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Abstract The prevalence of malignant cells in clinical specimens, or tumour purity, is affected by both intrinsic biological factors and extrinsic sampling bias. Molecular characterization of large clinical cohorts is typically performed on bulk samples; data analysis and interpretation can be biased by tumour purity variability. Transcription-based strategies to estimate tumour purity have been proposed, but no breast cancer specific method is available yet. We interrogated over 6000 expression profiles from 10 breast cancer datasets to develop and validate a 9-gene Breast Cancer Purity Score (BCPS). BCPS outperformed existing methods for estimating tumour content. Adjusting transcriptomic profiles using the BCPS reduces sampling bias and aids data interpretation. BCPS-estimated tumour purity improved prognostication in luminal breast cancer, correlated with pathologic complete response in on-treatment biopsies from triple-negative breast cancer patients undergoing neoadjuvant treatment and effectively stratified the risk of relapse in HER2+ residual disease post-neoadjuvant treatment.

Abstract Optical microscopy has long been the standard method for diagnosis in cytopathology. Whole slide scanners can image and digitize large sample areas automatically, but are slow, expensive and therefore not widely available. Clinical diagnosis of cytology specimens is especially challenging since these samples are both spread over large areas and thick, which requires 3D capture. Here, we introduce a new parallelized microscope for scanning thick specimens across extremely wide fields-of-view (54 × 72 mm2) at 1.2 and 0.6 μm resolutions, accompanied by machine learning software to rapidly assess these 16 gigapixel scans. This Multi-Camera Array Scanner (MCAS) comprises 48 micro-cameras closely arranged to simultaneously image different areas. By capturing 624 megapixels per snapshot, the MCAS is significantly faster than most conventional whole-slide scanners. We used this system to digitize entire cytology samples (scanning three entire slides in 3D in just several minutes) and demonstrate two machine learning techniques to assist pathologists: first, an adenocarcinoma detection model in lung specimens (0.73 recall); second, a slide-level classification model of lung smears (0.969 AUC).

The present research focused on the use of high temperatures to achieve fast separations in Liquid Chromatography (LC) and minimize the consumption of organic solvents. Particularly, superheated water was employed, by exploiting the decrease in dielectric constant of water when increasing the temperature. To realize this kind of application, the availability of LC columns resistant at high temperature is mandatory. Recently, porous graphitic carbon (PGC) columns have been proposed as stationary phases resistant up to 250 °C. Hardware modifications were also necessary to guarantee the efficient heating of the mobile phase prior of entering the column, as well as its rapid cooling before detection by photodiode array (PDA). Specifically, an LC system was interfaced to a GC oven, which hosted the LC column; to achieve a fast and efficient heating of the eluent coming from the autosampler prior to enter into the column, a pre-heating tube was interposed between the autosampler outlet and the column inlet. Instead, a cooling loop was connected between the column outlet and the PDA cell. A mixture of 1-nitroalkanes was used to evaluate the performance of the system in terms of dead volumes, band broadening and peak shape, as well as chromatographic efficiency and resolution. The use of higher temperatures allowed to perform analyses at higher flow rates in reduced analysis time and was also in terms of peak bandwidth resulting to increased efficiency for the most retained compound. Moreover, van't Hoff plots were obtained at different temperature, confirming that the most retained analyte greatly benefited of the temperature increase.The system was applied to the analysis of parabens in cosmetic and food products. The method combined eco-sustainable sample preparation with high-temperature LC separation on a PGC column and was fully validated in terms of linearity, precision, accuracy and detection and quantification limits. The developed method was critically evaluated by comparing the output of different metric tools implemented in the last decade for the quantitative assessment of the greenness.

