Ectoderm and endoderm tissues had been the right way patterned in many embryos treated with ClO starting hpf; only the expression of nodal was seriously disturbed. This suggests the presumptive stomodeum is specified but doesn’t differentiate thoroughly in embryos treated with ClO starting at hpf. In lots of of those embryos, the archenteron extended across the blastocoel and bent towards the cuboidal ectoderm from the presumptive oral discipline, indicating that the tip from the archenteron recognized this region of ectoderm as oral. Having said that, the archenteron tip failed to fuse together with the overlying oral ectoderm where presumptive stomodeal cells expressed bra. Thus the mouth formation defect doesn’t seem for being as a result of a failure of oral tissue specification. Furthermore, however OA polarity was restored, mouth formation was usually not rescued by addition of SO to ClO handled embryos .
Thus, ClO therapy prevents stomodeal invagination as well as oral tissue fusion occasion but undersulfation may not be direct the reason behind the observed mouth formation defect Developmental roles of sulfated GAGs Taken collectively, our experiments deliver proof that sulfation is important for your proper perform of GAGs and proteoglycans in establishment mTOR inhibitors selleckchem and or perception of the TGF beta signals launched by ectodermal cells resulting in usual OA patterning within the producing urchin embryo. Our final results display that proper nodal expression and Nodal signaling is dependent on continuing sulfation, quite possibly via an effect of sulfated GAGs over the diffusion of this TGF beta ligand. We propose that interaction of Nodal with sulfated GAGs is required to preserve an organizing center of Nodal signaling during the oral discipline at a enough community concentration to positively autoregulate its own expression and market the specification and differentiation of oral tissue and right patterning of aboral ectoderm. In addition, sulfation may well play necessary roles in convergent extension, tissue fusion and or cell adhesion occasions involved in gastrulation Experimental procedures Animal assortment, culture and solutions Grownup S.
purpuratus sea urchins from Vancouver Island, British Columbia, were induced to shed gametes by electrostimulation. Embryos were cultured in SW at C. Building embryos were handled with and mM NaClO at unique occasions submit fertilization. Embryos were also treated with mM Na Search engine marketing , mM NaSO , mM nitrophenyl beta D xylopyranoside , or . and . lM SB . Artificial Quizartinib selleckchem SW was prepared in accordance to Kester et al Low sulfate SW, containing only mM sulfate , was ready by replacing a part of the NaSO with NaCl Synthesis of riboprobes DNA fragments of a minimum of bp of coding sequence had been PCR amplified and cloned from the pBluescript II KS plasmid. RNA probes have been synthesized with T or T RNA polymerase , lg linear template DNA plus the DIG RNA labeling mix according to manufacturer instructions.

These misalignments were independent of mitotic checkpoint inactivity, as cells depleted of Mad had no trouble aligning all chromosomes . Analysis of chromosome movements in genuine time further exposed that of Mps depleted cells versus of manage cells showed misaligned chromosomes min immediately after entry into mitosis in the presence of MG . After hr, of Mps depleted cells nonetheless contained a single or far more chromosomes that had not reached the metaphase plate in contrast to of mock shRNA cells. Replacement of endogenous Mps by using a kinase dead mutant showed that chromosome alignment required Mps kinase action . In agreement with this, simultaneous treatment of prophase cells with MG and SP, a smaller molecule that inhibits Mps in mitotic human cells , brought about significant misalignments that persisted until eventually elimination with the inhibitor min soon after addition . Collectively, these data present that Mps action contributes to alignment of chromosomes over the metaphase plate in mitosis. Interactions concerning Kinetochores and Spindle Microtubules Are Secure in Cells Lacking Mps We following examined what procedure essential for chromosome alignment was defective in Mps depleted cells.
The following observations suggested that misalignments weren’t induced by general defects in spindle assembly or secure microtubule capture through the kinetochore. First, interkinetochore distances of aligned chromosomes in Mps depleted cells were just like people of handle cells , displaying that sufficiently solid attachments had been produced that can impose standard stress amongst sister centromeres. 2nd, no obvious distinctions in spindle morphology or density of cold secure Wortmannin datasheet kinetochore microtubules have been detected between mock and Mps depleted cells . As a handle, Nuf depleted cells showed a number of misaligned chromosomes that lacked apparent interactions with spindle microtubules . Third, alignment was maintained when SP was added following chromosomes had reached complete alignment . These 3 measurements excluded fundamental defects in spindle assembly and steady microtubule capture by kinetochores in cells lacking Mps.
