Fine-needle Aspiration Biopsy Material Research Articles

Uveal melanoma: the value of molecular genetic testing for early detection of metastatic disease. Clinical case

To date, the issue of not only timely diagnosis and treatment of patients with uveal melanoma (UM), but also the prediction of this pathology remains relevant. The technology of fine needle aspiration biopsy (FNAB) makes it possible to assess the risk of developing metastatic disease using tumor specimen in patients undergoing organ-preserving treatment by cytological and molecular genetic testing in the primary intraocular lesion. Here we present a case of newly identified locally advanced (T3a) UM, outline the possibilities of organ-preserving treatment using Gamma Knife stereotactic radiosurgery, and also demonstrate the feasibility of conducting a comprehensive prognostic testing on FNAB material, including cytological and molecular genetic and morphological prognostic factors. The results of this testing have made it possible to recommend a more frequent surveillance monitoring regime for metastatic disease, which ultimately led to the early detection of UM liver metastases, followed by their successful treatment.

Prognostic fine needle aspiration biopsy of uveal melanoma: Molecular and genetic factors of metastasis risk

Introduction. Molecular genetic testing is actively used for prognostication in patients with uveal melanoma (UM). Tissue for genetic analysis may be obtained either by surgical excision or through fine-needle aspiration biopsy (FNAB). Performing genetic testing and FNAB in each institution can differ in surgical techniques and laboratory methodologies.Aim. To present our own experience of performing FNAB-based molecular genetic testing for prognostication in patients with uveal melanoma.Materials and methods. Prognostic FNAB (n = 151) were combined with brachytherapy or stereotactic surgery. Genetic testing was performed by methods based on polymerase chain reaction (GNAQ, GNA11, EIF1AX and SF3B1 mutations) and fluorescence in situ hybridization (copy numbers of PPARG and MYC genes); cytology of FNAB material was also assessed.Results. Fine-needle aspiration biopsy material was informative in 91 % of cases. At the median follow-up of 36 months, 12 cases of distant metastases were detected. Occurrence of the assessed mutations and copy numbers were related to other representative studies. PPARG deletion was shown to be a significant prognostic factor for metastasis-free survival (p <0,01), which was demonstrated for the first time; EIF1AX and SF3B1 mutations, MYC amplification and cytological class were not shown to be significantly associated with survival in our study.Conclusion. FNAB-based molecular genetic testing for prognostication in patients with uveal melanoma was shown to be a reliable and highly informative approach. Some of the prognostic factors need to be evaluated further with longer follow-up.

Cribriform adenocarcinoma of the minor salivary glands: a case report

BackgroundPolymorphous adenocarcinoma is the third most common malignant salivary gland tumor. Within polymorphous adenocarcinoma, cribriform adenocarcinoma of salivary glands is a rare subtype and resembles papillary thyroid carcinoma histopathologically. Diagnostically, cribriform adenocarcinoma of salivary glands is challenging for pathologists and surgeons alike as initial presentation and cytologic nuclear features can be easily confused with papillary thyroid carcinoma arising from a thyroglossal duct remnant or lingual thyroid.Case presentationA healthy 64-year-old Caucasian woman presented to a community otolaryngologist with a 4-year history of progressive postnasal drip, globus sensation, and eventual dysphonia. Flexible fiberoptic laryngoscopy showed a large, smooth, vallecular lesion filling the oropharynx. Computed tomography imaging of the neck showed a rounded heterogeneous mass centered within the right aspect of the oropharynx measuring 4.2 × 4.4 × 4.5 cm. Fine needle aspiration biopsy was suspicious for papillary carcinoma due to microscopic findings of malignant cells, nuclear grooves, and a powdery chromatin pattern. In the operating room, the tumor was resected en bloc using a lateral pharyngotomy approach with partial resection of the right lateral hyoid. A limited cervical lymphadenectomy was performed to facilitate the lateral pharyngotomy approach and two out of three lymph nodes demonstrated regional metastatic disease. Nuclear grooves, nuclear membrane notching, and occasional intranuclear pseudoinclusions were identified, which are overlapping histopathological characteristics of papillary thyroid carcinoma and cribriform adenocarcinoma of salivary glands. It was negative for thyroglobulin and thyroid transcription factor-1, which was in keeping with cribriform adenocarcinoma of salivary glands rather than papillary thyroid carcinoma.ConclusionIt is difficult to distinguish cribriform adenocarcinoma of salivary glands from papillary thyroid carcinoma solely by cytology, and the distinct characteristics of regional lymph node metastasis coupled with nuanced histologic differences should be emphasized in the evaluation of patients presenting with neck lymphadenopathy and an unknown primary or tongue mass. If sufficient fine needle aspiration biopsy material is available, thyroid transcription factor-1, thyroglobulin, or molecular testing may prove useful in differentiating cribriform adenocarcinoma of salivary glands from papillary thyroid carcinoma. A misdiagnosis of papillary thyroid carcinoma may lead to inappropriate treatment including unnecessary thyroidectomy. Therefore, it is critical for both pathologists and surgeons to be aware of this uncommon entity to avoid misdiagnosis and subsequent mismanagement.

