Amplicon-based NGS test for assessing MLH1 promoter methylation and its correlation with BRAF mutation in colorectal cancer patients.

Detecting MLH1 promoter methylation is highly relevant to differentiate between possible Lynch syndrome patients or patients with sporadic causes of MLH1/PMS2 deficiency in colorectal (CRC) and endometrial cancers. Here, we aimed to develop a test for assessing MLH1 promoter methylation based in next generation sequencing (NGS), and to evaluate the concordance of MLH1 methylation and BRAF-V600 mutation status in CRC. For that, we performed a series of experiments with DNA from tumor, saliva and commercial control samples and our in house developed amplicon-based NGS test. In patients' samples, MLH1 methylation above 10% was only observed in tumors with MLH1/PMS2 loss. We confirmed the reproducibility and accuracy of MLH1 promoter analysis performing a serial dilution experiment with completely methylated and unmethylated control DNAs and a comparison between two NGS platforms (Ion Proton and Illumina). In MLH1/PMS2 deficient tumors, the MLH1 methylation status was concordant with the BRAF mutation status in 90% (18/20) of the cases. Our amplicon-based NGS test showed a great sensitivity and specificity for detecting MLH1 methylation in CRC samples, with a high agreement with the evaluation of BRAF mutation. This simple and affordable test could be used as a reflex test to identify patients with sporadic causes of MLH1/PMS2 deficiency in CRC, aiding to genetic test referral and identification of Lynch syndrome patients.

Should we wait 3 months for a repeat aspiration in non-diagnostic/indeterminate thyroid nodules? A cancer centre experience.

INTRODUCTION: The Bethesda System recommends repeat fine needle aspiration (rFNA) as a management option for nodules classified under the non-diagnostic (ND) and atypia of undetermined significance (AUS/FLUS) categories. We evaluated the impact of an rFNA in diagnostic resolution and the role of early (≤3 months) vs delayed (more than 3 months) rFNA of nodules initially diagnosed as ND and AUS/FLUS. METHODS: We retrospectively collected all thyroid FNA performed in a 4-year period with repeat aspiration. For cases initially signed out as ND or AUS/FLUS, diagnostic resolution was defined as a change to a Bethesda System category other than these two on rFNA. Comparison and regression models were fitted to identify the impact of time of rFNA on diagnostic resolution. RESULTS: In total, 184 cases were initially assigned as ND and 143 as AUS/FLUS, with overall diagnostic resolution rates for the reassessment of these nodules calculated at 70.1% and 62.9%, respectively. For ND cases, time of rFNA was not significantly associated with diagnostic resolution (P > .05). For AUS/FLUS nodules, however, repeat aspiration performed in more than 3 months after the initial diagnosis was 2.5 times more likely to achieve a resolution in diagnosis than early rFNA (P = .024). CONCLUSIONS: Repeat aspiration of ND and AUS/FLUS nodules helped define diagnosis for the majority of cases, being highly effective in determining correct patient management. For AUS/FLUS nodules, repeat aspiration performed more than 3 months after the initial diagnosis was associated with a higher diagnostic resolution.

Two-year study on the application of the Paris system for urinary cytology in a cancer centre.

INTRODUCTION: The Paris system for reporting urinary cytology (TPS) was published in order to provide clear cytomorphological criteria for urine cytology specimens, focusing on high-grade urothelial lesions. The aim of this study was to evaluate the impact of implementing TPS and to correlate with available concomitant histological samples, accessing overall sensitivity and specificity. METHODS: A retrospective analysis of urine cytology reports from 2017 to 2018 using TPS was carried out, with histological correlation to concomitant samples (up to 3 months). Statistical analysis was performed with calculation of sensitivity and specificity, positive and negative predictive values and risk of malignancy (ROM) for all TPS categories. RESULTS: A total of 1660 specimens were evaluated. Histological specimens were available for 611 (36.8%) cases. Urine cytology categorised by TPS had 2.4% non-diagnostic cases, 87.1% negative for high-grade urothelial carcinoma (HGUC), 4.6% atypical urothelial cells, 2.7% suspicious for HGUC, 2.7% HGUC and 0.5% cases of other malignancies. Sensitivity, specificity, negative predictive value and positive predictive value were 40.0%, 99.3%, 88.2% and 92.3%, respectively. ROM of each category was 0% for non-diagnostic, 11.1% for negative for HGUC, 32.4% for atypical, 64.9% for suspicious for HGUC and 87.9% for HGUC and other malignancies. CONCLUSION: Our findings indicated that implementation of TPS provided a high specificity and predictive positive value for the detection of high-grade urothelial lesions, with proportionally increasing ROMs as categories progress from negative to positive.

