Challenges to the effectiveness of next-generation sequencing in formalin-fixed paraffin-embedded tumor samples for non-small cell lung cancer.

INTRODUCTION: Next-generation sequencing (NGS) of Formalin-Fixed and Paraffin-Embedded (FFPE) specimens is routine in precision oncology practice. However, results are not always conclusive, and it is important to identify which factors may influence FFPE tumor sequencing success. MATERIALS AND METHODS: Here, we evaluated the influence of pre-analytical factors on 705 samples of non-small cell lung cancer specimens that underwent NGS testing. Factors such as tumor site, tumor cell percentage, fragment size, primary tumor or metastasis, presence of necrosis, DNA purity, DNA concentration, sample origin and year of testing. RESULTS: The overall NGS success rate was 84.9 % (n = 599). Bone site specimens had a very low success rate (42.1 %), differing from lung samples (79.8 %) (P < 0.05). Samples with tumor percentages <5 % (success rate of 44.4 %) represented 14.1 % of failed sequencings. Moreover, samples with tumor percentages >10 %-20 % (82 %) did not differ from those with >30 % (88.9 %) on sequencing outcomes (P = 0.086). Specimens that provided DNA concentrations >2.0 ng/uL, 1.0-2.0 ng/uL, 0.5-1.0 ng/uL and <0.5 ng/uL had success rates of 92 %, 77.1 %, 61.3 % and 20.4 %, respectively. Small fragments (≤0.2 cm2) had a success rate of 74.7 % and were more prevalent in the unsuccessful group (P < 0.05). CONCLUSIONS: Our results suggest that tumor percentage, fragment size, decalcified bone specimens, and DNA concentration are potential modifiers of NGS success rates. Interestingly, specimens with tumor percentages between 10 % and 20 % have the same sequencing outcome than specimens with >30 %. These results can strengthen the understanding of factors that lead to NGS success variability.

Acinic cell carcinoma of the oral and maxillofacial region: an international multicenter study.

The aim of this study was to describe the prevalence, clinicopathological, and prognostic features of acinic cell carcinoma (AciCC) of the oral and maxillofacial region. AciCC cases were retrospectively retrieved from 11 pathology centers of three different countries. Medical records were examined to extract demographic, clinical, pathologic, and follow-up information. A total of 75 cases were included. Females (65.33%) with a mean age of 45.51 years were mostly affected. The lesions usually presented as an asymptomatic (64.28%) nodule (95.66%) in the parotid gland (70.68%). The association of two histopathological patterns was the most common finding (48.93%) and the tumors presented mainly conventional histopathological grades (86.11%). Surgical treatment was performed in the majority of the cases (59.19%). Local recurrence was observed in 20% of the informed cases, regional metastasis in 30.43%, and distant metastasis in 12.50%. The statistical analysis showed that the cases with a solid histopathological pattern (p=0.01), high-grade transformation (p=0.008), recurrence (p=0.007), and regional metastasis (p=0.03) were associated with poor survival. In conclusion, high histopathological transformation, presence of nodal metastasis, and recurrence were prognostic factors for AciCC of the oral and maxillofacial region.

Acinic cell carcinoma of the oral and maxillofacial region: an international multicenter study

Abstract The aim of this study was to describe the prevalence, clinicopathological, and prognostic features of acinic cell carcinoma (AciCC) of the oral and maxillofacial region. AciCC cases were retrospectively retrieved from 11 pathology centers of three different countries. Medical records were examined to extract demographic, clinical, pathologic, and follow-up information. A total of 75 cases were included. Females (65.33%) with a mean age of 45.51 years were mostly affected. The lesions usually presented as an asymptomatic (64.28%) nodule (95.66%) in the parotid gland (70.68%). The association of two histopathological patterns was the most common finding (48.93%) and the tumors presented mainly conventional histopathological grades (86.11%). Surgical treatment was performed in the majority of the cases (59.19%). Local recurrence was observed in 20% of the informed cases, regional metastasis in 30.43%, and distant metastasis in 12.50%. The statistical analysis showed that the cases with a solid histopathological pattern (p=0.01), high-grade transformation (p=0.008), recurrence (p=0.007), and regional metastasis (p=0.03) were associated with poor survival. In conclusion, high histopathological transformation, presence of nodal metastasis, and recurrence were prognostic factors for AciCC of the oral and maxillofacial region.

