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Oral melanoma (OM) is the most common malignant oral tumour among dogs and shares similarities with human mucosal melanoma (HMM), validating the role of canine species as an immunocompetent model for cancer research. In both humans and dogs, the prognosis is poor and radiotherapy (RT) represents a cornerstone in the management of this tumour, either as an adjuvant or a palliative treatment. In this study, by means of RNA-seq, the effect of RT weekly fractionated in 9 Gray (Gy), up to a total dose of 36 Gy (4 weeks), was evaluated in eight dogs affected by OM. Furthermore, possible transcriptomic differences in blood and biopsies that might be associated with a longer overall survival (OS) were investigated. The immune response, glycosylation, cell adhesion, and cell cycle were the most affected pathways by RT, while tumour microenvironment (TME) composition and canonical and non-canonical WNT pathways appeared to be modulated in association with OS. Taking these results as a whole, this study improved our understanding of the local and systemic effect of RT, reinforcing the pivotal role of anti-tumour immunity in the control of canine oral melanoma (COM).

Induced pluripotent stem cell (iPSC) derived endothelial cells (iECs) have emerged as a promising tool for studying vascular biology and providing a platform for modelling various vascular diseases, including those with genetic origins. Currently, primary ECs are the main source for disease modelling in this field. However, they are difficult to edit and have a limited lifespan. To study the effects of targeted mutations on an endogenous level, we generated and characterized an iPSC derived model for venous malformations (VMs). CRISPR-Cas9 technology was used to generate a novel human iPSC line with an amino acid substitution L914F in the TIE2 receptor, known to cause VMs. This enabled us to study the differential effects of VM causative mutations in iECs in multiple in vitro models and assess their ability to form vessels in vivo. The analysis of TIE2 expression levels in TIE2 L914F iECs showed a significantly lower expression of TIE2 on mRNA and protein level, which has not been observed before due to a lack of models with endogenous edited TIE2 L914F and sparse patient data. Interestingly, the TIE2 pathway was still significantly upregulated and TIE2 showed high levels of phosphorylation. TIE2 L914F iECs exhibited dysregulated angiogenesis markers and upregulated migration capability, while proliferation was not affected. Under shear stress TIE2 L914F iECs showed reduced alignment in the flow direction and a larger cell area than TIE2 WT iECs. In summary, we developed a novel TIE2 L914F iPSC-derived iEC model and characterized it in multiple in vitro models. The model can be used in future work for drug screening for novel treatments for VMs., (© 2024. The Author(s).)

Analyzing the relations between Boolean functions has many applications in many fields, such as database systems, cryptography, and collision problems. This paper proposes four quantum algorithms that use amplitude amplification techniques to perform set operations, including Intersection, Difference, and Union, on two Boolean functions in O ( N ) time complexity. The proposed algorithms employ two quantum amplitude amplification techniques divided into two stages. The first stage uses the Younes et al. algorithm for quantum searching via entanglement and partial diffusion to prepare incomplete superpositions of the truth set of the first Boolean function. In the second stage, a modified version of Arima's algorithm, along with an oracle that represent the second Boolean function, is employed to handle the set operations. The proposed algorithms have a higher probability of success in more general and comprehensive applications when compared with relevant techniques in literature., (© 2024. The Author(s).)

Coronavirus disease 2019 (COVID-19) has a wide-ranging spectrum of clinical symptoms, from asymptomatic/mild to severe. Recent research indicates that, among several factors, a low vitamin D level is a modifiable risk factor for COVID-19 patients. This study aims to evaluate the effect of vitamin D on hospital and laboratory outcomes of patients with COVID-19.Five databases (PubMed, Embase, Scopus, Web of Science, and Cochrane Library) and clinicaltrials.gov were searched until July 2022, using relevant keywords/Mesh terms. Only randomized clinical trials (RCTs) that addressed the topic were included. The Cochrane tool was used to assess the studies' risk of bias, and the data were analyzed using the review manager (RevMan 5.4).We included nine RCTs with 1586 confirmed COVID-19 patients. Vitamin D group showed a significant reduction of intensive care unit (ICU) admission (risk ratio = 0.59, 95% confidence interval (CI) [0.41, 0.84], P = 0.003), and higher change in vitamin D level (standardized mean difference = 2.27, 95% CI [2.08, 2.47], P < 0.00001) compared to the control group. Other studied hospital and laboratory outcomes showed non-significant difference between vitamin D and the control group (P ≥ 0.05).In conclusion, vitamin D reduced the risk of ICU admission and showed superiority in changing vitamin D level compared to the control group. However, other outcomes showed no difference between the two groups. More RCTs are needed to confirm these results., (© 2022. The Author(s).)

