Examination for the Certificate in Advanced Practice in Cervical Cytology – the first year''s experience

Following the decision to establish an Advanced Biomedical Scientist Practitioner grade for senior biomedical scientists in the NHS Cervical Screening Programme, a conjoint examination board has been appointed by the Royal College of Pathologists and Institute of Biomedical Science to oversee the Certificate in Advanced Practice in Cervical Cytology examination. The examination consists of a multiple-choice paper, short-answer written questions and practical microscopy sections covering screening of unmarked slides, and more complex discussion cases. In the first year there were 58 entries with 29 successful candidates, a pass rate of 50%. The standard of performance in the examination showed a wide range, and some candidates appear to have underestimated the degree of preparation, knowledge or level of microscopy skill required.     

The Cardiff Cervical Cytology Study: prevalence of cytological grades and initial histological findings.

Among 45 266 women in the Cardiff Cervical Cytology Survey the peak prevalence of suspicious or positive smears was 11.2/1000 at age 45-50 years and of dyskaryosis 10.2/1000 at age 25-29. A suspicious or positive cytological picture at prevalence testing was associated with occult or clinical invasion in 24% of cases, and only 4% of patients with suspicious or positive smears were normal histologically. When dyskaryosis was detected in the prevalence test 20% had carcinoma in situ or microinvasion and 3% had occult or clinically invasive carcinoma. One hundred and twenty-nine (51%) women with dyskaryotic smears did not have a biopsy initially (that is, within two years of the prevalence test), but they were followed up at regular intervals. Subsequently 15 of the 129 gave smears consistently dyskaryotic or worse cytologically and were subjected to biopsy. Of these, two showed dysplasia, 12 carcinoma in situ, and one clinically invasive carcinoma. These findings emphasise the need for repeat cytological or histological examination in any woman with evidence of dyskaryosis in a cervical smear.     

Can older women be motivated to attend for their final Papanicolaou tests? The use of targeted and general personalised reminder letters

Background: Australian guidelines allow women to cease having Papanicolaou (Pap) tests at the age of 70, providing they have had two negative smears in the previous 5 years. In Victoria, a standard reminder letter system is in place and women receive one reminder letter 27 months after their last Pap test. Methods: A targeted letter informed by qualitative research was developed to specifically encourage older women to have a final Pap test. The Victorian Cervical Cytology Registry (VCCR) was used to identify 15,000 women aged 65–69 who had not had a Pap test for at least 3 years and up to 17 years. Women were randomly allocated to receive either no letter (control group), the targeted letter or a personally addressed general reminder letter. Results: At 11 weeks follow-up, 4.3% of women (CI 3.7–4.8%) who received the targeted letter had attended for a Pap test compared with 4.7% (CI 4.1–5.3%) of those receiving the general letter and 1.6% (CI 1.2–1.9%) of the control group. Effectiveness was limited to women whose test was no more than 10 years overdue, and was particularly strong for those whose test was 3–5 years overdue. Conclusions: This suggests that a reminder letter was effective, but that targeting the information in the letter did not further improve screening attendance. A second reminder letter sent to women before their Pap test is more than 5 years overdue has potential for increasing the number of women attending for screening.     

Cytology of Callus of Cryptomeria japonica

Callusing has been initiated from the cotyledons of 4-week-old seedlings of Cryptomeria japonica on Murashige and Skoog's medium supplemented with 22.8 μM IAA. Callus was grown successfully for more than 2 years on the same medium without any decline in growth vigour. The callus has shown differentiation of tracheids with reticulate thickenings but differentiation of root, shoot or whole plant has not yet been observed. Cytology of 2-month-old callus showed the presence of cells containing normal diploid chromosome complement of 2n = 22 and normal mitosis. However, the 6-month-old callus revealed majority of cells with normal diploid complement and the usual karyotype, but a few cells were noticed with aneuploid numbers ranging from 2n = 15–17. Anaphase bridges and lagging chromosomes have also been encountered.     

Primary Screening for Cervical Cancer Based on High-Risk Human Papillomavirus (HPV) Detection and HPV 16 and HPV 18 Genotyping,in Comparison to Cytology

Objectives

The objective of the present study is to assess the performance of a high-risk human papillomavirus (HR-HPV) DNA test with individual HPV-16/HPV-18 genotyping as a method for primary cervical cancer screening compared with liquid-based cytology (LBC) in a population of Greek women taking part in routine cervical cancer screening.

