Diallyl trisulfide-induced prostate cancer cell death is associated with Akt/PKB dephosphorylation mediated by P-p66shc

PURPOSE: P66Shc, an isoform of adaptor proteins, is known to mediate various signals including those leading to apoptosis or cell proliferation. Previously, we have shown that diallyl trisulfide (DATS)-induced prostate cancer cell death was mediated by increased ROS formation. In this study, we investigated the role of p66Shc protein and its serine 36 phosphorylation in DATS induced decrease in prostate cancer cell viability (PC-3). METHODS: PC-3 prostate cancer cells were used in this study. Stable cell lines expressing p66ShcS36A or an empty vector have been obtained. Cell viability, concentration of ROS, changes in P-p66Shc and P-Akt and DNA damage were determined. RESULTS: We observed that DATS treatment increased p66Shc phosphorylation at serine 36. Importantly, the phosphorylation was abolished by JNK inhibitor SP600125. Cells expressing plasmid-encoded variant of p66ShcS36A showed much higher resistance to DATS-induced cells death. In addition to that, we observed that DATS-induced ROS formation was completely abolished in cells expressing the p66ShcS36A variant. Interestingly, SP600125 proved to prevent DATS-induced Akt inactivation. In order to confirm that the observed effect is related to phosphorylation of p66Shc, we performed experiments on a stable cell line expressing p66ShcS36A. In such cells, DATS-induced Akt dephosphorylation was significantly reduced. On the other hand, hydrogen peroxide induced Akt activation in PC-3 cells, which was abrogated in cells expressing p66ShcS36A. CONCLUSIONS: Our results uncover a novel signaling pathway with p66Shc being indispensable for DATS-induced inactivation of Akt due to hypophosphorylation

NADH-generating substrates reduce peroxyl radical toxicity in RL-34 cells

There is general agreement that oxidative stress may induce apoptotic and necrotic cell death. Recently it has been shown that NADH can be considered an important antioxidant as it reacts with peroxyl and alkoxyl radicals under in vitro conditions. Therefore, in the present study we hypothesized that an increase in intracellular NADH using specific substrates will protect RL-34 cells against cytotoxicity of 2’-azobis (2-amidinopropane) dihydrochloride (AAPH), which is a peroxyl radical generating compound. Cells treated for 24 hours with 6.0 mM AAPH were severely damaged: mitochondria were vacuolated, and the level of free radicals significantly increased. Both apoptotic and necrotic cells were detected (11.1% and 11.4%, respectively) even after 5 hours of treatment. Pretreatment of the cells with substrates which increase the intracellular level of NADH, such as lactate, beta-hydroxybutyrate, and ethanol, distinctly inhibited AAPH-induced reactive oxygen species (ROS) formation and cell death. On the other hand, acetoacetate (AcA), which decrease the intracellular level of NADH, had opposite effects. Interestingly, NADH-generating substrates augment, while AcA reduced superoxide radical formation induced by AAPH. These results may suggest that although NADH generating substrates may exert some deleterious effects within a cell by inducing reductive stress, they diminish alkoxyl or peroxyl radical cytotoxicity. The protection is associated with a decrease in ROS formation measured by dichlorofluorescein, but with an increase in superoxide radical formation

PHEMTO: protein pH-dependent electric moment tools

PHEMTO (protein pH-dependent electric moment tools) is released in response to the high demand in protein science community for evaluation of electrostatic characteristics in relations to molecular recognition. PHEMTO will serve protein scientists with new advanced features for analysis of protein molecular interactions: Electric/dipole moments, their pH-dependence and in silico charge mutagenesis effects on these properties as well as alternative algorithms for electric/dipole moment computation—Singular value decomposition of electrostatic potential (EP) to account for reaction field. The implementation is based on long-term experience—PHEI mean field electrostatics and PHEPS server for evaluation of global and local pH-dependent properties. However, PHEMTO is not just an update of our PHEPS server. Besides standard electrostatics, we offer new, advanced and useful features for analysis of protein molecular interactions. In addition our algorithms are very fast. Special emphasis is given to the interface—intuitive and user-friendly. The input is comprised of the atomic coordinate file in Protein Data Bank format. The advanced user is provided with a special input section for addition of non-polypeptide charges. The output covers actually full electrostatic characteristics but special emphasis is given to electric/dipole moments and their interactive visualization. PHEMTO server can be accessed at http://phemto.orgchm.bas.bg/

