Management of thyroid cytological material, preanalytical procedures and bio-banking.

Thyroid nodules are common and are increasingly detected due to recent advances in imaging techniques. However, clinically relevant thyroid cancer is rare and the mortality from aggressive thyroid cancer remains constant. Fine needle aspiration cytology (FNAC) is a standard method for diagnosing thyroid malignancy and the discrimination of malignant nodules from goitre. As the examined nodules on thyroid FNAC are often small incidental findings, it is important to maintain a low rate of undetermined diagnoses requiring further clinical work up or surgery. The most important factors determining the accuracy of the cytological diagnosis and suitability for biobanking of thyroid FNACs are the quality of the sample and availability of adequate tissue for auxiliary studies. This article discusses technical aspects (preanalytics) of performing thyroid FNAC, including image guidance and rapid on-site evaluation, sample collection methods (conventional slides, liquid-based methods, cell blocks) and storage (bio-banking). The spectrum of special studies (immunocytochemistry on direct slides or liquid-based cytology, immunohistochemistry on cell blocks and molecular methods) required for improving the precision of the cytological diagnosis of the thyroid nodules is also discussed.

Guidelines for May-Grünwald-Giemsa staining in haematology and non-gynaecological cytopathology: recommendations of the French Society of Clinical Cytology (SFCC) and of the French Association for Quality Assurance in Anatomic and Cytologic Pathology (AFAQAP).

OBJECTIVE: Since the guidelines of the International Committee for Standardisation in Haematology (ICSH) in 1984 and those of the European Committee for External Quality Assessment Programmes in Laboratory Medicine (EQALM) in 2004, no leading organisation has published technical recommendations for the preparation of air-dried cytological specimens using May-Grünwald-Giemsa (MGG) staining. DATA SOURCES: Literature data were retrieved using reference books, baseline-published studies, articles extracted from PubMed/Medline and Google Scholar, and online-available industry datasheets. RATIONALE: The present review addresses all pre-analytical issues concerning the use of Romanowsky's stains (including MGG) in haematology and non-gynaecological cytopathology. It aims at serving as actualised, best practice recommendations for the proper handling of air-dried cytological specimens. It, therefore, appears complementary to the staining criteria of the non-gynaecological diagnostic cytology handbook edited by the United Kingdom National External Quality Assessment Service (UK-NEQAS) in February 2015.

Training and practice of cytotechnologists: a discussion forum focused on Europe.

OBJECTIVES: To discuss the role and training of cytotechnologists (CTs) in Europe, to identify areas of good practice and to provide an informed opinion to those providing guidelines for training and practice in Europe. METHODS: All members of the Editorial Advisory Board of Cytopathology were invited to take part in a 'discussion forum' for which six topics were circulated in advance concerning the roles of CTs with regard to: (1) pre-screening slides; (2) 'signing out' reports; (3) carrying out ancillary techniques; (4) supervising laboratory staff; (5) taking part in rapid on-site evaluation (ROSE) of fine needle aspirates (FNAs); and (6) whether CTs were trained specifically in cytopathology or in general histopathology. Notes of the meeting were circulated by email and a final report was agreed by 22 participants from 17 predominantly European countries. RESULTS: Training for CTs throughout Europe was variable, especially for non-gynaecological cytology, which was inconsistent with the range of activities required. The participants recommended graduate entry, preliminary training in general laboratory technology, and subsequent training to take account of the probability and, in some centres, the reality of primary cervical cancer screening changing from cytology to human papillomavirus (HPV) testing. They further recommended that CTs should perform HPV tests and take part in ROSE for FNAs, and they supported the European Federation of Cytology Societies developing guidelines for training and practice. CONCLUSION: With CT training added to a university-based education in laboratory or biomedical science, a career in cytotechnology should be an attractive option involving a diverse range of laboratory and clinically based activities.

SFE-AFCE-SFMN 2022 Consensus on the management of thyroid nodules : Recommendations in thyroid cytology: from technique to interpretation.

