“
“The membrane-bound alcohol dehydrogenase of Gluconacetobacter diazotrophicus contains one pyrroloquinoline quinone moiety (PQQ), one [2Fe-2S] cluster, and four c-type cytochromes. Here, we describe a novel and inactive enzyme. ADHi, similarly to ADHa, is a heterodimer of 72- and 44-kDa subunits

and contains the expected prosthetic groups. However, ADHa showed a threefold molecular mass as compared to ADHi. Noteworthy, the PQQ, the [2Fe-2S] and most of the cytochromes in purified ADHi is in the oxidized form, contrasting with RG7422 order ADHa where the PQQ-semiquinone is detected and the [2Fe-2S] cluster as well as the cytochromes c remained fully reduced after purification. Reduction kinetics of the ferricyanide-oxidized enzymes showed that while ADHa was brought back by ethanol to its full reduction state, in ADHi, only one-quarter of the total heme c was reduced. The dithionite-reduced ADHi was largely oxidized by ubiquinone-2, thus indicating that intramolecular electron transfer is not impaired in ADHi. The acidic pH of the medium might be deleterious for the membrane-bound ADH by causing conformational changes leading to changes in the relative orientation of heme groups and shift of corresponding redox potential to higher values. This would hamper electron transfer resulting in the low activity observed in ADHi. In Gluconacetobacter diazotrophicus,

the PQQ-dependent enzymes – ethanol dehydrogenase (ADH) EPZ015666 nmr and aldehyde dehydrogenase (ALDH) – are located in the cytoplasmic membrane and oriented toward the periplasmic space (Matsushita et al., 1992). ADH and ALDH catalyze the two sequential oxidation reactions that convert ethanol to acetic acid; both enzymes transfer electrons to membrane ubiquinone. The ethanol-oxidizing ability in acetic acid bacteria can be easily changed and sometimes lost during cultivation, especially in prolonged shaking Megestrol Acetate cultures

of Acetobacter aceti (Muraoka et al., 1982; Ohmori et al., 1982) and Acetobacter pasteurianus (Takemura et al., 1991). Under these conditions, spontaneous mutants unable to oxidize ethanol emerge with high frequency. In Gluconobacter suboxydans, genetic instability has not been detected (Matsushita et al., 1995); instead, a dramatic decay in ADH activity has been observed under particular cultivation conditions, such as low pH and/or with high aeration. The presence of an ADH with a very low enzyme activity level (named as inactive ADH) has been reported (Matsushita et al., 1995). Gómez-Manzo et al. (2008, 2010) have already purified and characterized a highly active ADH (ADHa) from N2-grown Ga. diazotrophicus, using forced aeration and physiological acidification caused by growth. In the present work, we purified and characterized an ADH with very low enzyme activity (ADHi). A comparative study of the molecular and catalytic properties of the active and inactive forms of ADH from Ga.

4-Aminobenzenesulfonate (4-ABS) is commonly used as intermediate in the manufacturing of dyes, brighteners and sulfa drugs. Degradation of 4-ABS is problematic due to poor permeability across the bacterial membrane (Hwang et al., 1989), high C–S bond stability (Wagner & Reid, 1931) and potential bacteriostatic effect (Brown, 1962). Constant exposure of bacteria to 4-ABS induces selection of enzymatic pathways necessary for the utilization of 4-ABS as an energy source. In

the last two decades, 4-ABS degradation has been described in the genus Hydrogenophaga, Sphingomonas, Agrobacterium and Pannonibacter (Feigel & Knackmuss, 1988; Perei et al., 2001; Singh et al., 2004; Wang et al., 2009). The first isolated 4-ABS degraders were two-membered co-cultures consisting of Hydrogenophaga intermedia S1 and Agrobacterium radiobacter ALK inhibitor S2 (Feigel & Knackmuss,

