Julia Cruz

Antimicrobial activity of benzophenones and extracts from the fruits of Garcinia brasiliensis.

The pericarp and seeds from fruits of Garcinia brasiliensis were subjected to extraction with hexane and ethanol. The pericarp hexane extract (PHE) and seed ethanol extract (SEE) were purified by silica gel column chromatography, which permitted isolation of the prenylated benzophenones 7-epiclusianone (1) and guttiferone-A (2), respectively. The antimicrobial activity of PHE, SEE, and compounds 1 and 2 were evaluated against Candida albicans, Staphylococcus aureus, Escherichia coli, and Bacillus cereus cultures. The minimum inhibitory concentration and minimum bactericidal concentration were established. The substances presented activity against S. aureus and B. cereus as follows: PHE, 4.0 microg/mL and 2.4 microg/mL; SEE, 10.0 microg/mL and 12.6 microg/mL; 7-epiclusianone, 1.2 microg/mL and 0.6 microg/mL; and guttiferone-A, 2.4 microg/mL and 2.4 microg/mL, respectively. The direct relationship between the lipophilic character of the structure and activity in Gram-positive bacteria was specifically observed. Therefore these extracts and prenylated benzophenones represent an interesting topic for further studies and open possibilities for an alternative control of diseases associated with Gram-positive bacteria.

Superficial EWSR1-negative undifferentiated small round cell sarcoma with CIC/DUX4 gene fusion: a new variant of Ewing-like tumors with locoregional lymph node metastasis

The present study describes a new case of EWSR1-negative undifferentiated sarcoma with CIC/DUX4 gene fusion. This case is similar to tumors described as primitive undifferentiated round cell sarcomas that occur mainly in the trunk and display an aggressive behavior. To our knowledge, this is the first report of such a tumor presenting locoregional lymph node metastasis. In view of previous studies that prove the existence of a particular variant of undifferentiated sarcoma with Ewing-like morphology and CIC/DUX-4 gene fusion, a search for this gene fusion in all undifferentiated round cell sarcomas should be considered if a conclusive diagnosis cannot be reached following other conventional studies. Although additional cases with more extensive follow-up studies are needed, we believe that EWSR1-negative undifferentiated small round cell sarcoma with CIC/DUX4 gene fusion should be added to the list of new sarcoma variants with the possibility of lymph node metastasis.

Superficial small round-cell tumors with special reference to the Ewing's sarcoma family of tumors and the spectrum of differential diagnosis

Superficial/cutaneous small round-cell tumors comprise a heterogeneous group of neoplasms including sarcoma, carcinoma, melanoma, and lymphomas. Among superficial sarcomas, the Ewings sarcoma family of tumors (ESFT) represents a poorly understood rare variant, having a behavioral difference characterized by a relative favorable prognosis. Several problems are still to be resolved in superficial ESFT, including the differential diagnosis between ESFT of bone (intraosseous or periosteal) with superficial infiltration and superficial ESFT with bone infiltration, especially in the fingers. Our aim is to review the most common types of small round-cell tumors included in the differential diagnosis of superficial ESFT, analyzing the histopathology, phenotype, and molecular alterations of each entity.

Pleomorphic hyalinizing angiectatic tumor: a report of 3 new cases, 1 with sarcomatous myxofibrosarcoma component and another with unreported soft tissue palpebral location.

Pleomorphic hyalinizing angiectatic tumor (PHAT) is an uncommon soft tissue tumor usually located in extremities or trunk. We report 3 new cases with histopathologic diagnosis of PHAT, one with recurrence and sarcomatous myxofibrosarcoma component and another with unreported soft tissue palpebral location. Clinical data, histopathology, immunohistochemistry, fluorescence in situ hybridization, and follow-up data are described. The histopathology showed a tumor with angiectatic blood vessel proliferation and perivascular hyaline material associated with focal pleomorphic cells. The recurrent tumor revealed a histopathologic pattern corresponding to a myxofibrosarcoma. Vimentin and CD99 were positive in tumor cells and CD34 was strongly positive in the tumor cells from the recurrence. Ki-67 was poor positive but with increased positivity in the recurrence. The positivity of p53 and chromosome 22 polysomy were detected in the recurrence. At present, the 3 patients are free of disease and no metastases have been detected. Indeed, the possibility that PHAT may represent a histopathologic pattern and not a true neoplastic entity with specific genetic alterations cannot be excluded at present, and further studies are required.

