Detection of BCG bacteria using a magnetoresistive biosensor: A step towards a fully electronic platform for tuberculosis point-of-care detection

Tuberculosis is one of the major public health concerns. This highly contagious disease affects more than 10.4 million people, being a leading cause of morbidity by infection. Tuberculosis is diagnosed at the point-of-care by the Ziehl-Neelsen sputum smear microscopy test. Ziehl-Neelsen is laborious, prone to human error and infection risk, with a limit of detection of 104 cells/mL. In resource-poor nations, a more practical test, with lower detection limit, is paramount. This work uses a magnetoresistive biosensor to detect BCG bacteria for tuberculosis diagnosis. Herein we report: i) nanoparticle assembly method and specificity for tuberculosis detection; ii) demonstration of proportionality between BCG cell concentration and magnetoresistive voltage signal; iii) application of multiplicative signal correction for systematic effects removal; iv) investigation of calibration effectiveness using chemometrics methods; and v) comparison with state-of-the-art point-of-care tuberculosis biosensors. Results present a clear correspondence between voltage signal and cell concentration. Multiplicative signal correction removes baseline shifts within and between biochip sensors, allowing accurate and precise voltage signal between different biochips. The corrected signal was used for multivariate regression models, which significantly decreased the calibration standard error from 0.50 to 0.03log10 (cells/mL). Results show that Ziehl-Neelsen detection limits and below are achievable with the magnetoresistive biochip, when pre-processing and chemometrics are used.Teresa Barroso thanks FCT for her PhD Grant SFRH/BD/33904/2009. Elisabete Fernandes acknowledges the Project N2020 -PE-Advancing Cancer (NORTE-01-0145-FEDER-000029).info:eu-repo/semantics/publishedVersio

Intravascular Large B-Cell Lymphoma Genomic Profile Is Characterized by Alterations in Genes Regulating NF-κB and Immune Checkpoints.

Intravascular large B-cell lymphoma (IVLBCL) is an uncommon lymphoma with an aggressive clinical course characterized by selective growth of tumor cells within the vessels. Its pathogenesis is still uncertain and there is little information on the underlying genomic alterations. In this study, we performed a clinicopathologic and next-generation sequencing analysis of 15 cases of IVLBCL using a custom panel for the detection of alterations in 68 recurrently mutated genes in B-cell lymphomagenesis. Six patients had evidence of hemophagocytic syndrome. Four patients presented concomitantly a solid malignancy. Tumor cells outside the vessels were observed in 7 cases, 2 with an overt diffuse large B-cell cell lymphoma. In 4 samples, tumor cells infiltrated lymphatic vessel in addition to blood capillaries. Programmed death-ligand 1 (PD-L1) was positive in tumor cells in 4 of 11 evaluable samples and in macrophages intermingled with tumor cells in 8. PD-L1 copy number gains were identified in a higher proportion of cases expressing PD-L1 than in negative tumors. The most frequently mutated gene was PIM1 (9/15, 60%), followed by MYD88L265P and CD79B (8/15, 53% each). In 6 cases, MYD88L265P and CD79B mutations were detected concomitantly. We also identified recurrent mutations in IRF4 , TMEM30A , BTG2 , and ETV6 loci (4/15, 27% each) and novel driver mutations in NOTCH2 , CCND3 , and GNA13 , and an IRF4 translocation in 1 case each. The mutational profile was similar in patients with and without evidence of hemophagocytic syndrome and in cases with or without dissemination of tumor cells outside the vessels. Our results confirm the relevance of mutations in B-cell receptor/nuclear factor-κB signaling and immune escape pathways in IVLBCL and identify novel driver alterations. The similar mutational profile in tumors with extravascular dissemination suggests that these cases may also be considered in the spectrum of IVLBCL

The entrepreneurial ecosystem of cultural and creative industries in Porto: A sub-ecosystem approach

It is still a matter of dispute whether entrepreneurial ecosystem (EE) frameworks can be confined to a single industry in isolation, let alone whether such a sub-ecosystem approach can be employed in a domain that is distinct from the highgrowth industries usually scrutinised in the literature. This article seeks to apply a systemic and dynamic EE perspective to the development of cultural and creative industries (CCIs) within an urban context, with a particular focus on how urban development interacts with the sub-ecosystem of this sector over time. An in-depth case study in the city of Porto (Portugal) revealed that existing EE frameworks are well-suited to research on creative sub-ecosystems. It also enabled us to flesh out associations with other entrepreneurial activities and policy domains within the city. We highlight the prominent roles of local culture and policies when the context is resource-constrained: policy led to an upward, positive spiral that moved Porto's EE in relation to CCIs into a growth stage, during which it began to interact with, and faced resource competition from, high-tech entrepreneurship. We argue that having an integrated view of the dynamics of entrepreneurial sub-ecosystems and urban affairs can improve what is understood of productivity and causality in entrepreneurship

