaryosis’ to replace the existing categories of moderate and severe dyskaryosis.

(2) Sub‐division of Borderline change into three categories.

Borderline change in glandular cells

Borderline change ?high grade

Borderline change, NOS

(3) The current grades of Mild Dyskaryosis and Borderline change with Koilocytosis to be merged.

Details of these proposals, together with illustrations and the evidence base for change will be presented.
THE CASE FOR CONTINUING PRIMARY CERVICAL SCREENING WITH CYTOLOGY RATHER THAN HPV TESTING
Authors: Duncan I. D.
Published: August 2006

HPV testing is used in several developed countries including the USA but has not yet found an established role in the UK cervical screening programmes. HPV testing is independent of the interpretive skills of the individual laboratory worker although the sensitivity of the test itself can be altered. New strains of the virus continue to be identified so the test can never be 100% accurate. It has been studied in women with mild or borderline dyskaryosis in their smear as a possible means of triage. If anything, however, HPV testing is too sensitive and lacks specificity. Its use resulted in too many women being referred for colposcopy putting a strain upon resources. Were it to be used in the same younger age groups as today's smears i.e. 20 onwards in Scotland, 25 and onwards in England then primary infections would be detected along with potential precancerous cases and again colposcopy clinics would be inundated. Thus, if HPV were to replace cytology in primary screening for cervical neoplasia it would be impractical before the age of 30 and the current common age for detection and treatment of cervical intraepithelial neoplasia (CIN) would have to be postponed, some early cases would be missed and some cancers would undoubtedly result. Girls are becoming sexually mature at an earlier age than their parents. Their sexual debut is also at an earlier age when lifelong monogamy is unlikely to result. Their opportunities for contact with genital strains of HPV are increased and cervical neoplasia may also develop sooner. Such cases would be missed. There would be further knock‐on effects if HPV testing were to replace cytology in primary screening. Cytology interpretive skills would be lost. It has been suggested that HPV testing be used for primary screening and cytology be confined to secondary examination of samples from HPV positive women. Although fewer personnel would be required to interpret those ‘smears’, they may have become a little less sharp. The beauty of HPV testing is the reliability of its negative predictive value. This could be put to good use combining it with LBC to get the best out of both worlds. Women could start 3 yearly cytological screening at 25 years. At 37 years, by which age most women would have developed viral immunity and entered a stable relationship, the LBC sample could also be tested for HPV. If both tests were negative the next tests could be postponed until 43 and then 49. It has been repeatedly shown in well‐screened populations that the incidence of CIN is extremely low in women from the age of 50 if they have been consistently negative until that point. As was the case with cytology and colposcopy which were shown to be complementary, cytology and HPV testing need not be mutually exclusive.
ETHICAL ISSUES IN CERVICAL SCREENING
Authors: Weller D.
Published: August 2006

This paper will provide an overview on current ethical dilemmas in cervical screening. Screening has led to very significant reductions in mortality from this disease. Yet it requires a major commitment from a wide range of health professionals and health sectors including pathology, primary care, public health and NHS cancer screening services. Despite its very significant achievements, cervical screening can be associated with adverse events and in contemporary cancer screening programmes, there is a need to provide patients with a balanced view on the benefits of participating. The following areas will be covered:

(1) Social inequalities: does cervical screening reinforce health and social inequalities, is it sufficiently available to lower socio‐economic groups and does it target the appropriate sub‐groups within the population?

(2) Informed choice: there is a change in focus in cancer screening programmes – until recently most effort has focused on promoting uptake. Informed choice is now seen as a worthy objective of screening programmes. GP practices have, in the past, been accused of overly coercive tactics in achieving their target rates for cervical screening. This has the potential to cause significant harm through such efforts. Ideally we want participants in cancer screening to have a very clear idea of both the risks and benefits of participation, yet this can involve a potentially lengthy process of information sharing between health care providers and invitees. How do we achieve an appropriate balance between informed choice and adequate levels of uptake?

(3) Opportunity costs of cervical screening: Screening is an expensive process, the health service is stretched and there is a significant opportunity cost. Should we be diverting resource to other diseases of higher prevalence and greater public health burden?

(4) When cervical screening goes wrong: what can we learn from instances in the past where QA systems and other aspects of cervical screening have failed and patients have been caused harm?

(5) New challenges posed by HPV testing and vaccination.

(6) Future directions. Where should cervical screening be heading, what will the future ethical challenges be?

The intention of the paper is to summarise current evidence and issues, and to prompt discussion on ways that health professionals, in conjunction with consumer groups, can move forward on these and other important ethical issues.