Stilbenoids remodel the DNA methylation patterns in breast cancer cells and inhibit oncogenic NOTCH signaling through epigenetic regulation of MAML2 transcriptional activity

DNA hypomethylation was previously implicated in cancer progression and metastasis. The purpose of this study was to examine whether stilbenoids, resveratrol and pterostilbene thought to exert anticancer effects, target genes with oncogenic function for de novo methylation and silencing, leading to inactivation of related signaling pathways. Following Illumina 450K, genome-wide DNA methylation analysis reveals that stilbenoids alter DNA methylation patterns in breast cancer cells. On average, 75% of differentially methylated genes have increased methylation, and these genes are enriched for oncogenic functions, including NOTCH signaling pathway. MAML2, a coactivator of NOTCH targets, is methylated at the enhancer region and transcriptionally silenced in response to stilbenoids, possibly explaining the downregulation of NOTCH target genes. The increased DNA methylation at MAML2 enhancer coincides with increased occupancy of repressive histone marks and decrease in activating marks. This condensed chromatin structure is associated with binding of DNMT3B and decreased occupancy of OCT1 transcription factor at MAML2 enhancer, suggesting a role of DNMT3B in increasing methylation of MAML2 after stilbenoid treatment. Our results deliver a novel insight into epigenetic regulation of oncogenic signals in cancer and provide support for epigenetic-targeting strategies as an effective anticancer approach

IL-6 Mediated Degeneration of Forebrain GABAergic Interneurons and Cognitive Impairment in Aged Mice through Activation of Neuronal NADPH Oxidase

BACKGROUND:Multiple studies have shown that plasma levels of the pro-inflammatory cytokine interleukin-6 (IL-6) are elevated in patients with important and prevalent adverse health conditions, including atherosclerosis, diabetes, obesity, obstructive sleep apnea, hypertension, and frailty. Higher plasma levels of IL-6, in turn, increase the risk of many conditions associated with aging including age-related cognitive decline. However, the mechanisms underlying this association between IL-6 and cognitive vulnerability remain unclear. METHODS AND FINDINGS:We investigated the role of IL-6 in brain aging in young (4 mo) and aged (24 mo) wild-type C57BL6 and genetically-matched IL-6(-/-) mice, and determined that IL-6 was necessary and sufficient for increased neuronal expression of the superoxide-producing immune enzyme, NADPH-oxidase, and this was mediated by non-canonical NFkappaB signaling. Furthermore, superoxide production by NADPH-oxidase was directly responsible for age-related loss of parvalbumin (PV)-expressing GABAergic interneurons, neurons essential for normal information processing, encoding, and retrieval in hippocampus and cortex. Targeted deletion of IL-6 or elimination of superoxide by chronic treatment with a superoxide-dismutase mimetic prevented age-related loss of PV-interneurons and reversed age-related cognitive deficits on three standard tests of spatial learning and recall. CONCLUSIONS:Present results indicate that IL-6 mediates age-related loss of critical PV-expressing GABAergic interneurons through increased neuronal NADPH-oxidase-derived superoxide production, and that rescue of these interneurons preserves cognitive performance in aging mice, suggesting that elevated peripheral IL-6 levels may be directly and mechanistically linked to long-lasting cognitive deficits in even normal older individuals. Further, because PV-interneurons are also selectively affected by commonly used anesthetic agents and drugs, our findings imply that IL-6 levels may predict adverse CNS effects in older patients exposed to these compounds through specific derangements in inhibitory interneurons, and that therapies directed at lowering IL-6 may have cognitive benefits clinically

Enterobactin-Mediated Delivery of β-Lactam Antibiotics Enhances Antibacterial Activity against Pathogenic Escherichia coli

The design, synthesis, and characterization of enterobactin–antibiotic conjugates, hereafter Ent-Amp/Amx, where the β-lactam antibiotics ampicillin (Amp) and amoxicillin (Amx) are linked to a monofunctionalized enterobactin scaffold via a stable poly(ethylene glycol) linker are reported. Under conditions of iron limitation, these siderophore-modified antibiotics provide enhanced antibacterial activity against Escherichia coli strains, including uropathogenic E. coli CFT073 and UTI89, enterohemorrhagic E. coli O157:H7, and enterotoxigenic E. coli O78:H11, compared to the parent β-lactams. Studies with E. coli K-12 derivatives defective in ferric enterobactin transport reveal that the enhanced antibacterial activity observed for this strain requires the outer membrane ferric enterobactin transporter FepA. A remarkable 1000-fold decrease in minimum inhibitory concentration (MIC) value is observed for uropathogenic E. coli CFT073 relative to Amp/Amx, and time-kill kinetic studies demonstrate that Ent-Amp/Amx kill this strain more rapidly at 10-fold lower concentrations than the parent antibiotics. Moreover, Ent-Amp and Ent-Amx selectively kill E. coli CFT073 co-cultured with other bacterial species such as Staphylococcus aureus, and Ent-Amp exhibits low cytotoxicity against human T84 intestinal cells in both the apo and iron-bound forms. These studies demonstrate that the native enterobactin platform provides a means to effectively deliver antibacterial cargo across the outer membrane permeability barrier of Gram-negative pathogens utilizing enterobactin for iron acquisition.Pacific Southwest Regional Center of Excellence for Biodefense and Emerging Infectious DiseaseKinship Foundation. Searle Scholars ProgramMassachusetts Institute of Technology. Department of Chemistr

