Amir Afrogheh

Impact of reclassifying noninvasive follicular variant of papillary thyroid carcinoma on the risk of malignancy in The Bethesda System for Reporting Thyroid Cytopathology.

Recent discussions have focused on redefining noninvasive follicular variant of papillary thyroid carcinoma (NI‐FVPTC) as a neoplasm rather than a carcinoma. This study assesses the potential impact of such a reclassification on the implied risk of malignancy (ROM) for the diagnostic categories of The Bethesda System for Reporting Thyroid Cytopathology (TBSRTC).

Is it oncologically safe to leave the ipsilateral submandibular gland during neck dissection for head and neck squamous cell carcinoma

AIM To investigate the incidence of metastasis to the submandibular gland in patients with head and neck squamous cell carcinoma. METHODS We retrospectively evaluated histological reports of neck dissections for upper respiratory tract carcinoma (performed 2002-2009), recording: primary tumour site, tumour-node-metastasis stage, level Ib involvement, previous radiotherapy, perineural invasion, lymphovascular invasion, extracapsular spread, and the presence of malignant disease in the submandibular gland. RESULTS We evaluated 107 cases. The most common primary site was the oral cavity (49 per cent) followed by the supraglottis (21 per cent), glottis (14 per cent), oropharynx (9 per cent) and hypopharynx (6 per cent). Forty-eight per cent of patients had advanced local disease, with 21 per cent at tumour stage 3 and 27 per cent at tumour stage 4. Fifty-six per cent had cervical lymph node metastasis, and 8 per cent received pre-operative radiotherapy. Forty-eight per cent had perineural invasion, 46 per cent lymphovascular spread, 27 per cent extracapsular spread and 8 per cent level Ib metastasis. Only one patient had submandibular gland involvement, due to direct spread (a case with prior radiotherapy and macroscopic submandibular gland involvement evident peri-operatively). CONCLUSION Submandibular gland metastasis from head and neck primary squamous cell carcinoma is extremely rare. Preservation of the ipsilateral submandibular gland during neck dissection is oncologically safe, except in patients with prior surgery or radiotherapy, or a primary tumour in close relation to the gland.

Evaluation for High-risk HPV in Squamous Cell Carcinomas and Precursor Lesions Arising in the Conjunctiva and Lacrimal Sac.

High-risk human papilloma virus (HR-HPV) is a well-established causative agent of oropharyngeal squamous cell carcinoma (SCC). In addition, HR-HPV has occasionally been reported to be present in dysplastic and malignant lesions of the conjunctiva and lacrimal sac, although its overall incidence and etiological role in periocular SCC are controversial. Sequential surgical samples of 52 combined cases of invasive SCC (I-SCC) and SCC in situ (SCCIS) from 2 periocular sites (conjunctiva and lacrimal sac) diagnosed over a 14-year period (2000 to 2014) were selected for evaluation, and relevant patient characteristics were documented. p16 immunohistochemistry was performed as a screening test. All p16-positive cases were further evaluated for HR-HPV using DNA in situ hybridization (DNA ISH), and a subset was also analyzed by polymerase chain reaction (PCR). Of 43 ocular surface squamous neoplasias (OSSNs), 30% (n=13; 8 SCCIS and 5 I-SCC cases) were positive for HR-HPV. HPV-positive OSSNs occurred in 8 men and 5 women with a mean age of 60 years (range, 39 to 94 y). HPV type-16 was detected in all conjunctival cases evaluated by PCR. All 5 conjunctival I-SCCs were nonkeratinizing (n=4) or partially keratinizing (n=1) and managed by simple excision. In contrast, HPV-negative conjunctival I-SCCs were predominantly keratinizing (11 keratinizing and 2 nonkeratinizing). Of 9 lacrimal sac I-SCCs (LSSCCs), 66.7% (n=6) were positive for HR-HPV by p16 and DNA ISH; HPV subtypes were HPV-16 (n=5) and HPV-58 (n=1). In addition, 2 p16-positive cases with negative DNA ISH results were HR-HPV positive (HPV-16 and HPV-33) when evaluated by PCR, suggesting that the rate of HR-HPV positivity among the LSSCCs may be as high as 89% (n=8). The combined group of HR-HPV-positive LSSCCs was seen in 4 men and 4 women with a mean age of 60 years (range, 34 to 71 y). Seven of the 8 HPV-positive LSSCCs (87.5%) had a nonkeratinizing or partially keratinizing histomorphology, whereas 1 case (12.5%) was predominantly keratinizing. The presence of HR-HPV in 30% of OSSNs and at least 66.7% of LSSCCs suggests the possibility of an etiologic role for HR-HPV at these sites.