1988; Contzen et al., 2000). Hydrogenophaga intermedia S1 can degrade 4-ABS as a pure culture when vitamins are added to the medium (Dangmann www.selleckchem.com/products/ipilimumab.html et al., 1996). To date, enzymes involved in the lower pathway of 4-ABS degradation in H. intermedia S1 have been characterized through heterologous expression in Escherichia coli host (Contzen et al., 2001; Halak et al., 2006; Halak et al., 2007). However, studies focusing on the upper pathway converting 4-ABS to 4-sulfocatechol have hitherto been scarce. Furthermore, the phenotype arising from the individual inactivation of 4-ABS-associated catabolic genes still remains unknown. To determine this and further elucidate the 4-ABS degradation pathway, it is necessary to perform genetic studies in the native microorganism. So far, the characterization of Hydrogenophaga strains involves 16S Miconazole rRNA

gene-based phylogenetic analysis, biochemical tests, DNA G+C content determination and DNA–DNA hybridization (Kampfer et al., 2005; Chung et al., 2007; Yoon et al., 2008). Although some strains show potential in the degradation of biphenyls and methyl-tert-butyl ether (Hatzinger et al., 2001; Lambo & Patel, 2006), the genetic aspects of the degradation pathway for these compounds are still unknown. Furthermore, there are no reports on in vivo genetic modification within the genus Hydrogenophaga. Hydrogenophaga sp. PBC is a Gram-negative bacterium isolated from textile wastewater for its ability to degrade 4-ABS (Gan et al., 2011). Similar to H. intermedia S1, strain PBC can degrade 4-ABS in the presence of vitamins. In this study, we describe the isolation and characterization of genes affecting 4-ABS biotransformation using a transposon mutagenesis approach. Hydrogenophaga sp. PBC was grown at 30 °C in nutrient broth (NB) containing 5 g L−1 peptone and 3 g L−1 beef extract, super optimal broth (SOB) (Hanathan, 1983) or phosphate-buffered minimal salt (PB) media containing 0.09 mM MgSO4, 0.042 mM KCl, 7.5 mM NaHPO4, 7.5 mM KHPO4, 15 mM KH2PO4, 0.0068 mM FeCl3, 0.1 mM CaCl2 and 0.001% w/v yeast extract. (NH4)2SO4, 2.5 mM, was included in PB medium to give PBN medium.

However, evidence of adrenal suppression has been documented in some neonates treated with lopinavir/ritonavir, particularly when preterm [228], in addition to case reports of cardiac, renal, and neurological toxicity, especially in, but

not restricted to, premature infants, and including one death during PEP with lopinavir/ritonavir [296]. No effects have been observed with maternal GDC-0068 mw lopinavir/ritonavir in the absence of neonatal dosing. It remains unclear whether these effects are related to lopinavir/ritonavir specifically or could be seen with other ritonavir-boosted PIs. The Writing Group therefore recommends that this PI should be avoided in routine infant PEP and should only be prescribed to preterm neonates in exceptional circumstances. Its use should only be considered after seeking expert advice and where there is multidrug resistance. Close metabolic monitoring in hospital should be undertaken. Nelfinavir, the only other PI with an infant-dosing regimen, BGB324 manufacturer will be withdrawn in the near future and will no longer be available for

prescription in the UK or elsewhere in Europe. See the CHIVA website for dosing updates (www.chiva.org.uk). In contrast to the PIs, nevirapine efficiently crosses the placenta (see below) and is well absorbed by the neonate [297]. Neonatal metabolism of nevirapine is induced where there has been antenatal in utero exposure [73, 75]; if this drug is given to the neonate when the mother has taken it for 3 or more days, the full dose of 4 mg/kg per day should be started at birth, rather than the induction dose of 2 mg/kg per day (Table 1). Owing to its long half-life, nevirapine should be stopped 2 weeks before co-prescribed antiretroviral drugs with shorter half-lives to reduce the risk of nevirapine monotherapy exposure and the development of NNRTI resistance should transmission have occurred. The only licensed ART available for i.v. use in sick and/or premature neonates, unable

to take oral medication, is zidovudine [284, 298]. Reduced oral and i.v. dosing schedules for premature infants are available (Table 1). most The fusion inhibitor, enfuvirtide does not cross the placenta. Although i.v. enfuvirtide (T20) has been given to a small number of infants born to mothers with multidrug resistant HIV, no formal neonatal pharmacokinetic studies for enfuvirtide have been conducted to date. The dose used has been adapted from a paediatric subcutaneous treatment study [299] and an adult i.v. dosing study [300]. For infants born to ART-naïve women, or where drug resistance is unlikely, zidovudine, lamivudine and nevirapine is the well-tolerated combination-therapy regimen with most experience (see Table 1 for dosing).