Nevertheless, as they have been principally focused within the aligned chromosomes, we couldn’t exclude the chance Abiraterone that the misaligned chromosomes had seasoned troubles in microtubule capture. To examine this, kinetochores were analyzed for the presence of CLIP . This microtubule binding protein localizes specifically to unattached kinetochores in a mitotic checkpoint independent manner and leaves the kinetochore on microtubule capture . Kinetochores of the two aligned and misaligned chromosomes in MG taken care of cells lacking Mps had no detectable CLIP , whereas the occasional misaligned chromosome in MG handled, mock shRNA transfected cells had recruited substantial amounts of CLIP on at least one of its kinetochores .

In the selection results, an apparent preference for proline containing peptides was noticed, often with a tryptophan residue adjacent to it in a PW motif. Interestingly, the choice final results didn’t correlate using the original Aurora A selection, as neither G or G appeared under the newer stringent wash ailments. Two in the most prevalent peptides, G and G, had been synthesized, purified, and assayed towards Aurora A. The two peptides had been identified for being just about the most potent peptide inhibitors of Aurora A evaluated therefore far with our variety technique, having IC values of and lM, respectively . Due to the fact the picked peptides themselves are very low micromolar inhibitors of equivalent potency despite the substantially various sequences, we subsequent sought to probe the mode of inhibition and selectivity. The picked peptide, G, was tested at 3 diverse concentrations of Kemptide and showed no appreciable adjust from the IC , which potentially recommended a noncompetitive mode of inhibition as we have previously observed when focusing on PKA.
Even further evaluations of the mode of inhibition in the chosen peptide by kinetic analysis, G also suggests noncompetitive inhibition with respect to peptide substrate as proven in Figure . Together with the peptides in hand and Tofacitinib the observation that the peptides are considerably hydrophobic which has a single Arg in one particular situation in addition to a Thr and Arg is the other case, we asked whether the observed inhibition was selective for Aurora A or irrespective of whether these peptides have been probably non exact protein kinase binders. For you to test this, each peptides had been assayed against the AGC kinase loved ones , the CMGC loved ones plus the target at lM. Gratifyingly, as Figure demonstrates, the Aurora A picked peptides inhibit their meant kinase at lM peptide concentration with negligible inhibition of either PKA or CLK. Hence these studies demonstrate that phage show solutions can be utilized to find out cyclic peptide inhibitors of therapeutically relevant kinases.
In summary, the application of our bivalent assortment approach to other kinases past PKA such as Aurora A has necessitated considerable adaptation and optimization to isolate kinase selective peptides despite the fact that avoiding background peptides. The improved protocol eliminates HPQ containing streptavidin binding compound libraries for drug discovery kinase inhibitor sequences, which have been totally characterized, whereas yielding a lot more potent peptide inhibitors of Aurora A. The peptides recognized in the program of this review are extra potent than that discovered by a latest helix loop helix displayed phage display technique against Aurora A, where the best peptide appended towards the helices, GRRVVVSFAWD, showed inhibition at a concentration of lM.

Cellular modifications for the duration of processes just like differentiation, ER stress responses or infections consequence within a profound remodeling from the ER with concomitant adjustments in Ca signaling. A equivalent remodeling also occurs for other organelles this kind of as the mitochondria plus the resulting ER mitochondria interactions. Phenotype switch in smooth muscle cells Vascular smooth muscle cells can undergo a phenotype switch from a contractile quiescent to a proliferative or synthetic phenotype. This plasticity is very well recognized as an very important mechanism for vascular repair while in injury or adaptation and it really is reversible beneath shear stress problems . This phenotype switch includes a profound rearrangement in the cellular Ca dealing with . With respect to intracellular Ca signaling there exists a loss of RyR Ca release channels plus a subsequent loss with the CICR mechanism . The switch to a proliferative cell type is characterized by a rise in expression on the IPR , which can be an important determinant of vascular smooth muscle proliferation .