A 2-year retrospective study on pleural effusions: A cancer centre experience.

BACKGROUND: Cytopathological examination of pleural effusions is a fast and minimally invasive method for verification of the presence of neoplastic cells. We report our 2-year experience using a categorised diagnostic system and reporting risks of malignancy (ROMs) for each defined category. METHODS: Cytological reports of patients between November 2016 and October 2018 were collected, with results primarily classified into a five-tiered classification scheme. Immunohistochemistry markers used in cytology and their results were also recorded. Final agreement to histology and overall test performance was calculated for cases with available concomitant (up to 3 months) pleural biopsies. RESULTS: A total of 519 samples from 385 patients were collected, being 29 (5.6%) classified as non-diagnostic, 291 (56%) as negative, 28 (5.4%) as atypical, 30 (5.8%) as suspicious and 141 (27.2%) as positive. Most requested markers were calretinin, TTF1, Ber-EP4 and Gata-3, being conclusive in 45 (76.3%) cases. Total cyto-histological agreement was achieved in 49 (80.3%) specimens, with an overall sensitivity and specificity of 69.4% and 93.3%, respectively. Positive predictive value was 96.2% and negative predictive value was of 56%. ROM for each diagnostic category was 50% for non-diagnostic, 44% for negative, 50% for atypical, 83.3% for suspicious and 96.2% for positive. CONCLUSIONS: Our 2-year retrospective study has shown a high specificity and positive predictive value for pleural cytology. The use of a five-tiered system has also shown to be highly effective, with a concordantly progressive higher ROM for the assigned diagnostic categories.

Geographic tongue and fissured tongue in 348 patients with psoriasis: correlation with disease severity.

Geographic tongue (GT) and fissured tongue (FT) are the more frequent oral lesions in patients with psoriasis. The aims of this study were to compare the prevalence of GT/FT between psoriasis group (PG) and healthy controls (HC) and investigate the correlation between GT/FT and psoriasis severity using the PASI and age of psoriasis onset. Three hundred and forty-eight PG and 348 HC were selected. According to the age of psoriasis onset, the individuals were classified as having early psoriasis and late psoriasis. The severity of vulgaris psoriasis was determined according to PASI. A follow-up was conducted in patients with psoriasis vulgaris (PV) with GT to evaluate the progression of oral and cutaneous lesions. The FT and GT were more frequent in PG than in HC. The incidence of GT was higher in patients with early psoriasis and that of FT in late-psoriasis. There is association between psoriasis intensity and GT; and a higher monthly decrease of PASI score in patients without GT. The presence of GT and FT is higher in PG than in the HC. GT is associated with disease severity and may be a marker of the psoriasis severity.

Value of videoroscopy in the detection of alterations of actinic cheilitis and the selection of biopsy areas.

BACKGROUND: To demonstrate the value of videoroscopy in identifying lesions and alterations not seen by oroscopy and to select the area for biopsy. MATERIAL AND METHODS: Eighty patients were subjected to anamnesis, physical exam, videoroscopy exam, toluidine blue test and biopsy. A diagram of the lips was created to record the exact location where the lesion was found. RESULTS: Physical exam identified 287 lesions, and videoroscopy identified 587 lesions; erythema and white lesions were the most common lesions associated with actinic cheilitis. Of the 59 performed biopsies, 32 (52.4%) cases were identified by videoroscopy that showed lesions that were not detected during physical examination. CONCLUSIONS: The establishment of a diagram of the lip permitted registration of the precise location of the lesion. Videoroscopy was effective in locating lesions not seen by oroscopy. Both videoroscopy and the diagram of the lips allowed for better and earlier diagnosis and better patient follow-up for those with actinic cheilitis.

