High-resolution profiling of histone h3 lysine 36 trimethylation in metastatic renal cell carcinoma.

Mutations in SETD2, a histone H3 lysine trimethyltransferase, have been identified in clear cell renal cell carcinoma (ccRCC); however it is unclear if loss of SETD2 function alters the genomic distribution of histone 3 lysine 36 trimethylation (H3K36me3) in ccRCC. Furthermore, published epigenomic profiles are not specific to H3K36me3 or metastatic tumors. To determine if progressive SETD2 and H3K36me3 dysregulation occurs in metastatic tumors, H3K36me3, SETD2 copy number (CN) or SETD2 mRNA abundance was assessed in two independent cohorts: metastatic ccRCC (n=71) and the Cancer Genome Atlas Kidney Renal Clear Cell Carcinoma data set (n=413). Although SETD2 CN loss occurs with high frequency (>90%), H3K36me3 is not significantly impacted by monoallelic loss of SETD2. H3K36me3-positive nuclei were reduced an average of ~20% in primary ccRCC (90% positive nuclei in uninvolved vs 70% positive nuclei in ccRCC) and reduced by ~60% in metastases (90% positive in uninvolved kidney vs 30% positive in metastases) (P<0.001). To define a kidney-specific H3K36me3 profile, we generated genome-wide H3K36me3 profiles from four cytoreductive nephrectomies and SETD2 isogenic renal cell carcinoma (RCC) cell lines using chromatin immunoprecipitation coupled with high-throughput DNA sequencing and RNA sequencing. SETD2 loss of methyltransferase activity leads to regional alterations of H3K36me3 associated with aberrant RNA splicing in a SETD2 mutant RCC and SETD2 knockout cell line. These data suggest that during progression of ccRCC, a decline in H3K36me3 is observed in distant metastases, and regional H3K36me3 alterations influence alternative splicing in ccRCC.

The Quantification of Representative Sequences pipeline for amplicon sequencing: case study on within-population ITS1 sequence variation in a microparasite infecting Daphnia.

Next generation sequencing (NGS) platforms are replacing traditional molecular biology protocols like cloning and Sanger sequencing. However, accuracy of NGS platforms has rarely been measured when quantifying relative frequencies of genotypes or taxa within populations. Here we developed a new bioinformatic pipeline (QRS) that pools similar sequence variants and estimates their frequencies in NGS data sets from populations or communities. We tested whether the estimated frequency of representative sequences, generated by 454 amplicon sequencing, differs significantly from that obtained by Sanger sequencing of cloned PCR products. This was performed by analysing sequence variation of the highly variable first internal transcribed spacer (ITS1) of the ichthyosporean Caullerya mesnili, a microparasite of cladocerans of the genus Daphnia. This analysis also serves as a case example of the usage of this pipeline to study within-population variation. Additionally, a public Illumina data set was used to validate the pipeline on community-level data. Overall, there was a good correspondence in absolute frequencies of C. mesnili ITS1 sequences obtained from Sanger and 454 platforms. Furthermore, analyses of molecular variance (amova) revealed that population structure of C. mesnili differs across lakes and years independently of the sequencing platform. Our results support not only the usefulness of amplicon sequencing data for studies of within-population structure but also the successful application of the QRS pipeline on Illumina-generated data. The QRS pipeline is freely available together with its documentation under GNU Public Licence version 3 at http://code.google.com/p/quantification-representative-sequences.

Manejo anestésico en una mujer con malformación de Arnold-Chiari tipo II residual / Anesthetic management in a woman with residual Arnold-Chiari malformation Type II

Background: The Arnold-Chiari malformation (ACM) is a group of congenital abnormalities of the hindbrain and the spinal cord and is characterized by herniation of the cerebellum, kinking of the medulla oblongata and hydrocephalus, the anesthetic management Is complicated due to the anatomic and physiologic alterations. Case description: A 23 years old woman with Arnold-Chiari type II malformation, and perforated appendicitis. Conclusions: The present case demonstrates that patients with partially corrected ACM type II, restrictive lung disease due to scoliosis and perforated appendicitis delivery require an interdisciplinary team approach, diligent preparation, and skilled physicians.