High-throughput phenotyping (HTP) provides new opportunities for efficiently dissecting the genetic basis of drought-adaptive traits, which is essential in current wheat breeding programs. The combined use of HTP and genome-wide association (GWAS) approaches has been useful in the assessment of complex traits such as yield, under field stress conditions including heat and drought. The aim of this study was to identify molecular markers associated with yield (YLD) in elite durum wheat that could be explained using hyperspectral indices (HSIs) under drought field conditions in Mediterranean environments in Southern Spain. The HSIs were obtained from hyperspectral imagery collected during the pre-anthesis and anthesis crop stages using an airborne platform. A panel of 536 durum wheat lines were genotyped by sequencing (GBS, DArTseq) to determine population structure, revealing a lack of genetic structure in the breeding germplasm. The material was phenotyped for YLD and 19 HSIs for six growing seasons under drought field conditions at two locations in Andalusia, in southern Spain. GWAS analysis identified 740 significant marker-trait associations (MTAs) across all the durum wheat chromosomes, several of which were common for YLD and the HSIs, and can potentially be integrated into breeding programs. Candidate gene (CG) analysis uncovered genes related to important plant processes such as photosynthesis, regulatory biological processes, and plant abiotic stress tolerance. These results are novel in that they combine high-resolution hyperspectral imaging at the field scale with GWAS analysis in wheat. They also support the use of HSIs as useful tools for identifying chromosomal regions related to the heat and drought stress response in wheat, and pave the way for the integration of field HTP in wheat breeding programs.

This study analyzes the productivity of grasslands in a Mediterranean oak-savanna ecosystem, focusing on its linkages with water availability. In these water-controlled ecosystems, grassland environmental preservation and sustainable management depend on quantitatively understanding these links. Productivity and water stress were modeled in southern Spain (2001–2018), integrating meteorological information and MODIS sensor data into a light-use efficiency model and a surface energy balance. The results provided valuable insights into how grasslands behaved during droughts at different spatiotemporal scales. During the most significant droughts, 2004/2005 and 2011/2012, aerial biomass production was reduced by 42 % and 67 %, respectively. The spatial analysis identified the central east side of the region, with low slopes and moderate tree cover, as the most productive area. The biomass production time series classification identified four distinct trends, all showing shifted relationships with similar slopes between production and anomalies of relative evapotranspiration. The seasonal analysis highlighted the importance of autumn, accounting for nearly 30 % of the annual biomass production, which was essential in years with spring water deficits. The proposed methodology provides on-farm grassland production curves depending on water availability (max-mean-min range with a mean error of 15.5 %). Together with weather forecast data, this could help farmers decide on the optimal level of management intensification and stocking rate. Although the regional specificity may limit the study’s direct applicability, this scheme offers valuable metrics that could be adapted to other areas under water scarcity conditions.

Abstract The pandemic of coronavirus disease 19 (COVID-19), caused by severe respiratory syndrome coronavirus 2 (SARS-CoV-2), had severe repercussions for breast cancer patients. Increasing evidence indicates that SARS-CoV-2 infection may directly impact breast cancer biology, but the effects of SARS-CoV-2 on breast tumor cells are still unknown. Here, we analyzed the molecular events occurring in the MCF7, MDA-MB-231 and HCC1937 breast cancer cell lines, representative of the luminal A, basal B/claudin-low and basal A subtypes, respectively, upon SARS-CoV-2 infection. Viral replication was monitored over time, and gene expression profiling was conducted. We found that MCF7 cells were the most permissive to viral replication. Treatment of MCF7 cells with Tamoxifen reduced the SARS-CoV-2 replication rate, suggesting an involvement of the estrogen receptor in sustaining virus replication in malignant cells. Interestingly, a metagene signature based on genes upregulated by SARS-CoV-2 infection in all three cell lines distinguished a subgroup of premenopausal luminal A breast cancer patients with a poor prognosis. As SARS-CoV-2 still spreads among the population, it is essential to understand the impact of SARS-CoV-2 infection on breast cancer, particularly in premenopausal patients diagnosed with the luminal A subtype, and to assess the long-term impact of COVID-19 on breast cancer outcomes.

In Mediterranean oak-savannas (known as dehesas in Spain), it is important to better understand the interactions between vegetation structure and local climate regulation at scales relevant to farm management and the dominant hydrological regime. This study evaluated the water use patterns of dehesa vegetation patches (open grasslands, lowland grasslands with high soil moisture, tree + grass, and riparian vegetation), estimating actual evapotranspiration (ET). We used different models, previously validated in the area, that integrate remotely sensed data. They apply (a) a soil water balance (Kc-FAO56), (b) a surface energy balance (ALEXI/DisALEXI, and SEBS), and (c) a sharpening algorithm (STARFM), obtaining products at multiple spatial resolutions (30 m, 1 km, 5 km). The conceptual and operational differences between the methodologies reinforce the idea of a combined application of models. We demonstrated the need for high spatial and temporal resolution for on-farm livestock management due to the importance of the grasslands layer. This scale is crucial to determine the grass's emergence/drying cycle, which is key for livestock feeding planning. In humid/denser areas that provide essential ecosystem services (e.g., refuge, pasture rotation), transpiration rates are higher throughout the year and were underestimated when coarser spatial scale data was used. Over the typical system (grass with dispersed trees), the ET maps at low spatial resolution reflected the water use trends, and all models correlated well. Higher differences were found when comparing the models' performance over open grasslands.

