P‐1DIRECT‐TO‐VIAL USE OF THE LIQUI‐PREP™ CYTOLOGY SYSTEM

Introduction: The authors initiated the use of Liqui‐PREP? (LGM International Inc., Fort Lauderdale, FL, USA) in August, 2005. Cytotechnologists received extensive (one month) training by cytopathologists experienced in Liquid‐based cytology. The Liqui‐PREP? direct‐to‐vial procedure (LP) was compared to the conventional Pap smears in a routine screening population. Methods: Data derived from 26 178 LP cervical‐vaginal (CV) specimens were compared to data derived from 218 548 conventional Pap smears (CS). Both data sets reflect patient samples collected concurrently (August–December, 2005) by 117 participating outpatient medical practices from a well‐defined geographic area. There were no significant personnel changes during the study period. The diagnostic results, classified according to Bethesda criteria were calculated. Results:

Atypical glandular cells in cervical smears: histological correlation and a suggested plan of management based on age of the patient in a low-resource setting

Objectives:  To perform an audit of all smears reported as atypical glandular cells (AGC) using the Bethesda system (TBS) 2001.
Methods:  A total of 18 376 cervical smears were screened from January 2005 to June 2007, of which 65 cases were reported as AGC. Follow-up histology was available in 31 cases (47.7%), in whom a detailed cytological/histological correlation was carried out.
Results:  AGC constituted 0.35% of all Pap smears. Follow-up histology was normal or benign in 20 cases, whereas a squamous or glandular abnormality was seen in 11 cases. Squamous abnormalities included one case each of cervical intraepithelial neoplasia (CIN)1, CIN2 and CIN3 and five cases of squamous cell carcinoma. All glandular epithelial abnormalities were endometrial in origin and included two endometrial adenocarcinomas and one uterine serous carcinoma. Neither in situ nor invasive adenocarcinoma of the endocervix was observed. Review of smears and reclassification as AGC, not otherwise specified and favour neoplasia revealed a higher proportion of abnormality in the latter group, reaffirming the utility of subtyping. The median age of women with AGC was 41 years. The outcome was analysed with respect to the median age. In women aged equal or more than 40 years, AGC reflected a high-grade squamous or glandular epithelial abnormality in 50% of cases compared with none in those less than 40 years old ( P  = 0.010).
Conclusion:  The age of the woman as well as the subtype of atypical glandular cells influences outcome and hence must be taken into consideration while formulating an acceptable management strategy in these women in a low-resource setting.     

Cytological diagnosis of basal cell carcinoma and actinic keratosis, using Papanicolaou and May–Grünwald–Giemsa stained cutaneous tissue smear

Objective:  Cytology may become the diagnostic method of choice with the advent of new non-invasive treatments for non-melanoma skin cancer, as the sampling technique for cytology entails little tissue disfiguration. The aim of this study was to compare and evaluate the diagnostic performance of scrape cytology using two different cytological staining techniques, and to evaluate additional touch imprint cytology, with that of histopathology of basal cell carcinoma (BCC) and actinic keratosis (AK).
Methods:  We investigated 50 BCC and 28 AK histologically verified lesions, from 41 and 25 patients, respectively. Two separate skin scrape samples and one touch imprint sample were taken from each lesion. The smears were stained with Papanicolaou (Pap) or May–Grünwald–Giemsa (MGG) stains. All cytological specimens were examined in random order by pathologists without knowledge of the histology. Cytodiagnostic results were compared with the histopathological report.
Results:  Scrape cytodiagnosis agreed with histopathology in 48 (Pap) and 47 (MGG) of the 50 BCC cases, and in 26 of 28 (Pap) and 21 of 26 (MGG) AK cases, yielding sensitivities of 96%, 94%, 93% and 81%, respectively. No significant difference in sensitivity between the two staining methods was found but a trend towards higher Pap sensitivity for AK was noted ( P  =   0.10). Touch imprint cytology confirmed histopathology in 38 of the 77 cases of BCC and AK.
Conclusion:  Cytological diagnosis with either Pap or MGG stain for BCC and AK is reliable, and differentiates well between BCC and AK. Imprint cytology proved to be non-diagnostic in half of the examined cases.     

The future of cytology laboratories

1. NHSCSP Annual Review 2006. Patnick J (ed.). Sheffield, UK, NHSCSP; 2007.
2. Dowie R, Stoykova B, Crawford D. et al . Liquid-based cytology can improve efficiency of cervical smear readers: evidence from timing surveys in two NHS cytology laboratories. Cytopathology 2006; 17(2):65–72.
3. Williams AR. Liquid-based cytology and conventional smears compared over two 12-month periods. Cytopathology 2006; 17(2):82–85.
4. Gregory LS, Dudding N, Smith JHF. The impact of introducing liquid based cytology into a routine screening laboratory. Cytopathology 2006; 17(S1):24.
5. Lord Carter of Coles. Report of the Review of NHS Pathology Services in England 2006. Department of Health.
6. Kardos TF. The focal point system: focal point slide profiler and focal point GS. Cancer 2004; 102(6):334–39.
7. Biscotti CV, Dawson AE, Dziura B. et al. Assisted primary screening using the automated ThinPrep Imaging System. Am J Clin Pathol 2005; 123(2):281–87.
8. Eltoum IA, Roberson J. Impact of HPV testing, HPV vaccine development and changing screening frequency on national Pap test volume: projections from the National Health Interview Survey (NHIS). Cancer 2007; 111(1):34–40.
9. Kohli M, Ferko N, Martin A. et al. Estimating the long-term impact of a prophylactic human papillomavirus 16/18 vaccine on the burden of cervical cancer in the UK. Br J Cancer 2007; 96(1):143–50.
10. Franco EL, Cuzick J, Hildesheim A, de Sanjose S. Chapter 20: issues in planning cervical cancer screening in the era of HPV vaccination. Vaccine 2006; 24 (Suppl. 3):S171–S177.
11. Peto J, Gilham C, Fletcher O, Matthews FE. The cervical cancer epidemic that screening has prevented in the UK. Lancet 2004; 364(9430):249–56.     