In serum stimulated vascular smooth muscle cells, proliferation is connected by using a 6 fold raise in IPR expression levels in the G S transition in the course of the ROCK inhibitors selleckchem cell cycle . Proliferating cultured myocytes from rat mesentery artery showed elevated resting cyt and an elevated IP sensitive retail outlet content .Moreover, SOCE and receptor operated Ca entry have been augmented and related to up regulated expression of TRPC and TRPC . On top of that, SERCAb, STIM and ORAI proteins were up regulated, displaying profoundly altered gene expression underlying the altered Ca handling all through vascular growth and remodeling . A short while ago, it had been demonstrated by RNAi targeting that STIM is actually a important regulator of in vitro and in vivo vascular smooth muscle cell proliferation Differentiation from the secretory apparatus It has due to the fact extended been recognized that the induction of substantial fee protein secretion for the duration of the differentiation plan of secretory cell styles, entails improved biogenesis of secretory apparatus organelles .
The adjust in biosynthetic capacity will involve induction of a broad spectrum of secretory pathway genes and an expansion of volume and location in the ER . In plasma cell differentiation, the GW-572016 transcription issue X box binding protein was identified to coordinate the alterations in cellular construction and perform . Also for that biogenesis with the secretory machinery of exocrine glands this kind of as exocrine pancreas and salivary gland, XBP is required, and its deletion severely impaired growth with the ER plus the expression of sure ER chaperones . XBP is now considered as the central player of an integration mechanism in between the necessities for ER membrane capability as well as the extent of protein processing as signaled from the UPR .

On this review, we summarize the roles that each with the Aurora kinases has in mitosis and cancer biology, and discuss the binding modes of Aurora A kinase inhibitors, the sizzling spot residues during the binding internet sites as well as the privileged inhibitor structures. Our examine within the reported chemical scaffolds of Aurora A kinase inhibitors and their binding modes could present a practical framework from which new design methods for inhibitors might be assessed or designed. Also, we analysis the principle preclinical and clinical scientific studies concerning Aurora kinase inhibitors at present beneath investigation. The in depth preclinical data supports the improvement of targeted Aurora kinase inhibitors in unique tumor forms. Aurora kinases The Aurora kinases certainly are a household of three hugely homologous Ser Thr protein kinases, which includes Aurora A, B and C. Since their discovery in along with the to start with observation of their expression in human cancer tissue in , these kinases have been the topic of extreme research in the two the academic and industrial oncology communities .
The 3 homologous Aurora kinases possess a conserved catalytic domain and an N terminal domain that varies in sequence and length, but are distinct in function . Nonetheless, all 3 are overexpressed in many kinds of cancer, through which polyploid cells containing numerous centrosomes are observed . Aurora A Aurora A localizes to your centrosomes, exactly where it can be required for his or her maturation and separation, therefore marketing mitotic Proteasome Inhibitor kinase inhibitor entry and spindle assembly. In mitosis, Aurora A associates with the spindle poles and it is involved in the two centrosomal assembly and acentrosomal spindle assembly . The gene encoding Aurora A lies within a region of chromosome q, which is amplified in many epithelial malignant tumors, which include breast, gastric, colon, ovarian and pancreatic cancers. On top of that, overexpression of an active mutant of Aurora A in rat cells induced neoplastic transformation, indicating that Aurora A is an oncogene . Overexpression of Aurora A contributes to genetic instability and tumorigenesis by disrupting the correct assembly of your mitotic checkpoint complicated.
Ectopic overexpression of Aurora A can transform rodent cells and also the resulting cells can induce tumor formation in nude mice. Additional help for its oncogenic role was supplied when Xenopus Sorafenib VEGFR inhibitor kinase inhibitor Aurora A transformed NIHT fibroblasts led for the improvement of tumors in mice . On top of that, this kinase is known as a major regulatory part within the p pathway and its overexpression prospects to a rise in p degradation, which yet again facilitates oncogenic transformation . Within the absence of Aurora A, centrosomes fail to assemble bipolar spindles, which might end result in mitotic arrest.

Taken together, these findings demonstrate that IP serves a pivotal function and is involved inside the reparative effect of iPSC CM on airway structural damage and oxygenation ability in VILI VILI is characterized by inflammation, enhanced alveolarcapillary membrane permeability, accumulation of protein wealthy pulmonary edema, eventually major to impaired gas exchange. Preceding research on an isolated, non perfused ALI model in mice have demonstrated that the silencing of PIK attenuates the functional and morphological disruption of VILI by means of the inhibition of its downstream Akt signaling . Uhlig and colleagues demonstrated that the PIK inhibitor, LY, prevents the expression of mechanical ventilation induced inflammatory mediators in alveolar macrophages and epithelial cells . We previously observed that iPSC or iPSC CM is helpful towards the recovery from the effects of endotoxin induced ALI . Even so, the mechanisms and mediators of iPSC dependent therapy are nevertheless unclear and must be evaluated in preclinical research.