The SFE-AFCE-SFMN 2022 consensus deals with the management of thyroid nodules, a condition that is a frequent reason for consultation in endocrinology. In more than 90% of cases, patients are euthyroid, with benign non-progressive nodules that do not warrant specific treatment. The clinician's objective is to detect malignant thyroid nodules at risk of recurrence and death, toxic nodules responsible for hyperthyroidism or compressive nodules warranting treatment. The diagnosis and treatment of thyroid nodules requires close collaboration between endocrinologists, nuclear medicine physicians and surgeons, but also involves other specialists. Therefore, this consensus statement was established jointly by 3 societies: the French Society of Endocrinology (SFE), French Association of Endocrine Surgery (AFCE) and French Society of Nuclear Medicine (SFMN); the various working groups included experts from other specialties (pathologists, radiologists, pediatricians, biologists, etc.). This section deals with the technique and interpretation of thyroid fine-needle aspiration biopsy (FNAB), a reference test for the analysis of thyroid nodules.

Immunocytochemistry in Europe: results of the European Federation of Cytology Societies (EFCS) inquiry.

OBJECTIVE: This report describes the results of the first chosen topic of the European Federation of Cytology Societies (EFCS) scientific committee, which concerns the application of immunocytochemistry (ICC) to cytological material. While ICC has become an important ancillary method, not only for diagnosis but also to assess prognostic and predictive factors on cytological material, there are many different methodologies used and the lack of standardization has been criticized. This inquiry aimed first to obtain an overview of the techniques used and second to suggest mechanisms for standardization and quality control. METHODS: We report the results of 28 replies from 13 countries to a web-based inquiry into types of specimens, preparations and technical methods. RESULTS: Conventional smears were the preparations most commonly used, followed by cytospins, cell blocks and liquid-based preparations, in that order. Avidin-biotin complex, labelled streptavidin-biotin or peroxidase anti-peroxidase were used in 61% and enhanced-polymer technology in 39%. Automated staining techniques were used by 64%. More than half used the same antibody dilutions as histopathology although only 20% used cell blocks. CONCLUSION: Reduction of variability by using automation, appropriate controls and customized dilution of antibodies according to different samples could improve the quality of ICC and standardize the techniques, along with quality control and quality assurance measures.

The Bethesda terminology for reporting thyroid cytopathology: from theory to practice in Europe.

OBJECTIVES: A 2007 conference held at the National Cancer Institute, Bethesda, Md., USA, proposed a new terminology for classifying the results of thyroid fine-needle aspiration (FNA) - The Bethesda System for Reporting Thyroid Cytology (TBSRTC). The need to standardize thyroid FNA terminology was emphasized during the 35th European Congress of Cytology in 2009. An interobserver review study to assess the new terminology for analyzing the results of thyroid FNA was organized by the scientific committee of the European Federation of Cytology Societies. STUDY DESIGN: Four experts in thyroid FNA examined and classified 116 FNAs according to the 6 levels of TBSRTC which are: nondiagnostic (ND); benign; atypia of undetermined significance/follicular lesion of undetermined significance (AUS/FLUS); follicular neoplasm/suspicious for a follicular neoplasm (FN/SFN), with those of Hürthle cell type reported as follicular neoplasm, Hürthle cell type/suspicious for a follicular neoplasm, Hürthle cell type (FNHCT/SFNHCT); suspicious (SUS), and malignant. RESULTS: The total consensus was 62.1%; the cytopathologists disagreed on 44 cases, including 8 cases of AUS/FLUS and 18 of FN/SFN; 59% of the cases had no consensus. They agreed on 73 and 80% of the cases classified as benign and malignant, respectively, and on 58.3% of the SUS cases. The percentage of no consensus for each expert was between 32 and 39%. CONCLUSIONS: Disagreement regarding the use of TBSRTC terminology for classifying the results of thyroid FNA mainly occurred in the most-often criticized categories of AUS/FLUS and FN/SFN.

Survey of medical training in cytopathology carried out by the journal Cytopathology.