The function of the cancer-associated lncRNA Malat1 during aging is as-of-yet uncharacterized. Here, we show that Malat1 interacts with Nucleophosmin (NPM) in young mouse brain, and with Lamin A/C, hnRNP C, and KAP1 with age. RNA-seq and RT-qPCR reveal a persistent expression of Malat1&#95; 2 (the 3'isoform of Malat1 ) in Malat1 Δ1 (5'-1.5 kb deletion) mouse retinas and brains at 1/4 th level of the full-length Malat1 , while Malat1&#95; 1 (the 5'isoform) in Malat1 Δ2 (deletion of 3'-conserved 5.7 kb) at a much lower level, suggesting an internal promoter driving the 3' isoform. The 1774 and 496 differentially expressed genes in Malat1 Δ2 and Malat1 Δ1 brains, respectively, suggest the 3' isoform regulates gene expression in trans and the 5' isoform in cis . Consistently, Malat1 Δ2 mice show increased age-dependent retinal oxidative stress and corneal opacity, while Malat1 Δ1 mice show no obvious phenotype. Collectively, this study reveals a physiological function of the lncRNA Malat1 3'-isoform during the aging process., Competing Interests: All the authors declare no competing interests., (© 2022 The Author(s).)

FOXA2 has been known to play important roles in liver functions in rodents. However, its role in human hepatocytes is not fully understood. Recently, we generated FOXA2 mutant induced pluripotent stem cell (FOXA2 -/- iPSC) lines and illustrated that loss of FOXA2 results in developmental defects in pancreatic islet cells. Here, we used FOXA2 -/- iPSC lines to understand the role of FOXA2 on the development and function of human hepatocytes. Lack of FOXA2 resulted in significant alterations in the expression of key developmental and functional genes in hepatic progenitors (HP) and mature hepatocytes (MH) as well as an increase in the expression of ER stress markers. Functional assays demonstrated an increase in lipid accumulation, bile acid synthesis and glycerol production, while a decrease in glucose uptake, glycogen storage, and Albumin secretion. RNA-sequencing analysis further validated the findings by showing a significant increase in genes associated with lipid metabolism, bile acid secretion, and suggested the activation of hepatic stellate cells and hepatic fibrosis in MH lacking FOXA2. Overexpression of FOXA2 reversed the defective phenotypes and improved hepatocyte functionality in iPSC-derived hepatic cells lacking FOXA2. These results highlight a potential role of FOXA2 in regulating human hepatic development and function and provide a human hepatocyte model, which can be used to identify novel therapeutic targets for FOXA2-associated liver disorders., (© 2022. The Author(s).)

Gene expression is controlled by epigenetic deregulation, a hallmark of cancer. The DNA methylome of canine diffuse large B-cell lymphoma (cDLBCL), the most frequent malignancy of B-lymphocytes in dog, has recently been investigated, suggesting that aberrant hypermethylation of CpG loci is associated with gene silencing. Here, we used a multi-omics approach (DNA methylome, transcriptome and copy number variations) combined with functional in vitro assays, to identify putative tumour suppressor genes subjected to DNA methylation in cDLBCL. Using four cDLBCL primary cell cultures and CLBL-1 cells, we found that CiDEA , MAL and PCDH17 , which were significantly suppressed in DLBCL samples, were hypermethylated and also responsive (at the DNA, mRNA and protein level) to pharmacological unmasking with hypomethylating drugs and histone deacetylase inhibitors. The regulatory mechanism underneath the methylation-dependent inhibition of those target genes expression was then investigated through luciferase and in vitro methylation assays. In the most responsive CpG-rich regions, an in silico analysis allowed the prediction of putative transcription factor binding sites influenced by DNA methylation. Interestingly, regulatory elements for AP2, MZF1, NF-kB, PAX5 and SP1 were commonly identified in all three genes. This study provides a foundation for characterisation and experimental validation of novel epigenetically-dysregulated pathways in cDLBCL.