Preoperative Diagnostics of Papillary Thyroid Carcinoma by Molecular Analysis of FNAB Materials

Papillary thyroid carcinoma (PTC) is the most common type of thyroid cancer. Molecular markers of papillary thyroid carcinoma (PTC) including the BRAFT1799A mutation, RET/PTC1,3 rearrangements, and differentially expressed SFTPB (up-regulated) and TFF3 (down-regulated) genes were prospectively assessed in FNAB (fine needle aspiration biopsy) material from thyroid nodules. We examined 73 cases, of them 59 PTC (papillary thyroid carcinoma), five nodular goiters, and nine follicular adenomas. Concordantly with cytology, molecular diagnosis of PTC by SFTPB and TFF3 expression levels was confirmed in 38/41 cases (92.7%); in them, 22/41 (53.6%) had BRAFT1799A mutation and 7/41 (17.1%) had RET/PTC rearrangements. For benign nodules, concordance was observed in 10/12 (83.3%) cases. Among 20 cases, including 19 suspicious for malignancy and one inadequate sample, 17 were positive for SFTPB/TFF3 test (85.0%). Of these, four were also positive for RET/PTC rearrangements, five were positive for BRAFT1799A and all resulted in PTC at histology. The remaining 3 cases were negative for three molecular markers. These nodules showed benign lesions in histology. Our results demonstrated that specific molecular tests improve the efficacy of preoperative diagnostics of PTC.

Diagnostic and prognostic significance of detecting mutations in the <i>BRAF, TERT, RAS, RET/PTC, PAX8/PPARG</i> in the material of fine needle aspiration biopsy thyroid nodules in the IV cytological group (Bethesda, 2017)

Introduction. Fine needle aspiration biopsy followed by cytological examination is the gold standard in the diagnosis of thyroid nodules. However, up to one third of cases represent an indeterminate result (Bethesda Thyroid Classification, 2017) III—V). Among such cases, category IV is the most common and most difficult to interpret (Bethesda, 2017). The study objective is to determination of the diagnostic and prognostic significance of the molecular genetic study of the fine needle aspiration biopsy material in patients with thyroid nodules with the cytological category Bethesda, IV.Materials and methods. The study included surgical thyroid samples obtained from patients whose cytological examination revealed pathology of cytological category IV according to the Bethesda classification (2017). group 1 included surgical samples from 143 patients with thyroid lesions, and group 2 - cytological material from 45 patients. Determination of the BRAF V600E mutation, mutations in the RAS genes (KRAS, HRAS, NRAS) was carried out using allele-specific polymerase chain reaction, and the RET / PTC1, RET / PTC3 and PAX8 / PPARG translocations were determined using reverse transcription polymerase chain reaction. Sanger sequencing was used to detect mutations in the promoter region of the TERT gene.Results. In group 1, an overall prevalence of the studied mutations in thyroid cancer was 35.1 %: 8.8 % of cases were mutation BRAF V600E, 24.6 % - mutations in the RAS genes, 1.8 % - mutation C228T in the TERT gene. The C228T mutation in the TERT gene was found in 1 case of widely invasive Hurtle cell carcinoma. The prevalence of mutations in benign formations was 4.7 %. mutations in RAS genes were also found in them in group 1, mutation BRAF V600E was associated with the presence of extrathyroid invasion (p = 0.024), vascular invasion (p = 0.018), and lymph node metastases (p = 0.018). In group 2, using the genetic panel sensitivity and specificity were equal: 36.4 and 93.9 %, respectively. positive and negative predictive values were 66.7 and 81.6 %, respectively. No RET / PTC and PAX8 / PPARG translocations were found in groups 1 and 2.Conclusion. The investigated molecular genetic panel, having a high specificity for carcinomas, will make it possible to supplement the cytological diagnostics of material in the category Bethesda, IV. BRAF V600E was associated with an aggressive morphological pattern.