Use of a liquid-based, thin-layer Pap test in a community hospital. Impact on cytology performance and productivity

OBJECTIVE: To evaluate the direct-to-vial efficacy of the SurePath Pap test (TriPath Imaging, Burlington, North Carolina, U.S.A.) in a community hospital laboratory and to assess its impact on productivity, as measured by Pap test turnaround times (TATs). STUDY DESIGN: A total of 8,771 SurePath Pap tests were compared to 5,055 conventional Pap smears collected and processed over the same 12-month period. SurePath histologic correlation rates were compared to historical correlation rates for conventional Pap smears. Pap test TATs for 3 months prior to implementing SurePath were compared to TATs for the last 3 months of the study, which included an approximate 70:30 ratio of SurePath to conventional Pap tests. Laboratory staffing was unchanged, and mean monthly accessions were relatively constant. RESULTS: SurePath showed statistically significant improvements in the detection of low grade (LSIL) (196%) and high grade (HSIL) (243%) squamous intraepithelial lesions (SILs) relative to conventional Pap smears. The atypical cells of undetermined significance (ASCUS) rate remained nearly constant with SurePath, while the ASCUS/SIL ratio decreased by 68%. Unsatisfactory rates declined 81%. Histologic correlation rates with SurePath increased over those of conventional Pap smears. Pap test mean TATs improved 73% with SurePath. CONCLUSION: Sure-Path detected significantly more cases of LSIL and HSIL than conventional smears without compromising specificity. The ASCUS/SIL ratio and unsatisfactory rate declined dramatically. Pap test TATs also improved markedly with SurePath.     

Higher diagnostic accuracy with the ThinPrep method in a simulated intraoperative environment

Objective:  To compare the accuracy of intraoperative fine needle aspiration cytology samples prepared by the ThinPrep method to conventional cytological methods. Specimen adequacy and turn around time (TAT) were also assessed.
Methods:  Fifty consecutive fresh tumours submitted for histological analysis were aspirated and each prepared as follows: (i) direct smear with H&E stain, (ii) direct smear with Pap stain, (iii) ThinPrep slide with H&E stain, and (iv) ThinPrep slide with Pap stain. The slides were randomly distributed to three cytopathologists for interpretation. The quality of the preparation, the diagnosis and the time needed for interpretation were recorded.
Results:  Accuracy was measured as the percentage of absolute agreement between the cytological and the histopathological diagnoses of the lesions. Histologically, there were 43 malignant and six benign lesions and one atypical lipoma. The TAT began when the slides/cytolyte specimens arrived at the lab and ended with the pathologist's diagnosis.
Conclusions:  In terms of accuracy and specimen adequacy, ThinPrep slides with Pap stain is the best procedure. This advantage however is offset by the longer testing time.     

P-13CERVICAL SMEAR REPORTING IN NORTHERN IRELAND SLE PATIENTS

Introduction:  SLE is an autoimmune disease with the potential for multi-organ involvement. It has been suggested that SLE patients may be at increased risk of cervical dysplasia. We studied a cohort of Northern Ireland Lupus patients and compared them to controls within the same geographical location assessing their cervical smear histories and performing an up-to-date cervical smear.
Methods:  A total of 141 SLE patients fulfilling the ACR criteria for lupus were enrolled into the study. They were compared to 138 control patients who were due to attend for a routine cervical smear within the same geographical location. Each patient gave written consent to be involved in the study including access to medical records and pathology data.
Results:   

     

Characterization of lactosporin, a novel antimicrobial protein produced by Bacillus coagulans ATCC 7050

Aims:  To characterize the antimicrobial protein produced by Bacillus coagulans used in the probiotic dietary supplement (Lactospore^® Probiotic, Sabinsa Corp., Piscataway, NJ, USA).
Methods and Results:  Bacillus coagulans ATCC 7050 was grown at 37°C for 18 h. The cell free supernatant was concentrated 10-fold (lactosporin preparation, LP). The antimicrobial activity of LP was confirmed against Micrococcus luteus ATCC 10420 in a well diffusion assay. The proteinaceous nature of LP was determined following exposure to different enzymes. The activity of LP was pH-dependent but stable to heat. The isoelectric point of LP was determined to be 3·5–4·0. PCR analyses showed no similarity between lactosporin and known antimicrobial proteins produced by the Bacillus spp.
Conclusions:  Lactosporin is a novel antimicrobial protein. Initial characterization indicates that it may fall outside of the conventional classification of class I and II bacteriocins. Loss of activity after exposure to a number of proteolytic enzymes and lipase suggest that lactosporin may posses a lipid moiety which contributes to its inhibitory activity.
Significance and Impact of the Study:  The unique characteristics of lactosporin, including its antimicrobial activity against pathogenic micro-organisms, indicate that it may have potential for application in foods and personal care products.     