Tumor cell heterogeneity is a crucial characteristic of malignant brain tumors and underpins phenomena such as therapy resistance and tumor recurrence. Advances in single-cell analysis have enabled the delineation of distinct cellular states of brain tumor cells, but the time-dependent changes in such states remain poorly understood. Here, we construct quantitative models of the time-dependent transcriptional variation of patient-derived glioblastoma (GBM) cells. We build the models by sampling and profiling barcoded GBM cells and their progeny over the course of 3 weeks and by fitting a mathematical model to estimate changes in GBM cell states and their growth rates. Our model suggests a hierarchical yet plastic organization of GBM, where the rates and patterns of cell state switching are partly patient-specific. Therapeutic interventions produce complex dynamic effects, including inhibition of specific states and altered differentiation. Our method provides a general strategy to uncover time-dependent changes in cancer cells and offers a way to evaluate and predict how therapy affects cell state composition., (© 2021 The Authors. Published under the terms of the CC BY 4.0 license.)

Bladder cancer, one of the most prevalent malignancies worldwide, remains hard to classify due to a staggering molecular complexity. Despite a plethora of diagnostic tools and therapies, it is hard to outline the key steps leading up to the transition from high-risk non-muscle-invasive bladder cancer (NMIBC) to muscle-invasive bladder cancer (MIBC). Carcinogen-induced murine models can recapitulate urothelial carcinogenesis and natural anti-tumor immunity. Herein, we have developed and profiled a novel model of progressive NMIBC based on 10 weeks of OH-BBN exposure in hepatocyte growth factor/cyclin dependent kinase 4 (R24C) (Hgf-Cdk4R24C) mice. The profiling of the model was performed by histology grading, single cell transcriptomic and proteomic analysis, while the derivation of a tumorigenic cell line was validated and used to assess in vivo anti-tumor effects in response to immunotherapy. Established NMIBC was present in females at 10 weeks post OH-BBN exposure while neoplasia was not as advanced in male mice, however all mice progressed to MIBC. Single cell RNA sequencing analysis revealed an intratumoral heterogeneity also described in the human disease trajectory. Moreover, although immune activation biomarkers were elevated in urine during carcinogen exposure, anti-programmed cell death protein 1 (anti-PD1) monotherapy did not prevent tumor progression. Furthermore, anti-PD1 immunotherapy did not control the growth of subcutaneous tumors formed by the newly derived urothelial cancer cell line. However, treatment with CpG-oligodeoxynucleotides (ODN) significantly decreased tumor volume, but only in females. In conclusion, the molecular map of this novel preclinical model of bladder cancer provides an opportunity to further investigate pharmacological therapies ahead with regards to both targeted drugs and immunotherapies to improve the strategies of how we should tackle the heterogeneous tumor microenvironment in urothelial bladder cancer to improve responses rates in the clinic., Competing Interests: SM is the founder and shareholder of Immuneed AB, Vivologica AB and Strike pharma AB and is the Chief Development Officer and shareholder of Ultimovacs ASA/AB. None of the mentioned companies have had any financial or working impact on the work presented herein. None of the other authors have any competing interests to declare. The above disclosure statement does not alter our adherence to PLOS ONE policies on sharing data and materials.

Background: Several coronavirus vaccine have been fast-tracked to halt the pandemic, the usage of immune adjuvants that can boost immunological memory has come up to the surface. This is particularly of importance in view of the rates of failure of seroconversion and re-infection after COVID-19 infection, which could make the vaccine role and response debatable. Peroxisome proliferator-activated receptors (PPARs) have an established immune-modulatory role, but their effects as adjuvants to vaccination have not been explored to date. It is increasingly recognized that PPAR agonists can upregulate the levels of anti-apoptotic factors such as MCL-1. Such effect can improve the results of vaccination by enhancing the longevity of long-lived plasma cells (LLPCs). The interaction between PPAR agonists and the immune system does not halt here, as T cell memory is also stimulated through enhanced T regulatory cells, antagonizing PD-L1 and switching the metabolism of T cells to fatty acid oxidation, which has a remarkable effect on the persistence of T memory cells. What is even of a more significant value is the effect of PPAR gamma on ensuring a profound secretion of antibodies upon re-exposure to the offending antigen through upregulating lipoxin B4, therefore potentially assisting the vaccine response and deterring re-infection., Short Conclusion: In view of the above, we suggest the use of PPAR as adjuvants to vaccines in general especially the emerging COVID-19 vaccine due to their role in enhancing immunologic memory through DNA-dependent mechanisms.

