Xanthohumol induces apoptosis in cultured 40-16 human colon cancer cells by activation of the death receptor- and mitochondrial pathway

Xanthohumol (XN) is one of the major prenylflavonoids found in hop cones (Humulus lupulus L.). In this study, we investigated the cell growth inhibitory potential of XN on cultured human colon cancer cells. Cell proliferation was measured by sulforhodamine B staining. Poly(ADP-ribose)polymerase (PARP) cleavage, activation of caspases-3, -7, -8, and -9, and Bcl-2 family protein expression were detected by Western blot analyses. XN significantly reduced proliferation of the HCT 116-derived colon cancer cell line 40--16. Half-maximal inhibitory concentrations decreased from 4.1 microM after 24 h treatment to 3.6 and 2.6 microM after 48 and 72 h incubation, respectively. Treatment with 15 microM XN for 48 h and with 5 microM for 72 h led to the detection of the cleaved 89 kDa fragment of 116 kDa PARP as an indication of apoptosis induction. Concomitantly, we observed activation and cleavage of the effector caspases-3 and -7, induced by activation of the initiator caspases -8 and -9. Expression of anti-apoptotic Bcl-2 was down regulated when the cells were treated with XN for 48--72 h. We conclude that induction of apoptosis by downregulation of Bcl-2 and activation of the caspase cascade may contribute to the chemopreventive or therapeutic potential of XN.     

Biphasic modulation of cell proliferation by sulforaphane at physiologically relevant exposure times in a human colon cancer cell line

Sulforaphane (SFN), a cancer chemopreventive compound derived from broccoli, is able to induce cell cycle arrest and apoptosis in various tumor cell lines. Here we show that cell growth inhibition by SFN follows a biphasic pattern: Transient exposure of 40-16 human colon carcinoma cells for up to 6 h resulted in reversible G(2)/M cell cycle arrest and cytostatic growth inhibition even at elevated concentrations, whereas a minimum continuous exposure time of 12 h was necessary for SFN to irreversibly arrest cells in G(2)/M phase and subsequently induce apoptosis. IC(50) values after 12 h of exposure followed by drug-free recovery up to 72 h (6.4-8.1 microM) were indistinguishable from those of chronic exposure for 24 to 72 h (5.4-6.6 microM). Low concentrations of SFN caused a transient decrease in glutathione (GSH) levels followed by GSH induction, which may be related to reversible G(2)/M arrest and cytostatic effects. Depletion of GSH does not seem to play a role in SFN-mediated apoptosis induction. Our data clearly contribute to a better understanding of the kinetics of antiproliferative activity of SFN.     

Effect of Cleaning Treatment on Adhesion of Streptococcus agalactiae to Milking Machine Surfaces

Streptococcus agalactiae is a common subclinical mastitis agent in dairy cattle. In this study, the influence of hygiene procedure time on the adhesion of S. agalactiae to rubber and silicone surfaces used in milking machines was evaluated. The effect of hygiene practices on the thermodynamic and microscopic features of both surfaces was also evaluated. The hydrophobicity of milking machines and bacterial surfaces was investigated by measuring the contact angle with a goniometer 0, 30, 90, and 180 days after a full hygiene procedure simulated using rubber and silicone coupons. As the time of cleaning procedures enhanced, there was a reduction in the adhesion of S. agalactiae to both surfaces. The rubber and silicone surfaces were hydrophobic in all treatments $ \left( {\Delta G_{\text{sws}}^{\text{TOT}}\; < \;0} \right) $ , while the bacterial surface presented hydrophilic behavior $ \left( {\Delta G_{\text{sws}}^{\text{TOT}}\; > \;0} \right) $ , which makes the adhesion process difficult. Photomicrographs showed rapid wear of both surfaces, pointing to damages caused by cleaning agents. However, the silicone was more resistant to cleaning and sanitizing treatments. Thus, this work shows that changes caused by hygiene procedures in the thermodynamic and in the morphology of milking surfaces have an enormous importance in the S. agalactiae adhesion and, consequently, in mastitis transmission between cattle herd.     

Modification of Polycarbonate and Glycol Modified Poly(ethylene terephthalate) by Addition of Silica‐Nanoparticles Grafted with SAN Copolymer Using “Classical” and ARGET ATRP

This study presents a nanocomposite manufacturing route starting from non‐functionalized nanosilica and SAN surface modification via “grafting from” ATRP up to processing of transparent polycarbonate and glycol modified poly(ethylene terephthalate) nanocomposites. Synthesis limitations of low molecular weight graft SAN derived from “classical” ATRP are overcome by employment of the emerging ARGET ATRP. Mechanical investigations of polycarbonate and glycol modified poly(ethylene terephthalate) nanocomposites present up to 80% enhancement of creep performance and additionally 70% enhancement against wear abrasion for glycol modified poly(ethylene terephthalate) nanocomposites. Strength and tensile modulus are only moderately influenced by the addition of nanoparticles. Morphological studies confirm that the present modification route is able to create uniform dispersions of single particles and small particle aggregates in polymer matrices.