Myositis Ossificans of the Breast Mimicking Breast Cancer: A Case Report.

Myositis ossificans (MO) is an uncommon tumor characterized by a rapidly growing mass following a history of local trauma. Few cases of MO affecting the breast have been reported, and some were misdiagnosed as primary osteosarcoma of the breast or metaplastic breast carcinoma. The following case report presents a patient with a growing breast lump whose core biopsy result was suspicious for breast cancer. MO was diagnosed after analysis of the mastectomy specimen. This case highlights the importance of MO as a differential diagnosis of a growing soft-tissue mass after trauma to avoid unnecessary overtreatment. Keywords: Myositis Ossificans, Osteosarcoma, Breast Cancer, Mastectomy, Heterotopic Ossification © RSNA, 2023.

Evaluation of the Th17 pathway in psoriasis and geographic tongue.

BACKGROUND: Psoriasis is a skin-articular disease with unclear etiopathogenesis. It has been suggested that the disease is immune-mediated by T-lymphocytes, predominantly Th17 cells. Similar to psoriasis, geographic tongue is an inflammatory disease with participation of Th17 cells and direct correlation with psoriasis. OBJECTIVE: To investigate and compare the inflammatory responses and the Th17 pathway in psoriasis and geographic tongue. METHODS: This was a cross-sectional study with 46 participants that were categorized into three groups: (A) patients with psoriasis vulgaris; (B) patients with geographic tongue and psoriasis; (C) patients with geographic tongue without psoriasis. All patients underwent physical examination, and a skin and oral biopsy for histopathological examination and immunohistochemical analysis with anti-IL6, anti-IL17, and anti-IL23 antibodies. RESULTS: Histological analysis of all lesions showed mononuclear inflammatory infiltrate. However, moderate intensity was prevalent for the patients with geographic tongue and psoriasis and geographic tongue groups. Immunopositivity for the antibodies anti-IL6, anti-IL17, and anti-IL23 revealed cytoplasmic staining, mainly basal and parabasal, in both psoriasis and geographic tongue. Regarding IL-6, in patients with geographic tongue and psoriasis cases the staining was stronger than in patients with geographic tongue without psoriasis cases. IL-17 evidenced more pronounced and extensive staining when compared to the other analyzed interleukins. IL-23 presented similar immunopositivity for both geographic tongue and psoriasis, demonstrating that the neutrophils recruited into the epithelium were stained. STUDY LIMITATION: This study was limited by the number of cases. CONCLUSION: The inflammatory process and immunostaining of IL-6, IL-17, and IL-23 were similar in geographic tongue and psoriasis, suggesting the existence of a type of geographic tongue that represents an oral manifestation of psoriasis.

Geographic tongue and psoriasis: clinical, histopathological, immunohistochemical and genetic correlation - a literature review.

Geographic tongue is a chronic, inflammatory, and immune-mediated oral lesion of unknown etiology. It is characterized by serpiginous white areas around the atrophic mucosa, which alternation between activity, remission and reactivation at various locations gave the names benign migratory glossitis and wandering rash of the tongue. Psoriasis is a chronic inflammatory disease with frequent cutaneous involvement and an immunogenetic basis of great importance in clinical practice. The association between geographic tongue and psoriasis has been demonstrated in various studies, based on observation of its fundamental lesions, microscopic similarity between the two conditions and the presence of a common genetic marker, human leukocyte antigen (HLA) HLA-C*06. The difficulty however in accepting the diagnosis of geographic tongue as oral psoriasis is the fact that not all patients with geographic tongue present psoriasis. Some authors believe that the prevalence of geographic tongue would be much greater if psoriatic patients underwent thorough oral examination. This study aimed to develop a literature review performed between 1980 and 2014, in which consultation of theses, dissertations and selected scientific articles were conducted through search in Scielo and Bireme databases, from Medline and Lilacs sources, relating the common characteristics between geographic tongue and psoriasis. We observed that the frequency of oral lesions is relatively common, but to establish a correct diagnosis of oral psoriasis, immunohistochemical and genetic histopathological analyzes are necessary, thus highlighting the importance of oral examination in psoriatic patients and cutaneous examination in patients with geographic tongue.