Inside the high ventilation induced mouse lung injury model, we discovered that iPSCs or iPSC CM suppressed high tidal volume induced VILI, Ruxolitinib as observed by decreased lung edema, microvascular permeability, neutrophil infiltration, and elevated PaO FiO ratio in bronchial epithelium in response to these therapies. iPSCs iPSC CM also inhibited PIK Akt signaling, suppressed production of MIP , nitrate nitrite, MDA, improved GSH content material and potentially restored the bronchial microstructure. This iPSC CM efficacy, similar to that of iPSCs, may be mimicked by PIK inhibitor LY or Akt heterozygous knockout, and either remedy didn’t in addition enhanced VILI in iPSC CM recipients. We also located that iPSC CM consists of high levels of chemokine IP that partially mediated the suppression of neutrophil infiltration and restoration of lung function in VILI. This report highlighted the therapeutic prospective of iPSC CM in VILI and also the predominant mechanism was via inhibition of PIK Akt signaling.
HMGB serves as a regulator of transcription and an extracellular inflammatory cytokine . HMGB can contribute for the release of cytokines; conversely, cytokines, for example PAI , can handle the additional release Resveratrol HMGB in to the extracellular space . PAI has been implicated in the fibrinolytic defect connected with diverse kinds of lung injury . An increase of HMGB and PAI is regularly observed in high stretch mechanical ventilation . Antibodies against HMGB or an anticoagulant that blocks PAI have already been shown to improve microvascular permeability, decrease neutrophil influx into the alveolar lumen, and inhibit the proinflammatory cytokines . Recently, an in vitro study in human vascular smooth muscle cells showed that PIK Akt is involved within the inflammation connected production of PAI .

The outcomes for Aquaporin and BCL had been discordant. The results of those experiments are summarized in SELLECKCHEM IngenuityTM pathways evaluation IngenuityTM pathway examination demonstrated many biological signaling pathways that have been deregulated in our ALCL samples. Genes which had been deregulated in both the NPM ALK positive and TPM ALK beneficial samples highlighted pathways concerned in cell cycle regulation, interleukin and IL signaling, antigen presentation, NFBB and PPAR signal transduction, and integrin signaling. The integrated biological relationships recognized from the genes that had been shared involving the NPM ALK positive and TPM ALK beneficial ALCsL applying IngenuityTM pathway evaluation is represented in SELLECKCHEM . The genes which had been deregulated only during the NPM ALK positive ALCL demonstrated disruption in the ERK MAPK pathway, B cell and T cell receptor signaling, p MAPK signaling, as well as IGF signaling pathways, although people inside the TPM ALK beneficial ALCL showed deregulation within the PIK AKT, Wnt Bcatenin, and estrogen receptor signaling pathways Discussion The huge vast majority of ALK constructive ALCLs carry the t , a translocation which has been very well characterized as a result of its utility being a disease marker for ALCL .
The molecular events related to the variant t translocation resulting in the TPM ALK fusion protein yet, are sizeable unknown. Within the present research, we implemented cDNA Temsirolimus microarray evaluation to characterize and compare the gene expression profiles of a normal t positive ALCL with one more containing the variant t translocation. A subset with the genes was chosen for validation by quantitative RT PCR with overall concordance fee of somewhere around . We recognized genes that were in popular to each forms of ALCL also as those who had been distinct. The biggest practical groups of genes recognized to get comparable in the two varieties of ALCLs were those concerned within the regulation of cell proliferation and programmed cell death. Multiple genes concerned in the G S cell cycle checkpoint had been identified in our experiments. The frequent deregulation of genes concerned in cell cycle manage suggests that cellular growth pathways really are a popular downstream target for ALK.