P-12A RETROSPECTIVE AUDIT OF FINE NEEDLE ASPIRATION CYTOLOGY OF PANCREAS

Background:  Fine needle aspiration cytology (FNAC) of pancreas is a widely accepted method of diagnosis of pancreatic mass lesions. We have performed a retrospective analysis of all radiological (CT/ultrasound) and endoscopic ultrasound guided procedures at our institution.
Aim:  (1) To review the results of FNAC of pancreas from January 2000 to April 2006. (2) To calculate the inadequate rate. (3) To account for discrepancies between the cytological and histological diagnoses. (4) To identify any false positive cases if present.
Method:  The results of all pancreatic FNAC reported at our institute from January 2000 to April 2006 were identified from the laboratory system. All results were classified as follows: Inadequate, inconclusive, benign, suspicious and malignant. The results were further categorised depending on whether they were CT/ultrasound guided or EUS guided. The histological diagnosis where available was used as the gold standard and where discrepancies were present the cytological preparations were reviewed.
Results:  Seventy-three patients underwent pancreatic FNAC during the study period. Table 1 illustrates our results.
 

     

10.

P-14CERVICAL DYSPLASIA AND RISK FACTORS IN NORTHERN IRELAND SLE PATIENTS     

M. T. McHenry  K. Cuschieri  C. Moore  H. A. Cubie  R. Clarke  N. Anderson  & A. L. Bell 《Cytopathology》2006,17(S1):25-26

Introduction:  An increased risk of cervical dysplasia has previously been reported in lupus. The role of traditional related risk factors are unclear. Persistent infection with certain genotypes of Human Papilloma Virus termed high risk (HR-HPV) is considered a prerequisite for the development of cervical cancer.
Methods:  A total of 141 SLE patients fulfilling the ACR criteria for lupus were enrolled into the study. They were compared to 138 control patients who were due to attend for a routine cervical smear. Each patient completed a cervical cancer risk factor questionnaire. A cervical smear was taken and residual material used to test for the presence of HR-HPV using a hybridisation microplate assay. Positive samples were then genotyped.
Results:   

     

11.

Cervical smear adequacy: cellularity references were found to increase both interobserver agreement and unsatisfactory rate     

D. Moore  D. Pugh-Cain  T. Walker 《Cytopathology》2009,20(3):161-168

Objectives:  To determine the degree of interobserver variation in the assessment of conventional cervical smear adequacy as defined by The Bethesda System (TBS) 2001, and to determine the effect of using reference images of known squamous cellularity when performing squamous adequacy assessments.
Methods:  Experimental pre-test/post-test design utilizing 70 conventionally prepared cervical smears. Sample smears containing scant squamous cellularity were independently rated on two occasions by six cytotechnologists. Time 1 was without the use of reference images, and Time 2 was aided by cellularity reference images. The κ statistic was used to compare rater agreement.
Results:  The level of agreement increased from an average κ of 0.26 (SD 0.10) for Time 1, to an average κ of 0.40 (SD 0.15) for Time 2. The difference in mean κ values at the two assessments was statistically significant ( t  = 3.71; P  = 0.002). Unanimous agreement among the raters was observed for 15 samples (21.42%) at Time 1 (only one of which was classified as unsatisfactory) and 21 samples (30.00%) at Time 2 (12 of which were classified as unsatisfactory).
Conclusion:  Interobserver agreement increased after cellularity reference images were implemented. Using TBS 2001 squamous adequacy criteria and images of known squamous cellularity as references resulted in a decreased number of smears reported as satisfactory.     

12.

P-27ATYPICAL GLANDULAR CELLS ON SMEAR – AGE RELATED IMPLICATIONS AND MANAGEMENT     

L. Tan  J. McCarthy  & D. Lyons 《Cytopathology》2006,17(S1):29-29

Introduction:  There is ongoing debate about the terminology used in the classification of dyskaryosis, including whether BNA smears should be classified according to whether koilocytes are present or not. We explored the effect of koilocytosis in the management experiences and clinical outcomes of women with a single BNA smear.
Methods:  This study includes 410 women aged 20–59 years, resident in Tayside who had an 'baseline' BNA smear between 31/10/1999 and 31/10/2002 who were eligible for the TOMBOLA trial but who did not participate. Recommended follow-up for these women was a repeat smear in six months. Up to three-years follow-up data was collected on subsequent cytological smears, colposcopy examinations and any related histology. The baseline smear was re-read for the presence or absence of koilocytes. Women were classified according to their clinical outcomes during their three years.
Results:  47% (192/410) of women were classified as having koilocytosis at baseline. The mean age was lower among these women (25 years) compared to those without koilocytosis (35 years). 55% (105/192) of women with koilocytosis at baseline had one or more negative smears and no further abnormal smears, compared to 63% (137/218) of those without koilocytosis, the difference was not statistically significant. 28% (53/192) of women with koilocytosis at baseline were referred to colposcopy after subsequent low-grade or high-grade smears compared to 19% (42/218) of those without koilocytosis; the difference was statistically significant. 14% (27/192) of women with koilocytosis at baseline were diagnosed with CIN2/3 or worse during follow-up compared to 10% (21/218) of women without koilocytosis; the difference was not statistically significant.
Discussion:  Our results suggest that the presence or absence of koilocytosis has little impact on the clinical outcomes of women with a BNA smear.     

13.

Utility of the in situ detection of HPV in Pap smears diagnosed as within normal limits.     