Scattering of Dirac particles from non-local separable potentials: the eigenchannel approach

An application of the new formulation of the eigenchannel method [R. Szmytkowski, Ann. Phys. (N.Y.) {\bf 311}, 503 (2004)] to quantum scattering of Dirac particles from non-local separable potentials is presented. Eigenchannel vectors, related directly to eigenchannels, are defined as eigenvectors of a certain weighted eigenvalue problem. Moreover, negative cotangents of eigenphase-shifts are introduced as eigenvalues of that spectral problem. Eigenchannel spinor as well as bispinor harmonics are expressed throughout the eigenchannel vectors. Finally, the expressions for the bispinor as well as matrix scattering amplitudes and total cross section are derived in terms of eigenchannels and eigenphase-shifts. An illustrative example is also provided.Comment: Revtex, 9 pages, 4 figures, published versio

Metal accumulation in tobacco expressing Arabidopsis halleri metal hyperaccumulation gene depends on external supply

Engineering enhanced transport of zinc to the aerial parts of plants is a major goal in bio-fortification. In Arabidopsis halleri, high constitutive expression of the AhHMA4 gene encoding a metal pump of the P1B-ATPase family is necessary for both Zn hyperaccumulation and the full extent of Zn and Cd hypertolerance that are characteristic of this species. In this study, an AhHMA4 cDNA was introduced into N. tabacum var. Xanthi for expression under the control of its endogenous A. halleri promoter known to confer high and cell-type specific expression levels in both A. halleri and the non-hyperaccumulator A. thaliana. The transgene was expressed at similar levels in both roots and shoots upon long-term exposure to low Zn, control, and increased Zn concentrations. A down-regulation of AhHMA4 transcript levels was detected with 10 μM Zn resupply to tobacco plants cultivated in low Zn concentrations. In general, a transcriptional regulation of AhHMA4 in tobacco contrasted with the constitutively high expression previously observed in A. halleri. Differences in root/shoot partitioning of Zn and Cd between transgenic lines and the wild type were strongly dependent on metal concentrations in the hydroponic medium. Under low Zn conditions, an increased Zn accumulation in the upper leaves in the AhHMA4-expressing lines was detected. Moreover, transgenic plants exposed to cadmium accumulated less metal than the wild type. Both modifications of zinc and cadmium accumulation are noteworthy outcomes from the biofortification perspective and healthy food production. Expression of AhHMA4 may be useful in crops grown on soils poor in Zn

Performance of Scanning Near-Field Optical Microscope Probes with Single Groove and Various Metal Coatings

We investigate the performance of a simple corrugated aperture scanning near-field optical microscope (SNOM) probe with various cladding metals. The probes have only one corrugation, however, they offer increased transmission over both uncorrugated probes and those with many grooves. Enhancement of light throughput results from excitation of surface plasmons at the corrugation at the core–cladding interface. We show how the choice of metal influences radiation properties of grooved probes

A role for accessory genes rI.-1 and rI.1 in the regulation of lysis inhibition by bacteriophage T4

Lysis inhibition (LIN) is a known feature of the T-even family of bacteriophages. Despite its historical role in the development of modern molecular genetics, many aspects of this phenomenon remain mostly unexplained. The key element of LIN is an interaction between two phage-encoded proteins, the T holin and the RI antiholin. This interaction is stabilized by RIII. In this report, we demonstrate the results of genetic experiments which suggest a synergistic action of two accessory proteins of bacteriophage T4, RI.-1, and RI.1 with RIII in the regulation of LIN

The Isoxazole Derivative of Usnic Acid Induces an ER Stress Response in Breast Cancer Cells that Leads to Paraptosis-like Cell Death