Genes concerned in lymphocyte activation, differentiation at the same time as cytoskeletal organization, adhesion and migration have been also popular to the two kinds of lymphomas. To date, the only normal signaling pathway which has been reported in between the two sorts Sorafenib selleckchem of fusion proteins is the PI K AKT pathway . A number of members with the NFkB pathway had been recognized in our IngenuityTM information analysis , suggesting its value while in the signaling cascades that arise being a consequence of ALK gene deregulation from the improvement of ALCL. Heme oxygenase , SA, interleukin receptorB, hematopoietic cell kinase, lymphotoxin beta receptor, TNF ligand superfamily member , CCAAT enhancer binding protein , IL receptor, and BCL have been overexpressed in both varieties of ALCL relative on the reactive lymph node.

Fraction affected , the concentration on the drug that made development inhibition along with the dose effect relationship in the level of IC were analyzed by CalcuSyn Software DNA material and apoptosis evaluation by movement cytometry Cell lines had been cultured in very well tissue plates under the conditions described over. Right after h of preincubation, cells had been exposed to expanding concentrations of PHA for h, washed with PBS and fixed in cold ethanol overnight at? ?C. Shortly prior to flow cytometry examination, cells have been rinsed with PBS, resuspended in PBS containing RNAse A and propidium iodide , and incubated for min on ice. Ten thousand cells had been analyzed in every sample Assessment of phosphorylation standing by intracellular flow cytometry After incubation with both M PHA or M IM for h or h, cells were collected, fixed in formaldehyde for min at ?C, chilled on ice for min and permeabilized with ice cold methanol for min on ice. cells per sample had been washed with ml incubation buffer . bovine serum albumin and centrifuged at rpm for min. Afterwards, cells were resuspended in l of incubation buffer with . l of either Phospho CrkL, Phospho Stat, Phospho c Abl or Phospho Histone H exact antibody and incubated at RT for min.
The washing stage was repeated twice and subsequently cells had been resuspended in l incubation buffer using the secondary antibody and incubated at RT for min from the dark followed by twowashing ways. Samples stained with Phospho Histone H particular antibody had been additionally stained with propidium iodide as described over. Movement cytometry acquisition was performed on FACS Calibur employing CellQuest for examination. The amount of phosphorylated proteins was established by calculating approved drug library differences while in the geometric imply fluorescence intensity as well as changes with the phosphorylation status were expressed as being a percentage of the untreated handle Success PHA preferentially inhibits BCR ABL favourable leukemic cell lines independent of their mutational standing To investigate the probable effects of PHA remedy on cellular proliferation, we carried out MTT assays having a panel of human and murine leukemic and manage cell lines.
PHA efficiently inhibited the proliferation of all examined cell lines with IC values ranging from .Mto .Min BCR ABL favourable and from . M to M in BCR ABL detrimental CCI-779 cell lines . This big difference points to a predominant impact in the compound on BCR ABL positive leukemic cells. Having said that, whereas expectedly considerable differences have been detected in IC values for IM involving BaF cells harbouring wild type rather than mutant BCR ABL, no this kind of distinctions had been observed for PHA . Taken with each other, these findings argue for exercise of the compound against Bcr Abl and that is unimpaired by mutations confering resistance to IM PHA induces anti proliferative effects in BCR ABL favourable leukemic cell lines which includes IM resistant BaF cells expressing MT, EK, and TI mutants So as to further characterize the effect within the BCRABL mutational standing on the anti proliferative results of PHA , we performed trypan blue exclusion assays with murine BaF and BaF p cells, as well as their IMresistant mutants MT, EK, and TI.

These neurosecretory granules have been evident inside the tumors we recognized in the zebrafish , strengthening their association with childhood neuroblastoma. The histopathological, immunohistochemical and ultrastructural features of neuroblastoma are proven in Figure E, to illustrate their similarities with individuals of neuroblastomas induced by MYCN overexpression in zebrafish . These findings support our utilization of this model to investigate activated ALK like a contributor to MYCN driven tumorigenesis. ALK Accelerates MYCN Induced Neuroblastoma We and many others have implicated activating mutations of ALK from the pathogenesis of neuroblastoma, which includes circumstances that also display MYCN amplification . To deal with no matter if ALK and MYCN genetically interact while in neuroblastoma induction, we generated a second secure transgenic zebrafish line that expresses the human ALK gene harboring the FL mutation, one of the most prevalent somatic activating mutations identified in neuroblastoma individuals and human cell lines .
