Notch activation induces endothelial cell cycle arrest and participates in contact inhibition: role of p21Cip1 repression

Although previous studies demonstrate that appropriate Notch signaling is required during angiogenesis and in vascular homeostasis, the mechanisms by which Notch regulates vascular function remain to be elucidated. Here, we show that activation of the Notch pathway by the ligand Jagged1 reduces the proliferation of endothelial cells. Notch activation inhibits proliferation of endothelial cells in a cell-autonomous manner by inhibiting phosphorylation of the retinoblastoma protein (Rb). During cell cycle entry, p21Cip1 is upregulated in endothelial cells. Activated Notch inhibits mitogen-induced upregulation of p21Cip1 and delays cyclin D-cdk4-mediated Rb phosphorylation. Notch-dependent repression of p21Cip1 prevents nuclear localization of cyclin D and cdk4. The necessity of p21Cip1 for nuclear translocation of cyclin D-cdk4 and S-phase entry in endothelial cells was demonstrated by targeted downregulation of p21Cip1 by using RNA interference. We further demonstrate that when endothelial cells reach confluence, Notch is activated and p21Cip1 is downregulated. Inhibition of the Notch pathway at confluence prevents p21Cip1 downregulation and induces Rb phosphorylation. We suggest that Notch activation contributes to contact inhibition of endothelial cells, in part through repression of p21Cip1 expression.     

The limits of log-ratios

Background  

DNA microarray assays typically compare two biological samples and present the results of those comparisons gene-by-gene as the logarithm base two of the ratio of the measured expression levels for the two samples.     

Genetic regulation of endotoxin-induced airway disease

To identify novel genes regulating the biologic response to lipopolysaccharide (LPS), we used a combination of quantitative trait locus (QTL) analysis and microarray-based gene expression studies of C57BL/6J x DBA/2J(BXD) F2 and recombinant inbred (RI) mice. A QTL affecting pulmonary TNF-alpha production was identified on chromosome 2, and a region affecting both polymorphonuclear leukocyte recruitment and TNF-alpha levels was identified on chromosome 11. Microarray analyses of unchallenged and LPS-challenged BXD RI strains identified approximately 500 genes whose expression was significantly changed by inhalation of LPS. Of these genes, 28 reside within the chromosomal regions identified by the QTL analyses, implicating these genes as high priority candidates for functional studies. Additional high priority candidate genes were identified based on their differential expression in mice having high and low responses to LPS. Functional studies of these genes are expected to reveal important molecular mechanisms regulating the magnitude of biologic responses to LPS.     

Influence of plant extracts and microbioagents on physiological traits of faba bean infected with<Emphasis Type="Italic">Botrytis fabae</Emphasis>

Laboratory and greenhouse experiments were conducted to assess the efficacy ofEucalyptus citriodora, Ipomoea carnea, Cuminum cyminum, Allium sativum and Hyoscyamus muticus leaf extracts, and Streptomyces exfloliatus (S) andTrichoderma harzianum (T) for controllingBotrytis fabae causing chocolate spot disease of faba bean. Laboratory study supported the use ofE. citriodora (Ex 1) andI. carnea (Ex 2) extracts than the others for controlling the growth ofB. fabae. S+T was the best for inhibiting spore germination followed by Ex 1 +Ex 2 after 8 h of testing, whereas Ex 1 +Ex 2 produced the lowest percent of germination after 16 h. After 4 days, the inhibiting order of the growth ofB. fabae was S+T > Ex 1 +Ex 2 > T > Ex 2 > Ex 1 =S. Greenhouse experiments showed the highest activities of peroxidase, catalase and pectinase in the infected plants. These activities were markedly reduced in healthy plants and widely changed by the biocontrol treatments. Applying biocontrol agents to the infected plants increased minerals (N, P, K and Mg), and both Chl biosynthesis and the photosynthetic activity, which in turn led to accumulation of metabolites. This served the plant to resist the detrimental effects ofB. fabae on the plant growth and yield. In this concern, the efficiency of test biocontrol agents seemed to be in the order:T+S > Ex 1 +Ex 2 >T >S > Ex 2 > Ex.     

Fasciola gigantica fatty acid binding protein (FABP) as a prophylactic agent against Schistosoma mansoni infection in CD1 mice

Although schistosomicidal drugs and other control measures exist, the advent of an efficacious vaccine remains the most potentially powerful means for controlling this disease. In this study, native fatty acid binding protein (FABP) from Fasciola gigantica was purified from the adult worm's crude extract by saturation with ammonium sulphate followed by separation on DEAE-Sephadex A-50 anion exchange chromatography and gel filtration using Sephacryl HR-100, respectively. CD1 mice were immunized with the purified, native F. gigantica FABP in Freund's adjuvant and challenged subcutaneously with 120 Schistosoma mansoni cercariae. Immunization of CD1 mice with F. gigantica FABP has induced heterologous protection against S. mansoni, evidenced by the significant reduction in mean worm burden (72.3%), liver and intestinal egg counts (81.3% and 80.8%, respectively), and hepatic granuloma counts (42%). Also, it elicited mixed IgG(1)/IgG(2b) immune responses with predominant IgG1 isotype, suggesting that native F. gigantica FABP is mediated by a mixed Th1/Th2 response. However, it failed to induce any significant differences in the oogram pattern or in the mean granuloma diameter. This indicated that native F. gigantica FABP could be a promising vaccine candidate against S. mansoni infection.     