In this research a miniaturized liquid-liquid extraction was developed for the determination of oxygen heterocyclic compounds in alcoholic beverages. Eight coumarins, twenty-one furocoumarins and seven polymethoxyflavones were identified and quantified, even at trace level, employing a fast and eco-friendly liquid chromatography, tandem mass spectrometry method, recently developed by our research group. Chromatographic separation was obtained in four minutes, using water and ethanol as mobile phases. LOQ values for all the analytes were in the ppb range. The miniaturized vortex-extraction developed was attained in about 12 minutes, employing only 1 mL of sample and of extractant. Furthermore, it allowed to extract more samples simultaneously. Extraction recovery was satisfactory with values ranging between 87 % and 96%. Moreover, the greenness of the proposed methodology was calculated employing the sample preparation method of sustainability metric, which allowed to weigh only the effect of the extraction techniques. Results, expressed in scores from 1-10, gave a value of 6.00, for the newly developed method against the 3.89 rated for a classic liquid-liquid extraction. All the sample analysed contained oxygen heterocyclic compounds in various concentrations with a minimum content of 38.8 µg L−1 and a maximum concentration of 49361 µg L−1.

Abstract Models derived from satellite image data are needed to monitor the status of terrestrial ecosystems across large spatial scales. However, a remote sensing‐based approach to quantify soil multifunctionality at the global scale is missing despite significant research efforts on this topic. A major constraint for doing so is the availability of suitable global‐scale field data to calibrate remote sensing indicators (RSI) and, to a lesser extent, the sensitivity of spectral data of available satellite sensors to soil background and atmospheric conditions. Here, we aimed to develop a soil multifunctionality model to monitor global drylands coupling ground data on 14 soil functions of 222 dryland areas from six continents to 18 RSI derived from a time series (2006–2013) Landsat dataset. Among the RSI evaluated, the chlorophyll absorption ratio index was the best predictor of soil multifunctionality in single‐variable‐based models (r = 0.66, P

The use of phenolic compounds, derived by plants, has recently emerged as a promising approach to prolong the lifespan by modulating metabolic pathways involved in aging. Phenolic compounds possess a broad spectrum of biochemical and pharmacological effects beneficial to human health such as modulating cellular senescence processes by interacting with molecular targets that regulate aging-related pathways. Phenolic compounds represent the major phytochemicals in our diet and possess several biological activities such as antioxidant and anti-inflammatory effects; protection against aging-related diseases (cancer, diabetes and cardiovascular diseases) with potential therapeutic applications and this could suggest that these compounds could be used as anti-aging nutraceutical support. In this review, we have considered the possible effects of some phenolic compounds in different aging pathways, to provide an overview of recent knowledge on their anti-aging mechanism of action.

Abstract Background Spinal anaesthesia complicates maternal hemodynamic and may expose the parturient to dangerous cardiovascular problems. Up to 7% to 89.2% of pregnant women can suffer from spinal anaesthesia-related hypotension. The aim of this study to compare the hemodynamic changes between preeclamptic and normotensive parturients who underwent caesarean section under spinal anaesthesia at North Showa Zone Public Hospitals, Oromia Region, from February 15 to May 15, 2022. Methods A prospective cohort study was conducted on a total of 140 parturients (70 in each group) who underwent cesarean delivery under spinal anesthesia. The study participants were chosen using a consecutive sampling technique. Data were collected from patient charts and intraoperative observations and entered into the Epi Data software version 4.6 and exported to the Statistical Package for the Social Sciences version 25 software. Hemodynamic change = (baseline value-current value/baseline value) * 100. The independent t-test, Mann–Whitney U test, two ways mixed ANOVA, chi-square, and Fisher's exact test was used to analyze the data as appropriate. A P

Oxygen heterocyclic compounds play a beneficial role in plants, and their presence in foods, such as Citrus fruits, cinnamon, carrots, and parsley, has been documented in recent years. Published research articles reported several extractions and chromatographic techniques for their determination. The aim of this review was to take into consideration the research articles published from 2016 to 2024 in which the authors developed extraction and chromatographic analysis methods of oxygen heterocyclic compounds in foods. The objective of this review was to assist researchers in choosing the best approach for their future work by identifying all the possible approaches to characterize coumarins, furocoumarins, and polymethoxyflavones in foodstuffs.

