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Lutein is a carotenoid that humans obtain primarily from green-leafy vegetables. However, due to generally low consumption of these foods, this bioactive is not sufficiently consumed. Therefore, there is growing interest in developing lutein-fortified foods to provide an additional source of lutein supplementation. In this study, we developed a lutein-fortified yoghurt and conducted a 72-hour single-dose bioavailability study in human subjects. Lutein remained stable in the yoghurt over shelf life at 14.6 mg of lutein per 100 g of yoghurt. Sensory evaluation yielded results similar to a lutein-free yoghurt. In the bioavailability study, consuming 100 g of lutein-fortified yoghurt significantly increased the average serum concentration of lutein by 1946.86 ± 1514.72 h μg·L−1 in human subjects, following a typical pharmacokinetic curve of absorption and decline. These findings confirm that our lutein-fortified yoghurt is suitable for commercial use and effectively enhances people’s serum lutein levels.

[Display omitted] • We produced a lutein-fortified yoghurt at a pilot-scale. • This yoghurt contains the maximum lutein allowed by EFSA for a dairy product. • It has excellent lutein stability and favourable flavour characteristics. • A single dose of this yoghurt significantly increases serum lutein in humans. Lutein is a carotenoid that humans obtain primarily from green-leafy vegetables. However, due to generally low consumption of these foods, this bioactive is not sufficiently consumed. Therefore, there is growing interest in developing lutein-fortified foods to provide an additional source of lutein supplementation. In this study, we developed a lutein-fortified yoghurt and conducted a 72-hour single-dose bioavailability study in human subjects. Lutein remained stable in the yoghurt over shelf life at 14.6 mg of lutein per 100 g of yoghurt. Sensory evaluation yielded results similar to a lutein-free yoghurt. In the bioavailability study, consuming 100 g of lutein-fortified yoghurt significantly increased the average serum concentration of lutein by 1946.86 ± 1514.72 h μg·L−1 in human subjects, following a typical pharmacokinetic curve of absorption and decline. These findings confirm that our lutein-fortified yoghurt is suitable for commercial use and effectively enhances people's serum lutein levels. [ABSTRACT FROM AUTHOR]

RMEL3 is a recently identified lncRNA associated with BRAFV600E mutation and melanoma cell survival. Here, we demonstrate strong and moderate RMEL3 upregulation in BRAF and NRAS mutant melanoma cells, respectively, compared to melanocytes. High expression is also more frequent in cutaneous than in acral/mucosal melanomas, and analysis of an ICGC melanoma dataset showed that mutations in RMEL3 locus are preponderantly C > T substitutions at dipyrimidine sites including CC > TT, typical of UV signature. RMEL3 mutation does not correlate with RMEL3 levels, but does with poor patient survival, in TCGA melanoma dataset. Accordingly, RMEL3 lncRNA levels were significantly reduced in BRAFV600E melanoma cells upon treatment with BRAF or MEK inhibitors, supporting the notion that BRAF‐MEK‐ERK pathway plays a role to activate RMEL3 gene transcription. RMEL3 overexpression, in immortalized fibroblasts and melanoma cells, increased proliferation and survival under serum starvation, clonogenic ability, and xenografted melanoma tumor growth. Although future studies will be needed to elucidate the mechanistic activities of RMEL3, our data demonstrate that its overexpression bypasses the need of mitogen activation to sustain proliferation/survival of non‐transformed cells and suggest an oncogenic role for RMEL3. [ABSTRACT FROM AUTHOR]

Sprouty proteins control length and intensity of the intracellular signal transduction cascade activated by mitogens in the cellular environment. As part of a negative feedback loop, their expression is supposed to be elevated by the same factors. In this report, Sprouty4 expression in response to serum and the underlying regulatory mechanisms were investigated. We verified that Sprouty4 expression is activated by serum addition in all tested cells independent of their origin. Strict correlation between Sprouty4 protein levels and promoter activity indicates mainly transcriptional regulation of Sprouty4 serum-responsiveness. Induction of the mitogen-activated protein kinase pathway is required for Sprouty4 promoter activation in the presence of serum. Nonetheless, signal transduction via this pathway is not sufficient to fully induce the Sprouty4 promoter. Instead, deletion and mutation analysis identified two annotated Specific protein 1 binding sites as the critical cis -elements responsible for conferring the serum induction of the promoter. Corroborating, repressed Specific protein 1 activity or levels result in constitutive lowered transcriptional activity of the Sprouty4 promoter. These data demonstrate that Specific protein 1 plays a crucial role in the regulation of Sprouty4 in response to serum. [ABSTRACT FROM AUTHOR]