The forthcoming evaluation of these tests in the field is keenly Selleck LDK378 awaited, since their introduction into clinical practice would represent an important improvement. The molecular diagnosis of HAT,

which has the great advantage of being highly specific, has evident constrains for field application. Only recently, with the development of the LAMP approach, has the translation of DNA amplification into a field test become feasible. One of the most fascinating staging approaches is polysomnography, probably due to its non invasiveness. It is unlikely that this method will become applicable for large-scale stage determination in rural areas, but as suggested by the same authors, it may find a niche application in paediatric cases, for which it would be preferable to avoid a lumbar puncture [119]. Great hopes currently rest on the immune-based detection of biomarkers, such as neopterin. Despite their

lack of specificity, these may prove to be very useful to replace WBC counts for the determination of stage, in combination with the detection of parasites in CSF. Furthermore, they could possibly be used as test-of-cure markers during post-therapeutic follow-up, thus extending their field of application. The translation of this type of molecule into immune-based lateral flow assays is underway, for the rapid determination of disease stage and/or the evaluation of post-treatment outcomes. For some of them, this has already been done for other applications [109]. Thanks to the disease control programmes and resolutions adopted over the last few years, HAT is currently considered selleck chemicals llc under control and complete elimination of the disease is no longer seen as a utopia [3]. However, to reach this goal and to not underestimate the disease, as has already happened in the past [37], patient management needs to be improved, above all in terms of diagnosis and treatment. Effective case detection and therapeutic intervention is essential to reduce disease transmission by decreasing the number of reservoirs. Huge efforts have been made Rho over the last 30 years to improve clinical practice with specific

regard to HAT patients by identifying biomarkers and developing new diagnostic tools. However, some widely used approaches for biomarker discovery in malignant conditions, including proteomics, have not been able to find clear application in sleeping sickness. A few published studies [66], [67] and [117] showed interesting results highlighting the potential utility of proteomics. It and other omics disciplines, by giving a global overview of the transcriptomic, proteomic and/or metabolic state of the samples analyzed, could help to achieve a better understanding of the mechanisms leading to the onset and the progression of sleeping sickness. Additionally, proteomics may also be useful in highlighting differences between the two forms of infecting parasites – T. b. gambiense and T. b. rhodesiense – at both host and parasite levels.

EVs are potential biomarkers for detection of diseases. Total numbers and/or numbers of certain subsets of EVs in body fluids may be used to predict the presence of a disease, or a risk factor TSA HDAC manufacturer of developing a disease. Recently, increased numbers of several types of EVs were shown to increase the Framingham risk score (FRS), a risk assessment tool to estimate a patient’s

10-year risk of developing CVD.[108], [109] and [110] These results are promising and imply that more prospective studies are needed to further investigate the prognostic value of EVs in individuals at risk for CVD. In cancer patients with VTE, the coagulant activity of TF associated with MVs isolated from platelet-poor plasma is markedly increased compared to the cancer patients without VTE.[13] and [98] These findings suggest that MVs associated with coagulant TF in cancer patients may predict thrombotic events in patients at risk

of developing VTE. EGFRvIII promotes the expression of the proangiogenic protein IL-8 through the NF-κB pathway.62 EGFRvIII mRNA was present not only in resected glioma tissue but also detectable in exosomes isolated from serum of 7 out of 25 glioblastoma patients.30 Thus, measuring EGFRvIII mRNA in vesicles may provide clinically relevant information on tumor presence, tumor progression, and response to therapy. Not only blood or fractions thereof, selleck but also other body fluids may be a useful source of vesicular biomarkers. For example, aquaporin-2, exposed by exosomes isolated from urine, may be a biomarker for renal and systemic disease.50 Exosomes isolated from urine were shown to contain SSR128129E the mRNA encoding two known prostate cancer biomarkers, PCA3 and TMPRSS2: ERG, and both mRNAs can be transferred to platelets.69 Thus, extraction of mRNA from urine or platelets may provide a useful means for prostate cancer diagnosis. Vesicles also offer therapeutic applications. For example, the adhesion of hematopoietic stem–progenitor cells (HSPC) to the endothelium is significantly improved in the presence of PMVs, thereby supporting engraftment after stem cell transplantation in lethally irradiated

mice.111 MVs derived from MSCs may provide a future (adjuvant) therapy for acute renal injury112 because intravenous administration of MSC-derived MVs improves the recovery of glycerol induced-acute renal injury in SCID mice.113 Exosomes from IL-10-treated immature DCs suppress inflammatory and autoimmune responses.114 This type of exosome may therefore become a suitable therapy for arthritis. Another interesting clinical application is exosome-based immunotherapy. The initial studies by using DC-derived exosomes (“dexosomes”) loaded with tumor peptides showed that “dexosomes” are capable of priming cytotoxic T cells and inducing tumor rejection in mice.115 Dexosomes also promote NK cell activation in immunocompetent mice and NK cell-dependent anti-tumor effects.