Peptidase activity of selected heat-treated Lactobacillus strains

Celem niniejszej pracy było określenie aktywności amino- i dipeptydaz wybranych kultur bakterii mlekowych (Lactobacillus casei, Lactobacillus acidophilus), poddanych działaniu temperatury 50 ÷ 75 °C, przez 1, 15 i 25 min. Badane kultury Lactobacillus syntetyzowały peptydazy o podobnej specyficzności substratowej, ale z różną aktywnością. Wykazywały one wyższą aktywność aminopeptydaz w porównaniu z aktywnością dipeptydaz. Średnia aktywność dipeptydaz Lb. acidophilus była wyższa o 45 % od średniej aktywności bakterii Lb. casei. Wyższą o 25 % aktywnością aminopeptydaz charakteryzował się szczep Lb. casei. Szczep ten wykazywał największą specyficzność względem substratów: Ala-Leu, Ala-Ala, Gly-Leu, natomiast Lb. acidophilus – względem Ala-Ala oraz Ala-pNa. Najwyższą aktywność amino- i dipeptydaz Lb. casei oraz dipeptydaz Lb. acidophilus stwierdzono po obróbce termicznej bakterii przez 15 min. W przypadku amino- i dipeptydaz Lb. casei ich aktywność wynosiła odpowiednio: 5,10 i 0,83 U·min⁻¹·mg⁻¹ oraz 1,66 U·min⁻¹·mg⁻¹ – w przypadku dipeptydaz Lb. acidophilus. Z kolei średnia aktywność aminopeptydaz Lb. acidophilus wzrastała wraz z wydłużaniem czasu ogrzewania – najwyższe jej wartości uzyskano po 25 min (3,89 U·min⁻¹·mg⁻¹). Wykazano, że wydłużenie czasu obróbki termicznej wpłynęło statystycznie istotnie (p < 0,05) na wzrost aktywności aminopeptydaz badanych kultur bakterii, co wskazuje na ich wysoką termostabilność.The objective of the research study was to determine the amino- and dipeptidase activity of selected lactic acid bacteria cultures (Lactobacillus casei, Lactobacillus acidophilus) subjected to heat- treatment at temperatures ranging between 50 and 75 °C for 1, 15, and 25 minutes. The analyzed Lactobacillus cultures synthesized peptidases that had a similar substrate specificity but different activities. They showed a higher activity of aminopeptidases compared to the activity of dipeptidases. The mean dipeptidase activity of Lb. acidophilus was 45% higher than that of Lb. casei. The Lb. casei strain was characterized by a 25% higher activity of aminopeptidases. This strain exhibited the highest specificity to the Ala-Leu, Ala-Ala, and Gly-Leu substrates, whereas the Lb. acidophilus strain – to the Ala-Ala and Ala-pNA substrates. The highest activity of the amino- and dipeptidases of Lb. casei and of the dipeptidases of Lb. acidophilus was reported after the bacteria were heat-treated for 15 min. As for the amino- and dipeptidases of Lb. casei, their activity was, respectively: 5,10, and 0.83 U/min/mg, and as for the dipeptidases of Lb. acidophilus, it was 1.66 U/min/mg. The mean aminopeptidase activity of Lb. acidophilus increased along with the increasing heat-treatment time; its highest values were reached after 25 min (3.89 U/min/mg). It was proved that the increasing of the heat-treatment time significantly impacted the growth of aminopeptidase activity of lactic acid bacteria; this fact confirms their high temperature resistance

Survival of Corynebacterium pseudotuberculosis within macrophages and induction of phagocytes death

Since C. pseudotuberculosis is a facultative intracellular pathogen the aim of this study was focused on evaluating mechanisms that allowed these bacteria to survive in macrophages and determining their influence on induction of cell death. The influence of Corynebacteria on the programmed cell death of macrophages was determined on the basis of induction the autophagy and apoptosis in the cultures of murine macrophage cell lines J774 infected with bacteria. Corynebacterium pseudotuberculosis strains could survive within macrophages more than 48 hours. During that time bacteria were released as a result of the process that lead to death of phagocytes. This property varied among studied strains. There was no increase of micro- tubule-associated protein I light chain 3 (MAP I LC3) activity in macrophages infected with examined strains comparing with uninfected cultures and cultures treated with autophagy inducer (rapamycin) that served as negative and positive controls, respectively. The study with confocal microscopy did not show the increasing of caspase-3 activity in the infected macrophages and their nucleus did not reveal the fragmentation

Nutritional habits of female students of the Medical University of Bialystok depending on the socioeconomic situation

Celem pracy była ocena wybranych zwyczajów żywieniowych studentek UMB w zależności od posiadanej sytuacji ekonomiczno-społecznej (SES). Badaniami objęto 251 studentek (18% posiadało sytuację ekonomiczno-społeczną dobrą, 56,5% przeciętną, a 25,5% złą). Niezależnie od sytuacji ekonomiczno-społecznej studentki najczęściej wybierały 4 - posiłkowy model żywienia. Do posiłków najczęściej pomijanych w jadłospisie należały drugie śniadanie i podwieczorek. Sytuacja ekonomiczno-społeczna nie była czynnikiem istotnie różnicującym częstotliwość spożycia wybranych grup produktów spożywczych. Sposób żywienia badanych studentek cechował się niską częstotliwością spożycia m. in. pieczywa razowego, kasz gruboziarnistych, mleka i jego przetworów, ryb, surowych warzyw i owoców.The aim of the study was the estimation of the selected nutritional habits of female students of the Medical University of Bialystok related to the socio-economic situation of families (SES). The investigation included 251 female students (18% of a higher SES, 56.5% students of an average SES and 25.5% students of a lower SES). The highest percentage of female students consumed four meals a day independently on the socio-economic status. Lunch and afternoon snack were the most frequently omitted meals. The socio-economic situation of families did not differ significantly the consumption frequency of selected groups of food products among the studied students. The diets analysed in this study were characterized by low intake of whole meal bread, coarse-grained barley, milk and dairy products, fish, raw vegetables and fruit