Combined evaluation of both WEE1 and phosphorylated cyclin dependent kinase 1 expressions in oral squamous cell carcinomas predicts cancer recurrence and progression.

Background/purpose: WEE1 is a mitotic inhibitor at G2 checkpoint of the cell cycle that negatively regulates cyclin-dependent kinase 1 (CDK1) through inhibitory phosphorylation. This study assessed whether the expressions of both WEE1 and phosphorylated CDK1 in specimens of oral squamous cell carcinoma (OSCC) might predict the OSCC recurrence and progression. Materials and methods: This study used immunohistochemistry to examine the expressions of WEE1 and phosphorylated CDK1 proteins in 75 specimens of OSCC and 30 specimens of normal oral mucosa (NOM). Results: The mean WEE1 labeling index (LI) was significantly lower in 75 OSCC samples than in 30 NOM samples (P < 0.001), whereas the mean phosphorylated CDK1 LI was significantly higher in 75 OSCC samples than in 30 NOM samples (P < 0.001). We found a significant association of low WEE1 LI (<21%) with OSCC recurrence (P = 0.047) and a significant association of low phosphorylated CDK1 LI (<10%) with larger tumor size (P = 0.011) and more advanced clinical stages (P = 0.021) of OSCC. Conclusion: Combined evaluation of WEE1 and phosphorylated CDK1 LI in specimens of OSCC may predict the OSCC recurrence and progression.

Chromophobe renal cell carcinoma: Novel molecular insights and clinicopathologic updates.

Chromophobe renal cell carcinoma (ChRCC) is the third most common renal cell carcinoma (RCC) subtype, which predominantly occurs in sporadic setting. ChRCCs are considered to originate from the intercalated cell of distal tubules with two main morphological variants, classic and eosinophilic. Most ChRCCs carry a favorable clinical outcome. Histology alone is limited in predicting the behavior of ChRCCs that do not have overtly aggressive morphologic findings such as necrosis and sarcomatoid features. Along with positive CD117 expression, classic ChRCCs generally express diffuse and uniform CK7, while eosinophilic variant demonstrates more heterogeneous CK7 expression (rare or patchy). Multiple losses of chromosomes 1, 2, 6, 10, 13, 17, and 21 are considered to be the genetic hallmarks of classic and eosinophilic ChRCCs, while chromosomal gains are known to be associated with sarcomatoid ChRCCs. TP53 and PTEN are the two most frequently mutated genes in ChRCCs. The major challenge in the differential diagnosis of ChRCCs includes considerations around the eosinophilic variant (of ChRCCs), where it may share overlapping features with oncocytoma or other recent emergent oncocytic tumors. Most eosinophilic ChRCCs share expression of the recently described biomarkers, LINC01187 and FOXI1, with classic ChRCCs, however, a subset of eosinophilic-like ChRCCs with lower biomarker expression have been demonstrated to harbor MTOR gene mutations. Overall, the morphologic features of ChRCCs and genetic profile with combinations of chromosomal losses and gains suggest this tumor entity to represent a distinct, yet heterogeneous group of renal neoplasms.

Immunoexpression of lactoferrin in triple-negative breast cancer patients: A proposal to select a less aggressive subgroup.

Triple-negative breast cancer (TNBC) indicates a subset of breast carcinomas that does not express estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor-2 (HER2). According to the literature, TNBCs are aggressive tumors, characterized by a high incidence of recurrence and a high risk of disease progression. Lactoferrin (LF) is a single-chain, iron-binding glycoprotein of ~700 amino acids, which is involved in a wide range of biological activities, including iron-trafficking and carcinogenesis. The present study aimed to assess LF expression in human TNBC samples and the possible correlation with clinico-pathological parameters associated with biological aggressiveness. LF immunohistochemical expression was investigated in formalin-fixed, paraffin-embedded samples of human TNBC. Cases were analyzed according to an intensity distribution (ID) score, and only those showing an ID score of >2 were considered as positive for LF. LF immunostaining was encountered in 26.15% cases. A significant correlation was found between LF expression and a low Ki-67 labeling index (P=0.040), the absence of recurrence (P=0.010) and alive status (P=0.020). LF may assist in identifying a subset of TNBC with less aggressive biological behavior. The meaning of LF expression in TNBC remains unclear and is controversial. The present findings indicated that LF expression is correlated with a low growth fraction in these tumors. Thus, it is possible that the inhibition of the LF axis may be a valid therapeutic target for TNBC, and this should be confirmed by future studies.