La denominada "diplomacia de la salud" ha adquirido un papel fundamental en las Relaciones Internacionales contemporaneas, especialmente en momentos de crisis sanitaria global, como ocurre con la pandemia de la Covid-19. Países como China e India se han destacado en este ámbito, principalmente a través de sus actividades de Cooperación Sur-Sur (CSS) con naciones del Sur Global. Este artí­culo tiene como objetivo analizar de manera compara-tiva la "diplomacia de la salud" implementada por China e India en África durante la pandemia de la Covid-19. La pregunta que guía este estudio es: "¿Cómo han utilizado estos países las polí­ticas de Cooperación Internacional para el Desarrollo (CID) en la región para combatir la pandemia?" Para responder a esta interrogante, examinamos cómo las prácticas de CID de estos países se relacionan con sus intereses geopolí­ticos y económicos en en la región. Los resultados del análisis indican que la actuación china se ha destacado a través de donaciones de vacunas, insumos y equipamiento hospitalario, además de acciones en el ámbito de la cooperación financiera, como el alivio de la deuda. El caso indio sobresale por la predominancia de la cooperación técnica, mediante la compartición de tecnologías para telecomunicaciones y teleconsultas, además del suministro de vacunas, medicamentos y equipamiento hospitalario

Durable remissions are observed in 10%–20% of treated patients with advanced metastatic melanoma but the factors associated with long-term complete clinical responses are largely unknown. Here, we report the molecular characteristics of tumor evolution during disease progression along a 9-year clinical course in a patient with advanced disseminated melanoma who received different treatments, including trametinib, ipilimumab, radiation, vemurafenib, surgical tumor debulking and a second ipilimumab course, ultimately achieving complete long-term disease remission.Longitudinal analyses of therapies-resistant metastatic tumors revealed the effects of different treatments on tumor’s microenvironment and immunogenicity, ultimately creating a milieu favorable to immunotherapy response. Monitoring of the temporal dynamics of T cells by analysis of the T cell receptor (TCR) repertoire in the tumor and peripheral blood during disease evolution indicated that T-cell clones with common TCR rearrangements, present at low levels at baseline, were maintained and expanded after immunotherapy, and that TCR diversity increased. Analysis of genetic, molecular, and cellular components of the tumor depicted a multistep process in which treatment with kinase inhibitors strongly conditioned the immune microenvironment creating an inflamed milieu converting cold into hot tumors, while ipilimumab impacted and increased the TCR repertoire, a requirement for tumor rejection.Since the optimal sequencing of treatment with antibodies targeting immune checkpoints and kinase inhibitors for advanced melanoma is still clinically debated, this case indicates that immunotherapy success is possible even after progression on targeted therapy.

Immune checkpoint inhibitors (ICIs) have revolutionized cancer therapy and now represent the mainstay of treatment for many tumor types, including triple-negative breast cancer and two agnostic registrations. However, despite impressive durable responses suggestive of an even curative potential in some cases, most patients receiving ICIs do not derive a substantial benefit, highlighting the need for more precise patient selection and stratification. The identification of predictive biomarkers of response to ICIs may play a pivotal role in optimizing the therapeutic use of such compounds. In this Review, we describe the current landscape of tissue and blood biomarkers that could serve as predictive factors for ICI treatment in breast cancer.The integration of these biomarkers in a “holistic” perspective aimed at developing comprehensive panels of multiple predictive factors will be a major step forward towards precision immune-oncology.

Ao nos debruçarmos sobre as edições das revistas Acrópole, Habitat e Módulo, publicadas entre a inauguração de Brasília e o Golpe Militar de 1964, nos damos conta de que, além das referências teóricas e projetuais, a leitura feita por arquitetos e urbanistas sobre seu objeto de trabalho informa e é informada por uma determinada maneira de contar a história do Brasil, e de encarar questões relacionadas à cultura, política e identidade, que conserva parte de sua estrutura até hoje. Nesse sentido, este artigo aborda três diferentes abordagens historiográficas, as quais, ao longo do período estudado, encontraram espaços de destaque nas revistas mencionadas. Nosso objetivo é examinar os termos empregados por aqueles que, durante um momento de intensa movimentação política, econômica e social no Brasil, procuravam escrever e reescrever a história do país.

