Interlaboratory reproducibility of atypical glandular cells of undetermined significance: a national survey

OBJECTIVE: The aim of this study was to evaluate the inter-laboratory reproducibility for atypical glandular cells (AGC) (The Bethesda System (TBS) 2001) of the laboratories involved in the screening programmes in Italy. METHODS: A set of 35 selected slides were circulated among 167 laboratories involved in local population-based cervical screening programmes. Each laboratory provided one single diagnosis per smear. The smears were read blind to the original diagnosis and to the diagnoses provided by other laboratories. A 'majority' diagnosis was defined for each case and assumed as the reference standard. The diagnosis provided from each laboratory was compared with the majority diagnosis. RESULTS: According to the majority report the 35 slides in the set were classified as negative in nine cases, AGC in eight, adenocarcinoma in eight, and squamous lesion or squamous + glandular lesion in 10. The crude agreement between all pairs of laboratories was 49.43%. K-values were 0.46, 0.21, 0.34, 0.36 and 0.32 for negative, AGC/AIS (adenocarcinoma in situ of endocervix), AdenoCa, Sq/Sq + Gl and all reporting categories respectively. Concordance according to overall K was moderate to substantial in 77% of the participating laboratories. CONCLUSIONS: The present study shows that the AGC category is not easily reproducible. The data confirmed the importance, in a screening scenario, of AGC/AIS diagnoses, but also presented difficulties in differentiating between the two diagnoses. In addition to the results obtained from the circulation of the slides, laboratories which had annually a low number of cervical smears were able to gain experience focused on particular morphological pictures.     

Test characteristics of various screening modalities for cervical cancer: a feasibility study to develop an alternative strategy for resource-limited settings

OBJECTIVE: To determine test characteristics - sensitivity, specificity, positive and negative predictive values - of different screening modalities to detect cervical precancerous and cancerous lesions in order to devise an effective alternative strategy for cervical cancer screening in resource-poor settings. METHODS: A total of 472 women presenting with nonspecific gynecologic symptoms were screened by cytology, visual inspection with acetic acid application (VIA), VIA with magnification (VIAM) and human papillomavirus (HPV)-DNA testing. Colposcopic examination was performed in all and on-site biopsy was taken if any grade I and above lesion was detected on colposcopy (230). On histopathological examination, 105 showed cervical intraepithelial neoplasia II and above lesions. Sensitivity, specificity and predictive values for each test were calculated taking colposcopy and or directed biopsy as the gold standard. Comparisons were made with cytology in order to assess the feasibility of alternative strategies in resource-poor settings. RESULTS: VIA was less sensitive (86.7% versus 91.4%) but more specific (90.7% versus 86.6%) than cytology at low grade squamous intraepithelial lesion (LSIL) threshold but the difference was not statistically significant (P > 0.01). HPV testing improved the sensitivity over cytology (97.1% versus 91.4%) but there was a nonsignificant loss of specificity (84.2% versus 86.6%). Results of VIAM were more or less similar to VIA. CONCLUSION: VIA can be used as a mass screening tool for cervical cancer in resource-poor settings.     

Cell blocks in cytopathology: a review of preparative methods,utility in diagnosis and role in ancillary studies

The cell block (CB) is a routine procedure in cytopathology that has gained importance because of its pivotal role in diagnosis and ancillary studies. There is no precise review in the published literature that deals with the various methods of preparation of CB, its utility in diagnosis, immunocytochemistry (ICC) or molecular testing, and its drawbacks. An extensive literature search on CB in cytology using internet search engines was performed for this review employing the following keywords: cell block, cytoblock, cytology, cytopathology, methods, preparation, fixatives, diagnostic yield, ancillary and molecular studies. Ever since its introduction more than a century ago, the CB technique has undergone numerous modifications to improve the quality of the procedure; however, the overall principle remains the same in each method. CBs can be prepared from virtually all varieties of cytological samples. In today's era of personalized medicine, cytological specimens, including CBs, augment the utility of cytological samples in analysing the molecular alterations as effectively as surgical biopsies or resection specimens. With the availability of molecular targeted therapy for many cancers, a large number of recent studies have used cytological material or CBs for molecular characterization. The various techniques of CB preparation with different fixatives, their advantages and limitations, and issues of diagnostic yield are discussed in this review.     