The association among dysmenorrhea, chronic pain, and conditions classified as central sensitivity syndromes (CSS) is largely unknown. We investigated the co-occurrence of dysmenorrhea with chronic pain and other CSS (e.g., fibromyalgia, migraines); and, whether severity of menstrual symptoms was associated with severity of chronic pain and of somatic symptoms. Women from a mid-sized Canadian university women (N = 248, M age = 21.52) completed measures of menstrual pain severity, chronic pain severity, somatic symptoms severity, and the presence of comorbid CSS. Pearson's correlations assessed the relationship between severity of dysmenorrhea, chronic pain, and somatic symptoms. MANOVA procedures assessed the interaction between dysmenorrhea and chronic pain and χ 2 analyses were used to test the frequency of CSS among women with dysmenorrhea. Higher ratings of menstrual symptom severity were associated with increased chronic pain severity ( r =.66, p <.001), and somatic symptom severity ( r =.66, p <.001). Women with dysmenorrhea were not more likely to experience chronic pain, but were more likely to report a CSS, χ 2 (1) = 5.12, p <.05. Dysmenorrhea symptoms may extend beyond the menstrual phase and be associated with more severe symptoms among women with comorbid pain and somatic concerns.

Glioblastoma (GBM) is a malignant brain tumor with few therapeutic options. The disease presents with a complex spectrum of genomic aberrations, but the pharmacological consequences of these aberrations are partly unknown. Here, we report an integrated pharmacogenomic analysis of 100 patient-derived GBM cell cultures from the human glioma cell culture (HGCC) cohort. Exploring 1,544 drugs, we find that GBM has two main pharmacological subgroups, marked by differential response to proteasome inhibitors and mutually exclusive aberrations in TP53 and CDKN2A/B. We confirm this trend in cell and in xenotransplantation models, and identify both Bcl-2 family inhibitors and p53 activators as potentiators of proteasome inhibitors in GBM cells. We can further predict the responses of individual cell cultures to several existing drug classes, presenting opportunities for drug repurposing and design of stratified trials. Our functionally profiled biobank provides a valuable resource for the discovery of new treatments for GBM., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.)

The increasing demand for more animal products put pressure on improving livestock production efficiency and sustainability. In this context, advanced animal nutrition studies appear indispensable. Here, the effect of grape pomace (GP), the polyphenol-rich agricultural by-product, was evaluated on Holstein-Friesian cows' whole-blood transcriptome, milk production and composition. Two experimental groups were set up. The first one received a basal diet and served as a control, while the second one received a 7.5% GP-supplemented diet for a total of 60 days. Milk production and composition were not different between the group; however, the transcriptome analysis revealed a total of 40 genes significantly affected by GP supplementation. Among the most interesting down-regulated genes, we found the DnaJ heat-shock protein family member A1 (DNAJA1), the mitochondrial fission factor (MFF), and the impact RWD domain protein (IMPACT) genes. The gene set enrichment analysis evidenced the positive enrichment of 'interferon alpha (IFN-α) and IFN-γ response', 'IL6-JAK-STAT3 signaling' and 'complement' genes. Moreover, the functional analysis denoted positive enrichment of the 'response to protozoan' and 'negative regulation of viral genome replication' biological processes. Our data provide an overall view of the blood transcriptomic signature after a 60-day GP supplementation in dairy cows which mainly reflects a GP-induced immunomodulatory effect., Competing Interests: The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Despite advances in the molecular exploration of paediatric cancers, approximately 50% of children with high-risk neuroblastoma lack effective treatment. To identify therapeutic options for this group of high-risk patients, we combine predictive data mining with experimental evaluation in patient-derived xenograft cells. Our proposed algorithm, TargetTranslator, integrates data from tumour biobanks, pharmacological databases, and cellular networks to predict how targeted interventions affect mRNA signatures associated with high patient risk or disease processes. We find more than 80 targets to be associated with neuroblastoma risk and differentiation signatures. Selected targets are evaluated in cell lines derived from high-risk patients to demonstrate reversal of risk signatures and malignant phenotypes. Using neuroblastoma xenograft models, we establish CNR2 and MAPK8 as promising candidates for the treatment of high-risk neuroblastoma. We expect that our method, available as a public tool (targettranslator.org), will enhance and expedite the discovery of risk-associated targets for paediatric and adult cancers.

