Voltammetric electronic tongue based on carbon paste electrodes modified with biochar for phenolic compounds stripping detection

Altres ajuts: Marta Bonet-San-Emeterio thanks to AGAUR, Generalitat de Catalunya and to European Social Fund, European Union for FI fellowship. Manel del Valle thanks the support from program ICREA Academia.Biochar is a charcoal produced from the biomass pyrolysis process that presents a highly porous and functionalized surface. In the present work an array of carbon paste electrodes (CPE) made of different forms of carbon (graphite, carbon nanotubes and activated biochar) was evaluated in the development of an electronic tongue for discrimination and stripping voltammetric determination of catechol (CAT), 4-ethylcatechol (4-EC) and 4-ethylguaiacol (4-EG) phenolic compounds. Morphological characterization of carbon materials and electrodes surfaces was performed by scanning electron microscopy (SEM) and semi-quantitative elemental composition by energy dispersive spectroscopy (EDS). Electrochemical Impedance Spectroscopy (EIS) measurements were used for electrochemical characterization of electrodes. Cyclic voltammetry measurements were performed for the phenolic compounds evaluated using different concentrations. Principal component analysis (PCA) was performed to evaluate the qualitative analysis. Quantitative data modeling was done using artificial neural networks (ANN). The proposed sensor array presented analytical potentiality allowing the distinction and determination of CAT, 4-EC and 4-EG by using chemometric processing. The method showed sensibility, reproducibility and a good linearity (R2>0.9940) for three compounds evaluated. Spontaneous preconcentration of three compounds was possible using all three sensors, which can allow the application of these as passive samplers for remote determinations of phenolic compounds in wine and food samples

Perspectives for cancer immunotherapy mediated by p19Arf plus interferon-beta gene transfer

While cancer immunotherapy has gained much deserved attention in recent years, many areas regarding the optimization of such modalities remain unexplored, including the development of novel approaches and the strategic combination of therapies that target multiple aspects of the cancer-immunity cycle. Our own work involves the use of gene transfer technology to promote cell death and immune stimulation. Such immunogenic cell death, mediated by the combined transfer of the alternate reading frame (p14ARF in humans and p19Arf in mice) and the interferon-b cDNA in our case, was shown to promote an antitumor immune response in mouse models of melanoma and lung carcinoma. With these encouraging results, we are now setting out on the road toward translational and preclinical development of our novel immunotherapeutic approach. Here, we outline the perspectives and challenges that we face, including the use of human tumor and immune cells to verify the response seen in mouse models and the incorporation of clinically relevant models, such as patient-derived xenografts and spontaneous tumors in animals. In addition, we seek to combine our immunotherapeutic approach with other treatments, such as chemotherapy or checkpoint blockade, with the goal of reducing dosage and increasing efficacy. The success of any translational research requires the cooperation of a multidisciplinary team of professionals involved in laboratory and clinical research, a relationship that is fostered at the Cancer Institute of Sao Paulo

Gene-based Interventions for Cancer Immunotherapy

Immunotherapy of cancer has deservedly gained much attention in the past few years and is likely to continue to advance and become a fundamental cancer treatment. While vaccines, chimeric antigen receptor (CAR) T cells and checkpoint blockade have received the lion’s share of the attention, an important direct role for gene transfer as an immunotherapy is emerging. For example, oncolytic viruses induce immunogenic cell death, thus liberating both antigens and the signals that are necessary for the activation of antigen-presenting cells, ensuring stimulation of an adaptive response. In another example, transfer of prodrug converting enzymes, such as the herpes simplex virus-thymidine kinase (HSV-tk) gene or the cytosine deaminase gene, has been shown to promote an immune response, thus functioning as immunotherapies. Alternatively, our own work involves the use of nonreplicating viral vectors for the simultaneous delivery of gene combinations that promote both cell death and an immune response. In fact, our gene transfer approach has been applied as a vaccine, immunotherapy or in situ gene therapy, resulting in immunogenic cell death and the induction of a protective immune response. Here, we highlight the development of these approaches both in terms of technical advances and clinical experience

Bayesian multivariate approach for growth, fertility and visual score traits of Brangus cattle herds