Methods

The study, conducted by the “HEllenic Real life Multicentric cErvical Screening” (HERMES) study group, involved the recruitment of 4,009 women, aged 25–55, who took part in routine cervical screening at nine Gynecology Departments in Greece. At first visit cervical specimens were collected for LBC and HPV testing using the Roche Cobas 4800 system. Women found positive for either cytology or HPV were referred for colposcopy, whereas women negative for both tests will be retested after three years. The study is ongoing and the results of the first screening round are reported herein.

Results

Valid results for cytology and HPV testing were obtained for 3,993 women. The overall prevalence of HR-HPV was 12.7%, of HPV-16 2.7% and of HPV-18 1.4%. Of those referred for colposcopy, cervical intraepithelial neoplasia grade 2 or worse (CIN2+) was detected in 41 women (1.07%). At the threshold of CIN2+, cytology [atypical squamous cells of undetermined significance (ASC-US) or worse] and HPV testing showed a sensitivity of 53.7% and 100% respectively, without change between age groups. Cytology and HPV testing showed specificity of 96.8% and 90.3% respectively, which was increased in older women (≥30) in comparison to younger ones (25–29). Genotyping for HPV16/18 had similar accuracy to cytology for the detection of CIN2+ (sensitivity: 58.5%; specificity 97.5%) as well as for triage to colposcopy (sensitivity: 58.5% vs 53.7% for cytology).

Conclusion

HPV testing has much better sensitivity than cytology to identify high-grade cervical lesions with slightly lower specificity. HPV testing with individual HPV-16/HPV-18 genotyping could represent a more accurate methodology for primary cervical cancer screening in comparison to liquid-based cytology, especially in older women.     

Human Papillomavirus Assays and Cytology in Primary Cervical Screening of Women Aged 30 Years and Above

In women aged ≥30 years, Human Papillomavirus testing will replace cytology for primary cervical screening. We compared Hybrid Capture 2 (HC2), cobas, CLART, and APTIMA HPV assays with cytology on 2869 SurePath samples from women undergoing routine screening at 30–65 years in Copenhagen, Denmark. Women with cytological abnormalities were managed according to routine recommendations, with 92% completeness. Those with cytology-normal/HPV-positive samples (on any of the four assays) were invited for repeated cytology and HPV testing in 1.5 year, and 58% had additional testing. HPV testing detected more ≥CIN3 than cytology (HC2: 35, cobas, CLART: 37, APTIMA: 34, cytology: 31), although statistically the differences were not significant. Cobas and CLART detected significantly more ≥CIN2 than cytology (cobas, CLART: 49, cytology: 39). The proportion of women with false-positive test results (positive test results without ≥CIN3) varied between 3.3% with cytology and 14.9% with cobas. All HPV assays led to significantly more false-positive tests, whereas compared to HC2 cobas and CLART were associated with a significantly higher and APTIMA with a significantly lower proportion. Detection of CIN1 was particularly increased for the three DNA assays. With APTIMA combined with cytological triage, about 20% more women were referred for colposcopy than with cytology screening. With the three DNA assays, the increase was ≥50%. The number of women with repeated testing was twice as high with APTIMA and almost five times as high with cobas compared to cytology. To our knowledge, Horizon was the only study set in routine practice that compared more than two HPV assays in the same women while also ascertaining the histological status of women with normal cytology/HPV-positive test results. HPV-based screening of Danish women aged 30–65 detected more high-grade CIN but decreased the screening specificity, and increased the demand for additional testing.     

Histology Verification Demonstrates That Biospectroscopy Analysis of Cervical Cytology Identifies Underlying Disease More Accurately than Conventional Screening: Removing the Confounder of Discordance

Background

Subjective visual assessment of cervical cytology is flawed, and this can manifest itself by inter- and intra-observer variability resulting ultimately in the degree of discordance in the grading categorisation of samples in screening vs. representative histology. Biospectroscopy methods have been suggested as sensor-based tools that can deliver objective assessments of cytology. However, studies to date have been apparently flawed by a corresponding lack of diagnostic efficiency when samples have previously been classed using cytology screening. This raises the question as to whether categorisation of cervical cytology based on imperfect conventional screening reduces the diagnostic accuracy of biospectroscopy approaches; are these latter methods more accurate and diagnose underlying disease? The purpose of this study was to compare the objective accuracy of infrared (IR) spectroscopy of cervical cytology samples using conventional cytology vs. histology-based categorisation.

Methods

Within a typical clinical setting, a total of n = 322 liquid-based cytology samples were collected immediately before biopsy. Of these, it was possible to acquire subsequent histology for n = 154. Cytology samples were categorised according to conventional screening methods and subsequently interrogated employing attenuated total reflection Fourier-transform IR (ATR-FTIR) spectroscopy. IR spectra were pre-processed and analysed using linear discriminant analysis. Dunn’s test was applied to identify the differences in spectra. Within the diagnostic categories, histology allowed us to determine the comparative efficiency of conventional screening vs. biospectroscopy to correctly identify either true atypia or underlying disease.