Oral and oropharyngeal diffuse large B-cell lymphoma and high-grade B-cell lymphoma: A clinicopathologic and prognostic study of 69 cases.

OBJECTIVE: The objective of this study was to describe the clinicopathological, molecular, and prognostic features of oral/oropharyngeal diffuse large B-cell lymphoma (DLBCL) and high-grade B-cell lymphoma. STUDY DESIGN: All cases were retrieved from 7 Brazilian institutions. Immunohistochemical reactions were performed to confirm the diagnoses and to categorize the tumors. In situ hybridization was used to detect Epstein-Barr virus (EBV) and fluorescence in situ hybridization was used to identify gene rearrangements. RESULTS: Most cases involved the oral cavity (76.8%). Males and females, with a mean age of 60 years, were evenly affected. Tumors mostly presented as painful swellings. Forty cases represented germinal center B-cell type (58%). Five cases presented double-hit translocation and 3 harbored rearrangement for MYC/BCL2/BCL6. EBV was detected in 3 cases (4.3%). The 5-year overall survival was 44.4%. Female sex, presence of pain and ulcer, microscopic "starry sky pattern" and necrosis, co-expression of c-Myc/Bcl2, and translocation of MYC were associated with a lower survival in univariate analysis (P = .05, P = .01, P = .01, P = .03, P = .05, P = .006, P = .05, respectively). CONCLUSION: Patients affected by oral/oropharyngeal DLBCL have a low survival rate. High-grade B-cell lymphoma (17.7%) and EBV-positive DLBCL, not otherwise specified (4.3%) account for a small number of cases.

Demographic and Clinicopathologic Features of Actinic Cheilitis and Lip Squamous Cell Carcinoma: a Brazilian Multicentre Study.

Data on the occurrence and clinicopathological characteristics of actinic cheilitis (AC) and lip squamous cell carcinoma (LSCC) are well studied; however, they are based on studies limited to a single centre. Herein, we described the frequency of AC and LSCC submitted to microscopic examination from representative geographic regions of Brazil. A retrospective multicentre study was performed on biopsies obtained from 1953 to 2018 at 10 Brazilian oral and maxillofacial pathology centres. A total of 198,709 biopsy specimens were surveyed. Sociodemographic data and clinicopathologic characteristics were analysed. A total of 2017 cases of ACs (1.0%) and 850 cases of LSCCs (0.4%) were recorded. A strong fair-skinned (> 87%) male (> 70%) predilection was observed in both conditions. The mean age was 54.8 ± 18.7 for individuals with AC and 57.8 ± 19.0 for individuals with LSCC. The most commonly affected site was the lower lip (> 90%). This is a large multicentre study of AC and LSCC from Brazil. The frequency and clinicopathological features of AC and LSCC were similar to those described worldwide. This study provides robust and representative epidemiological data of these conditions for the scientific community.

Expression of neurotrophins and their receptors in primary osteosarcoma. / Expressão de neurotrofinas e de seus receptores no osteossarcoma primário.

OBJECTIVE: to determine the expression of neurotrophins and their tyrosine-kinase receptors in patients with osteosarcoma (OS) and their correlation with clinical outcomes. METHODS: we applied immunohistochemistry to biopsy specimens of patients consecutively treated for primary OS at a single institution between 2002 and 2015, analyzing them for expression receptors of tyrosine kinase A and B (TrKA and TrKB), neural growth factor (NGF) and brain derived neurotrophic factor (BDNF). Independently, two pathologists classified the immunohistochemical markers as negative (negative or weak focal) or positive (moderate focal/diffuse or strong focal/diffuse). RESULTS: we analyzed data from 19 patients (10 females and 9 males), with median age of 12 years (5 to 17.3). Tumors' location were 83.3% in the lower limbs, and 63.2% of patients had metastases at diagnosis. Five-year overall survival was 55.3%. BDNF was positive in 16 patients (84%) and NGF in 14 (73%). TrKA and TrKB presented positive staining in four (21,1%) and eight (42,1%) patients, respectively. Survival analysis showed no significant difference between TrK receptors and neurotrophins. CONCLUSION: primary OS samples express neurotrophins and TrK receptors by immunohistochemistry. Future studies should explore their role in OS pathogenesis and determine their prognostic significance in larger cohorts.