Avocado (Persea americana Mill.) is a tropical fruit grown in different areas of the world. Avocado pulp is of interest to food companies while seed and peel are wasted. In 2021, about 25 thousand tons of avocado peel were produced as waste. In accordance with circular economy and green chemistry principles, avocado peel could be considered a source of bioactive molecules, specifically phenolic compounds, to be recovered using green methods. Deep eutectic solvents (DESs) are green solvents whose extractive efficiency for phenolic compounds from food and waste products of the agri-food chain has already been demonstrated. In this work, an ultrasound-assisted solid-liquid extraction, using DESs, was developed to recover phenolic compounds from avocado (Hass variety) peel. For this purpose, twelve DESs were tested as extraction solvents considering the provided total phenolic content. Then, in order to obtain the largest amount of phenolic compounds, the most relevant factors affecting solid-liquid extraction were evaluated. Matrix-to-solvent ratio of 1:30 (w/v), 25°C and 15 min as temperature and time of extraction, respectively, were found as optimal conditions to guarantee an extracted amount of phenolic compounds of 8.29 ± 0.07 g GAE/100g of dry avocado peel. Through high-performance liquid chromatography coupled with photodiode array and mass spectrometry detection, some flavonoids and phenolic acids were identified in the avocado peel extract.

Since antiquity, numerous advantages of olive oil and its by-products have been recognized in various domains, including cooking, skincare, and healthcare. Extra virgin olive oil is a crucial component of the Mediterranean diet; several of its compounds exert antioxidant, anti-proliferative, anti-angiogenic and pro-apoptotic effects against a variety of cancers, and also affect cellular metabolism, targeting cancer cells through their metabolic derangements. Numerous olive tree parts, including leaves, can contribute metabolites useful to human health. Olive mill waste water (OMWW), a dark and pungent liquid residue produced in vast amounts during olive oil extraction, contains high organic matter concentrations that may seriously contaminate the soil and surrounding waters if not managed properly. However, OMWW is a rich source of phytochemicals with various health benefits. In ancient Rome, the farmers would employ what was known as amurca, a mulch-like by-product of olive oil production, for many purposes and applications. Several studies have investigated anti-angiogenic and chemopreventive activities of OMWW extracts. The most prevalent polyphenol in OMWW extracts is hydroxytyrosol (HT). Verbascoside and oleuperin are also abundant. We assessed the impact of one such extract, A009, on endothelial cells (HUVEC) and cancer cells. A009 was anti-angiogenic in several in vitro assays (growth, migration, adhesion) and inhibited angiogenesis in vivo, outperforming HT alone. A009 inhibited cells from several tumors in vitro and in vivo and showed potential cardioprotective effects mitigating cardiotoxicity induced by chemotherapy drugs, commonly used in cancer treatment, and reducing up-regulation of pro-inflammatory markers in cardiomyocytes. Extracts from OMWW and other olive by-products have been evaluated for biological activities by various international research teams. The results obtained make them promising candidates for further development as nutraceutical and cosmeceutical agents or dietary supplement, especially in cancer prevention or even in co-treatments with anti-cancer drugs. Furthermore, their potential to offer cardioprotective benefits opens up avenues for application in the field of cardio-oncology.