04–2.74]) ( Table 3 and Fig. 2). In addition, cleaved caspase-8 was determined to

be an independent prognostic factor according to a multivariate analysis (P = 0.03, Table 3). In the glioblastomas, there were reasonable to good positive correlations between the expressions of FasL vs. Fas (r = 0.47, P < 0.0001) and between Fas vs. cleaved caspase-8 (r = 0.41, P < 0.0001) and poor positive correlations between Fas vs. cleaved caspase-3 (r = 0.26, P = 0.014), FasL vs. cleaved caspase-8 (r = 0.22, P = 0.0388), and cleaved caspase-8 and -3 (r = 0.31, P = 0.0026). No correlations were found among FasL, Fas, and cleaved caspase-8 and cleaved caspase-3 in normal nervous tissue. Both IDH1 and MGMT were negatively expressed in all 97 GBMs despite the positive controls used for immunohistochemistry. Deregulation of the normal mechanism for programmed cell death plays an important role in the pathogenesis and progression of gliomas [14], [16], [20] and [33]. Although evidence beta-catenin inhibitor has accumulated that gene mutations [22], microRNAs [11], [36] and [47], growth factors [17], [18] and [37], RNA-binding proteins [45], DNA-binding transcription

factors [23], Ca2+ binding proteins [31], signal transduction proteins [5] and [31], and DNA methylation [15] have critical roles in regulating cell apoptosis, the significance of the extrinsic apoptotic signaling pathway for glioblastomas remains unclear [19] and [26]. In this study, we used TMA technology and immunohistochemistry to

assess the expression of proteins involved in the extrinsic pathway. We looked at FasL, Fas, cleaved caspase-8, and cleaved caspase-3 in treatment-naïve human glioblastomas and normal Venetoclax mouse glial cells from control see more brains and examined these immunohistochemistry findings in the context of the clinicopathological data of the study patients. Death receptors of the tumor necrosis factor (TNF) family, including TNFR1, Fas (CD95/Apo-1), DR4/DR5, Apo-3 (DR3), and their respective cognate ligands TNF-α, FasL (CD95L/Apo-1L), TNF-related apoptosis-inducing ligand (TRAIL/Apo-2L), and Apo-3L can induce the extrinsic apoptotic pathway in the cytoplasm of tumor and normal glial cells [1]. Molecular assays of the Fas signaling pathway using yeast and eukaryotic cells have shown that after the binding of FasL to the Fas receptor, Fas binds directly to the adapter protein FADD (Mort1) and leads to apoptotic signal transduction. In turn, FADD interacts with caspase-8 through its death effector domain (DED), leading to DISC assembly and caspase-8 oligomerization, which drives its own activation in the cytoplasm through self-cleavage. Subsequently, cleaved caspase-8 molecules in the DISC activate downstream effector caspases, leading to the cleavage of caspase-3 and apoptosis [4], [7], [21] and [27]. We demonstrated that malignant glial cells of glioblastomas express Fas and FasL, an inducer of immunocyte cell death via the Fas-mediated pathway of apoptosis.

Neves is grateful to the Program to Disseminate Tenure Track System, University of Tsukuba, Japan, for the financial support. The author C. Prentice acknowledges for the financial support by the National Council for Scientific and Technological Development (CNPq) and the grants provided by the Coordination for the Improvement of Higher Education Personnel (CAPES) of Brazil. “
“Current Opinion in Food Science 2015, 1:13–20 This

review comes from a themed issue on Food chemistry and biochemistry Edited by Delia Rodriguez Amaya http://dx.doi.org/10.1016/j.cofs.2014.08.001 2214-7993/© 2014 Published by Elsevier Ltd. Although it is not possible to precisely determine the exact period when men mastered the use of fire, which might have happened in the Middle Paleolithic (400 000–200 000 years ago), it is unequivocal that its use for cooking was a major turning point in human evolution. Cooking hypoxia-inducible factor pathway roots and grains Selleck 5-Fluoracil allowed humans to retrieve more energy from available vegetable food and as a consequence, sufficient energy for hunting, which provided food with higher caloric density. This pattern of feeding was critical for the evolution of the species, once the development of a bigger brain required more available energy. Further, the use of heat allowed the development of food preservation technologies which substantially contributed to the decrease in food-borne