Antecedentes: La malformación de Arnold-Chiari (ACM) es un grupo de anomalías congénitas del cerebro posterior y la médula espinal que se caracteriza por la herniación del cerebelo, retorcimiento del bulbo raquídeo e hidrocefalia, el manejo anestésico se complica debido a la anatomía y alteraciones fisiológicas. Descripción del caso: Una mujer de 23 años con Arnold-Chiari tipo II malformación y apendicitis perforada. Conclusiones: El presente caso demuestra que los pacientes con corregido parcialmente ACM tipo II, enfermedad pulmonar restrictiva debido a la escoliosis y la entrega apendicitis perforada requieren un enfoque multidisciplinario en equipo, preparación diligente, y los médicos cualificados.

Human papillomavirus type 16 reduces the expression of microRNA-218 in cervical carcinoma cells.

Human papillomaviruses (HPVs) are involved in the pathogenesis of cancer of the cervix (CaCx). MicroRNA (miRNA) expression analysis using Ambion (Austin, TX, USA) arrays showed that three miRNAs were overexpressed and 24 underexpressed in cervical cell lines containing integrated HPV-16 DNA compared to the normal cervix. Furthermore, nine miRNAs were overexpressed and one underexpressed in integrated HPV-16 cell lines compared to the HPV-negative CaCx cell line C-33A. Based on microarray and/or quantitative real-time PCR and northern blot analyses, microRNA-218 (miR-218) was specifically underexpressed in HPV-positive cell lines, cervical lesions and cancer tissues containing HPV-16 DNA compared to both C-33A and the normal cervix. Expression of the E6 oncogene of high-risk HPV-16, but not that of low-risk HPV-6, reduced miR-218 expression, and conversely, RNA interference of E6/E7 oncogenes in an HPV-16-positive cell line increased miR-218 expression. We also demonstrate that the epithelial cell-specific marker LAMB3 is a target of miR-218. We also show that LAMB3 expression is increased in the presence of the HPV-16 E6 oncogene and this effect is mediated through miR-218. These findings may contribute to a better understanding of the molecular mechanisms involved in cervical carcinogenesis.

[Incidence and mortality due to cancer in Navarre, 1998-2002. Trends in the last 30 years]. / Incidencia y mortalidad por cáncer en Navarra, 1998-2002. Evolución en los últimos 30 años.

Between 1998-2002, 16,952 new cases of cancer were registered in Navarre. In men, the most frequently diagnosed cancers were in the following order: prostate, lung, colon and rectum, bladder and stomach, which accounted for 63.2%. In women, the sites were breast, colon and rectum, corpus uteri, stomach and ovary, which accounted for 57.6% of the cases. In the same period, 1998-2002, 4,127 men and 2,470 women died from cancer. Sixty percent of all deaths due to malign tumours in men were due to cancer of the lung, prostate, colon and rectum, stomach and bladder. In women this was due to cancers of colon and rectum, breast, stomach, pancreas and lung, which accounted for 49% of the cases. In men in Navarre there has been an increase in the incidence rates of cancer of the prostate, kidney and non-Hodgkin lymphoma. Avoidable cancers such as those related to smoking (lung, oral cavity and pharynx or pancreas) continue to rise, and represent a greater global risk of dying from cancer in the latest period studied than in the decades of the 1970s and 1980s. From 1995 up to the present, mortality due to cancer has moved from occupying the second place to become the first cause of death among men in Navarre. The global risk of death due to cancer in men is now equal to the first period studied, 1975-1977. Amongst women the global risk of death due to cancer fell by 25% between 1975 and 2002, basically at the cost of breast and stomach cancer. Tumours related to smoking increased both in mortality and in incidence and appear as a significant health problem amongst women in Navarre. Breast cancer has increased in incidence, with lower mortality figures than those of the first period 1975-1977. Invasive cancer of the cervix remains at very low rates in comparison with many European countries, including Spain. In both sexes colorectal and skin cancer has increased, while the incidence and mortality of stomach cancer continues to fall.