Conformity of Fine Needle Aspiration Biopsy (FNAB) and Core Needle Biopsy (CNB) in peripheral lung tumor patients: A cross-sectional study

BackgroundThe problem of establishing lung tumor diagnostics is a challenge for clinicians, especially pulmonologists, in determining a definitive diagnosis of a lung tumor. ObjectiveAnalyzing the conformity of anatomical pathology results between fine-needle aspiration biopsy (FNAB) and core needle biopsy (CNB) materials in peripheral lung tumors. MethodsA cross-sectional study was conducted from July 2019 to December 2020 with 66 participants. Participants were examined for CNB and FNAB, in which the results of these examinations were compared for conformity. Statistical analysis used the Kappa test with p < 0.05. ResultMost participants' tumor size was >70 mm, with FNAB results showing malignant category (39.5%), non-malignant (40.0%), and undiagnosed (38.9%; p = 0.757). Meanwhile, CNB examination showed a tumor size of >70 mm that was categorized into malignant (40.4%) and non-malignant (33.3%; p = 0.510). Most tumors were located in the right superior lobe that had FNAB results in the malignant (39.5%), non-malignant (30.0%) and undiagnosed (27.8%; p = 0.306) categories. The CNB examination also showed that most tumors were located in the right superior lobe, which had resulted in the category of malignant (34.4%), non-malignant (26.7%), and undiagnosed (75.0%; p = 0.240). Conformity of anatomical pathology results from FNAB and CNB subject such as malignancy category of 35 participants (74.5%), non-malignancy of 7 participants (53.8%) and undiagnosed of 4 participants (16.7%) with an accuracy of 69.69% (Κ = 0.43; p = 0.001). ConclusionThere is a conformity between the anatomical pathology results from FNAB and CNB materials for the diagnosis of lung tumors. CNB showed better results in the detection of anatomical malignancy and specimen adequacy.

The Role of Human U Three Protein 14a as a Predictor for Hepatocellular Carcinoma Recurrence After Microwave Ablation.

Human U three protein 14a (hUTP14a) is a nucleolar protein which promotes carcinogenesis by causing degradation of the tumor suppressor protein, p53. This study aimed to investigate hUTP14a expression in hepatocellular carcinoma (HCC) and its value as a predictor for HCC recurrence after treatment with microwave ablation (MWA). The hUTP14a expression was evaluated using immunohistochemistry on ultrasound-guided fine needle aspiration biopsy material from the tumor and the surrounding cirrhotic nontumor tissues. The relation between hUTP14a expression and clinic-pathologic variables was analyzed. Nuclear hUTP14a showed significant high expression in HCC tumor tissue compared with corresponding nontumor tissue (P<0.001). Tumoral hUTP14a expression was significantly higher in patients who experienced recurrence than those who were recurrence-free after MWA (P<0.001). We concluded that, hUTP14a has an oncogenic potential, as it is highly expressed in HCC tissues compared with surrounding nontumor cirrhotic tissues. Moreover, nuclear hUTP14a could be used as a promising prognostic biomarker for prediction of HCC recurrence after treatment with MWA.

Equine Sarcoid-histomorphological, Histochemical and Immunohistochemical Studies in A Thoroughbred Horse