Results

Conventional cytology-based screening results in poor sensitivity and specificity. IR spectra derived from cervical cytology do not appear to discriminate in a diagnostic fashion when categories were based on conventional screening. Scores plots of IR spectra exhibit marked crossover of spectral points between different cytological categories. Although, significant differences between spectral bands in different categories are noted, crossover samples point to the potential for poor specificity and hampers the development of biospectroscopy as a diagnostic tool. However, when histology-based categories are used to conduct analyses, the scores plot of IR spectra exhibit markedly better segregation.

Conclusions

Histology demonstrates that ATR-FTIR spectroscopy of liquid-based cytology identifies the presence of underlying atypia or disease missed in conventional cytology screening. This study points to an urgent need for a future biospectroscopy study where categories are based on such histology. It will allow for the validation of this approach as a screening tool.     

Cytology of lumpectomy specimens.

Residual microscopic disease after lumpectomy may cause significant local recurrence. This study evaluated the use of touch preparation cytology to assess lumpectomy margins. For 90 specimens, the findings on Diff-Quik-stained touch preparations (rendered intraoperatively within 15 minutes after the lumpectomy) were correlated with the gross findings, frozen-section results and, later, the findings in the permanent histologic sections. Three specimens were cytologically unsatisfactory, 68 yielded benign findings, and 19 were suspicious or diagnostic for malignancy. The margins showed tumor involvement in 5 lumpectomy samples by gross examination, in 13 by frozen-section evaluation and in 17 by the study of permanent sections. Touch preparation cytology was putatively falsely positive in two cases while frozen-section evaluation was falsely negative in four cases. Cytology had a sensitivity of 100%, a specificity of 97.1% and a diagnostic accuracy of 97.7%. These results demonstrate that touch preparation cytology rapidly and reliably evaluates lumpectomy margins and can overcome some sampling errors and artifacts related to frozen-section analysis. Touch preparation cytology is now being used to complement frozen-section evaluation of lumpectomy margins as part of a protocol aimed at reducing the local recurrence rate.     

Cytopathology laboratory accreditation, with special reference to the American Society of Cytology programs

The features of the Laboratory Accreditation Program of the American Society of Cytology that pertain to quality assurance in cytopathology are reviewed. The areas considered include: (1) specimen procurement and cytopreparation, (2) the role of the cytotechnologist in cytoscreening, evaluation and reporting, (3) the role of the cytopathologist and (4) quality control measures. Attention to the issues raised in these areas is essential to achieving the best possible cytopathology practice in the most efficient and economical manner.     

Estimating the Accuracy of Anal Cytology in the Presence of an Imperfect Reference Standard

Background

The study aim is to estimate sensitivity and specificity of anal cytology for histologic HSIL in analyses adjusted for the imperfect biopsy reference standard.

Methods and Principal Findings

Retrospective cohort study of an anal dysplasia screening program for HIV infected adults. We estimated the prevalence of histologic HSIL by concurrent cytology category and the associated cytology ROC area. Cytology operating characteristics for HSIL were estimated and adjusted for the imperfect reference standard by 3 methodologies. The study cohort included 261 patients with 3 available measures: (1) referral cytology; (2) HRA cytology; and (3) HRA directed biopsy. The prevalence of biopsy HSIL varied according to the concurrent HRA cytology result: 64.5% for HSIL or ASC-H, 12.6% for LSIL, 10.9% for ASCUS, and 6.3% for no abnormality. The cytology ROC area was 0.78. The observed prevalence of HSIL was 37% (referral cytology), 24% (HRA cytology), and 24% (HRA biopsy). Unadjusted estimates of sensitivity and specificity of cytology were 0.66 and 0.90, respectively. Adjusted estimates varied from 0.47–0.89 (sensitivity) and 0.89—1.0 (specificity).

Conclusions

Analysis of a single dataset yields widely different estimates of anal cytology operating characteristics that depend on difficult to verify assumptions regarding the accuracy of the imperfect reference standard.     

Nasseem Husain: homage to a pioneer of cytology automation

Dr Oliver Anthony Nasseem Husain, who died on 22 September 2014, aged 90 years, was one of the great names of European cytology, a pioneer of automated cervical screening and a founding member of both the British Society for Clinical Cytology (BSCC) and the European Federation of Cytology Societies (EFCS). The life of this one remarkable man involved much of the pioneering work, which is reviewed in this article, that has brought conventional cytology to the complex multimodal discipline it is today.     