DNA methylation paradigm shift: 15-lipoxygenase-1 upregulation in prostatic intraepithelial neoplasia and prostate cancer by atypical promoter hypermethylation.

Fifteen (15)-lipoxygenase type 1 (15-LO-1, ALOX15), a highly regulated, tissue- and cell-type-specific lipid-peroxidating enzyme has several functions ranging from physiological membrane remodeling, pathogenesis of atherosclerosis, inflammation and carcinogenesis. Several of our findings support a possible role for 15-LO-1 in prostate cancer (PCa) tumorigenesis. In the present study, we identified a CpG island in the 15-LO-1 promoter and demonstrate that the methylation status of a specific CpG within this island region is associated with transcriptional activation or repression of the 15-LO-1 gene. High levels of 15-LO-1 expression was exclusively correlated with one of the CpG dinucleotides within the 15-LO-1 promoter in all examined PCa cell-lines expressing 15-LO-1 mRNA. We examined the methylation status of this specific CpG in microdissected high grade prostatic intraepithelial neoplasia (HGPIN), PCa, metastatic human prostate tissues, normal prostate cell lines and human donor (normal) prostates. Methylation of this CpG correlated with HGPIN, PCa and metastatic human prostate tissues, while this CpG was unmethylated in all of the normal prostate cell lines and human donor (normal) prostates that either did not display or had minimal basal 15-LO-1 expression. Immunohistochemistry for 15-LO-1 was performed in prostates from PCa patients with Gleason scores 6, 7 [(4+3) and (3+4)], >7 with metastasis, (8-10) and 5 normal (donor) individual males. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to detect 15-LO-1 in PrEC, RWPE-1, BPH-1, DU-145, LAPC-4, LNCaP, MDAPCa2b and PC-3 cell lines. The specific methylated CpG dinucleotide within the CpG island of the 15-LO-1 promoter was identified by bisulfite sequencing from these cell lines. The methylation status was determined by COBRA analyses of one specific CpG dinucleotide within the 15-LO-1 promoter in these cell lines and in prostates from patients and normal individuals. Fifteen-LO-1, GSTPi and beta-actin mRNA expression in BPH-1, LNCaP and MDAPCa2b cell lines with or without 5-aza-2'-deoxycytidine (5-aza-dC) and trichostatin-A (TSA) treatment were investigated by qRT-PCR. Complete or partial methylation of 15-LO-1 promoter was observed in all PCa patients but the normal donor prostates showed significantly less or no methylation. Exposure of LNCAP and MDAPCa2b cell lines to 5-aza-dC and TSA resulted in the downregulation of 15-LO-1 gene expression. Our results demonstrate that 15-LO-1 promoter methylation is frequently present in PCa patients and identify a new role for epigenetic phenomenon in PCa wherein hypermethylation of the 15-LO-1 promoter leads to the upregulation of 15-LO-1 expression and enzyme activity contributes to PCa initiation and progression.

Cytokeratin profiles of male breast cancers.

AIMS: The prognostic factors and expression of molecular markers in male breast carcinomas are similar to those in female breast cancers. The identification of distinct cytokeratin (CK) profiles (basal as opposed to luminal cells) helps to identify subsets of tumours with different clinical behaviour. The aim of this study was to investigate CK expression in male breast cancer. METHODS AND RESULTS: Thirty-two cases of male breast cancer were studied. The panel of CKs studied by immunohistochemistry included: 5/6, 14, 17, 18 and 19. Pathological findings and CK expression were analysed in all cases. Histological patterns included ductal carcinoma in situ, invasive ductal carcinoma and mixed patterns. Four cases were positive for CK5/6 and CK14, identifying a basal-like phenotype. CK17 was negative in all but two cases. All cases expressing either CK5/6 or CK14 were invasive carcinomas of high nuclear and histological grade and were also larger compared with the tumours not expressing CK5/6 and CK14. All tumours except three (also negative for CK5/6) expressed CK18 and CK19. The four basal-like tumours were negative for Her-2 expression. CONCLUSIONS: Male breast carcinomas have a basal-like phenotype that is similar in frequency to that of female breast carcinomas. The expression of CK5/6 and CK14 identifies a subset of pathologically aggressive male breast cancers.