Prognostic factors in squamous cell lip carcinoma treated with high-dose-rate brachytherapy

The purpose of this study was for us to present our analysis of the results and prognostic factors in squamous lip carcinoma treated with high‐dose‐rate (HDR) brachytherapy.

Desmoplastic melanoma may mimic a cutaneous peripheral nerve sheath tumor. Report of three challenging cases.

Desmoplastic melanoma (DM) and cutaneous malignant peripheral nerve sheath tumors (MPNST) reveal histological and immunohistochemical similarities, including S100 positivity and negative staining for conventional melanocytic markers. We present 3 cases of cutaneous S100‐positive spindle cell tumors in elderly patients, in which first findings led to initial misdiagnoses as cutaneous MPNST and benign peripheral sheath nerve tumor (neurofibroma). The identification of adjacent atypical melanocytic hyperplasia in the overlying skin along with tumor cell proliferation, also in the superficial dermis, the neurotropic component and the absence of any relationship between the tumor and a major nerve, pre‐existing neural benign tumor or the existence of stigmata suggestive of neurofibromatosis raised consideration of a DM. Careful attention should be paid to the presence of a firm dermal nodule and atypical scar lesions especially in sun‐exposed areas (mainly head and neck region) in elderly patients associated with S100‐positive spindle cell proliferation, solar elastosis and adjacent atypical melanocytic proliferation. In such cases, the possibility of a DM should be excluded with caution, especially if the tumor reveals a paucicellular morphology resembling various non‐melanocytic neoplasms including malignant or benign peripheral sheath nerve tumors.

Malignant PEComa With Metastatic Disease at Diagnosis and Resistance to Several Chemotherapy Regimens and Targeted Therapy (m-TOR Inhibitor)

Perivascular epithelioid cell tumors (PEComas) are infrequent neoplasms with peculiar myomelanocytic differentiation. The aggressive abdominopelvic variant is rare, with only a small number of published cases. We present an additional case of this unusual variant, which showed an aggressive histologic and clinical behavior with multiple liver metastases and resistance to several therapies. We also discuss the histological and immunohistochemical profiles as well as the differential diagnosis.

Immunohistochemical evaluation of a novel clone of monoclonal anti-MYCN antibody B8.4B in neuroblastic tumours: a correlation with MYCN gene status