Pleomorphic adenoma of the breast

Pleomorphic adenoma (PA) is a common tumor of the salivary gland, but rarely occurs in the breast. PA of the breast is a benign tumor that usually presents as a periareolar nodule. Core-needle biopsies may yield misdiagnosis with complex fibroadenoma, phyllodes tumor and metaplastic breast cancer due to the mixture of stromal and epithelial elements. We present a case of PA of the breast suspected after core-needle biopsy, but confirmed after surgical excision. The importance to make a correct diagnosis consists in avoid extensive unnecessary surgery, such as mastectomy, since PA can be treated with local surgical resection.

Accuracy of ultrasound-guided core-needle biopsy confronted with pathological findings and comparison of its costs with vacuum-assisted biopsy's costs

Introduction: Breast cancer screening has enhanced early­stage diagnosis by detection of impalpable tumors which require histopathological evaluation. Main percutaneous biopsy types are core-needle biopsy (CNB) and vacuum-assisted biopsy (VAB). CNB is less invasive and related to less bleeding and pain. VAB allows larger tissue samples and permits metal clip placement in biopsy bed for posterior localization in case of surgery. Access to VAB is restricted in Brazil due to its high costs. Objectives: To evaluate the agreement between pathological results of ultrasound (US) guided CNB with metal clip placement and surgery and settle false negative rates (FNR), sensibility, specificity, and accuracy of this method, for breast lesions < 20 mm. Methods: 388 US-guided CNB were retrospectively reviewed. Results: Surgical excision was performed in 317 patients. Overall FNR was 9.8%, (5.2% for lesions 10­20 mm), sensibility 90.2% (94.8% for lesions 10­20 mm), specificity 94.9% (94.1% for lesions 10­20 mm), and accuracy 91.1% (94.7% for lesions 10­20 mm). Cost of VAB varies from 2.2 to 12.5 times US-guided CNB. With metal clip placement, VAB costs 1.95 to 5.2 times US-guided CNB. Conclusions: For lesions that can be identified in US, CNB with metal clip placement has high sensitivity, specificity, and accuracy, as well as low FNR.

Accuracy of ultrasound-guided core-needle biopsy confronted with pathological findings and comparison of its costs with vacuum-assisted biopsy's costs

Introduction: Breast cancer screening has enhanced early­stage diagnosis by detection of impalpable tumors which require histopathological evaluation. Main percutaneous biopsy types are core-needle biopsy (CNB) and vacuum-assisted biopsy (VAB). CNB is less invasive and related to less bleeding and pain. VAB allows larger tissue samples and permits metal clip placement in biopsy bed for posterior localization in case of surgery. Access to VAB is restricted in Brazil due to its high costs. Objectives: To evaluate the agreement between pathological results of ultrasound (US) guided CNB with metal clip placement and surgery and settle false negative rates (FNR), sensibility, specificity, and accuracy of this method, for breast lesions < 20 mm. Methods: 388 US-guided CNB were retrospectively reviewed. Results: Surgical excision was performed in 317 patients. Overall FNR was 9.8%, (5.2% for lesions 10­20 mm), sensibility 90.2% (94.8% for lesions 10­20 mm), specificity 94.9% (94.1% for lesions 10­20 mm), and accuracy 91.1% (94.7% for lesions 10­20 mm). Cost of VAB varies from 2.2 to 12.5 times US-guided CNB. With metal clip placement, VAB costs 1.95 to 5.2 times US-guided CNB. Conclusions: For lesions that can be identified in US, CNB with metal clip placement has high sensitivity, specificity, and accuracy, as well as low FNR.