Assessment of nutritional risk among in-school adolescents from Cantabria

Seminario “Promoción de hábitos saludables en adolescentes desde el ámbito educativo”.Objetivo: Evaluar el riesgo nutricional, por edad y sexo, que presentan los adolescentes escolarizados en la Comunidad Autónoma de Cantabria. Sujetos: Se realizó un estudio transversal, analizando una muestra de 1101 adolescentes, de los que 51,6% eran varones y 48,4% fueron mujeres de edades comprendidas entre los 10 y los 17 años, escolarizados en centros de enseñanza pública, mediante el cuestionario Krece Plus. Resultados: Se observa un elevado porcentaje de ado- lescentes que presentan un riesgo nutricional elevado (35%). Los varones presentan un riesgo nutricional alto en un porcentaje ligeramente superior a las mujeres (37,8 % vs 32,1%). Además, el riesgo nutricional alto sufre un notable incremento a medida que la edad de los jóvenes aumenta. Se aprecian diferencias estadísticamente signi- ficativas tanto en los grupos de edad de los varones (p = 0,024), de las mujeres (p < 0,001) como en el grupo global (p = 0,001). En los tres casos, la distribución del riego nutricional en los grupos de menor edad es muy similar (entre 35,2 y 35,8% en los h , entre 27,9 y 29,7% en las m , y entre 31,7 y 32,7% en el grupo total). Mientras que en el grupo de mayor edad estos valores prácticamente se duplican (57,1% en los h , 69,0% en las m , y 62,2 % en el grupo total). Conclusión: Los resultados obtenidos muestran una realidad preocupante debido, principalmente, al elevado porcentaje de adolescentes que presentan un riesgo nutri- cional elevado. Siendo los varones y los adolescentes de mayor edad los sectores en los que este riesgo nutricional elevado es superior.Objective: To analyse nutritional risk, by age and sex, among primary and secondary education adolescents from Cantabria. Methodology: a cross-sectional study was carried out, analysing a sample of 1101 adolescents: 568 (51.6%) were men and 533 (48.4%) were women, aged 12 to 17, attending 16 different primary and secondary education centres in Cantabria, by means of a Krece Plus questionnaire. Results: A high percentage of adolescents with a high nutritional risk (35%) can be observed. Men show a high nutritional risk slightly higher than women (37.8% h vs 32.1% m ). Moreover, the high nutritional risk expe riences a notable increase as young people get older. Significant statistical differences can be seen both in male and female groups, and as a global group. In all three cases, the nutritional risk distribution in the youngest group is very similar (35.2-35.8% in h , 27.9-29.7% in m , 31.7-32.7% in the global group); whereas in elder adolescents, those values are practically doubled (57.1% in h , 69.0% in m , y 62.2% in the global group). Conclusions: Results are alarming mainly given the high percentage of adolescents with a high nutritional risk. Men and older adolescents are the groups in which high nutritional risk is more evident

Prácticas alimentarias de los adolescentes de Cantabria

Objetivo. Analizar determinadas prácticas alimentarias en adolescentes escolarizados en centros de educación pública de Cantabria, participantes en el Proyecto "Promoción de Hábitos Saludables en Adolescentes desde el Ámbito Educativo". Sujetos. Se realizó un estudio transversal, analizando una muestra de 1.101 adolescentes: 568 (51,6%) varones y 533 (48,4%) mujeres, de edades comprendidas entre los 10 y los 17 años, escolarizados en dieciséis centros de enseñanza primaria y secundaria, mediante un cuestionario autocumplimentado. Resultados. Los adolescentes suelen realizar entre cuatro (41,5%) y cinco (31,6%) ingestas diarias. Durante los días de colegio, el 34% emplea entre diez y quince minutos en desayunar, y entre 30 y 35 minutos en comer (33,5%) y cenar (23%). Un elevado porcentaje (49,4%) de adolescentes desayunaba en soledad durante los días lectivos. Las principales ingestas alimenticias se realizan en el hogar. Las bebidas no alcohólicas (53,6%) y los dulces (42%) son los principales destinos de su dinero de bolsillo. En la casi totalidad de los hogares, es la madre la que se encarga de la compra de los alimentos, de la preparación de las comidas y de decidir tanto el almuerzo como la merienda. La pizza (72,6%) y las patatas fritas (70,8%) son los alimentos considerados más ricos entre los analizados, mientras que el perrito caliente (49,4%) y la hamburguesa (48,5%) son considerados como los menos sanos. El 58,6% de los encuestados cena viendo la televisión. Conclusión. En el estudio del comportamiento alimentario es necesario analizar la influencia de otros factores que, en muchas ocasiones, están detrás de las recomendaciones dietéticas y que casi siempre son ignorados. Prácticas alimentarias como las analizadas en el presente estudio, permiten, cuando estas se desarrollan de forma adecuada, una mejora sustancial en la salud alimentaria y nutricional de las personas