Assessing the wellbeing of cytoscreeners: experience in two NHS cytology laboratories

OBJECTIVE: Cervical screening programmes in England are in transition as the liquid-based cytology (LBC) method replaces conventional Papanicolaou screening and staff in NHS laboratories are trained to analyse LBC smears. Cytoscreeners and biomedical scientists undertake routine microscopy of slides, but the scientists usually have a wider professional role. Attitudinal surveys were carried out in laboratories where LBC was partially introduced. METHODS: Staff in two cytology laboratories in Greater Manchester were surveyed twice over 6 months. The questionnaire assessed work pressures using scales from the Measures of Work Characteristics instrument, work-related stress using the General Survey version of the Maslach Burnout Inventory, job intentions and job satisfaction. RESULTS: Cytoscreeners, many aged over 50 years, formed over 60% of respondents in both surveys (27/42 in the first survey), and biomedical scientists and doctors, 30%. Both groups were under moderate pressure from work demands in each survey, but cytoscreeners had significantly less autonomy over their working methods (P < 0.001). Although both groups experienced similar levels of exhaustion, cytoscreeners were much more cynical or indifferent towards work in the second survey (P = 0.008) and had lower expectations of being effective (P < 0.001). For the cytoscreeners, there were strong negative correlations in both surveys between cynicism and the work characteristics of influencing decisions and autonomy/control. CONCLUSIONS: The strength of the relationship between work performance and wellbeing serves to emphasize the importance of the new LBC technology in ameliorating low morale where it exists. Further attitudinal research involving larger samples of laboratories is warranted to assess the full impact of this innovation.     

Cell block as an adjunct to conventional Papanicolaou smear for diagnosis of cervical cancer in resource-limited settings

OBJECTIVE: To assess the utility of indigenously prepared cell blocks (CBs) as an adjunct to a conventional smear test in providing a reliable diagnosis of clinically suspicious cervical cancer in resource-limited settings. METHODS: Eighty-six clinically suspicious cervical cancer cases underwent a conventional smear test, CB preparation from residual cellular samples and biopsies at the same sitting. Correlations were performed between these modalities in order to derive the sensitivity and specificity of the CB technique to diagnose cervical cancer. OBSERVATION & RESULTS: Out of 86 clinically suspicious cervical cancers, 72 (83.7%), 70 (81.4%) and 67 (77.9%) cases were diagnosed as malignant on tissue biopsies, CBs and smears respectively. CB-biopsy agreement in the diagnosis of malignancy was feasible in 87.5% of the cases while CB-Pap smear agreement was feasible in 92.5% of the cases. Sensitivity and specificity of CB preparation to diagnose malignancy was 92.5% and 100%, respectively, when the smear was taken as the reference test (excluding the unsatisfactory smears). When biopsy was taken as the gold standard, the sensitivity and specificity of CBs were 87.5% and 100% respectively (excluding the unsatisfactory biopsies). In 8/19 cases where the smear diagnoses were either unsatisfactory or atypical squamous cells/atypical glandular cells, CBs picked up malignant lesions. CONCLUSION: CBs prepared from the residual cellular sample of conventional cervical scrapes augment the sensitivity of the smear test. When used as an adjunct to the smear, CBs aid in providing a reliable diagnosis of cervical cancer in the majority of the clinically suspected cases and thus the biopsy load can be reduced significantly in resource-poor settings.     