G Menezes  E Euscher  B Schwartz  F Catania  J Chancellor  G J Nuovo 《Acta cytologica》2001,45(6):919-926

OBJECTIVE: To determine the clinical significance in normal Pap smears of HPV detection as determined by Hybrid Capture (HC) and in situ hybridization analyses. STUDY DESIGN: We studied 135 consecutive Pap smears as well as 46 other smears from high-risk patients each initially diagnosed as within normal limits. RESULTS: The 135 "normal" Pap smears were rescreened, and 6 (4%) where found to be either ASCUS or SIL. In the remaining 129 cases, HPV DNA was detected in 0% and 9%, respectively, using in situ hybridization and HC I. Upon rescreening the high-risk patients, nine (20%) were reclassified as having SIL/ASCUS; each was in situ hybridization positive, and eight were HC positive; six (67%) of these women developed SIL on follow-up. In the 37 Pap smears in high-risk women still within normal limits after manual rescreening, HPV was detected in 2% by in situ hybridization and 46% by HC; 6% of the HC-positive women developed SIL on follow-up. CONCLUSION: In situ hybridization rarely detects HPV in Pap smears diagnosed as within normal limits after manual rescreening. In situ hybridization is very effective in detecting rare, atypical cells in Pap smears diagnosed as within normal limits and, in a high-risk population, is predictive of SIL on clinical follow-up.     

14.

Clostridia spore formation during aerobic deterioration of maize and sorghum silages as influenced by Lactobacillus buchneri and Lactobacillus plantarum inoculants     

E. Tabacco  S. Piano  L. Cavallarin  T.F. Bernardes  G. Borreani 《Journal of applied microbiology》2009,107(5):1632-1641

Aims:  The effect of the inoculation of maize and sorghum silages with Lactobacillus plantarum (LP) and Lactobacillus buchneri (LB) on the clostridia spore formation during aerobic deterioration has been studied.
Methods and results:  The crops were ensiled in 30 l jars, without a lactic acid bacteria inoculant (C), and with an LP or LB inocula (theoretical rate of 1 × 10⁶). After 90 days of conservation, the silages were analysed for the chemical and microbiological characteristics and subjected to an aerobic stability test, during which pH, temperature, nitrate, yeast, mould and clostridia spores were measured. Compared to the C and LP silages, yeasts were reduced in the LB silages, resulting in an increased aerobic stability. Clostridia spores, determined by most probable number (MPN) procedure, increased to 6 log₁₀ MPN g⁻¹ in the C and LP maize silages, whereas they reached 3 log₁₀ MPN g⁻¹ in C and LP sorghum silages.
Conclusions:  Clostridia spore count only slightly increased in the LB maize silages after 342 h (2·59 log₁₀ MPN g⁻¹), whereas it did not show any increase in the LB sorghum silages for the whole period of air exposure.
Significance and impact of the study:  The data indicated that clostridia spore outgrowth can take place during silo feedout in aerobic-deteriorated silages and that LB inoculation reduces the risk of clostridia outgrowth after silage opening by increasing the aerobic stability.     

15.

Evaluation of Papanicolaou stain for studying micronuclei in buccal cells under field conditions     

Ayyad SB  Israel E  El-Setouhy M  Nasr GR  Mohamed MK  Loffredo CA 《Acta cytologica》2006,50(4):398-402

OBJECTIVE: To compare Papanicolaou (Pap) and May-Grünwald Giemsa (MGG) stain as 2 techniques for staining for buccal mucosal cells to detect micronuclei (MN) infield studies. STUDY DESIGN: Eighty cytologic smears (2 per individual) were taken from the buccal mucosa of 40 cigarette smokers recruited at a rural village in Egypt. Forty smears were stained with Pap stain and 40 with MGG stain. All were assessed for cellularity and scored for MN. RESULTS: Pap stain was faster and easier to process and transport in the field study than was MGG stain. Regarding MGG smears, bacteria and cell debris masked the MN as compared to Pap smears, in which the fixative destroyed the bacteria and made the cell boundaries clearly demarcated. Using Pap stain, MN were seen easily in transparent cytoplasm. CONCLUSION: Pap stain is the preferred method infield studies for scoring and detecting MN in cells of buccal mucosa.     

16.

Altered expression of adhesion molecules in inflammatory cervical smears     

E. N. Politi  A. C. Lazaris  M. Kehriotis  T. G. Papathomas  E. Nikolakopoulou  H. Koutselini 《Cytopathology》2008,19(3):172-178

Objective:  The aim of the present study was to evaluate the expression of pan-cadherin and β-catenin in cervical smears with various types of infectious agents.
Patients and Methods:  Cervical smears obtained from 53 women, aged 21–65 years, with a diagnosis of specific inflammation were examined in our study. Eighteen subjects were infected by Candida albicans , 18 by Gardnerella vaginalis , nine by Bacteroides spp. and eight by Chlamydia trachomatis . All infectious agents found in the smears were at the same time confirmed by the microbiological laboratory methods. We performed a biotin–streptavidin-peroxidase immunocytochemical method using anti-β-catenin (Clone 12F7) and anti-pan-cadherin (pan, polyclonal) antibodies.
Results:  Aberrant expression of pan-cadherin was found in the cytoplasmic membrane of glandular, metaplastic, superficial and intermediate squamous cells in all types of infections. With regard to β-catenin, this was expressed in majority (90%) of glandular and metaplastic cells in all types of infections and in a small proportion (15%) of superficial and intermediate squamous cells in infections caused by C. albicans and G . vaginalis .
Conclusion:  Our data show that infectious agents may cause alterations in the expression and distribution of these adhesive molecules, which can be recognized in cervical smears. Additional studies in larger sets of patients should help clarify this issue further.     

