Lack of cytomegalovirus detection in human glioma.

Gliomas are the most common brain tumors and include a variety of histologic types and grades of malignancy. They arise from glial cells and represent approximately 70% of the primary brain tumors. According to the criteria of the World Health Organization (WHO), the majority of gliomas can be classified into four grades of malignancy (I-IV). Virus infection, especially by DNA viruses and retroviruses, which may cause insertion of viral DNA sequences into the host genome, often triggers the host defense mechanisms. Particularly, the DNA methylation machinery can be activated to cause the methylation of foreign movable viral sequences and, therefore, silence viral gene expression. Several studies have shown the presence of Human Cytomegalovirus (HCMV) in glioblastoma, suggesting that the virus may participate in tumor pathogenesis. But this relationship is controversial because many other studies did not detect HCMV in these tumors. This study aims to detect the presence of HCMV in several samples of human glioma (94 formalin-fixed, paraffin-embedded samples and 28 snap-frozen samples) by different sensitive techniques. We have been unable to detect HCMV DNA and proteins in glioma samples. Therefore, arguments used so far to conclude that HCMV is an oncomodulator virus in gliomas must be, in our view, seriously reconsidered.

Risk of cancer in cases of suspected lynch syndrome without germline mutation.

BACKGROUND & AIMS: Colorectal cancers (CRCs) with microsatellite instability (MSI) and a mismatch repair (MMR) immunohistochemical deficit without hypermethylation of the MLH1 promoter are likely to be caused by Lynch syndrome. Some patients with these cancers have not been found to have pathogenic germline mutations and are considered to have Lynch-like syndrome (LLS). The aim of this study was to determine the risk of cancer in families of patients with LLS. METHODS: We studied a population-based cohort of 1705 consecutive patients, performing MSI tests and immunohistochemical analyses of MMR proteins. Patients were diagnosed with Lynch syndrome when they were found to have pathogenic germline mutations. Patients with MSI and loss of MSH2 and/or MSH6 expression, isolated loss of PMS2 or loss of MLH1 without MLH1 promoter hypermethylation, and no pathogenic mutation were considered to have LLS. The clinical characteristics of patients and the age- and sex-adjusted standardized incidence ratios (SIRs) of cancer in families were compared between groups. RESULTS: The incidence of CRC was significantly lower in families of patients with LLS than in families with confirmed cases of Lynch syndrome (SIR for Lynch syndrome, 6.04; 95% confidence interval [CI], 3.58-9.54; SIR for LLS, 2.12; 95% CI, 1.16-3.56; P < .001). However, the incidence of CRC was higher in families of patients with LLS than in families with sporadic CRC (SIR for sporadic CRC, 0.48; 95% CI, 0.27-0.79; P < .001). CONCLUSIONS: The risk of cancer in families with LLS is lower that of families with Lynch syndrome but higher than that of families with sporadic CRC. These results confirm the need for special screening and surveillance strategies for these patients and their relatives.

SpadaHC: a database to improve the classification of variants in hereditary cancer genes in the Spanish population.

Accurate classification of genetic variants is crucial for clinical decision-making in hereditary cancer. In Spain, genetic diagnostic laboratories have traditionally approached this task independently due to the lack of a dedicated resource. Here we present SpadaHC, a web-based database for sharing variants in hereditary cancer genes in the Spanish population. SpadaHC is implemented using a three-tier architecture consisting of a relational database, a web tool and a bioinformatics pipeline. Contributing laboratories can share variant classifications and variants from individuals in Variant Calling Format (VCF) format. The platform supports open and restricted access, flexible dataset submissions, automatic pseudo-anonymization, VCF quality control, variant normalization and liftover between genome builds. Users can flexibly explore and search data, receive automatic discrepancy notifications and access SpadaHC population frequencies based on many criteria. In February 2024, SpadaHC included 18 laboratory members, storing 1.17 million variants from 4306 patients and 16 343 laboratory classifications. In the first analysis of the shared data, we identified 84 genetic variants with clinically relevant discrepancies in their classifications and addressed them through a three-phase resolution strategy. This work highlights the importance of data sharing to promote consistency in variant classifications among laboratories, so patients and family members can benefit from more accurate clinical management. Database URL: https://spadahc.ciberisciii.es/.