Abstract Background Autoimmune disorders, including Systemic Lupus Erythematosus (SLE), are associated with increased incidence of hematological malignancies. The matricellular protein osteopontin (OPN) has been linked to SLE pathogenesis, as SLE patients show increased serum levels of OPN and often polymorphisms in its gene. Although widely studied for its pro-tumorigenic role in different solid tumours, the role of OPN in autoimmunity-driven lymphomagenesis has not been investigated yet. Methods To test the role of OPN in the SLE-associated lymphomagenesis, the SLE-like prone Faslpr/lpr mutation was transferred onto an OPN-deficient background. Spleen from Faslpr/lpr and OPN-/-Faslpr/lpr mice, as well as purified B cells, were analysed by histopathology, flow cytometry, Western Blot, immunohistochemistry, immunofluorescence and gene expression profile to define lymphoma characteristics and investigate the molecular mechanisms behind the observed phenotype. OPN cellular localization in primary splenic B cells and mouse and human DLBCL cell lines was assessed by confocal microscopy. Finally, gain of function experiments, by stable over-expression of the secreted (sOPN) and intracellular OPN (iOPN) in OPN-/-Faslpr/lpr -derived DLBCL cell lines, were performed for further validation experiments. Results Despite reduced autoimmunity signs, OPN-/-Faslpr/lpr mice developed splenic lymphomas with higher incidence than Faslpr/lpr counterparts. In situ and ex vivo analysis featured such tumours as activated type of diffuse large B cell lymphoma (ABC-DLBCL), expressing BCL2 and c-MYC, but not BCL6, with activated STAT3 signaling. OPN-/-Faslpr/lpr B lymphocytes showed an enhanced TLR9-MYD88 signaling pathway, either at baseline or after stimulation with CpG oligonucleotides, which mimic dsDNA circulating in autoimmune conditions. B cells from Faslpr/lpr mice were found to express the intracellular form of OPN. Accordingly, gene transfer-mediated re-expression of iOPN, but not of its secreted isoform, into ABC-DLBCL cell lines established from OPN-/-Faslpr/lpr mice, prevented CpG-mediated activation of STAT3, suggesting that the intracellular form of OPN may represent a brake to TLR9 signaling pathway activation. Conclusion These data indicate that, in the setting of SLE-like syndrome in which double strand-DNA chronically circulates and activates TLRs, B cell intracellular OPN exerts a protective role in autoimmunity-driven DLBCL development, mainly acting as a brake in the TLR9-MYD88-STAT3 signaling pathway.

Introduction: Aging is a process characterised by a decline in physiological functions. Reactive species play a crucial role in the aging rate. Due to the close relationship between aging and oxidative stress, functional foods rich in phytochemicals are excellent candidates to neutralise age-related changes.Aim: This investigation aims to verify the potential protective role of bergamot (Citrus bergamia, Femminello cultivar) peel and juice extract in a model of aging represented by human red blood cells (RBCs) exposed to D-Galactose (DGal).Methods: Bergamot peel and juice extracts were subjected to RP-HPLC/PDA/MS for determination of their composition in bioactive compounds. Markers of oxidative stress, including ROS production, thiobarbituric acid reactive substances (TBARS) levels -a marker of lipid peroxidation, oxidation of total protein sulfhydryl groups, as well as the expression and anion exchange capability of band 3 and glycated haemoglobin (A1c) production have been investigated in RBCs treated with D-Gal for 24 h, with or without pre-incubation for 15 min with 5 μg/mL peel or juice extract. In addition, the activity of the endogenous antioxidant system, including catalase (CAT) and superoxide dismutase (SOD), as well as the diversion of the RBC metabolism from glycolysis towards the pentose phosphate pathway shunt, as denoted by activation of glucose-6-phosphate dehydrogenase (G6PDH), have been explored.Results: Data shown here suggest that bergamot peel and juice extract i) prevented the D-Gal-induced ROS production, and consequently, oxidative stress injury to biological macromolecules including membrane lipids and proteins; ii) significantly restored D-Gal-induced alterations in the distribution and ion transport kinetics of band 3; iii) blunted A1c production; iv) effectively impeded the over-activation of the endogenous antioxidant enzymes CAT and SOD; and v) significantly prevented the activation of G6PDH.Discussion: These results further contribute to shed light on aging mechanisms in human RBCs and identify bergamot as a functional food rich in natural antioxidants useful for prevention and treatment of oxidative stress-related changes, which may lead to pathological states during aging.

As most erb‐b2 receptor tyrosine kinase 2 (HER2)‐positive breast cancer (BC) patients currently receive dual HER2‐targeting added to neoadjuvant chemotherapy, improved methods for identifying individual response, and assisting postsurgical salvage therapy, are needed. Herein, we evaluated the 41‐gene classifier trastuzumab advantage risk model (TRAR) as a predictive marker for patients enrolled in the NeoSphere trial. TRAR scores were computed from RNA of 350 pre‐ and 166 post‐treatment tumor specimens. Overall, TRAR score was significantly associated with pathological complete response (pCR) rate independently of other predictive clinico‐pathological variables. Separate analyses according to estrogen receptor (ER) status showed a significant association between TRAR score and pCR in ER‐positive specimens but not in ER‐negative counterparts. Among ER‐positive BC patients not achieving a pCR, those with TRAR‐low scores in surgical specimens showed a trend for lower distant event‐free survival. In conclusion, in HER2‐positive/ER‐positive BC, TRAR is an independent predictor of pCR and represents a promising tool to select patients responsive to anti‐HER2‐based neoadjuvant therapy and to assist treatment escalation and de‐escalation strategies in this setting.