O‐110 YEAR AUDIT OF GLANDULAR CYTOLOGY REPORTS WITH SPECIAL REGARD TO THE EFFECT OF CONVERSION TO LBC

Objectives: A randomised trial in screening to improve cytology (ARTISTIC Trial) was set up to evaluate the effectiveness of human papilloma virus (HPV) testing in primary cervical screening. Design: Women attending the NHS Cervical Screening Programme in Greater Manchester were screened for HPV in addition to routine cytological testing with ThinPrep Liquid‐Based Cytology (LBC). 25 020 women aged 20–64 were screened at entry and 3 years later. Participants were randomised in a 3 : 1 ratio between those whose HPV test result was revealed and acted upon and those in whom it was concealed. Results: A total of 24 510 women (18 386 revealed and 6124 concealed) had adequate cytology and HPV results. Cytology results at entry were: 87% normal, 11% borderline or mild, 1.1% moderate and 0.6% severe dyskaryosis or worse. Prevalence of HPV decreased sharply with age, from 40% at age 20–24 to 12% at 35–39 and 7% or less above age 50, and increased with cytological grade, from 10% (normal cytology) to 31% (borderline), 70% (mild), 86% (moderate), and 96% (severe or worse). Only 60% of women with severe dyskaryosis showed HPV 16 and / or 18 infection. Conclusion: The ARTISTIC Trial has provided the largest collection of LBC samples with HPV typing in Greater Manchester population. It has shown that the HPV testing is practicable in routine screening in women aged over 30. HPV 16 and 18 are more predictive for underlying disease, but other HPV types account for 30% of high‐grade disease.     

Micellar effects upon the reaction of cobalt complexes with alkyl halides.

The reactions of the cobalozime Co (DH)-2 and the related Co(DOH)DOpn0 are strongly catalyzed by cationic micelles of cetyltrimethylammonium bromide (CTABr). The rate enhancements for the cobaloxime reaction and for that of Co (DOH)DOpn0 (in parentheses) are: EtBr approximately 1 (8.5); n-C5H11Cl, 220 (200); ClCH2CO-2, 420 (173); ClCH2CH2CO-2, 373. Anionic micelles of sodium lauryl sulfate inhibit the reaction of Co(DOH)DOpn0 with ClCH2CO-2, but do not affect that with n-C5H11Cl. The reactions of Co(DOH)DOpn0, like those of the cobaloxime are SN2 displacements and in the absence of surfactant in MeOH n-PrBr is more reactive than iso-PrBr.     

Cytology and Human Papillomavirus Co-Test Results Preceding Incident High-Grade Cervical Intraepithelial Neoplasia

Objective

High-risk HPV (hrHPV) and cytology co-testing is utilized for primary cervical cancer screening and for enhanced follow-up of women who are hrHPV-positive, cytology negative. However, data are lacking on the utility of this method to detect pre-cancer or cancer in community-based clinical practice. This study describes cytology and hrHPV results preceding high-grade cervical intraepithelial neoplasia, adenocarcinoma in situ, or cervical cancer (i.e., CIN2+) in an integrated health system employing routine co-testing among women aged 30 years and older.

Methods

We conducted a cross-sectional analysis of adult female members of Kaiser Permanente Northern California (KPNC) with incident CIN2+ between July 2008 and June 2009. The primary outcome was the proportions of cytologic diagnoses and hrHPV co-test results preceding a diagnosis of CIN2+. Cervical cytology and hrHPV testing results were abstracted from electronic medical records.

Results

Of 1283 CIN2+ cases among adult women, 880 (68.5%) were among women aged 30 years and older and 145/880 (16.5%, 95% CI 14.1–19.1) had only normal cytology during the 12 months prior to diagnosis. Furthermore, 133/880 (15.1%, 95% 12.9–17.7) were preceded by only normal cytology and persistent hrHPV infection (at least 2 positive hrHPV tests) during the 6–36 months preceding CIN2+ diagnosis.

Conclusions

Incident CIN2+ is frequently preceded by normal cytology and persistent hrHPV infection among women aged 30 years and older; screening strategies that employ HPV testing and cytology may improve the detection of CIN2+ compared with cytology alone.     

Micellar catalysis of the alkylation of mercuric ions by alkyl cobalt(III) complexes.