Fine needle aspiration cytology of the testis as the first-line diagnostic modality in azoospermia: a comparative study of cytology and histology

Objectives:  Male factors are responsible for about half of all infertility cases. Until recently, testicular biopsy was the standard method to ascertain the aetiology of azoospermia. Fine needle aspiration cytology has gained increasing popularity as a simple and minimally invasive procedure that can help in assessing testicular function accurately. This study was aimed at addressing the question whether testicular fine needle aspiration (FNA) may be used as a first-line diagnostic modality in azoospermia and to assess its usefulness in the diagnostic protocol.
Methods:  The FNA was performed in 78 consecutive azoospermic patients. To obviate sampling errors both testes were aspirated, except when contraindicated. Routine haematoxylin and eosin as well as Romanowsky staining was performed on the smears.
Results:  The smears were categorized on cytological examination into normal spermatogenesis in 35 (50%) patients, Sertoli cells only syndrome in 22 (31.4%) and maturation arrest at the spermatocyte/spermatid level was seen in 13 (18.4%) patients. There were eight (10.2%) cases with scant smears where cytological diagnosis could not be made. A good correlation between cytological smears and histological sections was found in 54 of 58 testes (93.1%) in which histopathological confirmation was available.
Conclusions:  Testicular FNA may be utilized as a first-line investigative modality in patients with azoospermia, provided the procedure is performed and interpreted by experts.     

Colposcopic management of high‐grade referral smears: a retrospective audit supporting ‘see and treat'?

OBJECTIVE: The National Health Service Cervical Screening Programme monitors the quality of colposcopy services through the annual KC65 returns. The 2002 returns demonstrated that Standard 7c, which specifies a biopsy rate > or = 90% at first colposcopy visit for high-grade referrals, was not met in the assessed 3-month period. This was investigated along with the other standards. METHODS: Retrospective colposcopy records were accessed for the 597 new referrals, excluding 10 pregnant patients, seen at the colposcopy clinic at the Royal Victoria Infirmary between 1 July 2001 and 31 December 2002, following an abnormal high-grade smear. Cytology and histopathology computer records were checked for confirmation. The results were assessed against the colposcopy standards applicable at that time and the revised standards (2004). RESULTS: Biopsies were taken from 94.47% (Standard > or = 90%) of women at index colposcopy visit including wire loop excision biopsies from 66.16% (87.97% of high-grade colposcopic appearances). Cervical intraepithelial neoplasia (CIN) on histology was found in 91.79% in the study group (Standard > or = 85%) and in 96.71% of index visit biopsies (Standard > or = 90%), meeting the applicable colposcopy standards. The revised 2004 standards specify a biopsy in > or = 95% of high-grade referrals and excision biopsies in 95% if colposcopic appearances are also high-grade, if colposcopy is low grade but the smear is severely dyskaryotic, or when the lesion extends into the canal. The positive predictive value of high-grade cytology for this entire group was 75.54% with CIN present in 90.95%. CONCLUSION: From this study it appears that high-grade cytology in this centre reliably indicates high-grade CIN. Therefore, in women referred for colposcopy following a high-grade smear, excision biopsies should be performed in a higher proportion at the first visit to comply with the revised standards.     

Atypical cells in the urinary sediment: a protocol for cytological analysis of the urinary sediment

Introduction:  The Department of Clinical Chemistry of the General Hospital of Móstoles (Madrid, Spain) has for many years diagnosed the presence of atypical cells in the urinary sediment (ACUS), but this finding only led to a comment in the final report of the urinalysis, informing the clinician of this fact in case he/she decided to perform further studies in these patients.
Methods:  Since 2004, the Clinical Chemistry Department has implemented a new protocol together with the Department of Surgical Pathology of the Hospital, according to which all the urinary sediment samples with ACUS are sent for cytological analysis. In this report, we comment on the results corresponding to 99 samples.
Results:  With this new strategy, we can directly diagnose if the ACUS correspond to a carcinoma or to another process, and this has allowed us to reduce time until diagnosis in our patients and also to avoid unnecessary studies in negative cases. We comment on our results with this new management strategy.     