O objetivo deste trabalho foi estimar parâmetros genéticos e tendências genéticas e fenotípicas de uma população da raça Brangus. As características peso, circunferência escrotal e escores visuais de conformação, precocidade, musculatura e umbigo, padronizadas para 550 dias de idade, foram avaliadas a partir de 6.789 registros de animais nascidos de 288 touros e 5.949 vacas, entre 1991 e 2001, em 49 fazendas localizadas nas regiões Centro‑Oeste, Sudeste e Sul do Brasil. Para a estimação dos parâmetros, das correlações e das tendências genéticas, foi adotado o modelo animal linear‑limiar hexacaracterística. As estimativas de herdabilidade direta foram de 0,39 e 0,27, para peso e circunferência escrotal, respectivamente, e de 0,22, 0,20, 0,23 e 0,33 para conformação, precocidade, musculatura e umbigo, o que indica considerável variação genética aditiva e que é possível obter ganho genético por meio da seleção. As correlações genéticas entre peso e circunferência escrotal com os escores de conformação, precocidade e musculatura mostram a possibilidade de resposta correlacionada. As tendências genéticas estimadas indicam grande contribuição de fontes de variação não genéticas para todas as características no período estudado, e apontam a necessidade de melhoria das condições ambientais, para que os animais expressem todo seu potencial genético.The objective of this work was to estimate genetic parameters, and genetic and phenotypic trends of a Brangus breed population. Weight, scrotal circumference, and visual scores of conformation, precocity, musculature, and navel, adjusted to 550 days of age, were evaluated from records of 6,789 animals sired by 288 bulls and 5,949 dams, born between 1991 and 2001 in 49 farms located in the Center‑West, Southeast, and South regions of Brazil. To estimate parameters, correlations, and genetic trends, a six‑trait, linear‑threshold animal model was adopted. Direct heritability estimates were 0.39 and 0.27 for weight and scrotal circumference, respectively, and 0.22, 0.20, 0.23, and 0.33 for conformation, precocity, musculature, and navel, indicating considerable additive genetic variation, and that it is possible to obtain genetic gain by selection. Genetic correlations between weight and scrotal circumference with the scores of conformation, precocity, and musculature show the possibility of correlated responses. Genetic trends indicate great contribution of non‑genetic sources of variation in the evaluated traits, which leads to the necessity of improving environmental conditions, in order for the animals to fully express their genetic potential

Transcriptional effects of 1,25 dihydroxyvitamin D3 physiological and supra-physiological concentrations in breast cancer organotypic culture

Background\ud Vitamin D transcriptional effects were linked to tumor growth control, however, the hormone targets were determined in cell cultures exposed to supra physiological concentrations of 1,25(OH)2D3 (50-100nM). Our aim was to evaluate the transcriptional effects of 1,25(OH)2D3 in a more physiological model of breast cancer, consisting of fresh tumor slices exposed to 1,25(OH)2D3 at concentrations that can be attained in vivo.\ud \ud Methods\ud Tumor samples from post-menopausal breast cancer patients were sliced and cultured for 24 hours with or without 1,25(OH)2D3 0.5nM or 100nM. Gene expression was analyzed by microarray (SAM paired analysis, FDR≤0.1) or RT-qPCR (p≤0.05, Friedman/Wilcoxon test). Expression of candidate genes was then evaluated in mammary epithelial/breast cancer lineages and cancer associated fibroblasts (CAFs), exposed or not to 1,25(OH)2D3 0.5nM, using RT-qPCR, western blot or immunocytochemistry.\ud \ud Results\ud 1,25(OH)2D3 0.5nM or 100nM effects were evaluated in five tumor samples by microarray and seven and 136 genes, respectively, were up-regulated. There was an enrichment of genes containing transcription factor binding sites for the vitamin D receptor (VDR) in samples exposed to 1,25(OH)2D3 near physiological concentration. Genes up-modulated by both 1,25(OH)2D3 concentrations were CYP24A1, DPP4, CA2, EFTUD1, TKTL1, KCNK3. Expression of candidate genes was subsequently evaluated in another 16 samples by RT-qPCR and up-regulation of CYP24A1, DPP4 and CA2 by 1,25(OH)2D3 was confirmed. To evaluate whether the transcripitonal targets of 1,25(OH)2D3 0.5nM were restricted to the epithelial or stromal compartments, gene expression was examined in HB4A, C5.4, SKBR3, MDA-MB231, MCF-7 lineages and CAFs, using RT-qPCR. In epithelial cells, there was a clear induction of CYP24A1, CA2, CD14 and IL1RL1. In fibroblasts, in addition to CYP24A1 induction, there was a trend towards up-regulation of CA2, IL1RL1, and DPP4. A higher protein expression of CD14 in epithelial cells and CA2 and DPP4 in CAFs exposed to 1,25(OH)2D3 0.5nM was detected.\ud \ud Conclusions\ud In breast cancer specimens a short period of 1,25(OH)2D3 exposure at near physiological concentration modestly activates the hormone transcriptional pathway. Induction of CYP24A1, CA2, DPP4, IL1RL1 expression appears to reflect 1,25(OH)2D3 effects in epithelial as well as stromal cells, however, induction of CD14 expression is likely restricted to the epithelial compartment.The authors would like to acknowledge the assistance of Dr. Sridar Chittur for helpful discussions on microarray data analysis and Dr. Igor Moysés Longo Snitcovsky for critical review and important suggestions to the manuscript.This work was supported by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP grants 07/04799-2; 09/10088-7; 08/51750-1) and Coordenação de Aperfeiçoamento de Pessoal de nível Superior (CAPES)