Anionic micelles of sodium lauryl sulfate, NaLS, catalyze the monoalkylation of Hg2+ in dilute acid by alkyl aquobis-(dimethyl glyoximato) cobalt (III), RCo(DH)2(H2O) degrees and the related propane derivatives RCo(DOH) DOpn (H2O)+, where R = Me, Et, n-C5H11. Nonionic micelles of Igepal do not catalyze the reaction. In the absence of micelles RCo(DH)2(H2O) degrees is considerably more reactive than RCo(DOH)DOpn(H2O)+, but this higher reactivity is offset in part by its higher basicity. Anionic micelles markedly increase the basicity of RCo(DOH)DOpn(H2O)+ and slightly increase that of RCo(DH)2(H2O) degrees. For reactions of the unprotonated Co(III) complexes the maximum rate enhancements by micelles of NaLS are: R = Me, 19(131); Et, 58 (65); n-C5H11, 46 (32). (The values in parentheses are for RCo(DOH)DOpn(H2O)+.)     

国产12种乌头属和18种翠雀属植物的细胞学研究

研究了12种乌头属Aconitum L．和18种翠雀属Delphinium L．植物的染色体。在12种乌头属植物中,除粗花乌头A．crassiflorum为四倍体(2n=4x=32)外,其他种类都为二倍体(2n=2x＝16),中甸乌头 A.piepunense中有B染色体存在,牛扁亚属Aconitum subgen．Lycoctonum的二倍体植物与乌头亚属Aconitum subgen．Aconitum 植物的染色体在大小和形态上有明显区别;所有18种翠雀属植物都为二倍体(2n＝2x=16),其染色体在大小和形态上极为相似,但与乌头亚属的染色体易于区别。翠雀属植物的核型不对称性程度明显高于乌头属植物,因此从染色体证据来看,翠雀属要比乌头属进化。     

新型冠状病毒相关靶点小分子抑制剂虚拟筛选

为确定治疗新型冠状病毒(SARS-CoV-2)感染的候选药物,开展了针对SARS-CoV-2的药物虚拟筛选研究。以SARS-CoV-2的刺突蛋白(S蛋白)和3CL蛋白酶(主蛋白酶)作为药物靶点,以美国食品药品监督管理局(FDA)批准上市的2 726个小分子药物作为候选,通过分子对接方法,筛选出了3种(阿巴瑞克(Abarelix)、西曲瑞克(Cetrorelix)、鞣酸(Tannic acid))与S蛋白具有较强结合能力的小分子药物,1种(戈舍瑞林(Goserelin))与3CL蛋白酶具有较好结合能力的小分子药物,它们理论上都具有抑制新型冠状病毒复制的效果。将靶向3CL蛋白酶的候选药物与辉瑞公司开发的药物Paxlovid进行比较,发现其作用位点均集中于3CL蛋白酶的第130-200位的残基周围,具有相似的结合位点与相互作用。此外也对候选药物的物理与化学性质及与基因相互作用进行了分析。本研究可为开发新型冠状病毒感染的治疗药物提供参考。     

Epilogue. Establishment of the Center for Chronic Disease Prevention and Health Promotion

The establishment of the Center for Chronic Disease Prevention and Health Promotion (CCDPHP) at the Centers for Disease Control (CDC) following the Conferences on the State of the Art in Quality Control Measures for Diagnostic Cytology Laboratories is briefly discussed. The CCDPHP is expected to play a major role in the CDC's cancer control program, including participation in establishing effective screening programs and assuring the quality of such methodologies as the Papanicolaou test and mammography.     

Cytology of pseudomyxoma peritonei

Cytology of pseudomyxoma peritonei
The cytology of five cases of pseudomyxoma peritonei is described. It is characterized by large masses of mucinous material, some of which is wrapped up in a network of elongated fibroblast-like cells. Irregular clusters of flat branching vimentin-positive fibroblast-like cells may be found as the disease progresses. Goblet or tumour cells were not found in the cytology preparations, although they were seen in the histological sections of the one case which was biopsied. The appendiceal origin of pseudomyxoma peritonei could not be traced in three of four male patients, and the cause of the disease remained undetermined. Autopsy of the fourth case demonstrated a complete replacement of the appendix by mucus.     

Pharmacodynamics and pharmacokinetics studies of phenoxazinium derivatives for antimalarial agent

In vivo antimalarial drug candidates screening test was carried out on a series of water-soluble 3,7-bis(dialkylamino)phenoxazin-5-ium derivatives. Among them, 3-(diethylamino)-7-(piperidin-1-yl)phenoxazin-5-ium chloride (SSJ-206) showing highest efficacy was chosen for further pharmcodynamics and pharmacokinetics study. It was supported from these data that the phenoxazinium salts, SSJ-206, would be one of hopeful candidates as an oral antimalarial drug.