The effects of iodine supplementation on the whole-transcriptome of dairy cow using RNA sequencing has been investigated in this study. Iodine did not influence the milk composition, while an improvement was observed in the immune response as well as in the quality of dairy product. Indeed, the iodine intake specifically influenced the expression of 525 genes and the pathway analysis demonstrated that the most affected among them were related to immune response and oxidative stress. As a consequence, we indirectly showed a better response to bacterial infection because of the reduction of somatic cell counts; furthermore, an improvement of dairy product quality was observed since lipid oxidation reduced in fresh cheese. Such findings, together with the higher milk iodine content, clearly demonstrated that iodine supplementation in dairy cow could represent a beneficial practice to preserve animal health and to improve the nutraceutical properties of milk and its derived products.

Canine malignant melanoma (MM) is a highly aggressive tumour with a low survival rate and represents an ideal spontaneous model for the human counterpart. Considerable progress has been recently obtained, but the therapeutic success for canine melanoma is still challenging. Little is known about the mechanisms beyond pathogenesis and melanoma development, and the molecular response to radiotherapy has never been explored before. A faster and deeper understanding of cancer mutational processes and developing mechanisms are now possible through next generation sequencing technologies. In this study, we matched whole exome and transcriptome sequencing in four dogs affected by MM at diagnosis and at disease progression to identify possible genetic mechanisms associated with therapy failure. According to previous studies, a genetic similarity between canine MM and its human counterpart was observed. Several somatic mutations were functionally related to MAPK, PI3K/AKT and p53 signalling pathways, but located in genes other than BRAF, RAS and KIT. At disease progression, several mutations were related to therapy effects. Natural killer cell-mediated cytotoxicity and several immune-system-related pathways resulted activated opening a new scenario on the microenvironment in this tumour. In conclusion, this study suggests a potential role of the immune system associated to radiotherapy in canine melanoma, but a larger sample size associated with functional studies are needed., (© 2019 John Wiley & Sons Ltd.)

The genomic landscape in human B-cell lymphoma has revealed several somatic mutations and potentially relevant germline alterations affecting therapy and prognosis. Also, mutations originally described as somatic aberrations have been shown to confer cancer predisposition when occurring in the germline. The relevance of mutations in canine B-cell lymphoma is scarcely known and gene expression profiling has shown similar molecular signatures among different B-cell histotypes, suggesting other biological mechanisms underlining differences. Here, we present a highly accurate approach to identify single nucleotide variants (SNVs) in RNA-seq data obtained from 62 completely staged canine B-cell lymphomas and 11 normal B-cells used as controls. A customized variant discovery pipeline was applied and SNVs were found in tumors and differentiated for histotype. A number of known and not previously identified SNVs were significantly associated to MAPK signaling pathway, negative regulation of apoptotic process and cell death, B-cell activation, NF-kB and JAK-STAT signaling. Interestingly, no significant genetic fingerprints were found separating diffuse large B-cell lymphoma from indolent lymphomas suggesting that differences of genetic landscape are not the pivotal causative factor of indolent behavior. We also detected several variants in expressed regions of canine B-cell lymphoma and identified SNVs having a direct impact on genes. Using this brand-new approach the consequence of a gene variant is directly associated to expression. Further investigations are in progress to deeply elucidate the mechanisms by which altered genes pathways may drive lymphomagenesis and a higher number of cases is also demanded to confirm this evidence., Competing Interests: The authors have declared that no competing interests exist.