Evaluation of the Th17 pathway in psoriasis and geographic tongue

Abstract Background: Psoriasis is a skin-articular disease with unclear etiopathogenesis. It has been suggested that the disease is immune-mediated by T-lymphocytes, predominantly Th17 cells. Similar to psoriasis, geographic tongue is an inflammatory disease with participation of Th17 cells and direct correlation with psoriasis. Objective: To investigate and compare the inflammatory responses and the Th17 pathway in psoriasis and geographic tongue. Methods: This was a cross-sectional study with 46 participants that were categorized into three groups: (A) patients with psoriasis vulgaris; (B) patients with geographic tongue and psoriasis; (C) patients with geographic tongue without psoriasis. All patients underwent physical examination, and a skin and oral biopsy for histopathological examination and immunohistochemical analysis with anti-IL6, anti-IL17, and anti-IL23 antibodies. Results: Histological analysis of all lesions showed mononuclear inflammatory infiltrate. However, moderate intensity was prevalent for the patients with geographic tongue and psoriasis and geographic tongue groups. Immunopositivity for the antibodies anti-IL6, anti-IL17, and anti-IL23 revealed cytoplasmic staining, mainly basal and parabasal, in both psoriasis and geographic tongue. Regarding IL-6, in patients with geographic tongue and psoriasis cases the staining was stronger than in patients with geographic tongue without psoriasis cases. IL-17 evidenced more pronounced and extensive staining when compared to the other analyzed interleukins. IL-23 presented similar immunopositivity for both geographic tongue and psoriasis, demonstrating that the neutrophils recruited into the epithelium were stained. Study limitation: This study was limited by the number of cases. Conclusion: The inflammatory process and immunostaining of IL-6, IL-17, and IL-23 were similar in geographic tongue and psoriasis, suggesting the existence of a type of geographic tongue that represents an oral manifestation of psoriasis.

Geographic tongue and psoriasis: clinical, histopathological, immunohistochemical and genetic correlation - a literature review

Abstract: Geographic tongue is a chronic, inflammatory, and immune-mediated oral lesion of unknown etiology. It is characterized by serpiginous white areas around the atrophic mucosa, which alternation between activity, remission and reactivation at various locations gave the names benign migratory glossitis and wandering rash of the tongue. Psoriasis is a chronic inflammatory disease with frequent cutaneous involvement and an immunogenetic basis of great importance in clinical practice. The association between geographic tongue and psoriasis has been demonstrated in various studies, based on observation of its fundamental lesions, microscopic similarity between the two conditions and the presence of a common genetic marker, human leukocyte antigen (HLA) HLA-C*06. The difficulty however in accepting the diagnosis of geographic tongue as oral psoriasis is the fact that not all patients with geographic tongue present psoriasis. Some authors believe that the prevalence of geographic tongue would be much greater if psoriatic patients underwent thorough oral examination. This study aimed to develop a literature review performed between 1980 and 2014, in which consultation of theses, dissertations and selected scientific articles were conducted through search in Scielo and Bireme databases, from Medline and Lilacs sources, relating the common characteristics between geographic tongue and psoriasis. We observed that the frequency of oral lesions is relatively common, but to establish a correct diagnosis of oral psoriasis, immunohistochemical and genetic histopathological analyzes are necessary, thus highlighting the importance of oral examination in psoriatic patients and cutaneous examination in patients with geographic tongue.

Diagnosis of secondary syphilis through oral lesionsin two patients with negative serology: case reports / Diagnóstico de sífilis secundária através daslesões orais em dois pacientes com sorologia negativa: relatos de caso

Syphilis is a sexually transmitted infection, and oral lesion can be the first manifestation. The serology test, such as Venereal Disease Research Laboratory test, is accepted as an effective testing strategy for detecting syphilis, although false-negative reaction can occur, and oral lesions may be pivotal to achievethe diagnosis. We report two cases of seronegative secondary syphilis, a human immunodeficiency virus positive patient and a no HIV positive patient,whose histopathological exams were pivotal to achieve the diagnosis of syphilis. The serology may be negative in secondary syphilis and the oral lesionsmay represent the unique method to diagnostic

A sífilis é uma doença sexualmente transmissível, e a lesão oral pode representar sua primeira manifestação. Testes sorológicos, como Venereal Disease Research Laboratory, são rotineiramente utilizados para detecção de sífilis, entretanto, em alguns casos, podem ocorrer resultados falso-negativos. Nesses casos, as lesões orais são essenciais para o diagnóstico. Relatamos dois casos de sífilis secundária com sorologia negativa, em um paciente HIV positivo e um paciente HIV negativo, que obtiveram o diagnóstico de sífilis a partir do exame histopatológico das lesões orais. Os testes sorológicos podem ser negativos na sífilis secundária, e as lesões orais podem representar o único método diagnóstico.