This report of the Editorial Advisory Board of Cytopathology gives the results of a survey of medical practitioners in cytopathology, which aimed to find out their views on the current situation in undergraduate and postgraduate training in their institutions and countries. The results show that training in cytopathology and histopathology are largely carried out at postgraduate level and tend to be organized nationally rather than locally. Histopathology was regarded as essential for training in cytopathology by 89.5% of respondents and was mandatory according to 83.1%. Mandatory cytopathology sections of histopathology were reported by 67.3% and specific examinations in cytopathology by 55.4%. The main deficiencies in training were due to its variability; there were insufficient numbers of pathologists interested in cytology and a consequent lack of training to a high level of competence. Pathologists without specific training in cytopathology signed out cytology reports according to 54.7% of responses, more often in centres where training was 3-6 months or less duration. Although 92.2% of respondents thought that specialist cytology should not be reported by pathologists without experience in general cytopathology, that practice was reported by 30.9%, more often in centres with small workloads. The survey report recommends that 6-12 months should be dedicated to cytopathology during histopathology training, with optional additional training for those wanting to carry out independent practice in cytopathology. Formal accreditation should be mandatory for independent practice in cytopathology. When necessary, temporary placements to centres of good practice should be available for trainees intending to practise independently in cytopathology. There should be adequate numbers of pathologists trained in cytopathology to a high level of competence; some of their time could be released by training cytotechnologists and trainee pathologists to prescreen cytology slides and assess adequacy of fine-needle aspiration samples when immediate diagnosis was not required. The survey demonstrated a clear need for European and international guidelines for training in cytopathology.

Diagnostic terminology for reporting thyroid fine needle aspiration cytology: European Federation of Cytology Societies thyroid working party symposium, Lisbon 2009.

A European Federation of Cytology Societies (EFCS) working party of 28 members from 14 European countries met at the European Congress of Cytology in Lisbon in September 2009, with two observers from the USA, to discuss the need for standardising thyroid FNA nomenclature in the light of the National Institute of Cancer (NCI) recommendations resulting from the State of the Science conference in Bethesda in 2007. The data were obtained through two questionnaires sent by email and a transcript of the live discussion at the congress, which is presented in full. The surveys and discussion showed that there were currently no national terminologies for reporting thyroid FNA in the different European countries except in Italy and the UK. Personal, 'local', surgical pathology and descriptive terminologies were in use. All but one of the working party members agreed that thyroid FNA reporting should be standardised. Whilst almost a third would adopt the NCI Bethesda terminology, which offers the advantages of a 'risk of cancer' correlation and is linked to clinical recommendations, more than half favoured a translation of local terminology as the first step towards a unified nomenclature, as has been done recently in the UK. There was some disagreement about the use of: a) the six-tiered as opposed to four or five-tiered systems, b) the use of an indeterminate category and c) the 'follicular neoplasm' category, which was felt by some participants not to be different from the 'suspicious of malignancy' category. The conclusions will be passed to the different national societies of cytology for discussion, who will be asked to map their local terminologies to the Bethesda classification, observe its acceptance by clinicians and audit its correlation with outcome.

Profiling and classification tree applied to renal epithelial tumours.

AIMS: Selection of the relevant combination from a growing list of candidate immunohistochemical biomarkers constitutes a real challenge. The aim was to establish the minimal subset of antibodies to achieve classification on the basis of 12 antibodies and 309 renal tumours. METHODS AND RESULTS: Seventy-nine clear cell (CC), 88 papillary (PAP) and 50 chromophobe (CHRO) renal cell carcinomas, and 92 oncocytomas (ONCO) were immunostained for renal cell carcinoma antigen, vimentin, cytokeratin (CK) AE1-AE3, CK7, CD10, epithelial membrane antigen, alpha-methylacyl-CoA racemase (AMACR), c-kit, E-cadherin, Bcl-1, aquaporin 1 and mucin-1 and analysed by tissue microarrays. First, unsupervised hierarchical clustering performed with immunohistochemical profiles identified four main clusters-cluster 1 (CC 67%), 2 (PAP 98%), 3 (CHRO 67%) and 4 (ONCO 100%)-demonstrating the intrinsic classifying potential of immunohistochemistry. A series of classification trees was then automatically generated using Classification And Regression Tree software. The most powerful of these classification trees sequentially used AMACR, CK7 and CD10 (with 86% CC, 87% PAP, 79% CHRO and 78% ONCO correctly classified in a leave-one-out cross-validation test). The classifier was also helpful in 22/30 additional cases with equivocal features. CONCLUSION: The classification tree method using immunohistochemical profiles can be applied successfully to construct a renal tumour classifier.