The dbh:EGFP and dbh:ALKFL constructs had been coinjected into zebrafish embryos in the onecell stage to make pan Syk inhibitor a transgenic line expressing both the EGFP and activated ALK transgenes, Tg , designated ALK in this post. EGFP was especially expressed by sympathoadrenal cells inside the interrenal gland in the ALK transgenic fish at weeks postfertilization , and ALK was coexpressed with EGFP through the exact same cells . This transgenic line was bred for the MYCN heterozygous transgenic line, plus the offspring had been monitored for evidence of tumors. Each of the expected genotypes were represented while in the offspring of this cross: MYCN; ALK; MYCN;ALK; and wild form AB fish lacking both transgene. A tumor view was carried out on a complete of , sorted offspring. The fish were isolated in personal tanks as soon as tumors appeared; and had been sacrificed for molecular and pathologic analyses when there was evidence of tumor progression. The primary tumors arose among weeks of age, and all had the compound transgenic genotype, MYCN;ALK .
The expression of MYCN and ALK proteins and ALK RNA was confirmed during the tumors of those compound transgenic supplier MLN9708 selleck chemicals fish by immunohistochemical and RT PCR analyses, respectively . Tumors continued to come up just after weeks of age in the two the MYCN only and the MYCN;ALK compound transgenic lines, but their price of induction was a great deal increased from the latter group . Tumor penetrance during the MYCN;ALK compound transgenic fish was also considerably greater versus . for that MYCN transgenic fish . Whilst germline mutations of ALK lead to hereditary neuroblastoma , tumors didn’t develop in fish expressing this transgene alone over the month monitoring period . Tumors while in the compound transgenic fish arose while in the interrenal gland, as did individuals while in the MYCN fish, and these tumors had been comparable histologically, immunohistochemically, and ultrastructurally to human neuroblastoma .

Bif endophilin B which binds to UVRAG and stimulates the activity of Vps and as a result autophagosome maturation . In addition, Ambra and Pink, two obligatory professional teins in mitophagy, can bind to Beclin and encourage autophagy of mitochondria . Beclin may also be recruited to dynein motor complicated by binding to Ambra or Bim . This translocation removes Beclin complex from autophagosome for mation however the phosphorylation of Ambra by ULK and Bim by JNK releases Beclin from cytoskeletal storage to facilitate autophagy and endocytosis. You can find emerging studies indicating the perform of Beclin complex is not really limited to autophagy but it could be concerned in phagocytosis and endocytosis. Joubert et al. demonstrated that CD, a very well acknowledged receptor recognizing a lot of micro organisms, recruited Beclin Vps complicated towards the cell sur face following the recognition of pathogen. CD was linked to Beclin through the scaffold protein GOPC. In addition they observed that measles virus induced autophagy through the CD GOPC pathway. Konishi et al. observed the Beclin Vps complex regulated the clearance of apoptotic cells into the two pro and nonprofes sional phagocytes.
Yet, you will find conflicting results about the purpose of Beclin complexes inside the regulation of endosomal pathway. Earlier observations exposed that UVRAG is involved in endo somal flux with C Vps complex but not with all the Beclin Vps complex . Improved UVRAG expression also acti vated Rab GTPase and promoted endosomal fusion. Recent research by Thoresen et al. have indicated the endocytosis of cell surface Proteasome Inhibitor selleck chemicals receptors and cytokinesis is mediated through the Beclin Vps UVRAG Bif complex. Ruck et al. demonstrated in C. elegans that the Beclin ortholog, BEC , is concerned in distinct endosomal functions, specifically inside the retrograde transport from endosomes on the trans Golgi network. They observed that BEC protein colocalized using the retromer complex proteins into the early endosomes but that BEC was not needed for membrane transport from multivesicular bodies to lysosomes.
It seems that Beclin complexes are involved from the early ways of autophago cytosis and endocytosis but no longer during the later on phases from the transport to autolysosomes. Not long ago, it was observed that additionally towards the canonical autophagy, there is a non canonical autophagy which can be Beclin independent . The initiation and nuclea tion ways induced through the ULK and Beclin Vps complexes might be bypassed and the phagophore is formed directly from exist ing cellular membranes, e.g. plasma membrane, Ubiquinone Golgi complex and mitochondria. For instance, hydrogen peroxide evokes a Beclin independent autophagic cell death in RAW cells . Grishchuk et al. observed that numerous pro apoptotic compounds, e.g. staurosporine and etoposide, induced Beclin independent autophagy and subsequently triggered apoptosis in rat cortical neurons.