OBJETIVO: determinar a expressão de neurotrofinas e seus receptores tirosina quinases em pacientes com osteossarcoma (OS) e sua correlação com desfechos clínicos. MÉTODOS: biópsias de tumores primários de pacientes com OS tratados em uma única instituição, consecutivamente, entre 2002 e 2015, foram analisados através de imuno-histoquímica para expressão de receptores de tirosina quinase A e B (TrKA e TrKB), fator de crescimento neural (NGF) e fator neurotrófico derivado do cérebro (BDNF). De forma independente, dois patologistas classificaram os marcadores de imuno-histoquímica como negativos (negativos e focais fracos) ou positivos (moderado focal/difuso ou forte focal/difuso). RESULTADOS: foram analisados dados de 19 pacientes (10 do sexo feminino e 9 do masculino) com mediana de idade de 12 anos (5 a 17,3 anos). Dos tumores, 83,3% estavam localizados em membros inferiores e 63,2% dos pacientes eram metastáticos ao diagnóstico. A sobrevida global em cinco anos foi de 55,3%. BDNF foi positivo em 16 pacientes (84%) e NGF em 14 pacientes (73%). TrKA e TrKB apresentaram coloração positiva em quatro (21,1%) e oito (42,1%) pacientes, respectivamente. A análise de sobrevida não demonstrou diferença significativa entre receptores TrK e neurotrofinas. CONCLUSÃO: amostras de OS primário expressam neurotrofinas e receptores TrK através de imuno-histoquímica. Estudos futuros podem auxiliar na identificação do papel das mesmas na patogênese do OS e determinar se há possível correlação prognóstica.

Expressão de neurotrofinas e de seus receptores no osteossarcoma primário / Expression of neurotrophins and their receptors in primary osteosarcoma

RESUMO Objetivo: determinar a expressão de neurotrofinas e seus receptores tirosina quinases em pacientes com osteossarcoma (OS) e sua correlação com desfechos clínicos. Métodos: biópsias de tumores primários de pacientes com OS tratados em uma única instituição, consecutivamente, entre 2002 e 2015, foram analisados através de imuno-histoquímica para expressão de receptores de tirosina quinase A e B (TrKA e TrKB), fator de crescimento neural (NGF) e fator neurotrófico derivado do cérebro (BDNF). De forma independente, dois patologistas classificaram os marcadores de imuno-histoquímica como negativos (negativos e focais fracos) ou positivos (moderado focal/difuso ou forte focal/difuso). Resultados: foram analisados dados de 19 pacientes (10 do sexo feminino e 9 do masculino) com mediana de idade de 12 anos (5 a 17,3 anos). Dos tumores, 83,3% estavam localizados em membros inferiores e 63,2% dos pacientes eram metastáticos ao diagnóstico. A sobrevida global em cinco anos foi de 55,3%. BDNF foi positivo em 16 pacientes (84%) e NGF em 14 pacientes (73%). TrKA e TrKB apresentaram coloração positiva em quatro (21,1%) e oito (42,1%) pacientes, respectivamente. A análise de sobrevida não demonstrou diferença significativa entre receptores TrK e neurotrofinas. Conclusão: amostras de OS primário expressam neurotrofinas e receptores TrK através de imuno-histoquímica. Estudos futuros podem auxiliar na identificação do papel das mesmas na patogênese do OS e determinar se há possível correlação prognóstica.