[Effects of air pollution on daily mortality in the city of Zaragoza, Spain, 1991-1995]. / Efectos de la contaminación atmosférica sobre la mortalidad diaria en la ciudad de Zaragoza, España, 1991-1995.

OBJECTIVE: To analyze the short-term impact of air pollution on daily mortality in the City of Zaragoza (Spain). MATERIAL AND METHODS: This ecologic study was conducted in Zaragoza, Spain, from 1991 to 1993. Poisson autoregressive models were constructed to assess the association between air particulate matter and sulphur dioxide SO2 and daily deaths. Four outcome variables were studied: overall mortality (excluding external deaths), mortality in subjects over 69 years of age, mortality due to respiratory disease, and mortality due to cardiovascular disease. A sample size was not obtained. Data analysis was conducted using the statistical software EGRET and SPSS. RESULTS: SO2 levels were significantly associated with cardiovascular deaths (RR = 1.018 IC 95%: 1.001-1.036) and particulate matter levels with respiratory deaths (RR = 1.028 IC 95%: 1.006-1.051). During the warm season, a significant relationship was found between particulate mater and cardiovascular deaths (RR = 1.020 IC 95%: 1.001-1.040). CONCLUSIONS: Low levels of air pollution were found in Zaragoza, with considerable variation in the concentration of air pollutant concentrations and daily respiratory and cardiovascular deaths, particularly during the warm season. The English version of this paper is available at: http://www.insp.mx/salud/index.html

Overexpression of p21WAF1/CIP1 and MDM2 characterizes serous borderline ovarian tumors.

Ovarian epithelial tumors are classically divided into benign, malignant, and borderline or of low malignant potential. It is controversial whether this last group of tumors should be considered benign or malignant. Expression of cell cycle markers has recently been linked to tumor behavior and response to treatment. It has been shown that one of the pathways through which the p53 gene controls the cell cycle is by transactivating p21WAF1/CIP1, a cyclin-dependent kinase (cdk) inhibitor. By inhibiting cdks, p21WAF1/CIP1 blocks the G-1 to S-phase transition in the cell cycle. p53 can be regulated by MDM2 (murine double minute-2) through direct inactivation or promotion of its cytoplasmic degradation. In an attempt to investigate the cell cycle checkpoint mechanisms of these tumors, we studied the expression of p53, Ki-67, MDM2, and p21WAF1/CIP1 by immunohistochemistry. We analyzed the expression of these proteins in 19 cystadenomas (8 serous and 11 mucinous), 40 borderline tumors (31 serous and 9 mucinous), and 18 serous carcinomas of the ovary. p21WAF1/CIP1 was expressed in 7 of 19 (37%) benign cystadenomas, 32 of 40 (80%) borderline tumors (93.5% of serous and 33% of mucinous), and in 9 of 18 (50%) serous carcinomas. Ki-67 was only weakly expressed in 8 of 19 (42%) benign cystadenomas, all borderline tumors showed Ki-67 staining in less than 50% of the cells, and 55% of serous carcinomas stained in more than 50% of tumor cells. p53 was absent in all but 1 of the cystadenomas, was expressed in 9 of 40 (22.5%) borderline tumors (25.8% of serous and 11% of mucinous), and in 10 of 18 (55%) carcinomas. All 11 implants of serous borderline tumors expressed p21WAF1/CIP1. Most serous borderline tumors expressed higher levels of MDM2 compared with the benign cystadenomas and carcinomas. Four of the serous borderline implants (40%) expressed MDM2. Coexpression of p21WAF1/CIP1 and MDM2 characterizes serous borderline tumors of the ovary and their implants, which suggests that these cell cycle control proteins are important in these tumors and may be related to tumor progression. Low expression of p53 protein in serous borderline tumors might be in part mediated by MDM2. This suggests that the p53 pathway is intact in most of these tumors, in contrast with carcinomas, in which high expression of p53 has been related to mutations of this gene.

