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[en] To identify new cadmium (Cd) hyperaccumulators, the artificially high soil Cd concentration method was used to screen six common farmland weeds. Among them, only Pterocypsela laciniata (Houtt.) C. Shih showed characteristics of a Cd hyperaccumulator and was selected for further studies. In pot experiments, soil Cd concentrations of 5, 10, and 25 mg kg−1 increased the biomass and photosynthetic pigment concentrations in P. laciniata when compared with the control, whereas 75 and 100 mg kg−1 decreased them (the maxima were at 10 mg kg−1 soil Cd). The antioxidant enzyme activities and the soluble protein concentrations of P. laciniata showed similar trends as biomass. The Cd concentrations in roots and shoots of P. laciniata increased as soil Cd concentration increased. When the soil Cd concentration was 50 mg kg−1, the Cd concentration in the shoots of P. laciniata was 116 mg kg−1 (the critical value for Cd hyperaccumulators is 100 mg kg−1). Both the root and shoot bioconcentration factors of P. laciniata were larger than 1.0, and the translocation factor exceeded 1.0 in almost all treatments. The Cd extractions by the shoots and whole plants of P. laciniata reached maxima at 208 and 375 μg plant−1, respectively. The Cd extractions by P. laciniata were different between two ecotypes. Therefore, P. laciniata is a Cd hyperaccumulator that could remediate Cd-contaminated soils, but the ecotypes should be considered when using P. laciniata for phytoremediation.
[en] The initial step in atherosclerosis is the adhesion of leukocytes to activated endothelial cells mediated by intercellular adhesion molecule-1 (ICAM-1). This study aimed to investigate the association of K469E polymorphism of the ICAM-1 gene and soluble ICAM-1 (sICAM-1) serum level with coronary heart disease (CHD) in Egyptian subjects. Patients and Methods: Using a case-control design, we studied 100 patients with CHD, including 73 patients with acute myocardial infarction (MI) and 27 with unstable angina (UA). The control group consisted of 50 healthy subjects with normal left ventricular function. All participants were genotyped for the ICAM-1 polymorphism by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Serum sICAM-1 was measured by enzyme-linked immunoassay (ELISA).In CHD patients, the frequencies of K genotype (KK and EK) were significantly higher when compared to controls (P<.001) and were associated with an increased risk of disease development (OR=3.8, 95% CI: 1.7 to 8.5; P=.001). K genotype frequencies in patients with MI showed no significant difference when compared to patients with UA (P= .121). Serum sICAM-1 levels were comparable between CHD patients and controls (P= .37) and between MI and UA patients (P=.23). There were no significant differences in sICAM-1 levels among patients with different genotypes (P=.532). Men presented with higher sICAM-1 levels than women (P=.004). Conclusion: ICAM-1 gene polymorphism in codon 469 is associated with a risk for CHD development in Egyptian subjects. Serum sICAM-1 is not influenced by this polymorphism and is not necessarily elevated in CHD (Author).
[en] The common soil arthropod Folsomia candida can survive well when fed only maize pollen and thus may be exposed to insecticidal proteins by ingesting insect-resistant genetically engineered maize pollen containing Bacillus thuringiensis (Bt) proteins when being released into the soil. Laboratory experiments were conducted to assess the potential effects of Cry1Ab/Cry2Aj-producing transgenic Bt maize (Shuangkang 12–5) pollen on F. candida fitness. Survival, development, and the reproduction were not significantly reduced when F. candida fed on Bt maize pollen rather than on non-Bt maize pollen, but these parameters were significantly reduced when F. candida fed on non-Bt maize pollen containing the protease inhibitor E-64 at 75 μg/g pollen. The intrinsic rate of increase (rm) was not significantly reduced when F. candida fed on Bt maize pollen but was significantly reduced when F. candida fed on non-Bt maize pollen containing E−64. The activities of antioxidant-related enzymes in F. candida were not significantly affected when F. candida fed on Bt maize pollen but were significantly increased when F. candida fed on non-Bt pollen containing E−64. The results demonstrate that consumption of Bt maize pollen containing Cry1Ab/Cry2Aj has no lethal or sublethal effects on F. candida. - Highlights: • A validated dietary exposure assay for assessing the effect of GE plant pollen on the springtails. • Cry1Ab/2Aj-containing maize pollen does not negatively affect on the fitness of F. candida. • The antioxidant-related enzyme activities in F. candida were not significantly affected. • Planting cry1Ab/2Aj-expressing maize will pose a negligible risk to the springtails.
[en] Pollution by heavy metals is a multi-element problem in many areas. The effect of growing L. polyrrhiza with different concentrations of Cd and Pb individually and in combination ranging from 1ppm to 20 ppm on growth rate, lipid peroxidation, different anti oxidative enzymes and metal accumulation was studied. 1ppm seemed to slightly enhance the growth of the aquatic angiosperm while higher concentrations of Cd and Pb, individually or in combination, decreased the growth and multiplication of fronds. Changes in malondialdehyde content and antioxidant enzyme activity were significantly (P<0.05) higher with Cd treatments as compared to Pb. Toxicity followed the pattern Cd > Cd + Pb > Pb while bioaccumulation was influenced by the presence of other metals in binary mixture. Also accumulation of Cd and Pb was concentration and time dependant. Antagonistic or synergistic bioaccumulations of individual metals were observed in mixtures depending on the different metal concentrations. (author)
[en] Highlights: • Cambinol inhibits spread of AD synaptosome derived tau seeds, from donor to recipient cells. • Cambinol inhibits nSMase2 through targeting the DK-switch in its active site. • Cambinol inhibits EV-mediated tau propagation. • Cambinol has low brain permeability and inhibition of nSMase activity in vivo. Targeting of molecular pathways involved in the cell-to-cell propagation of pathological tau species is a novel approach for development of disease-modifying therapies that could block tau pathology and attenuate cognitive decline in patients with Alzheimer's disease and other tauopathies. We discovered cambinol through a screening effort and show that it is an inhibitor of cell-to-cell tau propagation. Our in vitro data demonstrate that cambinol inhibits neutral sphingomyelinase 2 (nSMase2) enzyme activity in dose response fashion, and suppresses extracellular vesicle (EV) production while reducing tau seed propagation. Our in vivo testing with cambinol shows that it can reduce the nSMase2 activity in the brain after oral administration. Our molecular docking and simulation analysis reveals that cambinol can target the DK-switch in the nSMase2 active site.