Biofilm formation and lipopolysaccharide (LPS) are implicated in the pathogenesis of gastrointestinal (GI) diseases caused by Gram-negative bacteria. Grape seeds, wine industry by-products, have antioxidant and antimicrobial activity. In the present study, the protective effect of procyanidin-rich grape seed extract (prGSE), from unfermented pomace of Vitis vinifera L. cv Bellone, on bacterial LPS-induced oxidative stress and epithelial barrier integrity damage has been studied in a model of Caco-2 cells. The prGSE was characterized at the molecular level using HPLC and NMR. The in vitro activity of prGSE against formation of biofilm of Salmonella enterica subsp. enterica serovar Typhimurium and Escherichia coli was investigated. In vivo, prGSE activity using infected Galleria mellonella larvae has been evaluated. The results show that the prGSE, if administered with LPS, can significantly reduce the LPS-induced permeability alteration. Moreover, the ability of the extract to prevent Reactive Oxygen Species (ROS) production induced by the LPS treatment of Caco-2 cells was demonstrated. prGSE inhibited the biofilm formation of E. coli and S. Typhimurium. In terms of in vivo activity, an increase in survival of infected G. mellonella larvae after treatment with prGSE was demonstrated. In conclusion, grape seed extracts could be used to reduce GI damage caused by bacterial endotoxin and biofilms of Gram-negative bacteria.

Abstract Background Progression to stage IV disease remains the main cause of breast cancer-related deaths. Increasing knowledge on the hematogenous phase of metastasis is key for exploiting the entire window of opportunity to interfere with early dissemination and to achieve a more effective disease control. Recent evidence suggests that circulating tumor cells (CTCs) possess diverse adaptive mechanisms to survive in blood and eventually metastasize, encouraging research into CTC-directed therapies. Methods On the hypothesis that the distinguishing molecular features of CTCs reveal useful information on metastasis biology and disease outcome, we compared the transcriptome of CTCs, primary tumors, lymph-node and lung metastases of the MDA-MB-231 xenograft model, and assessed the biological role of a panel of selected genes, by in vitro and in vivo functional assays, and their clinical significance in M0 and M+ breast cancer patients. Results We found that hematogenous dissemination is governed by a transcriptional program and identified a CTC signature that includes 192 up-regulated genes, mainly related to cell plasticity and adaptation, and 282 down-regulated genes, involved in chromatin remodeling and transcription. Among genes up-regulated in CTCs, FADS3 was found to increases cell membrane fluidity and promote hematogenous diffusion and lung metastasis formation. TFF3 was observed to be associated with a subset of CTCs with epithelial-like features in the experimental model and in a cohort of 44 breast cancer patients, and to play a role in cell migration, invasion and blood-borne dissemination. The analysis of clinical samples with a panel of CTC-specific genes (ADPRHL1, ELF3, FCF1, TFF1 and TFF3) considerably improved CTC detection as compared with epithelial and tumor-associated markers both in M0 and stage IV patients, and CTC kinetics informed disease relapse in the neoadjuvant setting. Conclusions Our findings provide evidence on the potential of a CTC-specific molecular profile as source of metastasis-relevant genes in breast cancer experimental models and in patients. Thanks to transcriptome analysis we generated a novel CTC signature in the MDA-MB-231 xenograft model, adding a new piece to the current knowledge on the key players that orchestrate tumor cell hematogenous dissemination and breast cancer metastasis, and expanding the list of CTC-related biomarkers for future validation studies.