Immunohistochemical Expression of the Tumor Suppressor Protein p16 INK4a in Cervical Adenocarcinoma / Expressão imunhistoquímica da proteína de supressão tumoral p16 INK4a em adenocarcinoma cervical

ABSTRACT Objective: To evaluate the diagnostic utility of the p16ink4a protein expression as a marker for adenocarcinoma of the cervix. Methods: In a cross-sectional study, p16ink4a expression was evaluated in 30 cervical biopsies from patients diagnosed with invasive adenocarcinoma from 2 reference clinics in Brazil, and compared with 18 biopsies of endocervical polyps (control cases). The performance of the tests for p16ink4a was evaluated using a conventional contingency table, and the Kappa (k) index was used to evaluate the agreement of the marker with the tissue diagnosis. Results: In total, 66% of the invasive adenocarcinoma cases were positive for p16ink4a. All of the adenomatous polyps cases used as negative controls were shown to be negative for p16ink4a. The marker showed a high sensitivity and a high negative predictive value. The Kappa index was good for p16ink4a (k 1/4 0.6). Conclusion: Considering the strong association between the p16ink4a marker and the cervical adenocarcinoma, its use represents an important tool for reducing incorrect diagnoses of adenocarcinoma and thereby avoiding overtreatment.

RESUMO Objetivo: Avaliar a utilidade diagnóstica da expressão da proteína p16ink4a como marcador de adenocarcinoma do colo. Métodos: Em estudo transversal, a expressão de p16ink4a foi avaliada em 30 biópsias cervicais de pacientes diagnosticadas com adenocarcinoma invasivo de colo uterino provenientes de dois serviços de referência no Brasil, comparando com achados em 18 biópsias de pólipos endocervicais (grupo de controle). Para avaliar a performance do teste, foi utilizada tabela de contingência convencional, e para avaliar a concordância com o diagnóstico, foi aplicado o índice de Kappa (k). Resultados: No total, 66% dos casos de adenocarcinoma invasivo foram positivos para p16ink4a. Todos os pólipos adenomatosos foram negativos para p16ink4a. O marcador mostrou uma alta sensibilidade e alto valor preditivo negativo. O índice de Kappa foi bom para p16ink4a (k 1/4 0.6). Conclusion: Considerando a forte associação entre o marcador p16ink4a e o adenocarcinoma cervical, seu uso representa uma ferramenta importante para reduzir o risco de diagnóstico incorreto de adenocarcinoma e, por conseguinte, evitar o excesso de tratamentos.

Collagen IV and laminin expression in squamous cellcarcinomas of lower lip and tongue / Expressão de colágeno IV e laminina em carcinomas de células escamosas de lábio inferior e língua

In this study, the expression of extra cellular matrix proteins was immunohisto chemically studied and compared with the histological grading of squamous cell carcinomas of the lower lip and tongue. Material and Methods: The lower lip carcinomas (n =12) and the tongue carcinomas (n = 12) were histopathologically graded according to Bryne’smethod. The immunohisto chemical technique used specific antibodies for collagen IV and laminin. Histopathological and immunohisto chemical analysis were carried-out at the invasive tumor front. Results: Most of the lower lip carcinomas(91.7%) were classified with lower scores and all tongue carcinomas (100%) with high-grade malignant scores (p < 0.01). Collagen type IV expression was absent in the peri tumoral basement membrane in 50% of lower lip carcinomas and in66. 7% of tongue carcinomas (p = 0.09). Laminin expression was absent in the peritumoral basement membrane in 66.7% of lower lip carcinomas and in 58.3% of tongue carcinomas (p = 0.48). When these two glycoproteins were expressed, they showed alinear, thin and discontinuous pattern and a weak intensity of expression. Conclusion: The high gradem alignancy score of the tongue carcinomas was associated with the pattern of expression of the matrix proteins studied. This suggested that tonguesquamous cell carcinomas have more invasive potential and more aggressive biological behaviorthan the lower lip carcinomas...

Neste estudo, a expressão das proteínas da matriz extracelular foi estuda da imunoisto quimicamente e comparada com a classificação histológica dos carcinomas de células escamosas do lábio inferior e língua. Material e Métodos: Os carcinomas de lábio inferior (n = 12)e os carcinomas de língua (n = 12) foram graduados histopatologicamente de acordo com o método de Bryne. A técnica de imunoistoquímica utilizou anticorpos específicos para colágeno IV e laminina. Análises histopatológica e imunoistoquímica foram conduzidas na frente invasiva tumoral. Resultados: A maioria dos carcinomas de lábio inferior (91,7%) foi classificada em baixo grau e todos os carcinomas de língua (100%) em alto grau de malignidade (p < 0,01). A expressão de colágeno tipo IV estava ausente na membrana basal peri tumoral em 50% dos carcinomas de lábio inferior e em 66,7% dos carcinomas de língua (p = 0,09). A expressão de laminina estava ausente na membrana basal peri tumoral em 66,7%dos carcinomas do lábio inferior e em 58,3% dos carcinomas de língua (p = 0,48). Quando estas duas glicoproteínas foram expressas, mostraram-se comum padrão linear, fino e descontínuo e uma fraca intensidade de expressão. Conclusão: O alto grau de malignidade dos carcinomas de língua associou-se com o padrão de expressão das proteínas de matriz estudadas. Isso sugere que carcinomas de células escamosas de língua têm comportamento biológico mais agressivo e potencial mais invasivo do que os carcinomas de lábio inferior...