Autoimagen en las dos primeras fases de la adolescencia y factores relacionados

Se trata de la descripción de la imagen corporal en un amplio grupo de alumnos escolarizados en Cantabria (n=1179 adolescentes), de 10 a 17 años de edad (adolescencia temprana e intermedia) dentro de un estudio más amplio encaminado a evidenciar un estilo de vida saludable en estos adolescentes, llevado a cabo por profesores de universidad y profesores de educación física de los centros educativos. Los principales hallazgos consisten en que los adolescentes tienen, en general, una buena imagen de sí mismos y, aunque no reconocen la elevada prevalencia de sobrepeso y obesidad, desean adelgazar y el grado de satisfacción que tienen con su imagen corporal va empeorando conforme avanza la adolescencia, signifi cativamente más en las del sexo femenino. Esta insatisfacción debe ser tenida en cuenta en el abordaje de los adolescentes con obesidad

Stunned Silence: Gene Expression Programs in Human Cells Infected with Monkeypox or Vaccinia Virus

Poxviruses use an arsenal of molecular weapons to evade detection and disarm host immune responses. We used DNA microarrays to investigate the gene expression responses to infection by monkeypox virus (MPV), an emerging human pathogen, and Vaccinia virus (VAC), a widely used model and vaccine organism, in primary human macrophages, primary human fibroblasts and HeLa cells. Even as the overwhelmingly infected cells approached their demise, with extensive cytopathic changes, their gene expression programs appeared almost oblivious to poxvirus infection. Although killed (gamma-irradiated) MPV potently induced a transcriptional program characteristic of the interferon response, no such response was observed during infection with either live MPV or VAC. Moreover, while the gene expression response of infected cells to stimulation with ionomycin plus phorbol 12-myristate 13-acetate (PMA), or poly (I-C) was largely unimpaired by infection with MPV, a cluster of pro-inflammatory genes were a notable exception. Poly(I-C) induction of genes involved in alerting the innate immune system to the infectious threat, including TNF-alpha, IL-1 alpha and beta, CCL5 and IL-6, were suppressed by infection with live MPV. Thus, MPV selectively inhibits expression of genes with critical roles in cell-signaling pathways that activate innate immune responses, as part of its strategy for stealthy infection

The Chemotherapeutic Drug 5-Fluorouracil Promotes PKR-Mediated Apoptosis in a p53- Independent Manner in Colon and Breast Cancer Cells

The chemotherapeutic drug 5-FU is widely used in the treatment of a range of cancers, but resistance to the drug remains a major clinical problem. Since defects in the mediators of apoptosis may account for chemo-resistance, the identification of new targets involved in 5-FU-induced apoptosis is of main clinical interest. We have identified the ds-RNA-dependent protein kinase (PKR) as a key molecular target of 5-FU involved in apoptosis induction in human colon and breast cancer cell lines. PKR distribution and activation, apoptosis induction and cytotoxic effects were analyzed during 5-FU and 5-FU/IFNα treatment in several colon and breast cancer cell lines with different p53 status. PKR protein was activated by 5-FU treatment in a p53-independent manner, inducing phosphorylation of the protein synthesis translation initiation factor eIF-2α and cell death by apoptosis. Furthermore, PKR interference promoted a decreased response to 5-FU treatment and those cells were not affected by the synergistic antitumor activity of 5-FU/IFNα combination. These results, taken together, provide evidence that PKR is a key molecular target of 5-FU with potential relevance in the clinical use of this drug

Coronavirus Gene 7 Counteracts Host Defenses and Modulates Virus Virulence

Transmissible gastroenteritis virus (TGEV) genome contains three accessory genes: 3a, 3b and 7. Gene 7 is only present in members of coronavirus genus a1, and encodes a hydrophobic protein of 78 aa. To study gene 7 function, a recombinant TGEV virus lacking gene 7 was engineered (rTGEV-Δ7). Both the mutant and the parental (rTGEV-wt) viruses showed the same growth and viral RNA accumulation kinetics in tissue cultures. Nevertheless, cells infected with rTGEV-Δ7 virus showed an increased cytopathic effect caused by an enhanced apoptosis mediated by caspase activation. Macromolecular synthesis analysis showed that rTGEV-Δ7 virus infection led to host translational shut-off and increased cellular RNA degradation compared with rTGEV-wt infection. An increase of eukaryotic translation initiation factor 2 (eIF2α) phosphorylation and an enhanced nuclease, most likely RNase L, activity were observed in rTGEV-Δ7 virus infected cells. These results suggested that the removal of gene 7 promoted an intensified dsRNA-activated host antiviral response. In protein 7 a conserved sequence motif that potentially mediates binding to protein phosphatase 1 catalytic subunit (PP1c), a key regulator of the cell antiviral defenses, was identified. We postulated that TGEV protein 7 may counteract host antiviral response by its association with PP1c. In fact, pull-down assays demonstrated the interaction between TGEV protein 7, but not a protein 7 mutant lacking PP1c binding motif, with PP1. Moreover, the interaction between protein 7 and PP1 was required, during the infection, for eIF2α dephosphorylation and inhibition of cell RNA degradation. Inoculation of newborn piglets with rTGEV-Δ7 and rTGEV-wt viruses showed that rTGEV-Δ7 virus presented accelerated growth kinetics and pathology compared with the parental virus. Overall, the results indicated that gene 7 counteracted host cell defenses, and modified TGEV persistence increasing TGEV survival. Therefore, the acquisition of gene 7 by the TGEV genome most likely has provided a selective advantage to the virus