Single slide assessment: A highly effective cytological rapid on‐site evaluation technique for endobronchial and endoscopic ultrasound‐guided fine needle aspiration

No standardised, comprehensive approach to rapid on‐site evaluation (ROSE) of cytology samples currently exists. Recent meta‐analysis indicates variation in the effectiveness of ROSE, however, reviews commonly omit the details of how ROSE is conducted. This review demonstrates the clinical effectiveness of single slide assessment (SSA) for ROSE of cytology samples, providing a highly effective, standardised methodology, maximising cell yield and the diagnostic potential of samples obtained via endobronchial or endoscopic ultrasound. Advances in molecular testing and immunotherapy now allow patients to access sophisticated, targeted cancer treatments and, consequently, obtaining diagnostic material alone is no longer sufficient. SSA uses specific criteria, based on the morphological presentation, to ensure sufficient material is obtained through one procedure, allowing for all the molecular profiling and tumour expression testing required to provide the patient and clinicians with the optimal treatment options. In total, 450 endobronchial or endoscopic ultrasound procedures were conducted with ROSE SSA performed by a biomedical scientist between 2010 and 2017. In 97% of cases, ROSE SSA matched the final report (inadequate vs adequate—benign material vs malignancy). ROSE SSA provided sufficient material for immunocytochemistry in 200/208 cases (96%) and for additional molecular testing/tumour profiling in 92% (85/92) of cases. The median number of needle passes was three. ROSE SSA streamlines diagnostic pathways; minimising risk of complications to patients, reducing cost and delays to treatment associated with repeat or more invasive procedures. Using SSA, sufficient material for a comprehensive diagnosis can be obtained in one procedure.     

The value of immunohistochemistry in increasing diagnostic precision of undifferentiated tumours by the surgical pathologist

Summary The availability of polyclonal and monoclonal antibodies allows immunohistochemical staining (immunofluorescence and immunoperoxidase) procedures to be used by the surgical pathologist, so as to obtain a definite diagnosis in cases where light microscopic examination of tissue sections from biopsy and surgical specimens is inconclusive. Four cases of undifferentiated tumors are described in which only a differential diagnosis could be obtained using light microscopy but a definite diagnosis was achieved when additional information was provided by immunohistochemistry. A scheme is presented for the use of immunohistochemistry to increase diagnostic precision in assessing tumours.     

Breast metastasis from uterine leiomyosarcoma diagnosed by fine needle aspiration: a case report

BACKGROUND: Leiomyosarcomas of the breast are a rare subgroup of primary breast sarcomas. Even more rare is breast metastasis of an extramammary leiomyosarcoma. To date, only 4 cases have been reported in the literature. CASE: We report a case of breast metastatic leiomyosarcoma in a 58-year-old woman with a prior history of uterine leiomyosarcoma, resected 18 months earlier. The breast mass was palpable and a fine needle aspiration (FNA) was performed. The microscopic examination showed cellular smears composed of loosely structured clusters and tissue fragments of spindle-shaped and polygonal or rounded malignant cells in disorderly arrangement. The tumor cells were medium- or large-sized, with basophilic cytoplasm and enlarged, irregular, hyperchromatic nuclei with nucleoli. Tumor giant cells and multinucleation were also present. The morphologic features along with immunocytochemical positivity for desmin, muscle-specific actin and vimentin indicated the diagnosis of a metastatic leiomyosarcoma. CONCLUSION: FNA cytology can be a reliable method for the diagnosis of leiomyosarcoma. The morphologic criteria in combination Swith the clinical history and the immunocytochemical findings can indicate a definitive diagnosis and avoid additional painful and time-consuming diagnostic procedures for the appropriate patient's further clinical management.     

Clinical and diagnostic management of toxoplasmosis in the immunocompromised patient

With the advent of the highly active antiretroviral therapy (HAART), the natural course of HIV infection has markedly changed and opportunistic infections including toxoplasmosis have declined and modified in presentation, outcome and incidence. However, TE is a major cause of morbidity and mortality especially in resource-poor settings but also a common neurological complication in some countries despite the availability of HAART and effective prophylaxis. In most cases toxoplasmosis occurs in brain and toxoplasmic encephalitis (TE) is the most common presentation of toxoplasmosis in immunocompromised patients with or without AIDS. The need of a definitive diagnosis is substantial because other brain diseases could share similar findings. Rapid and specific diagnosis is thus crucial as early treatment may improve the clinical outcome. Classical serological diagnosis is often inconclusive as immunodeficient individuals fail to produce significant titres of specific antibodies. Polymerase chain reaction (PCR) has a high diagnostic value in the acute disease, but like many 'in-house' PCR assays, suffers from lack of standardization and variable performance according to the laboratory. Molecular diagnosis of toxoplasmosis can be improved by performing real-time PCR protocols. This article summarises the clinical manifestations, diagnostic procedures and management strategies for this condition.     