Detección del receptor tipo 2 del factor de crecimiento epidérmico humano (her2/neu): inmunohistoquímica en carcinomas de mama / Human epidermal growth factor receptor type 2 (her2/neu) immunohistochemical detection in breast carcinoma

Antecedentes: la incidencia y mortalidad por cáncer de mama han presentado un aumento neto. Uno de los tipos de esta heterogénea enfermedad se caracteriza por la amplificación y alta expresión del gen ERBB2, que codifica el receptor tipo 2 del factor de crecimiento epidérmico humano (HER2). Este receptor estimula procesos oncogénicos y dichos tumores se asocian a un peor pronóstico. El objetivo del estudio fue analizar las características de tumores HER-2+ en un grupo de pacientes con carcinoma de mama. Métodos: se estudió los casos de mujeres con biopsia diagnóstica de cáncer de mama registrados durante 2006 en los archivos del Servicio de Anatomía Patológica del Hospital San Juan de Dios; entre los que contaban con estudios inmunohistoquímicos, se estudió la expresión de HER-2 y se realizó análisis estadísticos. Resultados: se halló 34 tumores HER-2+ (15,7%), 24 con expresión fuerte (11,1%) y 10 con débil (4,6%). La edad media al diagnóstico de las pacientes con expresión fuerte fue de 46,9 años (42,5-51,1 IC 95%), y con expresión débil fue de 54,4 años (46,7-62,1 IC 95%); la edad promedio para los tumores HER-2- fue de 58,1 años (56,2-60,0 IC 95%). Las pacientes <50 años tuvieron un OR=3,477 de tener HER-2+. De 21 tumores, el 90,5% presentó un grado histológico alto, y de 16 casos, únicamente 3 tenían un tamaño <2 cm. Conclusión: se encontró una asociación del tipo HER-2+ con pacientes jóvenes, tumores de mayor tamaño y alto grado histológico. Los datos encontrados en este primer reporte son similares a las últimas estimaciones mundiales.

Background: The incidence and mortality due to breast cancer has suffered a net increase. One type of this heterogeneous disease is characterized by an amplification and higher expression of the ERBB2 gene that codifies for human epidermal growth factor receptor type 2 (HER-2). This receptor mediates oncogenic processes and such tumors have been associated with a worse prognosis. The aim of this study was to analyze the characteristics of HER-2+ tumors in a group of patients with breast cancer. Methods: This study included cases of breast cancer diagnosed through biopsy, registered during 2006 in the archives of the Pathology Department of the San Juan de Dios Hospital. The HER-2 expression was studied and statistical analyses were performed in cases that had immunohistochemical studies. Results: Thirty-four HER-2+ tumors were found (15.7 %), 24 with a strong expression (11.1%) and 10 with a weak expression (4.6%). The average age at diagnosis of patients with a strong expression was 46.9 years (42.5-51.1 IC 95%) and in those with a weak expression, 54.4 years (46.7-62.1 IC 95%). In the case ofHER-2- tumors, de mean age at diagnosis was 58.1 years (56.2-60.0 IC 95%). Patients <50 years had an OR=3.477 of having HER-2+. Out of 21 tumors, 90.5% showed a high histologic grade and out of 16 cases, only 3 measured <2 cm. Conclusion: The data found is similar to the latest global statistics. An association was found between HER-2+ type tumor and young patients, larger size tumors and higher histologic grade.

Expressions of p16 and p27 in urothelial carcinoma and their prognostic value.

Expressions of human p16 and p27 were tested for correlations with clinicopathologic features of urothelial carcinoma (UC). Tissue microarrays (TMA) constructed from paraffin-embedded specimens from 78 patients with UC were analyzed by immunohistochemical staining. In 49 of the 78 tumors (63%), high p16 expression was associated with absence of tumor invasiveness and low-grade carcinoma (p = 0.003 and p = 0.046, respectively). The p27 expression was high in 33 of the 78 tumors (42%) and showed a significant negative association with invasiveness, carcinoma grade, and tumor size (p = 0.016, p = 0.046, and p = 0.014, respectively). Kaplan-Meier analysis indicated that patients with high p27 levels had longer than average overall survival (p = 0.021). This study demonstrates that p16 and p27 are prognostic indicators of tumor stage and grade in UC and that they provide clinicians with the ancillary information needed for selecting suitable therapeutic strategies.