Amorphous tunable-size Co-B magnetic nanoparticles from the cobalt-catalyzed NaBH4 hydrolysis.

The cobalt-catalyzed hydrolysis of sodium borohydride (NaBH(4)) has become an attractive process in view of the possibilities of using the hydride for hydrogen storage material and also for the production of amorphous and tunable-size magnetic nanoparticles. This process in which the metallic catalyst transforms into a Co- and B-based magnetic by-product when in contact with NaBH(4) has been modified in order to control the mechanism of formation, tune the final size and study the particular magnetic behavior of the Co-B alloy nanoparticles provided.

Incisión de Spangaro en lesiones cardiacas penetrantes, reporte de casos.

INTRODUCCIÓN: El trauma en México es un problema mayor de salud pública, siendo una de las principales causas de mortalidad en personas jóvenes. La incidencia reportada de trauma cardiaco varía. El sitio primario de lesión miocárdica es la pared libre del ventrículo derecho. CASOS CLÍNICOS: Se reportan dos casos de pacientes lesionados con un instrumento punzocortante en el tórax, sufriendo lesión miocárdica que requirió tratamiento quirúrgico de urgencia. DISCUSIÓN: Existen diversas incisiones del tórax. La elección dependerá de la situación a tratar, el estado hemodinámico y el número de lesiones. Ante una lesión cardiaca, el abordaje de Spangaro es una prudente elección. INTRODUCTION: Trauma in Mexico is a major public health problem, being one of the main causes of mortality in young people. The reported incidence of cardiac trauma complications. The primary site of myocardial injury is the free wall of the right ventricle. CLINICAL CASES: Two cases of injured patients with a throbbing instrument in the chest are reported, suffering from myocardial injury that required emergency surgical treatment. DISCUSSION: There are various chest incisions. The choice will depend on the entity to be treated, the hemodynamic state, number of injuries. Faced with a heart injury, Spangaro's approach is a prudent choice.

Post-transcriptional regulation of P-glycoprotein expression in cancer cell lines.

The present study of inhibitors shows that the histone deacetylase-induced increase in P-glycoprotein (Pgp) mRNA (MDR1 mRNA) does not parallel either an increase in Pgp protein or an increase in Pgp activity in several colon carcinoma cell lines. Furthermore, studying the polysome profile distribution, we show a translational control of Pgp in these cell lines. In addition, we show that the MDR1 mRNA produced in these cell lines is shorter in its 5' end that the MDR1 mRNA produced in the MCF-7/Adr (human breast carcinoma) and K562/Adr (human erythroleukemia) cell lines, both of them expressing Pgp. The different size of the MDR1 mRNA is due to the use of alternative promoters. Our data suggest that the translational blockade of MDR1 mRNA in the colon carcinoma cell lines and in wild-type K562 cells could be overcome by alterations in the 5' end of the MDR1 mRNA in the resistant variant of these cell lines, as in the case of the K562/Adr cell line. This is, to our knowledge, the first report demonstrating that the presence of an additional 5' untranslated fragment in the MDR1 mRNA improves the translational efficiency of this mRNA.

A Label Free Disposable Device for Rapid Isolation of Rare Tumor Cells from Blood by Ultrasounds.

The use of blood samples as liquid biopsy is a label-free method for cancer diagnosis that offers benefits over traditional invasive biopsy techniques. Cell sorting by acoustic waves offers a means to separate rare cells from blood samples based on their physical properties in a label-free, contactless and biocompatible manner. Herein, we describe a flow-through separation approach that provides an efficient separation of tumor cells (TCs) from white blood cells (WBCs) in a microfluidic device, "THINUS-Chip" (Thin-Ultrasonic-Separator-Chip), actuated by ultrasounds. We introduce for the first time the concept of plate acoustic waves (PAW) applied to acoustophoresis as a new strategy. It lies in the geometrical chip design: different to other microseparators based on either bulk acoustic waves (BAW) or surface waves (SAW, SSAW and tSAW), it allows the use of polymeric materials without restrictions in the frequency of work. We demonstrate its ability to perform high-throughput isolation of TCs from WBCs, allowing a recovery rate of 84% ± 8% of TCs with a purity higher than 80% and combined viability of 85% at a flow rate of 80 µL/min (4.8 mL/h). The THINUS-Chip performs cell fractionation with low-cost manufacturing processes, opening the door to possible easy printing fabrication.