Objectives: Little is known about temporal trajectories of social support in adults with type 2 diabetes (T2D) and how they are associated with diabetes-related outcomes. This study identified and explored different trajectories of social support in a community sample of adults with T2D, as well as the extent to which different trajectories were prospectively associated with depressive symptoms and functional disability., Methods: Data came from five annual waves of the Evaluation of Diabetes Treatment study (N = 1077). Social support, depressive symptoms, and functional disability were assessed via self-report. Separate analyses were conducted to examine the associations between social support trajectories, depressive symptoms, and functional disability, adjusting for demographic characteristics, diabetes-related covariates, and baseline depressive symptoms and functional disability., Results: Latent class growth modeling identified four distinct social support trajectories. Trajectory Groups 1 and 2 comprised participants with persistently low and persistently moderate-low social support, respectively. Trajectory Groups 3 and 4 included participants with persistently moderate-high and persistently high social support, respectively. People with persistently low social support reported higher functional disability relative to those with persistently moderate-high and persistently high social support., Conclusions: The findings of the present study indicate that temporal patterns of social support are a predictor of future functional disability among adults with T2D., (© 2018 John Wiley & Sons, Ltd.)

Background: Heavy alcohol consumption in individuals with type 2 diabetes mellitus (T2DM) is related to increased risks of diabetes-related micro- and macrovascular complications. Depressive symptoms may be relevant to this relationship, because high depressive symptoms are associated with an increased risk of complications. This study investigated whether the interaction between depressive symptoms and alcohol frequency was positively related to the development of neuropathy, retinopathy, nephropathy, and coronary artery disease (CAD), such that those with high depressive symptoms and high alcohol frequency will be at increased risk of complications., Methods: Data were from five waves of the Evaluation of Diabetes Treatment annual survey including 1413 adults with T2DM in Quebec. Data on alcohol frequency (number of drinking occasions), depressive symptoms, and complications were collected annually. The development of each complication was investigated using multiple logistic regression analysis with generalized estimating equations., Results: After adjusting for sociodemographic, lifestyle, and diabetes-related covariates, the interaction between alcohol frequency and depressive symptoms was positively related to the incidence of neuropathy and CAD, such that those with high depressive symptoms who drank the most frequently had the highest risk of neuropathy (odds ratio [OR] 1.02, 95% confidence interval [CI] 1.00-1.04; P = 0.04) and CAD (OR 1.02, 95% CI 1.00-1.04; P = 0.04). This interaction was not significantly related to retinopathy or nephropathy., Conclusion: Individuals with high depressive symptoms and high alcohol frequency may have a particularly high risk of neuropathy and CAD. Future prevention efforts should examine both alcohol frequency and depressive symptoms when evaluating the risk of complications., (© 2018 Ruijin Hospital, Shanghai Jiaotong University School of Medicine and John Wiley & Sons Australia, Ltd.)

G-quadruplexes (G4) are secondary nucleic acid structures that have been associated with genomic instability and cancer progression. When present in the promoter of some oncogenes, G4 structures can affect gene regulation and, hence, represent a possible therapeutic target. In this study, RNA-Seq was used to explore the effect of a G4-binding anthraquinone derivative, named AQ1, on the whole-transcriptome profiles of two common cell models for the study of KIT pathways; the human mast cell leukemia (HMC1.2) and the canine mast cell tumor (C2). The highest non-cytotoxic dose of AQ1 (2 µM) resulted in 5441 and 1201 differentially expressed genes in the HMC1.2 and C2 cells, respectively. In both cell lines, major pathways such as cell cycle progression, KIT- and MYC-related pathways were negatively enriched in the AQ1-treated group, while other pathways such as p53, apoptosis and hypoxia-related were positively enriched. These findings suggest that AQ1 treatment induces a similar functional response in the human and canine cell models, and provide news insights into using dogs as a reliable translational model for studying G4-binding compounds.