17.

Performance of the AutoPap Primary Screening System in the detection of high-risk cases in cervicovaginal smears     

Alasio LM  Alphandery C  Grassi P  Ruggeri M  De Palo G  Pilotti S 《Acta cytologica》2001,45(5):704-708

OBJECTIVE: To assess the performance of the AutoPap Primary Screening System (APSS) (TriPath Imaging, Inc., Burlington, North Carolina, U.S.A.) for the detection of high grade cervical squamous intraepithelial lesions and invasive cervical cancer. STUDY DESIGN: A total of 14,779 consecutive conventional Pap smears were processed by the APSS. All slides designated as "Review" by the device were manually screened according to the Bethesda System. The ranking scores obtained from the device were compared with the cytologic interpretations in all cases and with the final histologic diagnoses in the cases with cytologic severe abnormalities. RESULTS: The device classified 10,349 slides as Review (78%) and 2,912 (22%) as "No Further Review." In the 78% Review cases, the samples were ranked in descending order of potential abnormality, broken into quintiles. The correlation between the slide quintile ranks and the manual cytologic diagnosis indicated that 90% of abnormal smears were categorized by the device as in the first and second quintile rank, and the correlation between the rank report of the device and the histologic diagnosis showed that all cases of HSIL or invasive carcinoma were in the top two ranks. No significant abnormalities were observed in any of the smears categorized as No Further Review. CONCLUSION: This study confirmed the effectiveness of APSS for the detection of Pap smears with severe abnormalities.     

18.

Rapid pre-screening of cervical smears as a method of internal quality control in a cervical screening programme     

S. B. do Nascimento Tavares  N. L. A. de Sousa  E. J. C. Manrique  Z. B. P. de Albuquerque  L. C. Zeferino  R. G. Amaral 《Cytopathology》2008,19(4):254-259

Objective:  To evaluate the performance of rapid pre-screening (RPS) as a method of internal quality control in the cytopathological examination of cervical smears for cervical cancer screening.
Methods:  The sample consisted of 6135 cervical smears submitted to RPS and routine screening (RS) methods. The smears classified as negative in RPS and RS were considered final diagnoses, and were not, therefore, submitted to any additional review. The smears identified as suspect or unsatisfactory according to RPS were analysed separately by two different cytologists irrespective of the diagnosis reached in RS. Smears considered abnormal or unsatisfactory at RS were also reviewed. When both cytologists issued concordant diagnoses, this was considered the final diagnosis. Discordant results were analysed by a third cytologist and a consensus meeting was held to define the final diagnosis.
Results:  Taking abnormalities detected by RS as the denominator, RPS had a sensitivity of 63.0% for the detection of all abnormal smears and 96.7% for high grade squamous intraepithelial lesion (HSIL). When compared with the final diagnosis, sensitivity of RPS for all abnormal smears was 74.9% and for HSIL 95.0%. Of the 529 abnormal smears confirmed in the final diagnosis, 2.15% were detected only by the RPS.
Conclusion:  RPS is an effective alternative method of internal quality control with high sensitivity for the detection of more severe lesions. It also permits monitoring of the laboratory rate of false-negative results, and allows constant evaluation of the performance both of the pre-screening and RS cytologists.     

19.

Decrease in numbers of glandular cell groups in post-LLETZ liquid-based cytology preparations     

A. Maguire  L. Turner  D. Magee  D. Gibbons 《Cytopathology》2008,19(1):44-47

Objective:  Large loop excision of the transformation zone (LLETZ) has become standard of care in the management of cervical squamous neoplasia and with cone biopsy glandular intraepithelial neoplasia. Controversy remains about the long-term effects of this traumatic procedure. The aim of this study was to count and compare the number of endocervical glandular cell groups in pre- and post-LLETZ cervical preparations using liquid-based cytology to establish a cyto-morphological correlate of destruction of the transformation zone.
Methods:  The cytology/histology correlation audit records of the Cytopathology Department of St Luke's Hospital in 2003 and early 2004 were used to select patients with a cytological diagnosis of high grade dyskaryosis followed by LLETZ. Only those cases with post-LLETZ cytological follow-up were selected. Cases using conventional smears were excluded. One hundred and twenty slides (60 pairs of slides) in total were retrieved. The cases underwent review and all groups of >3 glandular cells in each slide were counted by AM while blinded as to whether smears were pre- or post-LLETZ. Medians were compared using a Mann–Whitney U -test.
Results:  The median number of groups of endocervical glandular cells of the pre-treatment group was 5.5 and of the post-treatment group was 2.0. There were significantly fewer endocervical glandular cell groups in the post-LLETZ population ( P  = 0.03).
Conclusions:  The number of endocervical glandular groups in cervical cytological preparations decreases significantly following LLETZ procedure. This suggests that cytological follow-up may not be as useful in glandular neoplasia cases. Few or absent glandular cell groups in post-LLETZ preparations may have implications for adequacy assessment.     

20.