Reactions in the oral mucous membrane after exposure to CarisolvTM ‐ combined results from a clinical screening test in humans and an experimental study in rats

Objective: To evaluate reactions in the oral mucosa after direct contact with Carisolv^TM Setting: The Faculty of Odontology in Göteborg, Sweden. Subjects: 34 healthy persons for a clinical screening test and 35 Sprague Dawley rats for a histological study. Desing: Mixed Carisolv^TM or 0.5 % NaOCl were soaked in paper and applied to either side of the medial frenula of the lower lip of 34 persons. The solutions were left on the oral mucosa for three minutes. Inspection was made and photographs were taken immediately after exposure and also after 1 hour, 24 hours, and 72 hours. Mixed Carisolv^TM was applied in a similar manner as described above to 35 adult Sprague Dawley rats. The animals were killed and biopsies were taken immediately after Carisolv^TM exposure and also after 1 hour, 24 hours, and 48 hours. The biopsies were sectioned and prepared for histomorphometrical evaluation in light microscopy where cells were counted on regions from the epithelium layer deeper into the mucous membrane. Results: Some adverse reactions were detected on the oral mucosa of humans up to 24 hours after Carisolv^TM exposure for 3 minutes. The detected inflammatory reactions were slight and no patient felt any discomfort. The results of the histological study on rat did not show any statistically significant increase of the number of cells at any time after Carisolv^TM exposure. Conclusions: If the oral mucosa gets in direct contact with Carisolv^TM for 3 minutes no or only a weak inflammatory response may be expected.     

Proliferative activity in clinically healthy oral mucosa exposed to tobacco smoking and alcohol: a longitudinal study using the agNOR staining technique

OBJECTIVE: To evaluate cell proliferation in clinically healthy oral mucosa exposed to smoking and alcohol carcinogens over a period of 24 months using the AgNOR staining technique. STUDY DESIGN: Sixty patients were initially evaluated: 17 were control individuals, 25 were smokers and 18 were smokers and alcohol drinkers. Fifty-two of these patients were reevaluated. Specimens for cytology were obtained from swabs of lower lip mucosa, border of the tongue and floor of the mouth and underwent AgNOR staining for evaluation of mean number and mean area of AgNOR dots per nucleus and percentage of nuclei with > 3 and > 5 AgNOR dots. Student t and Kruskal-Wallis tests were used to compare values obtained. RESULTS: A statistically significant increase was found in mean number of AgNOR dots per nucleus in 2 groups. One group showed a tendency toward increase of these values. The results of the longitudinal evaluation (Kruskal-Wallis test) revealed a statistically significant difference in number and area of AgNOR dots in the cells of the lower lip. CONCLUSION: The increase of the variables suggests that the longitudinal evaluation of changes in cell proliferation in individuals exposed to smoking and alcohol carcinogens may be a useful monitoring tool.     

Genomic instability in blood cells is able to predict the oral cancer risk: an experimental study in rats

This study was undertaken to investigate the genomic instability on blood cells during 4-nitroquinoline 1-oxide (4NQO)-induced rat tongue carcinogenesis by means of single cell gel (comet) and micronucleus assays. Male Wistar rats were distributed into three groups of 10 animals each and treated with 50 ppm 4NQO solution through their drinking water for 4, 12, and 20 weeks. Ten animals were used as negative control. Although no histopathological abnormalities were induced in the epithelium after 4 weeks of carcinogen exposure, genetic damage was found in blood cells as depicted by the mean tail moment and an increase of micronucleated polychromatic erythrocytes. After 12 and 20 weeks treatment, the same picture occurred, being the strong effect observed in the micronucleus induction. These periods correspond to pre-neoplastic lesions and well-differentiated squamous cell carcinomas, respectively. Taken together, our results support the idea that genomic instability on blood cells appears to be associated with the risk and progression of oral cancer, being a reliable tool for detecting early systemic conditions of malignancy.     