[Genetic determinants of anatomical variants of the prostate]. / Facteurs génétiques déterminant les variations anatomiques de la prostate.

INTRODUCTION: The determinants of macroscopic and microscopic anatomical variants of the prostate during ageing are poorly defined. The authors evaluated the correlation between specific gene polymorphisms involved in androgen and oestrogen synthesis and gross (prostatic weight) and microscopic anatomy (stroma/epithelium ratio) of the prostate during ageing. METHODS: The prostatic weight and stromal surface area of an autopsy series of 85 men over the age of 50 were measured, then compared as a function of gene polymorphisms involved in androgen or oestrogen regulation. The following polymorphisms were studied: number of CAG repeats of the androgen receptor (AR), number of TA repeats and the V89L variant of the 5-alpha-reductase gene (SRD5A2) for androgens, and the A1A2 variant of 17-alpha-hydroxylase (CYP17) and number of TTTA repeats of the aromatase (CYP19) for oestrogens. RESULTS: No correlation was observed between the number of TA repeats of the SRD5A2 gene or TTTA repeats of the CYP19 gene and anatomical parameters of the prostate. A statistically significant positive correlation was observed between age and prostate weight (r=0.21, p=0.05) and a statistically significant negative correlation was observed between prostate weight and number of CAG repeats (r=-0.32, p=0.003). The group with less than 20 CAG repeats was associated with a higher prostate weight than the other group. The stromal surface area was greater in the [20-23] CAG repeat group (p=0.02), and in the A2A2 group of CYP17 (p=0.016) than in the other groups. CONCLUSION: A small number of CAG repeats is associated with a higher prostate weight. The mean number of CAG repeats of the androgen receptor and the A2A2 variant of the CYP17 gene are associated with a larger stromal surface area.

[Multidisciplinary consultation for patients with HPV-related invasive carcinoma or precancerous lesions]. / Consultation multidisciplinaire pour les patients atteints d'une pathologie tumorale (carcinome infiltrant ou lésion précancéreuse) liée à HPV : la consultation multidisciplinaire papillomavirus.

Given the recent increase in the number of human papillomavirus (HPV)-induced cancers in other locations than gynaecological, the number of patients with two cancers at distinct sites, and because of the lack of exhaustive data, we decided to create a multidisciplinary network around an HPV consultation at the Georges-Pompidou European Hospital (HEGP). This network aims to set up the best tools for detecting HPV-associated "multisite" precancerous lesions in order to determine the possible impact of dedicated care for this at-risk population. This monthly consultation was created at the HEGP in June 2014. It is currently organized around five consultations: gynaecological, ENT, urological, digestive and immunological. Every patient who has been diagnosed with HPV-related cancer and whose care is provided at the HEGP is offered this particular follow-up: systematically, once the initial lesion has been treated, the patient is convened annually for a day during which it benefits from the consultations mentioned above. A consultation with a psychologist is systematically proposed. Local samples are taken at each site: a cytological examination, the analysis of known predictive and prognostic virological markers are carried out. This study fits more broadly in a theme of clinical and fundamental research around cancers related to HPV.

The diagnostic value of fine-needle aspiration biopsy under ultrasonography in nonfunctional thyroid nodules: a prospective study comparing cytologic and histologic findings.

Fine-needle aspiration biopsy (FNAB) of thyroid nodules has been proposed as a preoperative screening method to reduce the number of patients with benign nodules referred for surgery. Only a few previously published studies have compared cytologic results with a systematic surgical control; furthermore, the role of ultrasound guidance for FNAB in obtaining representative material has been poorly evaluated. The authors tried to assess the diagnostic value of FNAB performed under ultrasonography in a series of patients with a cold nodule and a systematic surgical control. This prospective study included 132 consecutive patients, 19 men and 113 women, aged from 18 to 82 years. The rate of carcinomas was 13.8%. A very high rate of adequate material was obtained (96.2%). Sensitivity, specificity, and global accuracy were 95%, 87.7%, and 89%, respectively. These results, which are equivalent to those previously published but associated with a low level of insufficient material, emphasize the diagnostic value of thyroid FNAB.