The role of cervical cytology and colposcopy in detecting cervical glandular neoplasia     

A. Ullal  M. Roberts  J. N. Bulmer  M. E. Mathers  V. Wadehra 《Cytopathology》2009,20(6):359-366

Objective:  To determine the role of cervical cytology and colposcopy in the management of endocervical neoplasia.
Setting:  Colposcopy unit and cytology laboratory in a teaching hospital.
Sample:  Group 1 included 184 smears showing endocervical glandular neoplasia from 129 patients and group 2 included 101 patients with histology showing endocervical abnormalities in a 6-year period (1993–1998). Follow-up of 6–11 years to 2004 was available.
Methods:  Group 1 were identified from the cytology computer records. Group 2 were identified from histology records on the cytology database and a record of histology cases kept for audit purposes. The clinical records were examined retrospectively.
Results:  The positive predictive value (PPV) of abnormal endocervical cells in smears was 81.1% for significant glandular/squamous [cervical glandular intraepithelial neoplasia (CGIN)/cervical intraepithelial neoplasia grade2 (CIN2 or worse)] lesions. The PPV of colposcopy was 93.5% for significant glandular/squamous lesions of the cervix. The postcolposcopy probability of a significant lesion when colposcopy was normal was 87.5%. The sensitivity of colposcopy in detecting endocervical lesions was 9.8%. The sensitivity of cervical smears in detecting a significant endocervical abnormality (CGIN or worse) was 66.3%. The false negative rate for cytology of endocervical glandular lesions was 4.0%.
Conclusions:  Endocervical glandular neoplasia detected on cytology is predictive of significant cervical pathology even when colposcopy is normal, which supports excisional biopsy in the primary assessment of these smears. The high concomitant squamous abnormality rate justifies the use of colposcopy to direct biopsies from the ectocervix. Cervical cytology is the only current screening method for cervical glandular abnormalities but sensitivity is poor.     

P-14CERVICAL DYSPLASIA AND RISK FACTORS IN NORTHERN IRELAND SLE PATIENTS

Introduction:  An increased risk of cervical dysplasia has previously been reported in lupus. The role of traditional related risk factors are unclear. Persistent infection with certain genotypes of Human Papilloma Virus termed high risk (HR-HPV) is considered a prerequisite for the development of cervical cancer.
Methods:  A total of 141 SLE patients fulfilling the ACR criteria for lupus were enrolled into the study. They were compared to 138 control patients who were due to attend for a routine cervical smear. Each patient completed a cervical cancer risk factor questionnaire. A cervical smear was taken and residual material used to test for the presence of HR-HPV using a hybridisation microplate assay. Positive samples were then genotyped.
Results:   

     

Cervical smear adequacy: cellularity references were found to increase both interobserver agreement and unsatisfactory rate

Objectives:  To determine the degree of interobserver variation in the assessment of conventional cervical smear adequacy as defined by The Bethesda System (TBS) 2001, and to determine the effect of using reference images of known squamous cellularity when performing squamous adequacy assessments.
Methods:  Experimental pre-test/post-test design utilizing 70 conventionally prepared cervical smears. Sample smears containing scant squamous cellularity were independently rated on two occasions by six cytotechnologists. Time 1 was without the use of reference images, and Time 2 was aided by cellularity reference images. The κ statistic was used to compare rater agreement.
Results:  The level of agreement increased from an average κ of 0.26 (SD 0.10) for Time 1, to an average κ of 0.40 (SD 0.15) for Time 2. The difference in mean κ values at the two assessments was statistically significant ( t  = 3.71; P  = 0.002). Unanimous agreement among the raters was observed for 15 samples (21.42%) at Time 1 (only one of which was classified as unsatisfactory) and 21 samples (30.00%) at Time 2 (12 of which were classified as unsatisfactory).
Conclusion:  Interobserver agreement increased after cellularity reference images were implemented. Using TBS 2001 squamous adequacy criteria and images of known squamous cellularity as references resulted in a decreased number of smears reported as satisfactory.     

P-27ATYPICAL GLANDULAR CELLS ON SMEAR – AGE RELATED IMPLICATIONS AND MANAGEMENT

Introduction:  There is ongoing debate about the terminology used in the classification of dyskaryosis, including whether BNA smears should be classified according to whether koilocytes are present or not. We explored the effect of koilocytosis in the management experiences and clinical outcomes of women with a single BNA smear.
Methods:  This study includes 410 women aged 20–59 years, resident in Tayside who had an 'baseline' BNA smear between 31/10/1999 and 31/10/2002 who were eligible for the TOMBOLA trial but who did not participate. Recommended follow-up for these women was a repeat smear in six months. Up to three-years follow-up data was collected on subsequent cytological smears, colposcopy examinations and any related histology. The baseline smear was re-read for the presence or absence of koilocytes. Women were classified according to their clinical outcomes during their three years.
Results:  47% (192/410) of women were classified as having koilocytosis at baseline. The mean age was lower among these women (25 years) compared to those without koilocytosis (35 years). 55% (105/192) of women with koilocytosis at baseline had one or more negative smears and no further abnormal smears, compared to 63% (137/218) of those without koilocytosis, the difference was not statistically significant. 28% (53/192) of women with koilocytosis at baseline were referred to colposcopy after subsequent low-grade or high-grade smears compared to 19% (42/218) of those without koilocytosis; the difference was statistically significant. 14% (27/192) of women with koilocytosis at baseline were diagnosed with CIN2/3 or worse during follow-up compared to 10% (21/218) of women without koilocytosis; the difference was not statistically significant.
Discussion:  Our results suggest that the presence or absence of koilocytosis has little impact on the clinical outcomes of women with a BNA smear.     

Utility of the in situ detection of HPV in Pap smears diagnosed as within normal limits.