Selection of oocytes for in vitro maturation by brilliant cresyl blue staining： a study using the mouse model

Selecting oocytes that are most likely to develop is crucial for in vitro fertilization and animal cloning. Brilliant cresyl blue （BCB） staining has been used for oocyte selection in large animals, but its wider utility needs further evaluation. Mouse oocytes were divided into those stained （BCB＋） and those unstained （BCB-） according to their ooplasm BCB coloration. Chromatin configurations, cumulus cell apoptosis, cytoplasmic maturity and developmental competence were compared between the BCB＋ and BCB- oocytes. The effects of oocyte diameter, sexual maturity and gonadotropin stimulation on the competence of BCB＋ oocytes were also analyzed. In the large- and medium-size groups, BCB＋ oocytes were larger and showed more surrounded nucleoli （SN） chromatin configurations and higher frequencies of early atresia, and they also gained better cytoplasmic maturity （determined as the intracellular GSH level and pattern of mitochondrial distribution） and higher developmental potential after in vitro maturation （IVM） than the BCB-oocytes. Adult mice produced more BCB＋ oocytes with higher competence than the prepubertal mice when not primed with PMSG. PMSG priming increased both proportion and developmental potency of BCB＋ oocytes. The BCB＋ oocytes in the large-size group showed more SN chromatin configurations, better cytoplasmic maturity and higher developmental potential than their counterparts in the medium-size group. It is concluded that BCB staining can be used as an efficient method for oocyte selection, but that the competence of the BCB＋ oocytes may vary with oocyte diameter, animal sexual maturity and gonadotropin stimulation. Taken together, the series of criteria described here would allow for better choices in selecting oocytes for better development.     

Migration pattern, morphology and viability of cells suspended in or sealed with fibrin glue: a histomorphologic study

INTRODUCTION: We studied the migration pattern, morphology and viability of cells suspended in five different fibrin glues. Besides this, the behaviour of chondrocytes seeded on porous matrices comprising different collagen types sealed with fibrin glue was investigated. MATERIAL AND METHODS: In an experiment A, cell suspension (0.5x10(6) cells) was incubated with different fibrin glues. Experiment B was set up to evaluate chondrocytes migration either through a collagen I/III (Chondro-Gide, Geistlich Biomaterials, Switzerland) or collagen II matrix sealed with different fibrin glues in a perfusion chamber system. Analysis were performed by lightmicroscopy (Mayer's hematoxylin-eosin; Masson-Goldner; TUNEL test) and by transmission and scanning electron microscopy. All fibrin glues were measured for TGF-beta 1 and 2 with a specific ELISA. RESULTS: After incubation of cell suspension in autologous fibrin glue, the morphology of cells is chondrocyte-like. Spindly, process-bearing cells were seen in commercial fibrin glue. Cells suspended in commercial fibrin glue revealed a significant higher percentage of TUNEL positive cells compared to fibrin tissue adhesives mixed with autologous serum (p=0.006). The TGF-beta 1 and 2 concentration was significantly higher in partial autologous fibrin sealant (PAF) compared to their commercial counterparts (p=0.001). Cells seeded on the collagen I/III matrix retained their chondrocytic morphology, while in the type II collagen matrix the chondrocytes displayed a fibroblastic phenotype. The ratio of TUNEL positive cells for the collagen I/III matrix was significantly surpassed by the values, when a collagen II matrix was used (p=0.008). No ingrowth of cells was seen in any of the experimental conditions. CONCLUSION: Partial autologous fibrin glue and collagen I/III matrices are favourable in respect to migration pattern, morphology and viability, but definitive conclusions can only be drawn after in vivo studies. This will be addressed in future animal studies.     