Background: Equine sarcoid is a rare equine skin tumour seen in any age group of horses, usually in younger horses. Grossly, it appears as multinodular masses which may or may not ulcerate and pink to greyish white coloured. It was suggested that the papilloma virus is a causative organism along with predisposing factors like skin abrasions and wounds. The definitive diagnosis is based on histopathology and classified according to their gross appearance and clinical behavior. To find out the origin and proliferative nature, histochemical and immunohistochemical study was performed. The present communication reflects various patterns of sarcoid on histopathological examination and it was confirmed by histochemistry and immunohistochemistry. Methods: A four-year-old bay colt was presented to the Madras Veterinary College Teaching Hospital with nodular masses on the buccal cavity. Fine needle aspiration biopsy (FNAB), blood, serum and tumour biopsy materials were collected under local anesthesia. The collected samples were subjected to Leishman-Giemsa staining, haematological and biochemical analysis, histopathology, histochemical and immunohistochemical examination. Result: Histopathologically, there was mucosal hyperplasia and hyperkeratosis. The neoplastic cells were arranged as storiform or whorl-like pattern. Also, there was perpendicular orientation of fibroblasts towards the basement membrane (picket fence) at the dermo-epidermal junction, which was considered as characteristic feature of sarcoid. Histochemical examination with Picrosirius red revealed strong positivity characterised by deep red coloured mature collagen fibres. Immunohistochemically, the hyperplastic epithelial cells were positive for pan cytokeratin and fibroblast cells were strongly positive for vimentin. Based on the histopathology, histochemistry and immunohistochemistry, the condition was diagnosed as sarcoid which is rare in horses.

Tiroid Nodüllerinin Değerlendirilmesinde Bethesda Sistemi ve İnce İğne Aspirasyon Biopsisinin Yeri

Aim: In this study, the role of fine needle aspiration biopsy (FNAB) evaluated with the Bethesda System in the diagnosis of thyroid nodules was investigated. Materials and Methods: Total 2284 FNAB materials of 1846 patients were performed in the Pathology Department of Kutahya Health Sciences University, Evliya Celebi Traning and Research Hospital between 2016 and 2019 and reported according to the Bethesda category. Among these cases, diagnostic sensitivity, specificity, accuracy, positive and negative predictive values of FNAB were calculated by making cytohistopathologica comparison in 328 cases who underwent total thyroidectomy. Results: In the study, 1538 of the 1846 patients included were female (83.31%) and 308 (16.68%) were male. The average age was 49.92 years (Age range 19-87). The final cytological diagnoses of 2284 FNABs were as follows: 287 (12.57%) nondiagnostic or unsatisfactory, 1517 (66.42%) benign cytology, 316 (13.83%) atypia of undetermined significance/follicular lesion of undetermined significance, 63 (2.75%) follicular neoplasm or suspicious for a follicular neoplasm, 59 (2.59%) suspicious for malignancy and 42 (1.84%) malignant cytology. Histologically, 245 (74.70%) of the 328 cases were benign, while 83 (25.30%) were malignant. The number of true negative cases was 216 (65.85%), the number of true positive cases was 62 (19.90%), the number of false positive cases was 29 (8.84%) and the number of false negative cases was 21 (6.40%). In this case, diagnostic sensitivity, specificity, accuracy rates, positive and negative predictive values were 74.7%, 88.16%, 84.76%, 68.13% and 91.14%, respectively. Conclusion: In our study, it was observed that the sensitivity, specificity, accuracy rates, positive and negative predictive values of thyroid FNABs were compatible with the literature. We think that as the experience of the clinician and cytopathologist increases, false negative and positive results will decrease and at the same time, the confidence in thyroid FNABs will increase even more with the participation of some recent molecular methods (BRAF and RET/PTC, AX8/PPARG and RAS) applied to FNABs.

Intrapelvic Intestinal Leiomyoma in a Dog - Diagnostic and Therapeutic Challenges