OBJECTIVE: To determine the clinical significance in normal Pap smears of HPV detection as determined by Hybrid Capture (HC) and in situ hybridization analyses. STUDY DESIGN: We studied 135 consecutive Pap smears as well as 46 other smears from high-risk patients each initially diagnosed as within normal limits. RESULTS: The 135 "normal" Pap smears were rescreened, and 6 (4%) where found to be either ASCUS or SIL. In the remaining 129 cases, HPV DNA was detected in 0% and 9%, respectively, using in situ hybridization and HC I. Upon rescreening the high-risk patients, nine (20%) were reclassified as having SIL/ASCUS; each was in situ hybridization positive, and eight were HC positive; six (67%) of these women developed SIL on follow-up. In the 37 Pap smears in high-risk women still within normal limits after manual rescreening, HPV was detected in 2% by in situ hybridization and 46% by HC; 6% of the HC-positive women developed SIL on follow-up. CONCLUSION: In situ hybridization rarely detects HPV in Pap smears diagnosed as within normal limits after manual rescreening. In situ hybridization is very effective in detecting rare, atypical cells in Pap smears diagnosed as within normal limits and, in a high-risk population, is predictive of SIL on clinical follow-up.     

Clostridia spore formation during aerobic deterioration of maize and sorghum silages as influenced by Lactobacillus buchneri and Lactobacillus plantarum inoculants

Aims:  The effect of the inoculation of maize and sorghum silages with Lactobacillus plantarum (LP) and Lactobacillus buchneri (LB) on the clostridia spore formation during aerobic deterioration has been studied.
Methods and results:  The crops were ensiled in 30 l jars, without a lactic acid bacteria inoculant (C), and with an LP or LB inocula (theoretical rate of 1 × 10⁶). After 90 days of conservation, the silages were analysed for the chemical and microbiological characteristics and subjected to an aerobic stability test, during which pH, temperature, nitrate, yeast, mould and clostridia spores were measured. Compared to the C and LP silages, yeasts were reduced in the LB silages, resulting in an increased aerobic stability. Clostridia spores, determined by most probable number (MPN) procedure, increased to 6 log₁₀ MPN g⁻¹ in the C and LP maize silages, whereas they reached 3 log₁₀ MPN g⁻¹ in C and LP sorghum silages.
Conclusions:  Clostridia spore count only slightly increased in the LB maize silages after 342 h (2·59 log₁₀ MPN g⁻¹), whereas it did not show any increase in the LB sorghum silages for the whole period of air exposure.
Significance and impact of the study:  The data indicated that clostridia spore outgrowth can take place during silo feedout in aerobic-deteriorated silages and that LB inoculation reduces the risk of clostridia outgrowth after silage opening by increasing the aerobic stability.     

Evaluation of Papanicolaou stain for studying micronuclei in buccal cells under field conditions

OBJECTIVE: To compare Papanicolaou (Pap) and May-Grünwald Giemsa (MGG) stain as 2 techniques for staining for buccal mucosal cells to detect micronuclei (MN) infield studies. STUDY DESIGN: Eighty cytologic smears (2 per individual) were taken from the buccal mucosa of 40 cigarette smokers recruited at a rural village in Egypt. Forty smears were stained with Pap stain and 40 with MGG stain. All were assessed for cellularity and scored for MN. RESULTS: Pap stain was faster and easier to process and transport in the field study than was MGG stain. Regarding MGG smears, bacteria and cell debris masked the MN as compared to Pap smears, in which the fixative destroyed the bacteria and made the cell boundaries clearly demarcated. Using Pap stain, MN were seen easily in transparent cytoplasm. CONCLUSION: Pap stain is the preferred method infield studies for scoring and detecting MN in cells of buccal mucosa.     

Altered expression of adhesion molecules in inflammatory cervical smears

Objective:  The aim of the present study was to evaluate the expression of pan-cadherin and β-catenin in cervical smears with various types of infectious agents.
Patients and Methods:  Cervical smears obtained from 53 women, aged 21–65 years, with a diagnosis of specific inflammation were examined in our study. Eighteen subjects were infected by Candida albicans , 18 by Gardnerella vaginalis , nine by Bacteroides spp. and eight by Chlamydia trachomatis . All infectious agents found in the smears were at the same time confirmed by the microbiological laboratory methods. We performed a biotin–streptavidin-peroxidase immunocytochemical method using anti-β-catenin (Clone 12F7) and anti-pan-cadherin (pan, polyclonal) antibodies.
Results:  Aberrant expression of pan-cadherin was found in the cytoplasmic membrane of glandular, metaplastic, superficial and intermediate squamous cells in all types of infections. With regard to β-catenin, this was expressed in majority (90%) of glandular and metaplastic cells in all types of infections and in a small proportion (15%) of superficial and intermediate squamous cells in infections caused by C. albicans and G . vaginalis .
Conclusion:  Our data show that infectious agents may cause alterations in the expression and distribution of these adhesive molecules, which can be recognized in cervical smears. Additional studies in larger sets of patients should help clarify this issue further.     