Carcinoma of buccal mucosa in smokeless tobacco users: a preliminary study of the use of cytology for early detection

A type of smokeless tobacco, known as Maras powder, is widely used in lieu of cigarettes in the south-eastern region of Turkey. In the present study, we evaluated cytological smears obtained from the lesion of lower lip mucosa of 80 smokeless tobacco users. There were severe epithelial abnormalities in seven out of the 80 subjects. Punch biopsies from these cases disclosed dysplasia in three cases, carcinoma in situ in two, and carcinoma in the other two cases. There was mild dysplasia in two punch biopsies of the remaining 73 cases. Oral cancer in tobacco powder users appeared to be related to the length of use, as it was observed only in subjects with 15 or more years of exposure.     

A study on the effects of short-, medium- and long-term professional oral care in elderly persons requiring long-term nursing care at a chronic or maintenance stage of illness

Objective: The purpose of the present study is to clarify the frequency of provision of effective professional oral care by dental health workers in order to improve the oral hygiene state of elderly persons requiring long‐term care. Methods: In 105 patients requiring long‐term nursing care, 55 subjects with positive oral Candida albicans were divided into five groups according to the frequency of oral care intervention at intervals of 1, 2, 3, 4 and 6 weeks. The effects of professional oral care on the improvement of the oral hygiene state were investigated taking the frequency and duration of care into consideration. Results: The results showed that oral hygienic condition could be improved by performing professional oral care at intervals of 1 week for 12 consecutive weeks, and the improved condition could be maintained when professional oral care was continued at intervals of 1 week thereafter. Conclusion: It is necessary to continue professional oral care at intervals of 1 week for 12 consecutive weeks and at intervals of 2 weeks for more than 20 weeks. The oral hygiene state could be improved by professional intervention at intervals of three or four weeks in long‐term oral care, but this was limited to subjects whose Functional Independence Measure (FIM) scores were higher than 3.     

城市化进程中城市热岛景观格局演变的时空特征——以厦门市为例

热岛效应作为城市化过程中产生的特有环境问题,对其形成和演变规律的研究有助于人们提出有效的应对措施。以厦门市为研究对象,利用1987—2007年等时间间隔、同时相的5景Landsat TM/ETM+遥感影像数据进行地表温度反演,在此基础上使用景观格局指数分析厦门城市热岛景观格局随城市化进程演变的趋势。结果表明:随着厦门城市化进程加深,整个热岛景观在逐渐变得更加破碎化,高等级热岛景观斑块个数、类型面积和个体面积都增大;新的高等级热岛景观斑块都出现在原有高等级斑块附近,致使高等级类型的邻近度增加而各类型之间相互接触的程度也增加;景观总体的聚合度逐渐下降,而高等级热岛景观类型的聚合度则呈上升趋势;景观水平的蔓延度总体呈下降趋势,优势度高的低等级热岛景观所占的比重下降,优势度逐渐降低;多样性指数、均匀度指数总体呈上升趋势,各热岛景观面积在各类型间的分配逐渐趋于均匀;热岛景观斑块的转化方面,在20 a间低等级斑块类型(1、2、3级)向高等级斑块类型(4、5、6级)转化的面积总体上呈增加趋势,而高等级斑块类型向低等级斑块类型转化的面积总体上呈减小趋势,且等级升高的面积明显大于同期等级降低的面积;就高等级热岛景观斑块而言,他们与3级热岛景观斑块间的相互转化最容易发生,远比高等级斑块内部各类型之间的相互转化来得容易,尤其6类和5类的转化是最为困难的热岛景观变化之一;从空间上看,各高等级热岛景观斑块都经历了数量增加、面积扩大、等级升高三个方面的变化,形成了海沧、新阳、杏林、厦门岛西北港口区和机场5个高温组团。利用景观指数分析城市热环境,可探明热岛景观随城市化演变的趋势,并为有效的热岛效应减缓措施提供直接的理论依据。