Antibacterial cotton fabrics treated with core-shell nanoparticles

Multifinishing treatment of cotton fabrics was carried out using core-shell nanoparticles that consists of silver nanoparticles (Ag(0)) as core and chitosan-O-methoxy polyethylene glycol (CTS-O-MPEG) as shell. The synthesized (Ag(0)-CTS-O-MPEG) core-shell nanoparticle was applied to cotton fabrics using the conventional pad-dry-cure method. The finished fabrics were examined for their morphological features and surface characteristics by making use of scanning electron microscope (SEM-EDX), which reveals the well dispersion of (Ag(0)-CTS-O-MPEG) core-shell nanoparticles on cotton fabrics. Factors affecting the treatment such as core shell nanoparticles, citric acid (CA) concentration as well as curing temperature were studied. The treated fabrics, at optimum condition of 1% core shell nanoparticles, 5% citric acid, drying at 80°C, curing at 160°C for 2 min, showed excellent antibacterial activity against Gram-negative Escherichia coli (E. coli) and Gram-positive bacteria Staphylococcus aureus (S. aureus), even after 20 washing cycles in addition to an enhancement in crease recovery angles (CRA) along with a slight improvement in tensile strength (TS).     

Isolation and characterization of heavy-metal resistant microbes from roadside soil and phylloplane

Contamination by heavy metals is one of the major environmental problems in many countries and these contaminants reach from various sources such as traffic cars and other activities. Soil and phylloplane samples were collected from eight traffic and two non-traffic sites in Sohag city, Egypt. Heavy metal contents of Cd2?, Zn2? and Pb2? of soil and phylloplane samples were determined and revealed high levels of Zn2? and Pb2? in traffic samples. A total of 112 bacterial and 62 fungal isolates were obtained from soil and phylloplane. Bacterial isolates were characterized on the basis of morphological, physicochemical and biochemical characteristics; and 16S rRNA gene sequences. Fungal isolates were identified according to morphological characterization. Minimal inhibitory concentrations (MICs) of Cd2?, Zn2? and Pb2? for each isolate were detected. All bacterial and fungal isolates demonstrated resistance to lead with MICs >0.528 mM and >0.211, respectively. Moreover, the maximum MICs of cadmium and zinc for bacteria were 0.821 mM and 1.471 mM, respectively, where as, MICs for fungi were 0.328 mM and 0.588 mM, respectively. The most resistant bacterial and fungal isolates were Pseudomonas aeruginosa RA65 and Penicillium corylophyllum, respectively. Therefore, P. aeruginosa RA65 was selected for further investigations. Growth curve study showed that 0.264 mM lead had no efficiently effect on the growth of P. aeruginosa RA65. Plasmid isolation evidenced by transformation studies indicated that P. aeruginosa RA65 harbored a single plasmid (~9.5 kb) which mediated heavy meal resistance. Consequently, these microbial isolates could be potentially used in bioremediation of heavy metal-contaminated environment.     

Assessment of titanium dioxide nanoparticles toxicity via oral exposure in mice: effect of dose and particle size

Objective: The aim of the present work is to evaluate the toxicity of titanium dioxide nanoparticles (TiO₂NPs) according to their doses and particle sizes.

Materials and methods: The effect of five days oral administration of TiO₂NPs (21 and 80?nm) with different doses (50, 250 and 500?mg/kg body weight) was assessed in mice via measurement of oxidative stress markers; glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA) and nitric oxide (NO), liver function indices; aspartate and alanine aminotransferases (AST and ALT), chromosomal aberrations and liver histopathological pattern.

Results: The results revealed drastic alterations in all the measured parameters and showed positive correlation with the gradual dose increment. In addition, the smaller particle size of TiO₂NPS (21?nm) had more adverse effect in all the selected biochemical parameters, genetic aberrations and histological investigations.

Conclusions: Toxicity of TiO₂NPs increases in a dose-dependent manner and vice versa with particles size. The evaluated biomarkers are good indicators for TiO₂NPs toxicity. More detailed studies are required before the recommendation of TiO₂NP_S as food additives.     

Blimp1 Prevents Methylation of Foxp3 and Loss of Regulatory T Cell Identity at Sites of Inflammation

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