Angiomatoid melanoma: a novel pattern of differentiation in invasive periocular desmoplastic malignant melanoma.

A case of locally invasive, long-standing desmoplastic and amelanotic malignant melanoma is described in an 84-year-old man. Histologic examination of the involved periorbital tissue showed neoplastic foci exhibiting a novel pattern reminiscent of microvascular proliferation. These regions were characterized by malignant, S-100-positive tumor cells lining vessel-like spaces in transverse sections and forming tubuler-like structures in longitudinal sections. Recent data indicate that melanoma cells may express genes and patterns of differentiation in vitro akin to endothelial cells. Because angiosarcoma often involves facial and scalp skin of elderly individuals, awareness of angiomatoid differentiation in melanoma has important diagnostic implications. HUM PATHOL 31:1520-1522.

[The EMECAM project: the Spanish Multicenter Study on the Relationship between Air Pollution and Mortality. The background, participants, objectives and methodology]. / El proyecto EMECAM: Estudio Multicéntrico Español sobre la Relación entre la Contaminación Atmosférica y la Mortalidad. Antecedentes, participantes, objetivos y métodología.

In recent years, a growing number of studies suggests that increases in air pollution levels may have short-term impact on human health, even at pollution levels similar to or lower than those which have been considered to be safe to date. The different methodological approaches and the varying analysis techniques employed have made it difficult to make a direct comparison among all of the findings, preventing any clear conclusions from being drawn. This has led to multicenter projects such as the APHEA (Short-Term Impact of Air Pollution on Health. A European Approach) within a European Scope. The EMECAM Project falls within the context of the aforesaid multicenter studies and has a wide-ranging projection nationwide within Spain. Fourteen (14) cities throughout Spain were included in this Project (Barcelona, Metropolitan Area of Bilbao, Cartagena, Castellón, Gijón, Huelva, Madrid, Pamplona, Seville, Oviedo, Valencia, Vigo, Vitoria and Saragossa) representing different sociodemographic, climate and environmental situations, adding up to a total of nearly nine million inhabitants. The objective of the EMECAM project is that to asses the short-term impact of air pollution throughout all of the participating cities on the mortality for all causes, on the population and on individuals over age 70, for respiratory and cardiovascular design causes. For this purpose, with an ecological, the time series data analyzed taking the daily deaths, pollutants, temperature data and other factors taken from records kept by public institutions. The period of time throughout which this study was conducted, although not exactly the same for all of the cities involved, runs in all cases from 1990 to 1996. The degree of relationship measured by means of an autoregressive Poisson regression. In the future, the results of each city will be combined by means of a meta-analysis.

[The short-term effects of air pollution on mortality. The results of the EMECAM project in Saragossa, 1991-1995. Estudio Multicéntrico Español sobre la Relación entre la Contaminación Atmosférica y la Mortalidad]. / Efectos a corto plazo de la contaminación atmosférica sobre la mortalidad. Resultados del proyecto EMECAM en Zaragoza, 1991-1995.

BACKGROUND: To assess the short-term impact of pollution on the respiratory death rate in the city of Saragossa throughout the 1991-1995 period and to pinpoint whether any differences exists in terms of age and time of the year. METHODS: The relationship of daily concentrations of smog and SO2 to the daily deaths due to respiratory diseases (CIE-9 460-486) and chronic lung blockage disease and similar EPOC-EA (490-496) was analyzed using Poisson models in keeping with the EMECAM procedure. Possible differences in the impact on those below and over age 70 and according to the six-month period in question were researched. Relative risks (RR) and 95% confidence, intervals (CI95%) WERE CALCULATED FOR 10 micrograms/m3 rises in pollutant. RESULTS: A relationship was found to exist between the respiratory and smog death rate (RR 1.028 CI95% 1.006-1051), the highest risk being during the six-months period of warm weather. For those individuals over age 70, the relationship remained the same throughout this six-month period and was negative for those individuals under age 70. The RR's for the death rate based on EPOC-EA were, overall, 1.038 (CI95% 1.002-1075) and of 1.068 (CI95%: 1.004-1.137) for the six-month period of warm weather. The SO2 pollution showed a positive relationship to the respiratory death rate for the warm period for all ages, RR 1.093 (CI95%: 1.006-1.187) and for those under age 70 (RR 1.240 CI95%: 1.028-1.496). The impact was not conclusive for the cases of pneumonia. CONCLUSIONS: Low levels of air pollution can have a significant impact on the respiratory death rate, especially among the elderly and during the six-month period of warm weather.