KRAS and BRAF somatic mutations in colonic polyps and the risk of metachronous neoplasia.

BACKGROUND & AIMS: High-risk features of colonic polyps are based on size, number, and pathologic characteristics. Surveillance colonoscopy is often recommended according to these findings. This study aimed to determine whether the molecular characteristics of polyps might provide information about the risk of metachronous advanced neoplasia. METHODOLOGY: We retrospectively included 308 patients with colonic polyps. A total of 995 polyps were collected and tested for somatic BRAF and KRAS mutations. Patients were classified into 3 subgroups, based on the polyp mutational profile at baseline, as follows: non-mutated polyps (Wild-type), at least one BRAF-mutated polyp, or at least one KRAS-mutated polyp. At surveillance, advanced adenomas were defined as adenomas ≥ 10 mm and/or with high grade dysplasia or a villous component. In contrast, advanced serrated polyps were defined as serrated polyps ≥ 10 mm in any location, located proximal to the splenic flexure with any size or with dysplasia. RESULTS: At baseline, 289 patients could be classified as wild-type (62.3%), BRAF mutated (14.9%), or KRAS mutated (22.8%). In the univariate analysis, KRAS mutations were associated with the development of metachronous advanced polyps (OR: 2.36, 95% CI: 1.22-4.58; P = 0.011), and specifically, advanced adenomas (OR: 2.42, 95% CI: 1.13-5.21; P = 0.023). The multivariate analysis, adjusted for age and sex, also showed associations with the development of metachronous advanced polyps (OR: 2.27, 95% CI: 1.15-4.46) and advanced adenomas (OR: 2.23, 95% CI: 1.02-4.85). CONCLUSIONS: Our results suggested that somatic KRAS mutations in polyps represent a potential molecular marker for the risk of developing advanced neoplasia.

Histone deacetylase inhibitors induced caspase-independent apoptosis in human pancreatic adenocarcinoma cell lines.

The antitumor activity of the histone deacetylase inhibitors was tested in three well-characterized pancreatic adenocarcinoma cell lines, IMIM-PC-1, IMIM-PC-2, and RWP-1. These cell lines have been previously characterized in terms of their origin, the status of relevant molecular markers for this kind of tumor, resistance to other antineoplastic drugs, and expression of differentiation markers. In this study, we report that histone deacetylase inhibitors induce apoptosis in pancreatic cancer cell lines, independently of their intrinsic resistance to conventional antineoplastic agents. The histone deacetylase inhibitor-induced apoptosis is due to a serine protease-dependent and caspase-independent mechanism. Initially, histone deacetylase inhibitors increase Bax protein levels without affecting Bcl-2 levels. Consequently, the apoptosis-inducing factor (AIF) and Omi/HtrA2 are released from the mitochondria, with the subsequent induction of the apoptotic program. These phenomena require AIF relocalization into the nuclei to induce DNA fragmentation and a serine protease activity of Omi/HtrA2. These data, together with previous results from other cellular models bearing the multidrug resistance phenotype, suggest a possible role of the histone deacetylase inhibitors as antineoplastic agents for the treatment of human pancreatic adenocarcinoma.