Genotypic assignment of infection by Dirofilaria repens

Dirofilariasis is a parasitic disease, which if treated inappropriately due to misdiagnosis, can cause unwanted complications particularly when the infection is located in the breast. The numerous obstacles that can cause misdiagnosis of dirofilariases by standard morphological procedures prompted the development of a Dirofilaria repens-specific direct polymerase chain reaction (PCR)-based diagnostic approach using freshly infected dog blood. Reliable amplification of nematode DNA from formalin-fixed infected human specimens by this method is only possible from relatively fresh biological material, preserved in the fixative for up to 20 days. We report here our first case of dirofilariasis since the development of PCR genotyping, where the pathogen was morphologically unrecognizable and the diagnosis was based exclusively on DNA amplification. We complete our methodological contribution to the clinical laboratory diagnosis of dirofilariasis by presenting two more cases, where the primary genotypic assignment of infection by D. repens was further confirmed by conventional morphological means.     

Diagnostic value of lymph node fine needle aspiration cytology: an institutional experience of 387 cases observed over a 5-year period

Lymph node fine needle aspiration (LNFNA) cytology is valuable in solving the diagnostic problems of clinical adenopathy. The usefulness of the procedure in the staging and diagnosis of various malignant and lymphoproliferative tumours, as well as its role in distinguishing reactive hyperplastic lymph nodes from lymphoma, has been documented in the literature generally on an individual basis. We report our cumulative 5 year experience of LNFNA representing 387 cases. Approximately half (n = 182) were diagnosed as either metastatic carcinoma or melanoma; in 54 cases (30%) excisional biopsy or tissue study was performed to confirm the diagnosis; there was only one false-positive diagnosis of a metastatic squamous carcinoma rendered on a submandibular lymph node. Sixty-one lymphoma cases were successfully diagnosed via LNFNA with no false positives; concurrent flow cytometry was utilized in 51% (n = 31) of the 61 cases and supported the cytologic diagnosis of lymphoma in 27 of the 31 cases (87%). A benign or reactive lymph node process was also diagnosed via LNFNA alone or in combination with flow cytometry in 48 cases with only five false negatives, which included four cases of mantle cell lymphoma and one case of melanoma. Unsatisfactory cases accounted for 12%, and represented specimens obtained by 'Wang needle' or other emerging techniques. Our study demonstrates that LNFNA can be an accurate, economical and rapid diagnostic procedure.     

Diagnostic value of immunohistochemistry in lesions of the endocrine system

The aim of immunohistochemistry and immunocytochemistry is to reveal specific antigens in cells and tissue samples. Those techniques are based on an antigen-antibody reaction and visualization of its product in microscopic examination. The precursor of this new diagnostic procedure was an immunofluorescent reaction in frozen tissue samples performed by Albert Coons in 1940. Then the immunohistochemical techniques were perfected to increase sensitivity and specificity. Currently it is hard to imagine a modern pathological examination without immunohistochemistry. At the end of XXth century it was believed that 75% of cases is possible to be diagnosed due to immunohistochemical stains. Microscopic examination of endocrine glands tissue samples is extremely difficult because of coexistence of the presence of neoplasms and endocrine dysfunction. It is necessary to establish the type of hormones in the cells of the endocrine system lesions to make a proper diagnosis. Thanks to the use of antibodies against hormones and its precursors it becomes possible. At present most of the antigens are easily detected in both: formalin fixed paraffin embedded tissues samples and ethanol fixed cytological smears so immunohistochemocal and immunocytological stains can be a part of routine diagnostic procedures in pathology. However most of the biologically active substances are revealed in many organs and tissues and it is necessary to perform a satisfactory immunohistochemical panel to be sure the diagnosis. It is important to notice that there is no need to make a wide panel of antibodies in all of the cases and the economical aspect of examination is also important. Of course immunohistochemistry sometimes is the guarantee of proper diagnosis but in some cases too wide panel of antibodies can be a loss for the patient and for medical department. We discussed the proper diagnostic procedures and immunohistochemical profile in pathological lesions of endocrine system (thyroid and adrenal gland, adenohypophysis, neuroendocrine tumours and some hormones-secreting tumours of gonads).     