Background: Rectal tumors are uncommon in dogs and cats. The clinical signs result from intra- and extraluminal compression. Diagnosis and treatment of rectal tumors are challenging due to their intraplevic location. Owing to considerable bone superposition, computed tomography is the best exam to evaluate the tumor and plan surgery; however, poor availability and high costs may hinder its use. The objective of this case report is to describe the successful use of a combination of diagnostic techniques, namely transrectal ultrasound, transrectal fine-needle aspiration, and colonoscopy, for diagnosis and surgery planning in a case of intrapelvic intestinal leiomyoma in a dog.Case: A 13-year-old female mongrel dog with tenesmus, low stool production, and hematochezia for two months was presented for examination. During this two-month period, a symptomatic treatment was administered, but there was no clinical improvement. In the clinical evaluation revealed a painless mass on the left dorsolateral region, at a depth of around 4 cm, with considerable luminal reduction. Abdominal ultrasound revealed a mass close to the descending colon; however, bone superposition precluded identification of its origin or delimitation of its boundaries. The patient was subjected to transrectal ultrasound imaging, colonoscopy, and cytological examination of fine-needle aspiration biopsy material collected under general anesthesia. The mass was located at the final portion of the descending colon; it was extraluminal, and measured around 7 x 7 cm. The integrity of the intestinal wall was preserved. Next, radiographic examination of the thorax using three projections (ventrodorsal, left lateral, and right lateral) was performed to check for metastases, and no alteration was detected. Cytology suggested presence of leiomyoma. The patient underwent exploratory laparotomy with pubic osteotomy for intrapelvic access. The extraluminal mass was found adhered to the dorsal colorectal surface, whose serosa was compromised. The mass, which occupied around 80 to 90% of the pelvic canal, was completely removed and submitted to histopathological examination, which confirmed presence of proliferative neoplastic mesenchymal cells (intestinal leiomyoma). The patient's clinical picture evolved without intercurrences, and the patient was discharged 40 days after the pubic bone consolidation procedure.Discussion: The occurrence of leiomyomas in the colorectal segment of the intestine is rare in dogs. Neoplasms that develop in such a region of the intestine are usually more frequently found in elderly animals, such as the patient of this report. Leiomyomas are benign (non-invasive) neoplasms with slow growth. Consequently, clinical signs emerge when the mass exhibits a large size, which causes intra- or extraluminal compression, tenesmus, diminished production or absence of defecation, and hematochezia, as observed in the present case. Even though the clinical signs are similar in these cases, they are unspecific; consequently, for reaching a diagnosis, biopsy and histological investigation are required. In spite of the usefulness of these procedures for diagnosis, computed tomography is the exam of choice to investigate neoplasms in intrapelvic intestinal segments because it allows three-dimensional reconstruction of the affected structures and facilitates surgical planning. Unfortunately, computed tomography was not available for this case. Consequently, colonoscopy, transrectal ultrasound, and transrectal fine-needle aspiration biopsy were performed. When combined, these procedures allowed determination of the location, size and type of neoplasm, which were crucial pieces of information for the correct diagnosis and surgical planning, thus contributing for the successful management of the patient.

Оценка диссеминации увеальной меланомы по склеральному каналу при тонкоигольной аспирационной биопсии

Introduction. Fine needle aspiration biopsy (FNAB) of uveal melanoma (UM) is an invasive procedure. Many authors consider UM FNAB to be safe procedure, however extraocular extension of the tumor through the scleral tracts may occur. Current studies of scleral tracts after FNAB do not compare different biopsy techniques (cannula-assisted and without cannula) in terms of UM cells. Purpose. To assess morphology of scleral tracts for the presence of malignant cells when different biopsy techniques are approached. Material and methods. Forty-four scleral samples were analyzed after transvitreal FNAB performed in 22 enucleated eyes with UM. Cannula-assisted FNAB was performed in 22 cases. In the same eyes FNAB (n=22) was performed without cannula. Results. FNAB material was adequate for cytological examination in all cases. UM was confirmed in 22 eyes pathologically. UM cell implantation was detected in 5 scleral samples after FNAB without cannula. No signs of cell implantation were seen after cannula-assisted FNAB. The risk of scleral tract UM cell implantation was statistically lower in cannula-assisted FNAB technique (p=0.018, Pearson Chi-square test). Conclusions. UM FNAB performed directly through the sclera without cannula is associated with tumor cells implantation in scleral tract. Cannula-assisted FNAB significantly reduces the incidence of UM implantation in scleral tract. Key words: uveal melanoma, fine-needle aspiration biopsy, enucleation, morphology, ophthalmology.

Dual-color immunocytochemistry (Ki-67 with LCA) for precise grading of pancreatic neuroendocrine tumors with applicability to small biopsies and cell blocks.