Performance of the AutoPap Primary Screening System in the detection of high-risk cases in cervicovaginal smears

OBJECTIVE: To assess the performance of the AutoPap Primary Screening System (APSS) (TriPath Imaging, Inc., Burlington, North Carolina, U.S.A.) for the detection of high grade cervical squamous intraepithelial lesions and invasive cervical cancer. STUDY DESIGN: A total of 14,779 consecutive conventional Pap smears were processed by the APSS. All slides designated as "Review" by the device were manually screened according to the Bethesda System. The ranking scores obtained from the device were compared with the cytologic interpretations in all cases and with the final histologic diagnoses in the cases with cytologic severe abnormalities. RESULTS: The device classified 10,349 slides as Review (78%) and 2,912 (22%) as "No Further Review." In the 78% Review cases, the samples were ranked in descending order of potential abnormality, broken into quintiles. The correlation between the slide quintile ranks and the manual cytologic diagnosis indicated that 90% of abnormal smears were categorized by the device as in the first and second quintile rank, and the correlation between the rank report of the device and the histologic diagnosis showed that all cases of HSIL or invasive carcinoma were in the top two ranks. No significant abnormalities were observed in any of the smears categorized as No Further Review. CONCLUSION: This study confirmed the effectiveness of APSS for the detection of Pap smears with severe abnormalities.     

Rapid pre-screening of cervical smears as a method of internal quality control in a cervical screening programme

Objective:  To evaluate the performance of rapid pre-screening (RPS) as a method of internal quality control in the cytopathological examination of cervical smears for cervical cancer screening.
Methods:  The sample consisted of 6135 cervical smears submitted to RPS and routine screening (RS) methods. The smears classified as negative in RPS and RS were considered final diagnoses, and were not, therefore, submitted to any additional review. The smears identified as suspect or unsatisfactory according to RPS were analysed separately by two different cytologists irrespective of the diagnosis reached in RS. Smears considered abnormal or unsatisfactory at RS were also reviewed. When both cytologists issued concordant diagnoses, this was considered the final diagnosis. Discordant results were analysed by a third cytologist and a consensus meeting was held to define the final diagnosis.
Results:  Taking abnormalities detected by RS as the denominator, RPS had a sensitivity of 63.0% for the detection of all abnormal smears and 96.7% for high grade squamous intraepithelial lesion (HSIL). When compared with the final diagnosis, sensitivity of RPS for all abnormal smears was 74.9% and for HSIL 95.0%. Of the 529 abnormal smears confirmed in the final diagnosis, 2.15% were detected only by the RPS.
Conclusion:  RPS is an effective alternative method of internal quality control with high sensitivity for the detection of more severe lesions. It also permits monitoring of the laboratory rate of false-negative results, and allows constant evaluation of the performance both of the pre-screening and RS cytologists.     

Decrease in numbers of glandular cell groups in post-LLETZ liquid-based cytology preparations

Objective:  Large loop excision of the transformation zone (LLETZ) has become standard of care in the management of cervical squamous neoplasia and with cone biopsy glandular intraepithelial neoplasia. Controversy remains about the long-term effects of this traumatic procedure. The aim of this study was to count and compare the number of endocervical glandular cell groups in pre- and post-LLETZ cervical preparations using liquid-based cytology to establish a cyto-morphological correlate of destruction of the transformation zone.
Methods:  The cytology/histology correlation audit records of the Cytopathology Department of St Luke's Hospital in 2003 and early 2004 were used to select patients with a cytological diagnosis of high grade dyskaryosis followed by LLETZ. Only those cases with post-LLETZ cytological follow-up were selected. Cases using conventional smears were excluded. One hundred and twenty slides (60 pairs of slides) in total were retrieved. The cases underwent review and all groups of >3 glandular cells in each slide were counted by AM while blinded as to whether smears were pre- or post-LLETZ. Medians were compared using a Mann–Whitney U -test.
Results:  The median number of groups of endocervical glandular cells of the pre-treatment group was 5.5 and of the post-treatment group was 2.0. There were significantly fewer endocervical glandular cell groups in the post-LLETZ population ( P  = 0.03).
Conclusions:  The number of endocervical glandular groups in cervical cytological preparations decreases significantly following LLETZ procedure. This suggests that cytological follow-up may not be as useful in glandular neoplasia cases. Few or absent glandular cell groups in post-LLETZ preparations may have implications for adequacy assessment.     

The role of cervical cytology and colposcopy in detecting cervical glandular neoplasia

Objective:  To determine the role of cervical cytology and colposcopy in the management of endocervical neoplasia.
Setting:  Colposcopy unit and cytology laboratory in a teaching hospital.
Sample:  Group 1 included 184 smears showing endocervical glandular neoplasia from 129 patients and group 2 included 101 patients with histology showing endocervical abnormalities in a 6-year period (1993–1998). Follow-up of 6–11 years to 2004 was available.
Methods:  Group 1 were identified from the cytology computer records. Group 2 were identified from histology records on the cytology database and a record of histology cases kept for audit purposes. The clinical records were examined retrospectively.
Results:  The positive predictive value (PPV) of abnormal endocervical cells in smears was 81.1% for significant glandular/squamous [cervical glandular intraepithelial neoplasia (CGIN)/cervical intraepithelial neoplasia grade2 (CIN2 or worse)] lesions. The PPV of colposcopy was 93.5% for significant glandular/squamous lesions of the cervix. The postcolposcopy probability of a significant lesion when colposcopy was normal was 87.5%. The sensitivity of colposcopy in detecting endocervical lesions was 9.8%. The sensitivity of cervical smears in detecting a significant endocervical abnormality (CGIN or worse) was 66.3%. The false negative rate for cytology of endocervical glandular lesions was 4.0%.
Conclusions:  Endocervical glandular neoplasia detected on cytology is predictive of significant cervical pathology even when colposcopy is normal, which supports excisional biopsy in the primary assessment of these smears. The high concomitant squamous abnormality rate justifies the use of colposcopy to direct biopsies from the ectocervix. Cervical cytology is the only current screening method for cervical glandular abnormalities but sensitivity is poor.