Diagnosis of Brain Tumors—A Comparison of Photoscanning and Neuroradiological Techniques

Results of radioisotope photoscans and diagnostic neuroradiological procedures performed in 167 patients with suspected brain tumors led to the conclusion that correct identification of the intracranial process can be obtained in a high proportion of cases by the proper selection of available diagnostic techniques. The radioisotope photoscan had a relative diagnostic accuracy nearly equal to that of the neuroradiological techniques. The accuracy, taken with the relative ease and safety of the method, make it an ideal screening procedure in cases in which intracranial pathologic change is suspected.     

Fine needle aspiration cytology of late-stage callus in stress fracture. A case report

BACKGROUND: Fine needle aspiration cytology (FNAC) is effective in the diagnosis of bone lesions when combined with careful radiologic and clinical evaluation. The cytologic features of callus have not been described before in the English-language literature. CASE: An 18-year-old female presented with a pain in the right lower leg that had been present for two months. Clinical and radiologic findings suggested either stress fracture or periosteal osteosarcoma. The aspiration specimen showed individually scattered, oval cells with moderate amounts of pale pink cytoplasm. The cells contained a single eccentrically located nucleus with evenly distributed, fine chromatin. Osteoclastic giant cells were scattered in the smears. A cytologic diagnosis of benign bone-forming lesion, compatible with callus in fracture, was made. The diagnosis of late-stage callus was confirmed by subsequent histologic examination. CONCLUSION: Typical cases of stress fracture do not need histologic examination, but some cases may be confused with benign and malignant bone tumors. The typical and unique cytologic features of late-stage callus combined with clinical and radiologic findings may prevent the use of more invasive diagnostic procedures and can be a choice for management.     

Cytologic features of pheochromocytoma and retroperitoneal paraganglioma: a morphologic and immunohistochemical study of 13 cases

OBJECTIVE: To review the cytologic features and potential pitfalls of pheochromocytoma and retroperitoneal paraganglioma and to evaluate complications of the aspiration procedure and the diagnostic utility of immunocytochemistry. STUDY DESIGN: We reviewed 15 cytologic specimens from 12 patients with 13 tumors (1 bilateral case). Ten were adrenal (pheochromocytomas) and 3 extraadrenal paragangliomas. Eleven specimens were from fine needle aspiration (FNA) procedures that were performed in collaboration with radiologists using 23-25-gauge needles. In 3 patients the cytologic material was obtained during intraoperative diagnosis. Immunocytochemistry was performed on alcohol-fixed smears. RESULTS: Two aspirates were hypocellular, while the remainder were cellular. Cells were distributed singly or formed discohesive groups. When present, cytoplasm was abundant and ill defined. Most cells had an eccentric nucleus and plasmacytoid morphology. Nuclear pleomorphism, binucleation and multinucletaion, naked nuclei and intranuclear preudoinclusions were common findings. In 2 cases a lipid background was seen focally. Evident cytoplasmic immunoexpression of synaptophysin or chromogranin was detected in the 10 cases analyzed. One patient developed a hypertensive episode during the FNA procedure. It was controlled medically without complications. CONCLUSION: When adequate cytologic material is present, the recognition of pheochromocytoma and extraadrenal paraganglioma is possible. Together with morphology, immunocytochemical studies allow a specific preoperative diagnosis. Scarce material can be a source of diagnostic errors. FNA of pheochromocytomas is not necessarily contraindicated. When analytic data are not diagnostic, FNA may follow. Aspiration must be performed in an area equipped with the therapeutic tools necessary to control a pheochromocytoma crisis.     

Telediagnosis of transbronchial fine needle aspirations--a feasibility study.

AIM: To analyze the diagnostic accuracy of telediagnostic procedures for transbronchial fine needle aspirations. MATERIAL AND METHODS: A double blind study was performed on cytological slides of 54 randomly selected cases with transbronchial fine needle aspirations. The slides were digitized using a Leica digital camera DC100 mounted on a microscope Leica Laborlux S, and analyzed by an experienced pathologist without knowing the definite diagnosis or any additional clinical data. The diagnoses stated by analyzing the digital images were compared to the final conventional diagnoses. In addition, the duration of the digital diagnosis, used magnifications, and difficulties for correct sampling were documented. RESULTS: The "digital" diagnoses of the 54 cases were all in general agreement with the definite diagnoses. No wrong positive or wrong negative case in respect to malignant/non-malignant (31/23) or to small cell/non-small cell (9/22) occurred. The performance of a digital diagnosis lasted for 115 s at average (15-260 s), and is significantly longer compared to that of conventional fine needle aspiration judgement (20 s). The screening magnification was commonly set to x2.5, that for definite diagnostic analysis x40. Benign diseases (2 tuberculosis and 3 sarcoidosis cases) were correctly classified. CONCLUSIONS: Telepathology systems can probably be used for fine needle aspiration analysis without major diagnostic errors. Their use can improve the endoscopic sampling and avoid second anesthesia when missing the lesion of request during the first examination.     