[en] The present study was planned to observe the activity of cefuroxime, a second generation cephalosporin after combining it with a beta-lactamase inhibitor calvulanic acid. The study was conducted to evaluate the restoration or increase in sensitivity of beta-lactamase producing isolates of Staphylococcus aureus. Staphylococcus aureus were identified by standard procedures. For beta-lactamase detection chromogenic Nitrocefin impregnated sticks were used. The sensitivity of the bacteria to the antibiotic disks was measured by disk diffusion method using standard zone diameter criteria given by National Committee of Clinical Laboratory Standards. The disks of cefuroxime with clavulanic acid had developed larger zones of inhibition. The activity of cefuroxime against Staphylococcus areus was significantly increased by clavulanic acid. Clavulanic acid if used in combination with cefuroxime, can improve the antimicrobial activity of cefuroxime against beta - lactamase producing Staphylococcus aureus. (author)
[en] The Nipah virus fusion (F) protein is proteolytically processed to F1 + F2 subunits. We demonstrate here that cathepsin L is involved in this important maturation event. Cathepsin inhibitors ablated cleavage of Nipah F. Proteolytic processing of Nipah F and fusion activity was dramatically reduced in cathepsin L shRNA-expressing Vero cells. Additionally, Nipah virus F-mediated fusion was inhibited in cathepsin L-deficient cells, but coexpression of cathepsin L restored fusion activity. Both purified cathepsin L and B could cleave immunopurified Nipah F protein, but only cathepsin L produced products of the correct size. Our results suggest that endosomal cathepsins can cleave Nipah F, but that cathepsin L specifically converts Nipah F to a mature and fusogenic form
[en] Under nitrogen starvation, Ustilago maydis forms lipid droplets (LDs). Although the dynamics of these organelles are known in the literature, the identity of the lipases implicated in their degradation is unknown. We determined lipase activity and identified the intracellular lipases expressed during growth under nitrogen starvation and YPD media by zymograms. The results showed that cytosolic extracts exhibited higher lipase activity when cells were grown in YPD. Under nitrogen starvation, lipase activity was not detected after 24 h of culture, resulting in lipid accumulation in LDs. This suggests that these lipases could be implicated in LD degradation. In the zymogram, two bands, one of 25 and the other of 37 kDa, presented lipase activity. The YPD extracts showed lipase activity in olive and almond oils, which contain triacylglycerols with mono and polyunsaturated fatty acids. This is the first report about U. maydis cytosolic lipases involved in LD degradation.
[es]En ausencia de nitrógeno, Ustilago maydis forma cuerpos lipídicos (LDs). Aunque se conoce la dinámica de estos organelos, se desconoce la identidad de las lipasas implicadas en su degradación. En este estudio se determinó la actividad de lipasa, y se identificaron las lipasas intracelulares expresadas durante el crecimiento sin nitrógeno y en YPD mediante zimogramas. Los extractos citosólicos de células en YPD exhibieron mayor actividad de lipasa. En medio sin nitrógeno no se detectó actividad de lipasa después de 24 h de cultivo, presentándose acumulación de lípidos en los LDs. Esto sugiere que estas lipasas podrían implicarse en la degradación de los LDs. En el zimograma bandas de 25 y 37 kDa, presentaron actividad de lipasa. Los extractos de células cultivadas en YPD hidrolizaron triacilgliceroles compuestos de ácidos grasos mono y poliinsaturados. Este es el primer reporte sobre lipasas citosólicas de U. maydis implicadas en la degradación de LDs.
[en] Enzyme-linked immunoassays based on an antibody-antigen interaction are widely used in biological and medical sciences. However, the conjugation of an enzyme to antibodies needs an additional chemical process, usually resulting in randomly cross-linked molecules and a loss of the binding affinity and enzyme activity. Herein, we present the development of an alkaline phosphatase-fused repebody as a new format of immuno-reagent for immunoassays. A repebody specifically binding to human TNF-α (hTNF-α) was selected through a phage display, and its binding affinity was increased up to 49 nM using a modular engineering approach. A monomeric alkaline phosphatase (mAP), which was previously isolated from a metagenome library, was genetically fused to the repebody as a signal generator, and the resulting repebody-mAP fusion protein was used for direct and sandwich immunoassays of hTNF-α. We demonstrate the utility and potential of the repebody-mAP fusion protein as an immuno-reagent by showing the sensitivity of 216 pg mL"−"1 for hTNF-α in a sandwich immunoassay. Furthermore, this repebody-mAP fusion protein enabled the detection of hTNF-α spiked in a serum-supplemented medium with high accuracy and reproducibility. It is thus expected that a mAP-fused repebody can be broadly used as an immuno-reagent in immunoassays. - Highlights: • A human TNF-α (hTNF-α)-specific repebody was selected using a phage display. • A monomeric alkaline phosphatase (mAP) was genetically fused to the repebody. • mAP-fused repebody enabled detection of hTNF-α with high sensitivity and accuracy. • mAP-fused repebody can be widely used as a new immuno-reagent in immunoassays.