diseases, to the decrease of under-nutrition,

by making food available which, in turn, contributed to the drastic changes in life style and population distribution (rural and urban areas) all around the world in the last century. Different reactions take place during thermal processing of foods, some of them are desirable and relate to the sensory properties that increase their acceptance, while some of them must be avoided as they generate harmful substances to human health, such as acrylamide and nitrosamines. Lipid oxidation, sugar caramelization, enzyme inactivation, protein denaturation are some examples of modifications that heat can provoke in foods. Food reactions that initiate with the condensation of a carbonyl group and an amine group, producing, at the final stage, brown pigments, were first studied and described by the French biochemist Louis-Camille Maillard from 1912 to 1917 and, Abiraterone clinical trial therefore, are known as Maillard reaction. Maillard was able to predict, working on peptide synthesis by heating free amino acids in glycerol, that the amine-carbonyl compounds reactions could lead to nutrients loss during heat processing, to the abiotic generation of humic substances in soil and to protein modification in vivo and, yet, his work was put aside for almost 35 years. Robert et al. [1] provide an interesting analysis of the scientific scenario at the time of Maillard’s discoveries and why his work was overlooked for so long.

Sharing the same basic body shape, their weight ranged from 0.055 to 5.2 g (Table 3). Basal energy turnover diminished with increasing body mass also in locusts (Harrison et al., 2010) and in honeybee larvae (Petz et al., 2004). Niven and Scharlemann (2005) came to similar findings comparing resting metabolism of many flying insects. If also non-flying Selleck Volasertib arthropod species are included, the decrease of mass specific resting metabolism

with body mass is smaller (Fig. 8). Nonetheless there is an enormous variation in (resting) metabolism measurements of even closely related taxa of arthropods (compare Fig. 7 and Fig. 8). There are several hypotheses concerning this variation. The evolutionary trade-off hypothesis tries to explain the relationship between resting metabolic rate and ambient temperature, and the cause of variation on all taxonomic levels (order, family, inter- as well as intra-species; e.g. Clarke, 2006 and Riveros and Enquist, 2011). The aerobic capacity hypothesis (developed for mammals by Hayes and Garland, 1995) states that the higher the maximal metabolic rates that can

be achieved by animals the higher the resting metabolism. Transferring this hypothesis to insects with a similar energetic capacity than mammals, species with a highly energetic life-style (see Riveros and Enquist, 2011) like yellowjackets and learn more honeybees should have a higher mass-specific resting metabolism than insects with a more settled way of life like Eupsilia

sp. ( Heinrich, 1987) and P. dominulus ( Kovac et al., 2009 and Weiner et al., 2009). Our findings support this hypothesis (see Fig. 7 and Fig. 8). Another explanation for differences in medroxyprogesterone resting metabolism is provided by the life-style hypothesis (Reinhold, 1999 and Riveros and Enquist, 2011). If one compares the tachinid fly Nowickia sp. ( Chappell and Morgan, 1987) and the winter flying cuculinid moth Eupsilia sp. ( Heinrich, 1987 and Heinrich and Mommsen, 1985) which weigh 0.130 g and 0.155 g, respectively, they differ highly in resting metabolism – and also in way of life ( Table 2; Fig. 7 and Fig. 8, No. 10 Nowickia sp. and No. 11 Eupsilia sp.). The fly with the higher metabolism lives “on the wing” whereas the moth is rather inactive and sits still most of the day. However, Terblanche and Anderson (2010) showed that the resting metabolic rate in the hawkmoth Macroglossum trochilus and the long-proboscid fly Moegistorhynchus longirostrus differs despite a similar size and life-style (in this case foraging behavior).