An evaluation of the Shandon Papspin liquid-based oral test using a novel cytologic scoring system.

OBJECTIVES The aim of this study was to evaluate a cost-effective oral liquid-based cytology screening test, Shandon Papspin (PS). STUDY DESIGN We compared the diagnostic accuracy of PS with surgical biopsy in 69 patients. Transepithelial cytology specimens were obtained with the use of cervical cytobrushes. Cytology specimens were graded and scored by using a novel oral cytologic grading and scoring system. RESULTS Histologic diagnosis of dysplasia or malignancy was made in 51/69 cases, and cytology identified 49/51 cases, with a sensitivity of 96% and specificity of 100%. The best cutoff value for distinguishing reactive/mildly dysplastic lesions from high grade/invasive squamous cell carcinoma was determined to be a cytologic score of 3, representing a sensitivity of 95% and specificity of 96%. CONCLUSIONS PS appears to be an accurate and economical test for detection of high-risk dysplasias and cancers, but the real significance of this method will be its diagnostic accuracy in studies focusing strictly on lesions with a low level of clinical concern.

Diffuse Staining for Activated NOTCH1 Correlates With NOTCH1 Mutation Status and Is Associated With Worse Outcome in Adenoid Cystic Carcinoma

NOTCH1 is frequently mutated in adenoid cystic carcinoma (ACC). To test the idea that immunohistochemical (IHC) staining can identify ACCs with NOTCH1 mutations, we performed IHC for activated NOTCH1 (NICD1) in 197 cases diagnosed as ACC from 173 patients. NICD1 staining was positive in 194 cases (98%) in 2 major patterns: subset positivity, which correlated with tubular/cribriform histology; and diffuse positivity, which correlated with a solid histology. To determine the relationship between NICD1 staining and NOTCH1 mutational status, targeted exome sequencing data were obtained on 14 diffusely NICD1-positive ACC specimens from 11 patients and 15 subset NICD1-positive ACC specimens from 15 patients. This revealed NOTCH1 gain-of-function mutations in 11 of 14 diffusely NICD1-positive ACC specimens, whereas all subset-positive tumors had wild-type NOTCH1 alleles. Notably, tumors with diffuse NICD1 positivity were associated with significantly worse outcomes (P=0.003). To determine whether NOTCH1 activation is unique among tumors included in the differential diagnosis with ACC, we performed NICD1 IHC on a cohort of diverse salivary gland and head and neck tumors. High fractions of each of these tumor types were positive for NICD1 in a subset of cells, particularly in basaloid squamous cell carcinomas; however, sequencing of basaloid squamous cell carcinomas failed to identify NOTCH1 mutations. These findings indicate that diffuse NICD1 positivity in ACC correlates with solid growth pattern, the presence of NOTCH1 gain-of-function mutations, and unfavorable outcome, and suggest that staining for NICD1 can be helpful in distinguishing ACC with solid growth patterns from other salivary gland and head and neck tumors.

Application of liquid‐based transepithelial flexible brush cytology in the detection of high‐grade laryngeal mucosal lesions