Grape pomace (GPO), the main by-product of the wine making process, is a rich source of polyphenols with potent antioxidant properties. Recently, GPO has emerged as a potential feed additive in livestock nutrition, with several reports describing its beneficial effects on animals' overall health status or production traits. However, little is known about it from a molecular biology standpoint. In the present study, we report the first RNA sequencing-based whole-transcriptome profiling of Friesian calves fed with a GPO-supplemented diet. We identified 367 differentially expressed genes ( p < 0.05) in the GPO-supplemented calves ( n = 5), when compared with unsupplemented control group ( n = 5). The pathway analysis showed that 'cholesterol lipid biosynthesis' was the most negatively-enriched ( p < 0.001) pathway in the GPO-supplemented animals. In specific terms, five important genes coding for cholesterol biosynthesis enzymes, namely the Farnesyl-diphosphate Farnesyltransferase 1 (FDFT-1), Squalene Epoxidase (SQLE), NAD(P)-dependent Steroid Dehydrogenase-like (NSDHL), Methylsterol Monooxygenase (MSMO)-1, and Sterol-C5-desaturase (SC5D), two major transcription factors (the Sterol Regulatory Element-binding Transcription Factor 1 and 2), as well as the Low-Density Lipoprotein Receptor (LDLR), were all downregulated following GPO supplementation. Such an effect was mirrored by a reduction of blood cholesterol levels ( p = 0.07) and a lowered ( p < 0.001) Malondialdehyde (lipid oxidation marker) level in carcasses. We provide evidence on the effects of GPO-supplemented diets on the whole-transcriptome signature in veal calves, which mainly reflects an antioxidant activity.

Although heavy alcohol consumption is associated with diabetes-related complications, little is known about patterns of alcohol use among people with diabetes. Moreover, heavy drinking is more common among individuals with major depressive disorder (MDD), bipolar disorder (BD), and generalized anxiety disorder (GAD) than in the general population, and these disorders are often comorbid with diabetes. The present study tested the hypothesis that mental disorders moderate the association between diabetes status and alcohol consumption. A total of 14,302 adult participants aged 40-79 were included from the cross-sectional 2012 Canadian Community Health Survey-Mental Health (1,698 with diabetes). Data were analyzed using hierarchical linear regression models. MDD and BD, but not GAD, significantly moderated the association between diabetes status and alcohol quantity, such that the presence of diabetes was strongly and negatively associated with alcohol quantity if individuals had MDD or BD. There was no interaction between diabetes status and any of the mental disorders and alcohol frequency. This study suggests that among individuals with diabetes, those with comorbid MDD or BD drink less than those without MDD or BD. Further investigation of this association is needed and could help inform future alcohol-related interventions among individuals with diabetes.

Bovine primary cultured hepatocytes (CHs) are widely used in vitro models for liver toxicity testing. However, little is known about their whole-transcriptome profile and its resemblance to the normal liver tissue. In the present study, we profiled - by microarray - the whole-transcriptome of bovine CHs (n=4) and compared it with the transcriptomic landscape of control liver samples (n=8), as well the Madin-Darby bovine kidney (MDBK) cells (n=4). Compared with liver tissue, the bovine CHs relatively expressed (fold change >2, P<0.05) about 2155 and 2073 transcripts at a lower and higher abundance, respectively. Of those expressed at a lower abundance, many were drug biotransformation enzyme-coding genes, such as the cytochrome P450 family (CYPs), sulfotransferases, methyltransferases, and glutathione S-transferases. Also, several drug transporters and solute carriers were expressed at a lower abundance in bovine CHs. 'Drug metabolism', 'PPAR signaling', and 'metabolism of xenobiotics by CYPs' were among the most negatively-enriched pathways in bovine CHs compared with liver. A qPCR cross-validation using 8 selected genes evidenced a high correlation (r=0.95, P=0.001) with the corresponding microarray results. Although from a kidney origin, and albeit to a lower extent compared to bovine CHs, the MDBK cells showed a basal expression of many CYP-coding genes. Our study provides a whole-transcriptome-based evidence for the bovine CHs and hepatic tissue resemblance. Overall, the bovine CHs' transcriptomic profile might render it unreliable as an in vitro model to study drug metabolism., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