The existence of bilateral neural connections between the two SON was suggested by electrophysiological and in vivo studies, thus supporting our results that both SON are involved in the mediation of the cardiovascular response to the microinjection of carbachol into the BST. Takano Adriamycin order et al. (1990) reported that one-third of the vasopressin-containing neurons tested in the SON were excited by electric stimulation

of the contralateral SON. In the same study, those authors reported that vasopressin neurons tested in the SON were not antidromically activated by a contralateral SON stimulation, thus suggesting that neural connections between the bilateral SON are mainly polysynaptic. It was also reported that antidiuretic effect associated with noradrenaline microinjection into the SON was inhibited either by a lesion of the contralateral SON or its pretreatment

with adrenoceptor antagonists (Tsushima et al., 1996), indicating the existence of bilateral adrenergic neural connections between selleck chemical supraoptic nuclei. Because the pressor response to the microinjection of carbachol into the BST was inhibited by the blockade of either the ipsilateral or the contralateral SON, it is possible that carbachol administration into the BST activates a pathway from the BST to the ipsilateral SON, in relation to BST microinjection site, which would stimulate neuron(s) that project to contralateral SON, thus suggesting that carbachol responses would depend on a bilateral SON cross-talking. Therefore, activation of vasopressinergic neurons in the contralateral SON in relation to BST stimulation site would mediate pressor response to carbachol administration into the BST. A schematic representation sketching the mechanism by which carbachol microinjection

into the BST evokes a vasopressin-mediated pressor response is presented next in Fig. 9. The pathway for the neural connection between bilateral SON is not totally understood. Moos and Richard (1989) concluded that the supraventricular gray commissura is important for interconnection of oxytocin-containing neurons in the SON, because synchronization of oxytocin-containing neurons in the bilateral SON disappeared after an inter hemisphere sectioning (including the supraventricular gray commissura and the corpus callosum), but persisted after a superficial interhemisphere sectioning that was limited to the corpus callosum. Therefore, the supraventricular gray commissura is a possible pathway for interconnections between bilateral SON vasopressin-containing neurons. Also, other connections between bilateral supraoptic nuclei, through the medulla oblongata and pons, have been suggested to exist (Tsushima et al., 1996), thus indicating alternative pathways for a bilateral SON cross-talking.

Supporting this speculation is the result that survivin was detectably increased by ANE in OC2 cells (Fig. S6). ANE also obviously induced HIF1α, the master regulator of hypoxia adaptation, via activating ERK (Fig. S6). In addition,

activation of NF-κB appeared to favor cell survival during ANE treatment in spite of the potential side effect, cell cycle retardation. As a cyclin-dependent kinase (CDK) inhibitor, p21 is well known as a negative regulator of cell proliferation [36]. However, increasing evidence www.selleckchem.com/products/Metformin-hydrochloride(Glucophage).html has suggested that nuclear p21 may not simply induce cell cycle arrest. Accumulation of p21 in the nucleus has been shown to be correlated with poor prognosis and disease progress in OSCC [37] and [38]. Interestingly, p21 may facilitate

G1/S transition after assembling into CCND1/Cdk2/p21/PCNA complex unless cyclin E/Cdk2 is sequestered by excessive p21 proteins ([39] and [40]). Given that ANE-induced p21 retards cell cycle, cells may continue proliferation once areca nut is removed after chewing. In surviving cells, ANE possibly triggers transformation via mechanisms besides the ROS-mediated DNA damage. In the shown examples, E7080 mw however, it is unclear how and why EGFR and Akt were downregulated by ANE at lower serum concentration. Although Akt is commonly known as an oncoprotein, accumulating evidence has suggested that like Ras, overactivated Akt may induce senescence under specific circumstances HSP90 [41]. Because Akt could sensitize cells to ROS-mediated apoptosis, downregulation of Akt activity might facilitate early carcinogenesis induced by ANE [42]. Once nutrients and serum are sufficiently available especially after angiogenesis, activation of EGFR and Akt signaling possibly accelerates the progression of OSCC. Taken together, by manipulating FBS concentration we discovered that ANE differentially determined cellular destiny, thus delineating a possible progression

of ANE-mediated oral carcinogenesis (Fig. 5). Without the interference from exogenous growth factors, the effects of ANE on epithelial-mesenchymal transition are also easier to observe in cells supplemented with less FBS. Our results give a potential model for the simulation of ANE-mediated pathogenesis in culture cells. WT, Ji initiated this project, executed most of the experiments, and wrote the manuscript. YC, Chuang provided related resources. HP, Chen was responsible for morphology photos. CC, Lee provided the comments of clinical observations. Jeff YF, Chen conceived the plan and corrected the manuscript. SR, Yang was responsible for independent Western blot and morphology photos. JH, Chen and CJ, Wang were responsible for RT-PCR and reporter assay, respectively. HR, Chen conceived the plan and corrected the manuscript. All authors read and approved the final manuscript. [43] This work was partially supported by National Science Council (97-2311-B-194-001-MY3) and no additional external funding was received for this study.