Ki-67 (MIB-1) immunostaining to quantify the proliferative index of neuroendocrine tumors (NETs) has been recommended (especially for small biopsies). However, this has a number of challenges with nonrepresentative Ki-67 index due to interference by Ki-67 immunoreactive proliferating lymphocytes infiltrating the tumor and also some proliferating stromal cells including endothelial cells in the background. Our pilot project showed that dual-color immunostaining with inclusion of leukocyte common antigen (LCA) (Ki-67: nuclear brown; LCA: cytoplasmic red) can facilitate the weeding out of lymphocyte interference. We analyzed the results with 23 surgical cases of pancreatic NETs. This was followed by poststudy examination of 11 cases of endoscopic ultrasound-guided fine-needle aspiration of the pancreatic NETs (PanNETs) to evaluate the findings of the study. Dual-color immunostaining for Ki-67 with LCA increased the precision of quantifying Ki-67 index, due to ability to exclude LCA immunoreactive lymphocytes. Other nontumor Ki-67 immunoreactive cells such as endothelial and stromal cells could be distinguished morphologically. Digital methods were also attempted, but this approach could not distinguish infiltrating lymphocytes and other cells in sections resulting in erroneous results. This study demonstrated that grading of PanNET can be performed with increased precision with dual-color Ki-67 immunostaining protocol standardized in this study. As evaluated on a few cytopathology cases, this protocol is especially useful for the evaluation of small biopsies and cell block sections of fine-needle aspiration biopsy material where 50 high-power fields cannot be evaluated but have >500 tumor cell nuclei.

Teaching and learning FNA biopsy: An update for the modern audience

Teaching and learning FNA biopsy: An update for the modern audience

Immunocytochemical characteristics ofthyrocytes in radioiodine refractory metastases ofpapillary thyroid cancer.

The ICC research was carried out on a fine needle aspiration biopsy material of 104metastases that were found after thyroidectomy and RI therapy, i.e. in postoperative period (79- radioiodine-refractory metastases, 25- radioiodine-uptake metastases). The ICC analysis of TPO and Tg expression in PTC was performed with the use of monoclonal antibodies against TPO and Tg. RI-refractory (RIRM) and RI-uptake metastases of PTC significantly differ by the percentage of cells expressing TPO and Tg (p < 0.05, and p < 0.05respectively). The effectiveness of RI therapy was different for patients with different percentages of TPO-positive cells. A statistically significant difference was demonstrated in the expression of TPO in a punctates of RI-resistant and RI-sensitive metastatic TPC, based on which a new method for preoperative prediction of RI resistance and RI therapy efficiency was developed. It was shown that ICC determination of Tg expression in metastases is effective in preoperative monitoring of RI resistance of PTC if the part of Tg-positive cells in punctates is lower than 56%. The comprehensive study of the ICC profile of thyrocytes in RI-uptake metastases punctates allows us to develop a personified approach to prediction, monitoring and therapy of patients with PTC.

Surgical tactics optimization for treatment of BRAF positive papillary thyroid cancer

Backgraund. Papillary thyroid cancer (PTC) has a favorable course and low mortality rates. However, the incidence of morbidity and mortality continues to increase, especially in the high-risk group. BRAF mutation is a marker of the PTC aggressive course.&#x0D; Aims. Тo study the relationship between surgical treatment tactics and the BRAF positive PTC prognosis.&#x0D; Materials and methods. Our prospective study included 80 patients with BRAF positive PTC, operated from 2009 to 2016 y. Surgical tactics were determined by existing clinical guidelines. To the patients with BRAF positive thyroid cancer it is proposed to perform thyroidectomy with central compartment lymph node dissection. BRAF status was determined by the PCR method in fine needle aspiration biopsy material (FNAB).&#x0D; Results. Recurrence was detected in 7 of the 18 patients of the hemi/thyroidectomy group. Among 62 patients from the group of thyroidectomy with central compartment lymph node dissection recurrence was detected only in 3 cases. The mean recurrence time also differed significantly, and in the group of patients with (hemi)thyroidectomy without central compartment lymph node dissection is detected earlier (53 months and 59.61 months). Also, organ-preserving operations without central compartment lymph node dissection in BRAF positive tumor has worse result, regardless of T stage.&#x0D; Conclusions. In the presence of BRAF-positive PTC thyroidectomy with central compartment lymph node dissection is indicated.