French gynecologic cytology.

Diagnostic schema may be developed on a national or global level, but may be varied depending on the local conditions. In this article, the differences between French and North American gynecologic cytology are discussed. This article emphasizes the training, diagnostic, technologic, and regulatory differences that exist in France. By studying such differences, local practices can learn potential benefits and why their currently used systems may be optimal.

Ureteral and bladder lesions after ballistic, ultrasonic, electrohydraulic, or laser lithotripsy.

Four techniques of intracorporeal lithotripsy are now available: ballistic, ultrasonic, electrohydraulic, and laser. Their therapeutic efficacies have generally been evaluated and compared, but very few data have been available on their relative risks of iatrogenic trauma to the urothelial wall. We conducted a comparative analysis of this risk by testing the pig ureteral and bladder wall with the EMS Lithoclast, Olympus ultrasonic lithotripter, Walz Lithotron EL 23, and Versa Pulse Ho:YAG Coherent Laser. We measured the number of shockwaves or the energy required to perforate the ureter and bladder by delivering shocks perpendicular to the walls. Ureteral perforation was impossible with the 1.0-mm Lithoclast transducer and the 1.5-mm ultrasound transducer. Perforation was induced after 250 shocks with the 0.8-mm Lithoclast transducer, after 110 shocks with the 3F electrohydraulic electrode, and after 0.02 kJ with the laser. Bladder perforation was impossible with the 2.0-mm Lithoclast device and the 3.4-mm ultrasound transducer but was induced after 0.04 kJ had been delivered with the laser. We evaluated the iatrogenic risk under normal conditions of use by delivering the shocks tangentially to the ureteral wall and perpendicular to the bladder wall. We sacrificed animals on days 0, 1, and 6. The immediate histologic lesions induced by the Lithoclast and the ultrasound lithotripter were similar, consisting of a moderate reduction of the epithelial layers or intraepithelial detachments. Electrohydraulic shocks induced almost complete abrasion of the urothelium, and the laser induced extensive lesions of partial or complete necrosis of the urothelial wall.(ABSTRACT TRUNCATED AT 250 WORDS)

Experimental study of Wallstent self-expandable metal stent in ureteral implantation.

In regard to the risk of stent obstruction, there are persistent controversies concerning the use of the Wallstent self-expandable metal stents in the ureter. Only a few clinical trials have been published to date, and, from a technical point of view, no success predictors have been identified. In an experimental study using a pig model, we have shown that neither the length of the stent (5 or 10 cm) nor an initial 1-month intubation had any influence on the outcome of the stents. Among the eight implantations studied (four 5-cm and four 10-cm stents), only one 5-cm stent was perfectly patent at 35 days. In all the remaining stents, after 35 or 90 days, constant failure was related to distal narrowing at the edge of the stent, with no difference between the 5- and 10-cm stents or between the intubated and the nonintubated stents. This distal narrowing, probably attributable to a functional discrepancy between an adynamic stented ureter and a normal underlying ureter, demands extreme caution in human applications when the ureter is normally peristaltic.

Laparoscopic fibrin glue ureteral anastomosis: experimental study in the porcine model.