ABSTRACT Objective: to determine the expression of neurotrophins and their tyrosine-kinase receptors in patients with osteosarcoma (OS) and their correlation with clinical outcomes. Methods: we applied immunohistochemistry to biopsy specimens of patients consecutively treated for primary OS at a single institution between 2002 and 2015, analyzing them for expression receptors of tyrosine kinase A and B (TrKA and TrKB), neural growth factor (NGF) and brain derived neurotrophic factor (BDNF). Independently, two pathologists classified the immunohistochemical markers as negative (negative or weak focal) or positive (moderate focal/diffuse or strong focal/diffuse). Results: we analyzed data from 19 patients (10 females and 9 males), with median age of 12 years (5 to 17.3). Tumors' location were 83.3% in the lower limbs, and 63.2% of patients had metastases at diagnosis. Five-year overall survival was 55.3%. BDNF was positive in 16 patients (84%) and NGF in 14 (73%). TrKA and TrKB presented positive staining in four (21,1%) and eight (42,1%) patients, respectively. Survival analysis showed no significant difference between TrK receptors and neurotrophins. Conclusion: primary OS samples express neurotrophins and TrK receptors by immunohistochemistry. Future studies should explore their role in OS pathogenesis and determine their prognostic significance in larger cohorts.

Trk inhibition reduces cell proliferation and potentiates the effects of chemotherapeutic agents in Ewing sarcoma.

Ewing sarcoma (ES) is a highly aggressive pediatric cancer that may arise from neuronal precursors. Neurotrophins stimulate neuronal devlopment and plasticity. Here, we found that neurotrophins nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF), as well as their receptors (TrkA and TrkB, respectively) are expressed in ES tumors. Treatment with TrkA (GW-441756) or TrkB (Ana-12) selective inhibitors decreased ES cell proliferation, and the effect was increased when the two inhibitors were combined. ES cells treated with a pan-Trk inhibitor, K252a, showed changes in morphology, reduced levels of ß-III tubulin, and decreased mRNA expression of NGF, BDNF, TrkA and TrkB. Furthermore, combining K252a with subeffective doses of cytotoxic chemotherapeutic drugs resulted in a decrease in ES cell proliferation and colony formation, even in chemoresistant cells. These results indicate that Trk inhibition may be an emerging approach for the treatment of ES.

A TP53 founder mutation, p.R337H, is associated with phyllodes breast tumors in Brazil.

A few studies have reported phyllodes tumors (PT) of the breast with germline TP53 mutations. Given this potential association and the high frequency of the TP53 p.R337H in southern and southeastern Brazil, the aim of this study was to assess whether p.R337H occurs among women diagnosed with such rare tumors in this region. Benign, borderline, and malignant breast PT were retrieved from eight pathology laboratories, and DNA was extracted from tumor tissue to perform p.R337H analysis. Overall, 128 cases classified as benign, 7 as borderline, and 13 as malignant PT were included in the study. The TP53 p.R337H mutation was identified in tumor cells of eight (5.4 %) cases. Analysis of DNA from non-tumoral tissue was possible in two of these, and both were p.R337H carriers in the germline. In addition, haplotype analysis was done in these two p.R337H carriers showing the presence of the founder haplotype previously reported in Brazilian mutation-positive individuals. Mutation frequency was significantly higher among malignant (3 of 13; 23 %) compared to benign tumors (5 of 128; 3.4 %) (p = 0.004). Mean age at PT diagnosis was not significantly different between mutation carriers and non-carriers. However, when subgroups where analyzed, the difference in age at diagnosis of carriers versus non-carriers within the group of benign tumors reached borderline significance. Our findings reinforce previous evidence that TP53 mutations have an important role in the development of both benign and malignant PT of the breast.

["Epidemiological profile of chronic hepatitis C virus patients and his early virological response to pegylated interferon plus ribavirin therapy. Gastroenterology Service, HNCASE EsSALUD Arequipa"]. / "Perfil epidemiológico de los pacientes con hepatitis crónica por virus C y su respuesta virológica temprana al tratamiento con interferón pegilado más ribavirina. Servicio de Gastroenterología, HNCASE ESSALUD Arequipa 2004-2006".