[Ki-1+ large-cell anaplastic lymphoma with a leukemic appearance. Study of a case]. / Linfoma anaplásico de célula grande Ki-1 (+) con expresión leucémica. Estudio de un caso.

Ki-1 anaplastic large cell lymphoma is a well-described subtype of non-Hodgkin's lymphoma with distinctive characteristics from the cytological, immunohistochemical and clinical points of view. One of the clinical behavior characteristic is that it rarely evolves into a leukaemic phase. We report the case of a 72-year-old man in which the appearance of tumor cells in peripheral blood was one of the most revealing information. The patient showed B-symptoms, bicytopenia and bone marrow involvement, together with hepatosplenomegaly and right axilar adenopathy, which after biopsied lead to Ki-1 anaplastic large cell lymphoma's diagnosis (stage IV-B). As far as the treatment and evolution are concerned, we choose a polychemotherapy (ACOP-B) because of the patient's age. Up to now clinical and analitical course is excellent and the patient is now in remission.

[Dendriform diffuse pulmonary ossification. Report of 2 cases]. / Osificación pulmonar difusa ramificada. Presentación de dos casos.

Two cases of diffuse dendriform pulmonary ossification are reported which was diagnosed postmorten. Both patients had a previous history of chronic obstructive pulmonary disease with pneumonic complications. The radiological study was interpreted as pulmonary fibrosis in the first case, an entity which ranks first in the differential diagnosis. In some cases the obtention of highly contrasted X-ray films can be useful for recognizing this are entity.

[Bronchioalveolar adenocarcinoma associated with a chylous ascites. An unusual case]. / Adenocarcinoma bronquioloalveolar asociado a ascitis quilosa. Un caso inusual.

A case of bronchioloalveolar carcinoma with long evolution associated to chylous is reported. The natural clinical history of the adenocarcinoma follows with an unusual dissemination as an atypical manifestation of lymphatic metastasis.

[Phospholipase A2 and premature membrane rupture]. / Fosfolipasa A2 y ruptura prematura de membranas.

The phospholipase A2 activity in young amniotic fluids: 1.782, was determined. In the term amniotic fluids, the activity was 10.693. The detected activity in fluids from premature rupture of membranes was 29.077 in the group with infection, add 12.357 for the samples without infection. It was seen that young amniotic fluid has less activity. The group with infection showed the greatest activity. The significant increase of phospholipase A2 activity in term amniotic fluids and with PRM, in comparison with what is seen in young fluids suggests the active participation of the soluble form of this enzyme as mediator in the increase of Prostaglandin E2 seen in the amniotic fluid of the two first groups. Accordingly, one may think that PPLA2 could be a central mediator in membranes rupture mechanism in normal labor, as in PRM.

[Unavoidable epistaxis in the nasal infestation of leeches]. / Epistaxis debida a infestición nasal por sanguijuela.

An adult man is presented with a case of epistaxis due to leech (Limnatis linotica) intranasal parasitation. The geographical area where infection was performed is in the province of Teruel, northwest Spain. The only presented symptom was an incoercible nasal haemorrhage with low flow which didn't cease by anterior tamponade. 7 days after the process began he was heeded at the Emergency Service, Alcañiz Hospital, where the exam of left nasal fossa showed the presence of a olive-green long mass, very mobile. The treatment consisted on the leech extraction with a dissection forceps. Haemorrhage stopped immediately after the mild intervention and, reexamination the nasal fossa 15 days after, it was no wound or infectious complication observed. The patient was infested when drinking water from a fountain placed in rural area.