To the Editor, Amplification of the MYCN protooncogene is recognised as a reliable, established and an independent prognostic factor in neuroblastic tumours (NTs), identifying children with rapidly progressive disease often refractory to conventional therapy [5]. Detection of MYCN amplification is routinely carried out in the clinical staging of NTs; however, very few studies investigating the clinical significance of MYCN expression were reported. The biological value of over-expression of MYCN RNA or its protein remains controversial. Some reports have indicated a significant correlation between MYCN protein overexpression and poor prognosis [2, 4], whilst others deny its prognostic value [3]. The aim of this report was to test a novel clone of mouse monoclonal anti-human MYCN antibody B8.4B for detecting the MYCN protein on formalin-fixed paraffin embedded specimens and to compare the MYCN protein expression determined by this marker with the MYCN gene status in a series of 210 neuroblastic tumours. The 210 neuroblastic tumours included in the study were classified according to the International Neuroblastoma Pathology Classification criteria. Forty-six of the 210 archival specimens were assembled in a tissue microarray comprising duplicate tissue cores, and the remaining 164 were analysed using conventional tissue sections. Hematoxylin and eosin sections were examined to estimate the percentage of tumour cells. Antigen retrieval was performed by boiling in pressure cooker for 3 min in 10 mM citrate buffer (pH 6.0). Immunohistochemical staining was carried out using primary mouse monoclonal anti-human N-Myc antibody (clone B8.4.B, BD PharMingen, Heidelberg, Germany) at 1:100 dilution using the DAKO ENVISION system (Dako Diagnostics, Denmark). The clone B8.4B recognises an epitope in the N terminus of human MYCN. Nuclear positivity in more than 5% of the tumour cells was considered as positive staining. Although this cutoff percentage of 5% was low, majority of the tumours showed more than 25% positivity, and only few cases displayed around 5 to 25% positivity. The topographical distribution and intensity of the staining were considered. Staining pattern was classified as focal when only one positive focus is present in the tumour sections examined, heterogeneous when multiple scattered positive foci are present with intervening negative areas and homogenous if the staining is uniform and diffuse. The range of the tumour cells positive for MYCN was grouped as 5–25%, 26–75% and more than 75%, and the number of cases under each group was calculated. The staining intensity was classified as mild, moderate and intense. Dual colour fluorescence in situ hybridization (FISH) analysis was performed according to standard protocol on touch preparations of all the 210 specimens using a cocktail DNA probe (Q-BIOgene) consisting of 2p24 MYCN red/D2Z green probes. The number of signals was analysed in at R. Noguera (*) . M. Piqueras . M. Subramaniam . J. Cruz . A. L. Bosch . S. Navarro Department of Pathology, Medical School, University of Valencia, Avda. Blasco Ibañez, 17, 46010 Valencia, Spain e-mail: rnoguera@uv.es Tel.: +34-96-3864146 Fax: +34-96-3864173

Angiomiolipoma epitelioide/PEComa hepático e hiperplasia nodular focal en una paciente con antecedentes de melanoma cutáneo

Hepatic perivascular epithelioid cell tumors (PEComas) are uncommon mesenchymal neoplasms. PEComas concurrent with other hepatic lesions is a very rare occurrence, with only two previously reported cases. We report a primary hepatic PEComa associated with focal nodular hyperplasia in a patient with a previous history of cutaneous melanoma. Diagnostic imaging studies suggested a hepatic adenoma and the patient underwent a segmentectomy. The tumor was mainly composed of epithelioid cells, adipose tissue and smooth muscle fibers intermixed with blood vessels. The neoplastic cells were diffusely immunoreactive for HMB-45, Melan-A and smooth muscle actin, but not for Hepatocyte, S100, MITF or BRAF. Molecular studies were negative for BRAFV600 mutation. The final diagnosis was hepatic epithelioid angiomyolipoma/PEComa. The differential diagnosis of hepatic PEComa is discussed.

Gene expression analyses determine two different subpopulations in KIT-negative GIST-like (KNGL) patients

Introduction There are limited findings available on KIT-negative GIST-like (KNGL) population. Also, KIT expression may be post-transcriptionally regulated by miRNA221 and miRNA222. Hence, the aim of this study is to characterize KNGL population, by differential gene expression, and to analyze miRNA221/222 expression and their prognostic value in KNGL patients. Methods KIT, PDGFRA, DOG1, IGF1R, MIR221 and MIR222 expression levels were determined by qRT-PCR. We also analyzed KIT and PDGFRA mutations, DOG1 expression, by immunohistochemistry, along with clinical and pathological data. Disease-free survival (DFS) and overall survival (OS) differences were calculated using Log-rank test. Results Hierarchical cluster analyses from gene expression data identified two groups: group I had KIT, DOG1 and PDGFRA overexpression and IGF1R underexpression and group II had overexpression of IGF1R and low expression of KIT, DOG1 and PDGFRA. Group II had a significant worse OS (p = 0.013) in all the series, and showed a tendency for worse OS (p = 0.11), when analyzed only the localized cases. MiRNA222 expression was significantly lower in a control subset of KIT-positive GIST (p < 0.001). OS was significantly worse in KNGL cases with higher expression of MIR221 (p = 0.028) or MIR222 (p = 0.014). Conclusions We identified two distinct KNGL subsets, with a different prognostic value. Increased levels of miRNA221/222, which are associated with worse OS, could explain the absence of KIT protein expression of most KNGL tumors.