The present descriptive, retrospective and longitudinal study was designed to determine the epidemic profile of patients infected by hepatitis C virus and chronic hepatitis, as well as to describe the eligibility criteria for treatment with pegilated interferon plus ribavirina and its early virological answer to the treatment. We studied 20 patients treated at Gastroenterology Service of Carlos Seguín Escobedo Hospital of EsSalud between 2004 and 2006. The diagnosis of HVC infection was confirmed by detection of viral RNA with PCR, and the viral load by counting number of RNA copies. The eligibility criteria for antiviral treatment were determined, and also the METAVIR score to determine hepatic fibrosis. After 12 weeks of treatment the patients with eligibility criteria and received treatment were evaluated, and the effectivity of treatment was evaluated with a new determination of viral load. There were more female (15; 75%) than male patients (5; 25%), with ages between 50 and 59 years. The more frequent risk antecedent was blood transfusion (45%), surgery (35%), and traffic accident (10%). The hematological, hepatic, renal and hormonal parameters were in normal range; and the viral load at the moment of diagnosis was of 580 thousand copies mean (between 4100 to 2 millions copies). The most frequent viral genotypes were 1a and 1b. Up to 40% of patients did not fulfilled eligibility criteria for treatment. Nine of 12 patients with eligibility criteria received treatment (75%), 6 of them were evaluated for early virologic response and of them 100,0% had an early virologic response with decrease of post-therapeutic viral load in all but one cases until non-detectable levels. In conclusion our study described the profile of patients infected by HCV with chronic hepatitis in gastroenterology ward of our hospital, and it has been determined that is suggestive that antiviral treatment is highly effective in patients with eligibility criteria for treatment, related to early virologic response.

Perfil epidemiológico de los pacientes con Hepatitis Crónica por virus C y su respuesta virológica temprana al tratamiento con Interferón Pegilado más Ribavirina. Servicio de Gastroenterología, HNCASE ESSALUD Arequipa 2004-2006 / Epidemic profile of the patients with Chronic Hepatitis for virus C and its early virological answer to the treatment with Pegilated Interferon more Ribavirina. Service of Gastroenterology, HNCASE ESSALUD Arequipa 2004-2006

Se realizó el presente estudio descriptivo retrospectivo y longitudinal para determinar el perfil epidemiológico de los pacientes infectados por virus de hepatitis C con hepatitis crónica, así como la evaluación de sus criterios de legibilidad para recibir tratamiento con Interferón Pegilado más Ribavirina y su respuesta virológica temprana al tratamiento. Se estudiaron 20 pacientes atendidos en el Servicio de Gastroenterología del Hospital Carlos Seguín Escobedo de EsSalud entre los años 2004 y 2006. El diagnóstico de infección por HVC se confirmó mediante la detección de DNA viral mediante PCR, y de la carga viral mediante el conteo de número de copias. Se determinaron los criterios de legibilidad para tratamiento antiviral; se empleó además el score METAVIR para determinar la presencia de fibrosis hepática. Luego de 12 semanas de tratamiento se evaluó a los pacientes que cumplieron criterios de elegibilidad y que accedieron al tratamiento y se evaluó su efectividad mediante una nueva determinación de la carga viral. Hubo más pacientes mujeres (15; 75 por ciento) que varones (5; 25 por ciento), con edades entre 50 y 59 años. El antecedente de riesgo más frecuente fue transfusión sanguínea (45 por ciento), cirugía (35 por ciento), y accidentes de trabajo (10 por ciento). Los valores de parámetros hematológicos, renal y hormonal estuvieron dentro de límites normales, los parámetros de función hepática (bilirrubinas, aminotransferasas, fosfatasa alcalina y gamaglutamil transpeptidasa) estuvieron por encima de lo normal; la carga viral al momento del diagnóstico fue en promedio de 580 mil copias (entre 4100 a 2 millones de copias). Los genotipos virales más frecuentes fueron 1a y 1b. Hasta 40 por ciento de pacientes no tuvieron criterios de elegibilidad para recibir tratamiento antiviral. Sólo 9 a 12 pacientes que tuvieron criterios de tratamiento lo recibieron (75 por ciento), en 6 de ellos se evaluó la respuesta virológica temprana y de ellos...