Derivatives of usnic acid (UA), a secondary metabolite from lichens, were synthesized to improve its anticancer activity and selectivity. Recently we reported the synthesis and activity of an UA isoxazole derivative, named 2b, against cancer cells of different origins. Herein, the molecular mechanisms underlying its activity and efficacy in vivo were tested. The viability of breast cancer or normal cells has been tested using an MTT assay. Cell and organelle morphology was analyzed using light, electron and fluorescence microscopy. Gene expression was evaluated by RNAseq and protein levels were evaluated by Western blotting. In vivo anticancer activity was evaluated in a mice xenograft model. We found that 2b induced massive vacuolization which originated from the endoplasmic reticulum (ER). ER stress markers were upregulated both at the mRNA and protein levels. ER stress was caused by the release of Ca2+ ions from the ER by IP3R channels which was mediated, at least partly, by phospholipase C (PLC)-synthetized 1,4,5-inositol triphosphate (IP3). ER stress led to cell death with features of apoptosis and paraptosis. When applied to nude mice with xenografted breast cancer cells, 2b stopped tumour growth. In mice treated with 2b, vacuoli-zation was observed in tumour cells, but not in other organs. This study shows that the antiprolif-erative activity of 2b relates to the induction of ER stress in cancer, not in healthy, cells and it leads to breast cancer cell death in vitro and in vivo

Erratum: Synthesis of Usnic Acid Derivatives and Evaluation of Their Antiproliferative Activity against Cancer Cells (Journal of Natural Products (2019) 82:7 (1768-1778) DOI: 10.1021/acs.jnatprod.8b00980)

The (+)-enantiomer of compound 1 was incorrectly assigned as the S-configuration where it is in fact the Rconfiguration. As a consequence, compounds 2b, 3b, 3d, 3f, 3h, 3j, 3l, and 3n are also the R-configuration. Page 1769: The table within Scheme 2 should be substituted with the following:. The compound names within the Supporting Information (pp 2-6) need to be altered as follows: • (R)-8-Acetyl-5,7-dihydroxy-3,4a,6-trimethyl-1-phenyl- 1,4a-dihydro-4Hbenzofuro[3,2-f ]indazol-4-one (3d) • (R)-8-Acetyl-5,7-dihydroxy-1-(4-methoxyphenyl)- 3,4a,6-trimethyl-1,4a-dihydro4H-benzofuro[3,2-f ]- indazol-4-one (3f) • (R)-8-Acetyl-1-(4-fluorophenyl)-5,7-dihydroxy-3,4a,6- trimethyl-1,4a-dihydro-4Hbenzofuro[3,2-f ]indazol-4- one (3h). • (R)-8-Acetyl-1-(3,4-dichlorophenyl)-5,7-dihydroxy- 3,4a,6-trimethyl-1,4a-dihydro4H-benzofuro[3,2-f ]- indazol-4-one (3j) (Table Presented). • (R)-8-Acetyl-1-(4-chlorophenyl)-5,7-dihydroxy-3,4a,6- trimethyl-1,4a-dihydro4H-benzofuro[3,2-f ]indazol-4- one (3n) Accordingly, the structures of compounds 2b, 3b, 3d, 3f, 3h, 3j, 3l, and 3n in the Supporting Information have been revised. Additionally, the Abstract is revised as follows: Moreover, they induced massive cytoplasmic vacuolization, which was associated with elevated dynamin-dependent endocytosis, a process that has not been reported for usnic acid and indicates a novel mechanism of action of its synthetic derivatives. The authors sincerely apologize for any inconvenience caused by these errors

A renormalization group study of a class of reaction-diffusion model, with particles input

We study a class of reaction-diffusion model extrapolating continuously between the pure coagulation-diffusion case ($A+A\to A$) and the pure annihilation-diffusion one ($A+A\to\emptyset$) with particles input ($\emptyset\to A$) at a rate $J$. For dimension $d\leq 2$, the dynamics strongly depends on the fluctuations while, for $d >2$, the behaviour is mean-field like. The models are mapped onto a field theory which properties are studied in a renormalization group approach. Simple relations are found between the time-dependent correlation functions of the different models of the class. For the pure coagulation-diffusion model the time-dependent density is found to be of the form $c(t,J,D) = (J/D)^{1/\delta}{\cal F}[(J/D)^{\Delta} Dt]$, where $D$ is the diffusion constant. The critical exponent $\delta$ and $\Delta$ are computed to all orders in $\epsilon=2-d$, where $d$ is the dimension of the system, while the scaling function $\cal F$ is computed to second order in $\epsilon$. For the one-dimensional case an exact analytical solution is provided which predictions are compared with the results of the renormalization group approach for $\epsilon=1$.Comment: Ten pages, using Latex and IOP macro. Two latex figures. Submitted to Journal of Physics A. Also available at http://mykonos.unige.ch/~rey/publi.htm