Background The aim of this study was to evaluate the effectiveness of liquid-based transepithelial flexible brush cytology (LBTFBC) in the detection of high-grade laryngeal mucosal lesions. Methods Diagnostic accuracies of LBTFBC and flexible biopsy (FB) were compared with the gold standard of biopsy under general anaesthesia (BUA) in 49 and 46 patients, respectively. Using a flexible laryngoscope, transepithelial cytology and biopsy specimens were obtained with the aid of flexible brushes and biopsy forceps. Cytology specimens were graded and scored using a recently proposed oral cytologic grading and scoring system. Results Cytology showed 97, 29% sensitivity, 100% specificity, 97.9% accuracy, and FB disclosed 77.1% sensitivity, 100% specificity, and 82.2% accuracy when compared with BUA. The best cutoff value for discriminating reactive/mildly dysplastic lesions from high-grade dysplasias/invasive squamous cell carcinomas (SCCs) was determined as a cytologic score of 3, with sensitivity and specificity of 100%. Conclusion LBTFBC is a simple office-based procedure, which in combination with the newly proposed classification scheme appears to be an accurate technique in the detection of high-grade laryngeal mucosal lesions. LBTFBC is more effective than FB owing to the enhanced range of sampling and ease of application. It effectively eliminates the need for general anaesthesia, and thus reducing theatre costs and the number of hospital admissions. LBTFBC is ideal for patients who require regular clinical examinations, where repeated biopsies may lead to significant vocal morbidity. Diagn. Cytopathol. 2015;43:21–27.

The Development of a Novel Oral Cytologic Grading System

While the Bethesda system represents one of the great success stories in cervical cytology, there has been little appetite for the adoption of a universal grading system for oral cytology. This could be explained by the general lack of interest in oral cytology due to a high percentage of false negative diagnoses, a great variation in technical quality and cellularity of oral smears as well as the use of inadequate sampling procedures. The lack of a standardized method for reporting oral cytology adversely affects proper management of patients with oral lesions. The emergence of Liquid-Based Cytology (LBC) with dramatic improvements in technical quality and cellularity of the cytology specimens has provoked a new interest in using this diagnostic modality for suspicious oral mucosal lesions. This chapter describes the adequacy criteria and minimum cellularity specifications of oral cytologic specimens, and proposes an oral cytologic grading system analogous to the Bethesda System for reporting cervical cytology based on LBC techniques. Using this classification, the terminology for reporting results obtained by oral cytology examination of class I and class II oral mucosal lesions is discussed with ample illustrations of the morphologic criteria and diagnostic categories. These include normal, reactive changes, changes including probably atypical reactive/low-grade lesions, low-grade squamous intraepithelial lesions, atypical probably high-grade changes, high-grade squamous intraepithelial lesion and invasive squamous carcinoma. While still at its infancy, this grading system provides a standardised and uniform method of reporting for the practising pathologist. To further validate the newly proposed classification scheme and discover the best cut-off value for distinguishing reactive/low grade lesions from high grade/squamous cell carcinoma, a simple and easy scoring method based on nine cytologic characteristics is proposed. This may well increase the specificity of the oral cytology test in a manner similar to that of the robust Papanicolaou test.

Liquid-based trans-epithelial flexible brush cytology of high-grade laryngeal mucosal lesions

Laryngeal cytology, which was popular in the 1950s and 60s, has lost its appeal due to difficulties in obtaining adequate specimens and frequent false negative results which are often associated with smears of suboptimal quality. Flexible laryngoscope and brushes, and liquid-based cytology (LBC) allow for improved collection and quality of laryngeal mucosal samples. In LBC the cells are optimally fixed and significantly retrieved. The samples are virtually devoid of air drying artefact and obscuring elements such as blood, mucus, necrotic debris and inflammation. The diagnostic accuracies of liquid-based transepithelial flexible brush cytological (LBTEFBC) samples and flexible biopsies (FB) were compared with those of biopsy under general anaesthesia (BUA) in 49 and 46 patients respectively. Cytology specimens were graded and scored using a recently proposed oral cytological grading and scoring system. When compared with BUA, the cytology showed 97% sensitivity and 100% specificity, and FB yielded 77.1% sensitivity and 100% specificity. Reactive/mildly dysplastic lesions can be discriminated from high-grade dysplasias/invasive squamous cell carcinomas with 100% sensitivity and specificity. LBTFBC is a simple, cost-effective, relatively atraumatic office-based procedure, and with the recently proposed classification, it can be accurate in the detection of high-grade laryngeal lesions.

Oral Cytology Techniques

A significant proportion of oral squamous cell carcinomas (OSCC) develop from premalignant lesions. Histological examination of tissue remains the gold standard for diagnosis and identification of pre-malignant and malignant oral lesions, but has many disadvantages. Exfoliative cytology is a simple and noninvasive technique which is based on the concept that malignant cells adhere much less to each other and thus are easier to harvest by scraping or brushing a suspicious tissue surface in a relatively atraumatic way.