Liquid Biopsies: Emerging role and clinical applications in solid tumours

Late detection and lack of precision diagnostics are the major challenges in cancer prevention and management. Biomarker discovery in specific cancers, especially at the pre-invasive stage, is vital for early diagnosis, positive treatment response, and good disease prognosis. Traditional diagnostic measures require invasive procedures such as tissue excision using a needle, an endoscope, and/or surgical resection which can be unsafe, expensive, and painful. Additionally, the presence of comorbid conditions in individuals might render them ineligible for undertaking a tissue biopsy, and in some cases, it is difficult to access tumours depending on the site of occurrence. In this context, liquid biopsies are being explored for their clinical significance in solid malignancies management. These non-invasive or minimally invasive methods are being developed primarily for identification of biomarkers for early diagnosis and targeted therapeutics. In this review, we have summarised the use and importance of liquid biopsy as significant tool in diagnosis, prognosis prediction, and therapeutic development. We have also discussed the challenges that are encountered and future perspective.     

Gastrin--a normal and pathologic regulator of gastric function.

Fine needle aspiration biopsy (FNAB) is an underused diagnostic procedure in children, particularly in the evaluation of superficial masses. A total of 54 FNABs of superficial masses were performed in children aged 1 month to 15 years. Adequate material for diagnosis was obtained in 50 attempts. The cytologic diagnosis increased clinical understanding and provided a guide for treatment in 46 of the 50 cases. The cytologic diagnosis was confirmed in 15 of 19 patients who underwent an operation. Surgical intervention was obviated in 31 patients. There was one false-positive diagnosis of cancer. We describe the role of FNAB in children and its technique, accuracy, and diagnostic problems.     

Computer-assisted phenotype characterization for genetic research in psychiatry

Psychiatric disorders differ from other complex phenotypes in their lack of objectively assessable biological markers that contribute to the establishment of a research diagnosis for genetic studies. To nevertheless allow for the delineation of genetically meaningful diagnostic entities for psychiatric genetic research, comprehensive phenotype characterization procedures are required. It is widely agreed that these should include the standardized assessment of life-time clinical symptomatology, sociodemographic, and environmental factors. Data should be based on several sources, i.e. diagnostic interviews with probands and their relatives as well as a thorough review of medical records, and final assignment of diagnosis should follow robust algorithms (i.e. best-estimate procedures, consensus diagnosis). Here, we outline a practical implementation of such a phenotype characterization strategy, including patient recruitment, study enrolment procedures, comprehensive diagnostic assessment, and data management. We argue that successful psychiatric phenotype characterization requires flexible tools. For this purpose, we have developed a computer-assisted phenotype characterization inventory, built around the backbone of a relational database. It allows for the straightforward assessment of symptoms, automated error checks and diagnostic assignment, easily manageable data storage and handling, and flexible data transfer between various research centers even across language barriers, while at the same time keeping up with the highest standards for the protection of sensitive patient data.     

High diagnostic accuracy of cytologic smears of central nervous system tumors. A 15-year experience based on 4,172 patients

OBJECTIVE: To investigate the diagnostic accuracy and current role of intraoperative cytologic smears of central nervous system tumors. STUDY DESIGN: Retrospective analysis of 4,172 patients operated on during 1985-1999, with 3,541 intraoperative smears performed during open procedures and 631 during stereotactic biopsies. RESULTS: Complete correlation with the final diagnosis was achieved in a mean of 89.8% (range, 83-93.7% per year). Diagnostic accuracy increased to 95% on average (range, 91.5-96.7% per year) when cases of partial correlation, mainly due to grading deviations, were included. The most accurate intraoperative diagnoses were obtained in cases of meningioma (97.9%), metastasis (96.3%) and glioblastoma (95.7%). A significant reduction in diagnostic accuracy was observed in cases of oligodendroglioma (80.9%) and ependymoma (77.7%). Besides diagnosis and grading, smear cytology provided resection guidance in cases of well-delineated tumors. CONCLUSION: Intraoperative smears in neurosurgery are easy to obtain and inexpensive and have high diagnostic accuracy. In addition to stereotactic biopsy procedures, intraoperative smears permit reliable intraoperative guidance during lesion targeting and resection.     