Bethesda Made It Clearer: A Review of 542 Patients in a Single Institution

Background: It is essential to interpret fine needle aspiration biopsy (FNAB) material correctly to create a common language among pathologists and surgeons, leading to a uniform approach to thyroid nodule management. We aimed to compare FNAB reports of patients at our institution who were treated with total thyroidectomy, before and after the Bethesda classification system. Patients and Methods: Patients who underwent total thyroidectomy for thyroid nodules are reviewed. 226 patients who underwent total thyroidectomy before the Bethesda era (2006-2009) were classified as Group-I, and 316 patients in whom total thyroidectomy was performed after the Bethesda classification system was introduced (2010-2014) were classified as Group-II. Results: Before Bethesda, ‘nondiagnostic' or ‘benign' lesions were reported in 16.4 and 45% of patients, respectively, which then significantly decreased to 4.7 and 32.9% as the Bethesda classification criteria came into use. In Group-II, the actual malignancy rates were 13.3, 2.8, 7.3, 15.5, 85.4, and 96.5% for Bethesda I, II, III, IV, V, and VI, respectively. Conclusion: Our experience confirms that the Bethesda classification system leads to a significant reduction in lesions that used to be reported as ‘benign' without compromising the actual rates of malignancy. It ensures better classification of so-called suspicious lesions, and allows for more accurate predictions of suspicious or malignant lesions.

Role of fine needle aspiration biopsy cytology in the diagnosis of infections.

The role of fine needle aspiration biopsy (FNAB) cytology in diagnosing infections has expanded due to the increase in the number of immune compromised patients and the increasing role of FNAB in the developing world where infection is a major cause of illness. FNAB has become the first procedural test in cases where the clinical and imaging findings suggest an infectious lesion or where there is a differential diagnosis of infection or metastatic or primary tumor. This applies to FNAB of palpable or image directed or deep seated lesions accessed by EUS and EBUS. This article details a recommended approach and technique for FNAB of infectious lesions, and discusses the role of rapid on site evaluation and the application of ancillary testing including the rapidly expanding array of molecular tests based on FNAB material. The utility of recognizing suppurative and granulomatous infectious patterns in FNAB direct smears, and the specific cytomorphological features on routine Papanicolaou and Giemsa stains and on special stains of FNAB smears is described for a large number of bacterial, fungal, viral, parasitic, and protozoan infections. The role of cytopathologists is to now train cytopathologists in sufficient numbers to provide FNAB services, teach trainee cytopathologists and cytotechnologists, and to encourage our clinical colleagues to use FNAB in the diagnosis of infections and other lesions to the benefit of patients and the medical system. Diagn. Cytopathol. 2016;44:1024-1038. © 2016 Wiley Periodicals, Inc.

Improvement of accuracy and diagnostic significance of breast tumor fine-needle aspiration biopsy by miRNA analysis of material isolated from cytological smears

Relevance. Breast cancer is the most commonly diagnosed cancer in women. Tumor biopsy, a key diagnostic approach, is required to evaluate the nature of tumor and to determine the therapeutic strategy. In clinical practice methods are being applied: trepan-biopsy and fine needle aspiration biopsy (FNAB). The latter is less traumatic however is used less often because it provides with less information. Moreover, dependence from quality of biopsy and qualification of morphologist are attributes of both techniques. A possibility to use biopsy material for farther analysis of tumor-markers would open a perspective to obtain more information and to improve objectivity of traditional diagnostic approaches, including FNAB. MicroRNAs (miRNAs), regulatory molecules involved in control of virtually all physiologic and pathologic process, emerged as promising tumor markers. Malignant transformation of mammary gland epithelia is associated with specific alterations of cellular miRNAs profile. Analysis of these alterations is of great diagnostic potency. Objective. Development of method for miRNAs analysis in cytological smears material and evaluation of its practical applicability. Material and methods. Archived cytological material (smears on the glass slides) from patients with benign tumor and breast cancer was used. Analysis of miRNAs expression was performed by reverse transcription followed by quantitative PCR. Results of reverse transcription polymerase chain reaction were analyzed with use of relevant cytological and morphological data. Results. Method of miRNAs analysis in material of cytological smears was developed. Expression of 9 miRNAs in 80 samples was evaluated. Statistically significant expression difference between benign and malignant tumor samples was found for 5 miRNAs: miR-21, miR-205, miR-125b, miR-200a, miR-221. MiR-125b exhibited most prominent expression dysregulation: malignant transformation of mammary epithelium is associated with 500 fold decrease of miR-125b expression level. Expression alterations of several miRNAs were revealed to correlate with clinically relevant characteristics of tumors. Conclusions. Results of our study indicated the possibility of miRNAs expression analysis in the FNAB material and the applicability of this method as additive approach to the convention cytological examination. Application of this method will allow to specify diagnosis and to optimize choice of the therapeutic strategy.