We investigated the transcriptomic signature of some anabolic steroids in cattle. Our main objective was to evaluate the effect of a combined trenbolone acetate (TBA, 200mg) and estradiol-17β (E2, 40 mg) implant (Revalor-XS®, REV) on the transcriptome of muscle (target tissue for anabolic steroids) and liver (main biotransformation site). Transcriptomic profiling was performed on 60 samples (30 per tissue) representing 2 groups of animals: REV (sustained release implant for 71 days, n=15), and a control group (CTR, n=15). The analyses (REV vs. CTR) evidenced the differential expression of 431 (down-regulated) and 503 transcripts (268 up-regulated and 235 down-regulated) in muscle and liver tissues, respectively. Functional annotation showed the enrichment of several ion transport systems (cation, metal ion and potassium ion transport) in muscle, while revealing the enrichment of carbohydrate, protein and glycoprotein metabolism and biosynthesis mechanisms in the liver. Both tissues had 20 genes commonly expressed in-between. Seven randomly-selected genes showed positive correlation with their corresponding microarray data upon a qPCR cross-validation step. In muscle, but not the liver, Principal Component Analysis (PCA) on the microarray data resulted in the separation of treated animals from the untreated ones (first 2 components=97.87%.). Overall, the identification of different genes, pathways and biological processes has illustrated the distinctive transcriptomic profile of muscle and liver in response to anabolic steroids. Moreover, it is becoming more clear that anabolic steroids are working through a complex interaction of numerous pathways and processes incorporating different tissues., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Indole-3-carbinol (I3C) and diindolylmethane (DIM), found in cruciferous vegetables, have chemopreventive and anticancer properties. In the present study, 14 substituted indoles were tested for activity against SW480 colon cancer cells. Among these, 3-(2-bromoethyl)-indole, named BEI-9, showed the greatest inhibition. The effects of BEI-9 on cancer cells were analyzed by MTS and CellTiter-Glo assays for effects on cell viability, by microscopy for phenotypic changes, by scratch wound assays for effects on migration, by flow cytometry for changes in the cell cycle, by immunoblotting for cyclin D and A to assess effects on cell cycle regulation, and by NF-κB reporter assays for effects on basal and drug-induced NF-κB activation. BEI-9 inhibited the growth of SW480 and HCT116 colon cancer cells at concentrations of 12.5 and 5 µM, respectively. BEI-9 also inhibited cell motility as determined with scratch wound assays, and reduced the levels of cyclin D1 and A. Furthermore, in reporter cells, BEI-9 (0.8 µM) inhibited basal and induced NF-κB activation and increased cell death when combined with the cytokine TNFα or the drug camptothecin (CPT), both of which activate NF-κB. Preliminary experiments to identify a safe dose range for immunodeficient mice showed that BEI-9, administered intraperitoneally, was tolerable at doses below 10 mg/kg. Thus, BEI-9 and other indole derivatives may be useful in chemoprevention or as chemosensitizers. Since NF-κB activation is implicated in carcinogenesis and in reducing sensitivity to anticancer drugs, BEI-9 should be investigated in combination with drugs such as CPT, which activate NF-κB.

Growth promoters (GPs) such as the glucocorticoid dexamethasone (DEX) and the β-adrenergic agonist clenbuterol (CLEN) are still used abusively in beef cattle production. Transcriptomic markers for indirect detection of such GPs have been discussed in either experimentally treated animals or commercial samples separately. In the present study we examine the transcriptomic signature of DEX alone or in combination with CLEN in skeletal muscle of experimentally treated beef cattle, and, furthermore, compare them with previously screened commercial samples from a field-monitoring study, as well as with proteomics data representing the same set of samples. Using DNA microarray technology, transcriptomic profiling was performed on 12 samples representing three groups of animals: DEX (0.75 mg/animal/day, n = 4), a combination of DEX (0.66 mg/animal/day) and CLEN (from 2 to 6 mg/animal/day, n = 4) and a control group (n = 4). Analyses showed the differential expression of 198 and 39 transcripts in DEX and DEX-CLEN groups, respectively. Both groups had no common modulated genes in between, neither with the proteomics data. Sixteen candidate genes were validated via qPCR. They showed high correlation with the corresponding microarray data. Principal component analysis (PCA) on both the qPCR and normalised microarray data resulted in the separation of treated animals from the untreated ones. Interestingly, all the PCA plots grouped the DEX-positive samples (experimental or commercial) apart from each other. In brief, this study provides some interesting glucocorticoid-responsive biomarkers whose expression was contradictory to what is reported in human studies. Additionally, this study points out the transcriptomic signature dissimilarity between commercial and experimentally treated animals.