乳腺浸润性导管癌96例影像诊断体会

目的：探讨全数字化乳腺摄影与彩色多普勒相结合对乳腺癌的诊断价值。方法：搜集经X线摄影及超声检查并经病理证实为乳腺浸润性导管癌的96例进行回顾性分析。结果：96例乳腺癌中,单发病灶83例,多发病灶13例。采用全数字化X线摄影单独诊断乳腺癌80例,诊断符合率83.33%,采用超声单独诊断乳腺癌87例,诊断符合率90.63%。结论：乳腺X线与超声相结合,诊断乳腺癌95例,诊断符合率99.06%,此二种检查综合诊断,可明显提高乳腺癌的诊断符合率,减少漏诊和误诊。     

Comparative validation of c-kit exon 11 mutation analysis on cytology samples and corresponding surgical resections of gastrointestinal stromal tumours

Objective:  Studies have shown that c-kit mutation analysis of gastrointestinal stromal tumours (GISTs) obtained by endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) can be routinely performed. We validated c-kit exon 11 mutational analysis on cell block material obtained from fine needle aspiration cytology (FNAC) for diagnostic purposes and compared it with the same analysis in formalin-fixed paraffin-embedded full sections of the corresponding resection specimens.
Methods:  c-kit mutation analysis was done on cell block material obtained from ten cases encountered in our department from 1999 to 2008 on which FNAC was attempted pre-operatively. The findings were compared with analysis on full paraffin section of the corresponding resected tumours in seven cases where patients opted for resection. c-kit exon 11 was examined via bidirectional nucleic acid sequencing.
Results:  Our results showed 100% concordance for the presence and type of exon 11 mutation in the resected and aspirated tumours in all seven cases. These mutations had diagnostic value when compared with other neoplasms that are part of the cytomorphological differential diagnosis, such as leiomyosarcoma or gastric adenocarcinomas.
Conclusion:  Molecular cytopathology is a powerful tool that can complement morphology and immunohistochemical assessment of cytological material in routine practice for the diagnosis and prognostication of GISTs. We briefly discuss the advantages and limitations of the fine needle method of obtaining tissue for the diagnosis and prognostication of GISTs, and its current therapeutic strategies.     

Applications of statistical analysis in diagnostic histopathology and cytopathology

Corresponding to the rapid increase in the amount of data available for use in clinical diagnoses, there is an increased need for procedures that can provide the diagnostician with meaningful statistical summaries of data and with statements concerning the statistical significance associated with a diagnostic evaluation. It has been demonstrated that multivariate statistical assessment of clinical material can provide consistent, reliable and highly sensitive diagnostic clues, even in instances in which trained personnel are unable to see any change. Several examples of applications of statistical analyses in diagnostic cytology and histopathology are given in this paper. The examples were chosen to be illustrative of the different types of problems for which statistical analyses have been found useful. These problems differ with respect to the extent of the statistical methods thus far developed and the difficulty involved in developing further analyses. For many problems, appropriate statistical analyses are readily available; other problems require definition of custom-made test statistics and, in some cases, also definition of new statistical distributions. The problems discussed here are only a small sample of the existing problems, but they provide at least an indication of the scope of the role that statistics plays in cytopathologic and histopathologic diagnosis.     

Fine needle aspiration biopsy for preoperative workup of pancreatic cystic neoplasms: report of 4 cases

BACKGROUND: Cystic neoplasms of the pancreas comprise a pathologically heterogeneous group of lesions that usually present with similar, nonspecific clinical features. Based on the diagnosis, treatment varies from watchful observation of the lesion to total surgical resection of the pancreas. Therefore the importance of a precise and accurate diagnosis on fine needle aspiration (FNA) biopsy cannot be overemphasized from the patient management standpoint. There is debate regarding the accuracy of FNA diagnosis of cystic lesions of the pancreas. We report 4 cases and review the literature to explore and highlight the cytologic findings and diagnostic pitfalls that may help the cytopathologist accurately distinguish mucinous cystic neoplasm (MCN), intraductal papillary mucinous neoplasm (IPMN), serous cystadenoma (SCA) and ductal adenocarcinoma (DAC). CASES: We present 4 cases of patients with abdominal masses who underwent either computed tomography (CT)-guided or endoscopic ultrasound (EUS)-guided FNA biopsy as preoperative workup. Based on the cytologic diagnosis, the patients underwent surgery. CONCLUSION: Our cases illustrate the cytologic criteria that help the cytopathologist distinguish among MCN, IPMN, SCA and DAC. Correlation with clinical and radiologic findings is strongly advocated for accurate diagnosis. We describe the diagnostic pitfalls frequently encountered in these cases and how to avoid them.