Association of pocket epithelial cell proliferation in periodontitis with TLR9 expression and inflammatory response.

BACKGROUND/PURPOSE: Inflammatory response is triggered after recognition of microbial ligands by innate receptors such as Toll-like receptors (TLRs) and Nucleotide oligomerization domain (NOD)-like receptors (NLRs). In this study, we examined serial frozen sections of gingival biopsies from patients with gingivitis or periodontitis by immunohistochemical analysis for the topographic expression patterns of selected innate receptors and their association with cell proliferation in clinically healthy and diseased gingival tissues. METHODS: A total of 19 gingival biopsies were collected from patients at the School of Dentistry, National Taiwan University Medical Center according to approved protocol and with informed consent. The specimens were assigned to either the gingivitis group or periodontitis group after clinical evaluation using gingival index. Frozen sections of gingival biopsies were stained with hematoxylin and eosin for histological evaluation. Serial sections of the same samples were stained with a panel of antibodies for immunohistochemical analysis. Expression of each protein marker was compared in the oral versus the sulcular epithelium of the same section. RESULTS: Expression of cytokeratin 19 (CK19) was markedly increased in the basement membranes of the oral epithelium and in all layers of the pocket epithelium where it caused evident cell proliferation and migration of sulcular epithelial cells into the lamina propria of periodontitis tissue. TLR4 and the cytoplasmic NLRP3 were expressed in all sections examined regardless of disease state. However, expression of TLR9-, CK19- and collagenolytic matrix metalloproteinase-13 and activated NF-κB subunit p65 was more commonly found in periodontitis tissues than in gingivitis tissues. CONCLUSION: Activation of TLR9 signaling in the pocket epithelium was highly associated with periodontal inflammation and possibly with loss of tissue integrity. Further studies of mechanisms by which TLR9 signaling is activated in the periodontal epithelium may lead to new strategies for treating periodontitis.

High TMPRSS4 expression is a predictor of poor prognosis in cervical squamous cell carcinoma.

OBJECTIVE: The aim of this study was to clarify the clinical role of TMPRSS4 expression in cervical squamous cell carcinoma (CSCC) and to investigate the role of TMPRSS4 in predicting outcomes of patients with CSCC. METHODS: The retrospective study enrolled 87 patients diagnosed with CSCC between 2004 and 2006. TMPRSS4 expression in CSCC was assessed by immunohistochemistry, and data on clinical variables were collected by retrospective chart review. The impact of TMPRSS4 expression on 5-year disease-free survival (DFS) and 5-year overall survival (OS) was assessed by Kaplan-Meier analysis and Cox proportional hazards modeling. RESULTS: The high expression of TMPRSS4 was 63.2% in 87 patients with CSCC, and 17.5% in 40 patients with benign cervical disease (P<0.001). High TMPRSS4 expression was significantly associated with tumor grade (P=0.005), lymph node metastasis (P=0.004), and deep cervical stromal invasion (P=0.025). Patients with high expression of TMPRSS4 had shorter OS and DFS than those with low expression (P=0.0205 and P=0.0318, respectively). In multivariate Cox regression analysis, high expression of TMPRSS4 was a potential prognostic indicator for OS (P=0.041) and DFS (P=0.015). CONCLUSION: Our findings suggest that TMPRSS4 might play an important role in the progression of CSCC. TMPRSS4 could be a potential prognostic marker of CSCC.

Differential expression of Rab27A/B correlates with clinical outcome in hepatocellular carcinoma.