Influence of the interaction between nodal fibroblast and breast cancer cells on gene expression

Our aim was to evaluate the interaction between breast cancer cells and nodal fibroblasts, by means of their gene expression profile. Fibroblast primary cultures were established from negative and positive lymph nodes from breast cancer patients and a similar gene expression pattern was identified, following cell culture. Fibroblasts and breast cancer cells (MDA-MB231, MDA-MB435, and MCF7) were cultured alone or co-cultured separated by a porous membrane (which allows passage of soluble factors) for comparison. Each breast cancer lineage exerted a particular effect on fibroblasts viability and transcriptional profile. However, fibroblasts from positive and negative nodes had a parallel transcriptional behavior when co-cultured with a specific breast cancer cell line. The effects of nodal fibroblasts on breast cancer cells were also investigated. MDA MB-231 cells viability and migration were enhanced by the presence of fibroblasts and accordingly, MDA-MB435 and MCF7 cells viability followed a similar pattern. MDA-MB231 gene expression profile, as evaluated by cDNA microarray, was influenced by the fibroblasts presence, and HNMT, COMT, FN3K, and SOD2 were confirmed downregulated in MDA-MB231 co-cultured cells with fibroblasts from both negative and positive nodes, in a new series of RT-PCR assays. In summary, transcriptional changes induced in breast cancer cells by fibroblasts from positive as well as negative nodes are very much alike in a specific lineage. However, fibroblasts effects are distinct in each one of the breast cancer lineages, suggesting that the inter-relationships between stromal and malignant cells are dependent on the intrinsic subtype of the tumor

Goodbye Hartmann trial: a prospective, international, multicenter, observational study on the current use of a surgical procedure developed a century ago

Background: Literature suggests colonic resection and primary anastomosis (RPA) instead of Hartmann's procedure (HP) for the treatment of left-sided colonic emergencies. We aim to evaluate the surgical options globally used to treat patients with acute left-sided colonic emergencies and the factors that leading to the choice of treatment, comparing HP and RPA. Methods: This is a prospective, international, multicenter, observational study registered on ClinicalTrials.gov. A total 1215 patients with left-sided colonic emergencies who required surgery were included from 204 centers during the period of March 1, 2020, to May 31, 2020. with a 1-year follow-up. Results: 564 patients (43.1%) were females. The mean age was 65.9 ± 15.6 years. HP was performed in 697 (57.3%) patients and RPA in 384 (31.6%) cases. Complicated acute diverticulitis was the most common cause of left-sided colonic emergencies (40.2%), followed by colorectal malignancy (36.6%). Severe complications (Clavien-Dindo ≥ 3b) were higher in the HP group (P < 0.001). 30-day mortality was higher in HP patients (13.7%), especially in case of bowel perforation and diffused peritonitis. 1-year follow-up showed no differences on ostomy reversal rate between HP and RPA. (P = 0.127). A backward likelihood logistic regression model showed that RPA was preferred in younger patients, having low ASA score (≤ 3), in case of large bowel obstruction, absence of colonic ischemia, longer time from admission to surgery, operating early at the day working hours, by a surgeon who performed more than 50 colorectal resections. Conclusions: After 100 years since the first Hartmann's procedure, HP remains the most common treatment for left-sided colorectal emergencies. Treatment's choice depends on patient characteristics, the time of surgery and the experience of the surgeon. RPA should be considered as the gold standard for surgery, with HP being an exception