Abstract: Human mitochondrial transcription termination factor 2 (MTERF2) is a member of the mitochondrial transcription termination factors (MTERFs) family and a cell growth inhibitor. To create a specific mouse monoclonal antibody against human MTERF2, the full-length His-tag MTERF2 protein (1–385 aa) was expressed in Escherichia coli, and purified recombinant protein was injected into three BALB/c mice to perform an immunization procedure. Eight stable positive monoclonal cell lines were screened and established. ELISA results demonstrated that all antibody light chains were kappa, while the heavy chains displayed three subtypes IgG1, IgG2a, and IgG2b respectively. The sensitivity and specificity of the monoclonal antibodies against human MTERF2 were determined using immunoblotting, immunoprecipitation and immunofluorescence analyses. Furthermore, serum regulation of human MTERF2 protein expression levels in human glioma U251 cells was examined with these monoclonal antibodies and the results demonstrated that the expression level of MTERF2 protein was dramatically inhibited by the addition of serum to serum-starved cells. Taken together, our results demonstrate the functionality of these mouse anti-human MTERF2 monoclonal antibodies, which may provide a useful tool to elucidate the role of MTERF2 in human mitochondrial transcription as well as other potential activities. To our knowledge, this is the first report on the preparation and characterization of mouse monoclonal antibodies against human MTERF2. [Copyright &y& Elsevier]

Background: Keloids are benign tumors that occur only in response to injury, for which there is no effective treatment. We demonstrated previously that keloid keratinocytes (KKs) promote fibroblast proliferation more than normal keratinocytes (NKs) and that transforming growth factor (TGF)-β is a component of that signal. We used the transcriptional response to serum stimulation to examine how TGF-β expression is stimulated in KKs. Materials and methods: Quiescent KKs and NKs were stimulated using serum; harvested using RNA at 0, 1, 6, 12, and 24 h; and analyzed usingquantitative real-time polymerase chain reaction. TGF-β activity in the conditioned medium was measured with an MLEC/PAI-luciferase assay. Inhibition of ERK1/2, p38 kinase, and JNK pathways was performed with PD98059, SB203580, and SP600125, respectively. Results: Increased transcription of TGF-β2 occurs within 1 h of serum stimulation in KKs but not in NKs. In contrast, TGF-β3 transcription was suppressed in KKs compared with NKs. No significant differences were observed in the transcriptional response of TGF-β1. Increased TGF-β2 mRNA correlated with increased TGF-β biological activity in the conditioned medium. Inhibition of the ERK, p38 kinase or JNK signal transduction pathways blocked the transcriptional up-regulation of TGF-β2, TβR1, and TβR2 in KKs. Conclusions: KKs produce more TGF-β2 mRNA than NKs in response to serum stimulation, resulting in increased TGF-β activity in conditioned medium. Combining these results with our previous data lead us to propose a model of keloid formation characterized by an exaggerated response to cellular stress and abnormal epithelial-mesenchymal signaling promoting keloid formation. [Copyright &y& Elsevier]

Serum stimulation leads to activation of the serum response factor (SRF)-mediated transcription of immediate-early genes such as c-fos via various signal transduction pathways. We have previously reported that promyelocytic leukemia protein (PML) is involved in the transcriptional regulation by SRF. PML is one of the well-known substrates for modification by small ubiquitin-related modifier-1 (SUMO-1) and several SUMO-1-modified proteins associate with PML. Here, we report that SRF is modified by SUMO-1 chiefly at lysine147 within the DNA-binding domain. Substitution of this target lysine for alanine did not affect the translocation of SRF to PML-nuclear bodies. The SRF mutant augmented the transcriptional activity under Rho A-stimulated condition but not under serum-starved condition, suggesting that activated SRF is suppressed by its sumoylation. These data support the transcriptional role of SUMO-1 conjugating system in cellular serum response. [Copyright &y& Elsevier]

Serum responses to oral cholera vaccines were assessed in three paediatric vaccine trials, two in Leo´n, Nicaragua and one in Stockholm, Sweden. A calibrated anti-cholera toxin B subunit (CTB) IgA ELISA was used together with an assay for vibriocidal antibodies. Swedish children had lower pre-vaccination levels of antibody, but serum responses were more pronounced in Swedish children than in Nicaraguan children. Post-vaccination levels of anti-toxin antibody were generally above those found after natural infections with enterotoxigenic Escherichia coli, that cross-reacts serologically with Vibrio cholerae. Adverse events seen after vaccination were generally mild and of little clinical significance. [Copyright &y& Elsevier]

CDK8 belongs to a group of cyclin-dependent kinases involved in transcriptional regulation from yeast to mammals. CDK8 associates with the Mediator complex, but functions outside of Mediator are also likely. Historically, CDK8 has been described mostly as a transcriptional repressor, but a growing body of research provides unequivocal evidence for various roles of CDK8 in gene activation. Several transcriptional programs of biomedical importance employ CDK8 as a co-activator, including the p53 network, the Wnt/β-catenin pathway, the serum response network, and those governed by SMADs and the thyroid hormone receptor, thus highlighting the importance of further investigation into this enigmatic transcriptional regulator.