AIM: To investigate the association of Rab27A and Rab27B expression with clinicopathological characteristics and prognosis of hepatocellular carcinoma (HCC). METHODS: We used reverse transcription polymerase chain reaction (RT-PCR), real-time PCR, and Western blotting to detect Rab27A and Rab27B mRNA and protein expression in 5 human HCC lines and the immortalized hepatic HL-7702 cell line. We further examined 148 primary HCC samples matched with adjacent normal tissue and 80 non-HCC specimens by immunohistochemistry to evaluate the correlation of Rab27A and Rab27B expression with clinicopathological features and prognosis. RESULTS: Our data showed that Rab27A and Rab27B were differentially expressed in cell lines and primary HCC tumors. Rab27A mRNA and protein were detected in 67% (4/6) of human cell lines and 80% (4/5) of HCC cell lines, while Rab27B was found in 50% (3/6) of human lines and 40% (2/5) of HCC lines. Rab27A expression was higher in primary HCC (46.2%, 66/143) than in matched adjacent tissue (24.3%, 33/136, P < 0.001), whereas immunopositivity for Rab27B was lower in primary HCC (57.4%, 81/141) than in matched adjacent tissue (87.5%, 119/136, P < 0.001). Analysis of clinicopathological characteristics of 148 HCC specimens revealed significant correlations between Rab27A and Rab27B expression and tumor tumor-node-metastasis (TNM) classification (P = 0.046 and P = 0.027, respectively), and between strong Rab27A expression and tumor differentiation grade (P = 0.008). Survival analyses revealed that patients with Rab27A(+) or Rab27B(+) tumors had significantly reduced overall survival compared with that of patients with Rab27A(-) or Rab27B(-) tumors (P = 0.015 and P = 0.005, respectively). Risk analyses revealed that Rab27B(+) and TNM III-IV were independent poor prognosis factors associated with a 3.36- and 3.37-fold higher relative risk of death, respectively. CONCLUSION: Rab27A and Rab27B expression were closely correlated with tumor progression and can be valuable prognostic indicators for HCC patients.

Pneumonia enzoótica em javalis (Sus scrofa) / Enzootic pneumonia in wild boars (Sus scrofa)

O objetivo desse estudo foi descrever o quadro clínico e epidemiológico, os achados patológicos, bacteriológicos e imuno-histoquímicos de um surto de pneumonia em uma granja de Javalis do Distrito Federal, Brasil. Em um período de cinco meses, morreram 90 javalis. Desses, 63 tinham lesões pulmonares. Clinicamente apresentavam atraso no desenvolvimento corporal, diminuição do apetite, letargia, tosse e dificuldade respiratória, principalmente quando movimentados. Constatou-se elevação da temperatura, 40ºC em média. Na auscultação, havia crepitações e estertores pulmonares de intensidade moderada. As alterações macroscópicas nos pulmões analisados eram típicas de broncopneumonia lobular. As lesões caracterizavam-se por consolidação crânio-ventral na maioria dos pulmões. A coloração variava de difusamente vermelho-escuro a um padrão mosaico (lóbulos vermelho-escuros intercalados por lóbulos cinzas) ou difusamente acinzentados. Na maioria dos pulmões observou-se exsudato mucopurulento na luz dos brônquios e fluindo do parênquima. Histologicamente, as alterações eram de broncopneumonia purulenta e histiocitária com focos de necrose. Em alguns animais havia também hiperplasia do BALT e, na maioria dos animais, infiltração linfocítica perivascular e peribronquial. Bordetella bronchiseptica e Streptococcus spp. foram as principais bactérias isoladas. A imuno-histoquímica demonstrou a bactéria Mycoplasma hyopneumoniae no epitélio bronquiolar e bronquial e o DNA desta bactéria foi detectado pela PCR. Este é o primeiro relato de broncopneumonia em Javalis associado à infecção por M. hyopneumoniae.

The aim of this paper is to describe the clinical, epidemiological, pathological, bacteriological and immunohistochemical aspects of a pneumonia outbreak in a wild pig farm in the Distrito Federal, Brazil. Ninety wild pigs died in a period of five months, and 63 of these had pulmonary lesions. Clinically, the pigs presented reduced growth rate, anorexia, lethargy, cough and dyspnea, especially after they were moved. High body temperature (40ºC in average) was verified in some animals. Auscultation revealed moderate pulmonary crepitation and stertors. Pulmonary gross lesions were typical of lobular bronchopneumonia. Lung lesions were characterized by ventral-cranial consolidation in the majority of the cases. The color of affected pulmonary areas varied from diffuse dark red to mosaic pattern (dark red lobule intercalate by grayish lobule) or diffusely grayish. The majority of the lungs had mucopurulent exsudate in the bronchial lumen that also drained from the parenchyma cut surface. Upon microscopy, the changes were characterized by purulent and histiocytic bronchopneumonia with necrotic foci. In some animals, there was BALT hyperplasia associated with perivascular and peribronchial plasma cells and lymphocytes infiltration in most of these cases. Bordetella bronchiseptica and Streptococcus spp. were the most frequently isolated bacteria. Immunohistochemistry evaluation demonstrated Mycoplasma hyopneumoniae on the luminal surface of bronchial and bronchiolar epithelial cells, and the DNA of bacteria was detected by PCR. This is the first report of bronchopneumonia in wild boars associated with M. hyopneumoniae infection.