Microcalorimetría: un método de diagnóstico precoz de infecciones bacterianas / Microcalorimetry contributions to early diagnosis of bacterial infections

Microcalorimetry is a highly sensitive experimental technique that determines heat changes in any process or transformation. All organisms produce heat due to their metabolism. Rate of heat flow is an adequate measure of metabolic activity of living beings and their component parts. Microorganisms produce small amounts of heat: 1-3 pW per cell. Although the heat produced by bacteria is very small, their exponential reproduction in a culture medium permits heat detection through microcalorimetry. A thermal conductivity calorimeter of the Calvet type was used. The inside of the calorimeter contains two stainless steel cells (experimental and reference) with a screw on Teflon cap with a hole in the centre. Experiments were carried out with final concentrations of the order of 10 , 10 , 10 and 10 UFC/ml. These were kept at a constant temperature of 309.65 K. The plot of change in heat voltage vs. time enables us to obtain the characteristic growth curve for each bacterial strain. Thermograms were analyzed mathematically allowing us to calculate the constant growth, generation time and the amount of heat exchanged over the culture time

Microcalorimetry is a highly sensitive experimental technique that determines heat changes in any process or transformation. All organisms produce heat due to their metabolism. Rate of heat flow is an adequate measure of metabolic activity of living beings and their component parts. Microorganisms produce small amounts of heat: 1-3 pW per cell. Although the heat produced by bacteria is very small, their exponential reproduction in a culture medium permits heat detection through microcalorimetry. A thermal conductivity calorimeter of the Calvet type was used. The inside of the calorimeter contains two stainless steel cells (experimental and reference) with a screw on Teflon cap with a hole in the centre. Experiments were carried out with final concentrations of the order of 10 , 10 , 10 and 10 UFC/ml. These were kept at a constant temperature of 309.65 K. The plot of change in heat voltage vs. time enables us to obtain the characteristic growth curve for each bacterial strain. Thermograms were analyzed mathematically allowing us to calculate the constant growth, generation time and the amount of heat exchanged over the culture time (AU)

La microcalorimetría es una técnica experimental que permite determinar con alta sensibilidad la energía expuesta como consecuencia de cualquier proceso o transformación. Todos los organismos producen calor debido a su metabolismo. La tasa de calor es una medida adecuada de la actividad metabólica de los organismos y sus partes constituyentes. Los microorganismos producen pequeñas cantidades de calor, del orden de 1-3 pW por célula. A pesar de la baja cantidad de calor producido por las bacterias, su exponencial replicación en un medio de cultivo permite su detección mediante microcalorimetría. Se ha utilizado un calorímetro de conducción calorífica tipo Calvet, en cuyo recinto interior se sitúan dos células (experimental y testigo) de acero inoxidable con tapón roscado de teflón perforado en el centro. Las experiencias se llevaron a cabo a concentraciones finales de 106, 105, 103 y 10 UFC/ml y se mantuvo una temperatura constante de 309,65 K. La representación de la diferencia de potencial calorífico frente al tiempo permite obtener las curvas de crecimiento características de cada bacteria. Los termogramas se analizaron matemáticamente permitiendo calcular la constante de crecimiento, el tiempo de generación y la cantidad de calor intercambiado

Circulating tumour cell analysis as an early marker for relapse in stage II and III colorectal cancer patients: a pilot study.

INTRODUCTION: Recent studies have identified both the prognostic and predictive utility of determining the number of circulating tumour cells (CTC) in patients with solid cancers. MATERIAL AND METHODS: In the present pilot study we evaluated the ability of two different methods to isolate CTC in combination with two strategies to enumerate CTC from patients with stages II and III surgically treated colorectal cancer (CRC). First, we used two systems for tumour cell enrichment (differential centrifugation and immunomagnetic beads), combined with two methods to enumerate CTC (real-time PCR and fl ow cytometry), to determine the most efficient combination. These experiments were performed in a model system using serial dilutions of HT29 tumour cell lines with lymphocytes. Then, CTC analysis using the technical approach selected before was performed in 109 blood samples from 16 stage II and III CRC patients during chemotherapy treatment and follow-up. RESULTS: Immunomagnetic beads followed by flow cytometry was the most efficient combination (ED=60.53; p=0.5). Two cases out of 16 patients analysed had clinical tumour relapse. In both, we detected a significant increase of CTC five and six months, respectively, before the relapse was clinically evidenced. An increase of CTC was also observed in another case without clinical evidence of relapse. The remaining cases (13) had very few or no detectable CTC and no clinical evidence of relapse (p=0.029). CONCLUSIONS: Changes in CTC numbers during follow-up might predict tumour relapse. Further evaluation of CTC prognostic and predictive value in patients with early CRC is warranted.