Abstract Triple negative breast cancer (TNBC) is characterized by clinical aggressiveness, lack of recognized target therapy, and a dismal patient prognosis. Several studies addressed genomic changes occurring during neoadjuvant chemotherapy (NAC) focusing on somatic variants, but without including copy number alterations (CNAs). We analyzed CNA profiles of 31 TNBC primary tumor samples before and after NAC and of 35 single circulating tumor cells (CTCs) collected prior, during and after treatment by using next-generation sequencing targeted profile and low-pass whole genome sequencing, respectively. In pre-treatment tissue samples, the most common gains occurred on chromosomes 1, 2 and 8, and SOX11 and MYC resulted the most altered genes. Notably, amplification of MSH2 (4/4 versus 0/12, p

BackgroundEpilepsy remains one of the world's most common neurological diseases, but it appears to be widely misunderstood, particularly in under-resourced countries like Ethiopia. Improving individuals' knowledge and attitude toward epilepsy is critical for reducing the multifaceted impacts of epilepsy. Therefore, in this study, we sought to estimate the pooled levels of good knowledge and a favorable attitude toward epilepsy and also identify the associated factors using available data collected from different segments of the population.MethodsArticles were searched in international electronic databases. A standardized Microsoft Excel spreadsheet and STATA software version 16 were used for data extraction and analysis, respectively. The Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) checklist was used to write this report. The random-effect meta-analysis model was used to estimate Der Simonian-Laird's pooled effect. Statistical heterogeneity of the meta-analysis was checked via Higgins and Thompson's I2 statistics (0–100%), and Cochran's Q test at P < 0.10. Subgroups, based on the study regions, and sensitivity analyses were also performed. Publication bias was examined subjectively using funnel plots and objectively using the nonparametric rank correlation test of Begg and the regression-based test of Egger for small study effects with P < 0.05 considered to indicate potential publication bias. Furthermore, the Trim-and-fill method of Duval and Tweedie was used to explore sources of publication bias for the favorable level of attitudes toward epilepsy.ResultA total of 12 studies with 6,373 study participants and 10 studies with 5,336 study participants were included to estimate the pooled level of good epilepsy knowledge and favorable attitudes respectively. The overall estimated levels of good epilepsy knowledge and favorable attitudes toward epilepsy among Ethiopians were 47.37% [(95% CI: 35.00, 59.74), I2 = 99.2, P < 0.001] and 46.83%[(95% CI: 32.75, 60.90), I2 = 99.2, P < 0.001] respectively. Subgroup analysis revealed that the pooled level of good epilepsy knowledge was 48.51% [(95% CI: 38.95, 58.06), I2 = 95.6%, P < 0.001] in the Amhara region.ConclusionIn the current review, we found out that there is a huge knowledge gap and an unfavorable level of attitudes towardepilepsy, which demand immediate public health action as well as a targeted policy intervention.

Normal development of the immune system is essential for overall health and disease resistance. Bony fish, such as the zebrafish (Danio rerio), possess all the major immune cell lineages as mammals and can be employed to model human host response to immune challenge. Zebrafish neutrophils, for example, are present in the transparent larvae as early as 48 hours post fertilization and have been examined in numerous infection and immunotoxicology reports. One significant advantage of the zebrafish model is the ability to affordably generate high numbers of individual larvae that can be arrayed in multi-well plates for high throughput genetic and chemical exposure screens. However, traditional workflows for imaging individual larvae have been limited to low-throughput studies using traditional microscopes and manual analyses. Using a newly developed, parallelized microscope, the Multi-Camera Array Microscope (MCAM™), we have optimized a rapid, high-resolution algorithmic method to count fluorescently labeled cells in zebrafish larvae in vivo. Using transgenic zebrafish larvae, in which neutrophils express EGFP, we captured 18 gigapixels of images across a full 96-well plate, in 75 seconds, and processed the resulting datastream, counting individual fluorescent neutrophils in all individual larvae in 5 minutes. This automation is facilitated by a machine learning segmentation algorithm that defines the most in-focus view of each larva in each well after which pixel intensity thresholding and blob detection are employed to locate and count fluorescent cells. We validated this method by comparing algorithmic neutrophil counts to manual counts in larvae subjected to changes in neutrophil numbers, demonstrating the utility of this approach for high-throughput genetic and chemical screens where a change in neutrophil number is an endpoint metric. Using the MCAM™ we have been able to, within minutes, acquire both enough data to create an automated algorithm and execute a biological experiment with statistical significance. Finally, we present this open-source software package which allows the user to train and evaluate a custom machine learning segmentation model and use it to localize zebrafish and analyze cell counts within the segmented region of interest. This software can be modified as needed for studies involving other zebrafish cell lineages using different transgenic reporter lines and can also be adapted for studies using other amenable model species.