Detection of PIK3CA mutations, including a novel mutation of V344G in exon 4, in metastatic lung adenocarcinomas: A retrospective study of 115 FNA cases.

Phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) mutations and amplification are detected in 1% of primary lung adenocarcinomas (ADCs) and in 38% of primary lung squamous cell carcinomas. Alterations of PIK3CA in metastatic non-small cell lung carcinoma (NSCLC), however, are still not fully understood. This study investigated PIK3CA alterations in metastatic ADCs and correlated the findings with those for other commonly tested molecular abnormalities via fine-needle aspiration (FNA) and small-core biopsy materials. This study identified 115 FNA cases of metastatic lung ADC with standard lung cancer panel analysis by targeted next-generation sequencing and fluorescence in situ hybridization at the Johns Hopkins Medical Institute over a 12-month period. The panel included mutational analysis of PIK3CA, AKT, BRAF, EGFR, ERBB2, KRAS, and NRAS genes and tests of rearrangements for ALK and ROS1 genes. A PIK3CA mutation was detected in 7 of 115 cases of metastatic ADC (6.1%). The majority of the mutations were located in exon 9 or exon 20; however, a mutation in exon 1 was seen in 1 case. Furthermore, p.V344G in exon 4 was detected in 2 cases. Among cases with PIK3CA mutations, 4 had coexisting EGFR mutations, whereas 2 had a coexisting BRAF or KRAS mutation. Several common mutations as well as a novel mutation in the PIK3CA gene were observed in metastatic NSCLC (particularly ADC). The unique role, however, of PIK3CA mutations in metastatic NSCLC and the clinical implications need to be further investigated. Cancer Cytopathol 2016;124:485-92. © 2016 American Cancer Society.

KRAS G12D Mutation Subtype Is A Prognostic Factor for Advanced Pancreatic Adenocarcinoma.

Objectives:There is no molecular biomarker available in the clinical practice to assess the prognosis of advanced pancreatic carcinoma. This multicenter prospective study aimed to investigate the role of KRAS mutation subtypes within the primary tumor to determine the prognosis of advanced pancreatic cancer.Methods:The exon-2 KRAS mutation status was tested on endoscopic ultrasound-guided fine-needle aspiration biopsy material (primary tumor; restriction fragment-length polymorphism plus sequencing and TaqMan allelic discrimination) of patients with proven locally advanced and/or metastatic pancreatic ductal carcinoma. We used the Kaplan–Meier method, log-rank test, and Cox's model to evaluate the impact of KRAS status on the overall survival (OS), adjusting for age, stage of disease, clinical performance status, CA 19-9 levels, and treatment.Results:A total of 219 patients (men: 116; mean age: 67±9.4 years) were included: 147 harbored a codon-12 KRAS mutation (G12D: 73; G12V: 53; G12R: 21) and 72 had a wild-type KRAS. There was no difference in the OS between patients with a mutant KRAS (8 months; 95% confidence interval (95% CI): 8.7–12.3) and the wild-type (9 months; 95% CI: 8.7–12.8; hazard ratio (HR): 1.03; P=0.82). However, the patients with a G12D mutation had a significantly shorter OS (6 months; 95% CI: 6.4–9.7) compared with the other patients (OS: 9 months; 95% CI: 10–13; HR: 1.47; P=0.003; i.e., wild type: 9 months, G12V: 9 months, G12R: 14 months). Similar results were observed in the subgroup of 162 patients who received chemotherapy (HR: 1.66; P=0.0013; G12D (n=49): 8 months, wild type (n=56): 10 months, G12V (n=38): 10 months, G12R (n=19): 14 months). Multivariate analyses identified KRAS G12D as an independent predictor for worse prognosis within the entire series (HR: 1.44; P=0.01) and in the subgroup of patients that received chemotherapy (HR: 1.84; P=0.02).Conclusions:The KRAS G12D mutation subtype is an independent prognostic marker for advanced pancreatic ductal carcinoma. Codon and amino-acid-specific mutations of KRAS should be considered when evaluating the prognoses as well as in trials testing drugs that target RAS and downstream RAS pathways.