Laparoscopic suturing is still difficult and time-consuming. The aim of this study, using the porcine model, was to evaluate the feasibility, safety, and efficacy of laparoscopic fibrin glue ureteral anastomosis without stay sutures for approximating the ureteral ends. In five pigs, after bilateral cystoscopic insertion of a 7F ureteral catheter, each upper ureter was laparoscopically dissected free and transected with scissors. The ureteral ends were then approximated with two atraumatic grasping forceps, and the fibrin glue was applied over the anastomotic site using a specially designed catheter (Duplocath). After waiting 5 minutes for the sealant to adhere, the forceps were removed, and the anastomotic site was examined for any early disruption. The ureteral stents were then pulled back to the distal ureter, and retrograde ureteropyelography was accomplished bilaterally in order to assess the immediate patency of the anastomoses. The animals were sacrificed and the ureteral anastomoses surgically removed for histologic examination. The operative time after insertion of the trocars averaged 15 minutes for each anastomosis, and no early disruption was observed after withdrawal of the grasping forceps. Immediate ureteral fluoroscopic patency was achieved in all 10 ureteral anastomoses, without leakage in 8 and with minimal leakage in 2. Histologic examination revealed a mild inflammatory reaction in the serosa with no modifications of the mucosa or the muscularis. Subsequently, two pigs were subjected to the same procedure bilaterally and not sacrificed. These two animals died with enormous urinomas on postoperative days 6 and 8. In each case, the anastomotic site was completely disrupted on one side, while the other side remained grossly patent. However, histologic examination of these latter anastomoses revealed no real coaptation of the ureteral ends, while demonstrating complete eversion of the mucosa. In conclusion, fibrin glue ureteroureterostomies, although easy to accomplish, are not safe enough to be used without stay sutures in laparoscopic surgery.

Renal chromophobe cell carcinoma and oncocytoma. A comparative morphologic, histochemical, and immunohistochemical study of 124 cases.

BACKGROUND: Renal oncocytoma has several features that overlap with other renal neoplasms, including the eosinophilic subtype of chromophobe cell carcinoma. In fact, strict criteria for renal oncocytoma have not been well defined and remain a matter of controversy. Ultrastructural studies or sophisticated methods such as flow cytometry and cytogenetic techniques can be of great use in distinguishing the two tumors, but are difficult to propose as routine methods because of their limited availability. OBJECTIVE: To further characterize the histologic criteria of these tumors, we undertook a retrospective study to define the utility of routinely available histochemical and immunohistochemical techniques. DESIGN AND SETTING: Twenty-one cases of chromophobe cell carcinoma, eosinophilic subtype, and 103 cases of oncocytoma were tested with histochemical (Perls, periodic acid-Schiff, and Hale's colloidal iron) and immunohistochemical (peanut agglutinin antigen and UEA-1 for lectins; cytokeratin KL1, epithelial membrane antigen, vimentin, S100 protein, and lysozyme) staining. RESULTS: The antibodies tested and the histochemical staining using Hale's colloidal iron allowed eosinophilic chromophobe cell carcinoma to be distinguished by its characteristic reaction pattern. Seventy-six percent of the chromophobe cell carcinomas showed a microvacuolated pattern, and 89% of the renal oncocytomas showed an apical positivity with Hale's colloidal iron staining (P < .01). Peripheral cell accentuation reactivity for cytokeratin KL1 was observed in 66% of the chromophobe cell carcinoma cases, and apical cytoplasmic positivity was observed in 37% of the renal oncocytoma cases (P = .01). Significant patterns were observed with anti-epithelial membrane antigen and anti-peanut agglutinin antigen antibodies (P = .05 and P = .01, respectively). Positive reactions for vimentin, S100 protein, lysozyme, and UEA-1 were not significant characteristics. CONCLUSION: Our study demonstrated that a precise morphologic description associated with simple histochemical and immunohistochemical techniques provides sufficient criteria for a high level of discrimination between the eosinophilic subtype of chromophobe cell carcinoma and renal oncocytoma.

Retroperitoneal lymph node aspiration biopsy in staging of pelvic cancer: a cytological study of 228 consecutive cases.

Knowledge of the status of the pelvic lymph nodes is important for accurate staging and adequate treatment of patients with genitourinary tract cancer (bladder and prostatic carcinoma, testicular tumors, and uterine carcinoma). A total of 228 consecutive patients underwent preoperative evaluation of the lymph node status by lymphangiography combined with fine-needle aspiration biopsy. A lymphadenectomy was performed in 94 patients. The overall diagnostic accuracy was 93%. There were 5% false-negative results and no false-positive diagnoses. Fine-needle aspiration biopsy is an inexpensive method with which to detect the presence of metastatic lymph nodes visualized by bipedal lymphangiography. It is also a safe and well-tolerated method, with a low morbidity and no mortality. The various cytological features are described as are some possible pitfalls.