     

Erythropoietin Modulates Autophagy Signaling in the Developing Rat Brain in an In Vivo Model of Oxygen-Toxicity

Autophagy is a self-degradative process that involves turnover and recycling of cytoplasmic components in healthy and diseased tissue. Autophagy has been shown to be protective at the early stages of programmed cell death but it can also promote apoptosis under certain conditions. Earlier we demonstrated that oxygen contributes to the pathogenesis of neonatal brain damage, which can be ameliorated by intervention with recombinant human erythropoietin (rhEpo). Extrinsic- and intrinsic apoptotic pathways are involved in oxygen induced neurotoxicity but the role of autophagy in this model is unclear. We analyzed the expression of autophagy activity markers in the immature rodent brain after exposure to elevated oxygen concentrations. We observed a hyperoxia-exposure dependent regulation of autophagy-related gene (Atg) proteins Atg3, 5, 12, Beclin-1, microtubule-associated protein 1 light chain 3 (LC3), LC3A-II, and LC3B-II which are all key autophagy activity proteins. Interestingly, a single injection with rhEpo at the onset of hyperoxia counteracted these oxygen-mediated effects. Our results indicate that rhEpo generates its protective effect by modifying the key autophagy activity proteins.     

The influence of incompatibility and dielectric contrast on the electric field-induced orientation of lamellar block copolymers

We investigate the influence of incompatibility and dielectric contrast on the reorientation kinetics of concentrated solutions of lamellar block copolymers in the presence of an external DC electric field. We study solutions of AC diblock copolymer and ABC triblock terpolymers. The inclusion of a short, polar middle block B is used to tailor both the degree of incompatibility and the dielectric contrast between the two majority phases. In situ synchrotron radiation small-angle X-ray scattering is used to monitor the reorientation process. For the AC diblock copolymer sample only weak electric field induced reorientation could be achieved, following a very slow kinetics, whereas for the ABC triblock terpolymer, reorientation is observed above a threshold value of 0.3 kV/mm. The orientation kinetics is well described by a single exponential with characteristic time constants varying between a few seconds and several minutes depending on the polymer concentration and the electric field strength. We identify a narrow concentration window, in which the interplay between chain mobility and gain in free energy in the electric field allows the preparation of highly anisotropic bulk polymer samples by exposure to an electric field. The results are compared to free energy calculations revealing a distinct difference in the driving force for reorientation as a consequence of an increase in dielectric contrast and chain mobility upon introduction of the B middle block.     

Deformation and orientation behavior of polystyrene-b-polybutadiene-b-poly(methyl methacrylate) triblock terpolymers: Influence of polybutadiene microstructures and the molar masses

Morphological changes caused by deformation and orientation of different domains of polystyrene-b-polybutadiene-b-poly(methyl methacrylate), SBM, triblock terpolymers were investigated using in-situ small angle X-ray scattering (SAXS), tensile testing, and transmission electron microscopy (TEM). Two sets of SBM triblock terpolymers with similar weight fractions of the three blocks were studied. The two sets differ in terms of their molecular weights. Each set contained two SBM differing in their polybutadiene isomers (1,2- and 1,4-B). Results showed that for 1,2-B based SBMs the polybutadiene block forms cylindrical domains which coalesce in the glassy lamellar matrix of the two glassy outer blocks whereas the lower molar mass 1,4-B based SBM forms mixed S/M and the higher one forms well segregated long range ordered lamellae. These morphological differences indicate that the deformation and the orientation behavior of the polymers' domains differ. In 1,2-B based SBMs yielding at high stresses was followed by a stress drop after the yield point. The other polymer type showed homogeneous deformation of the lamellar domains at their yield point. 2D-SAXS during deformation of the triblock terpolymers showed an anisotropic deformation pattern in the 1,2-B SBMs, whereas four point SAXS patterns were found for the 1,4-B SBMs. Further studies showed that the fragmented lamellar planes of the lower molar mass 1,2-SBM oriented randomly whereas the orientation of the lamellar planes of the higher molar mass 1,2-B SBM was parallel to the strain direction. The alignment of the lamellar planes of 1,4-B SBMs were randomly distributed in the unstretched state.