The present descriptive, retrospective and longitudinal study was designed todetermine the epidemic profile of patients infected by hepatitis C virus and chronic hepatitis, as well as to describe the eligibility criteria for treatment with pegilated interferon plus ribavirina and its early virological answer to the treatment. We studied 20 patients treated at Gastroenterology Service of Carlos Seguín Escobedo Hospital of EsSalud between 2004 and 2006. The diagnosis of HVC infection was confirmed by detection of viral RNA with PCR, and the viral load by counting number of RNA copies. The eligibility criteria for antiviral treatment were determined, and also the METAVIR score to determine hepatic fibrosis. After 12 weeks of treatment the patients with eligibility criteria and received treatment were evaluated, and theeffectivity of treatment was evaluated with a new determination of viral load.There were more female (15; 75 per cent) than male patients (5; 25 per cent), with ages between 50 and 59 years. The more frequent risk antecedent was blood transfusion (45 per cent), surgery (35 per cent), and traffic accident (10 per cent). The hematological, hepatic, renal and hormonal parameters were in normal range; and the viral load at the moment of diagnosis was of 580 thousand copies mean (between 4100 to 2 millions copies). The most frequent viral genotypes were 1a and 1b. Up to 40 per cent of patients did not fulfilled eligibility criteria for treatment. Nine of 12 patients with eligibility criteria received treatment (75 per cent), 6of them were evaluated for early virologc response and of them 100,0 per cent had an early virologic response with decrease of post-therapeutic viral load in all but one cases until non-detectable levels. In conclusion our study described the profile of patients infected by HCV with chronic hepatitis in gastroenterology ward of our hospital, and it has been determined that is suggestive that antiviral treatment is highly...

High-dose chemotherapy and autologous peripheral blood stem cell rescue in a patient with pleuropulmonary blastoma.

Pleuropulmonary blastoma (PPB) is a rare and aggressive malignant tumor of the lung. Approximately 80 cases of PPB have been published, and in only three cases high-dose chemotherapy with autologous hematopoietic stem cell transplantation (HSCT) was applied. A 5-year-old girl presenting with cough, fever, and shortness of breath was referred to the authors in March 1999. A computed tomography scan of the chest showed a tumor mass in the left hemithorax. The lesion was biopsied and the histopathologic report suggested the diagnosis of PPB. The patient received chemotherapy comprising vincristine, actinomycin D, and cyclophosphamide with only a minor response, and treatment was switched to ifosfamide, carboplatin, and etoposide, which produced a partial response. Tumor resection was performed, but margins were positive for PPB. Due to the high risk of recurrence, the authors elected to administrate high-dose chemotherapy using melphalan, etoposide, and carboplatin, followed by autologous HSCT. The patient achieved complete hematologic recovery, and reimaging after HSCT showed no evidence of disease. She relapsed 4 months later and died about 9 months after the completion of high-dose therapy. The role of high-dose chemotherapy and autologous HSCT is likely to be limited in PPB.

Primary splenic angiosarcoma: case report and literature review

É relatado um caso de angiossarcoma primário de baço em um homem de 29 anos, fazendo-se a revisäo dos achados clínicos, diagnóstico anatomopatológico, tratamento e prognóstico. Angiossarcomas correspondem a menos de 1 porcento dos sarcomas de tecidos moles, e somente uma pequena fraçäo surge no baço. Esses tumores freqüentemente apresentam-se como um desafio diagnóstico para o patologista, tendo em vista as variaçöes na histologia, e o desenvolvimento de novos marcadores em painéis de imuno histoquímica, como o CD31, ajudam a descartar outros diagnósticos. Devido à raridade desse tumor näo existe um consenso em relaçäo ao tratamento quimioterápico e, via de regra, o prognóstico é ruim.