Eating Disorders and Internet: Content Analysis of Croatian Web Pages

U posljednje vrijeme internet postaje sve popularniji način komunikacije koji omogućava pojedincima stvaranje web stranica na kojima mogu izraziti svoja razmišljanja i vlastitu ličnost. Pro-anorexia websites su web stranice koje teže promicanju, podržavanju i diskutiranju o anoreksiji (Dias, 2003). Takve stranice prikazuju anoreksiju u pozitivnom svjetlu i obično ju definiraju kao životni stil, a ne kao oboljenje. Autori su tih stranica najčešće mlade djevojke, a cilj im je stvoriti zajednicu koja će redefinirati anoreksiju, pri čemu stvaraju i vlastiti rječnik. Na ovim se stranicama promiče uporaba lijekova za mršavljenje te pružaju štetni savjeti o načinima postizanja i održavanja vrlo niske tjelesne težine. Budući da ovakve stranice postoje i na hrvatskom jeziku, te činjenica da do sada nisu podvrgnute detaljnijim analizama, zanimalo nas je ispitati kakvi su sadržaji koji se na njima nude. Analizirani su internetski članci (eng. postovi) objavljeni na 31 elektroničkom dnevniku (eng. blogovi) čiji se sadržaj odnosio na podupiranje anoreksije, bulimije i općenito poremećaja hranjenja. Provedene su analize pokazale da su sudionici na ovim stranicama najčešće adolescentne djevojke, a glavne teme o kojima raspravljaju uključuju prejedanje, povraćanje, dijetu, post, vježbanje i zloporabu lijekova. Uz to, često se daju i traže savjeti te pruža socijalna podrška u cilju gubitka i održavanja niske tjelesne težine. Negativne emocije kao tuga, mržnja i ljutnja mnogo su češće zastupljene negoli pozitivne emocije. Dvije trećine analiziranih elekroničkih dnevnika sadrži slike i pjesme čiji je cilj ojačati motivaciju i ponašanja usmjerena ka postizanju ideala mršavosti. S obzirom na potencijalno negativne, ali još uvijek nepoznate efekte ovakvih stranica, svakako je potrebno planirati dodatne analize neophodne za izradu plana intervencije.The Internet has of late become more and more popular as a means of communication, providing individuals with the opportunity to create websites and web pages on which they can express their own opinions, thoughts and personality. Pro-anorexia websites are those which seek to promote, encourage and discuss anorexia (Dias, 2003). Websites like these portray anorexia in a positive light and usually define it as a lifestyle, not an ailment. The authors of the web pages are usually young girls whose goal is to create an anorexia-redefining community and, in the process, draw up their own vocabulary. These websites promote the use of weight-loss medicaments and are a source of harmful tips on how to attain and maintain a very low body weight. Given the fact that there are also Croatian pro-anorexia web pages and they have not, as yet, been the subject of any detailed analyses, we were interested in examining their contents. We have analyzed internet posts published on 31 blogs, whose contents referred to encouraging anorexia, bulimia, and eating disorders in general. The analyses conducted for this study show that the participants of these websites are usually adolescent girls, and the main subjects of their discussions are binge eating, regurgitation, dieting, fasting, exercising and drug abuse. In addition, pro-anorexia websites are also a means of seeking and giving advice as well as providing social support needed to attain and maintain a low body weight. Negative emotions such as sadness, hatred and anger are far more common than positive ones. Two thirds of blogs that were subject to analysis also contain images and poems intended to reinforce motivation and behavior directed towards attaining the ideal of thinness. Given the potentially negative, and as yet unknown, effects of these web pages, it is necessary to conduct additional analyses in order to draw up a plan of intervention

Neurodegenerative Diseases and Autophagy

Most neurodegenerative diseases are characterized by the accumulation of aggregated proteins within neurons. These aggregate-prone proteins cause toxicity, a phenomenon that is further exacerbated when there is defective protein clearance. Autophagy is an intracellular clearance pathway that can clear these protein aggregates and has been shown to be beneficial in the treatment of neurodegenerative diseases in a variety of model systems. Here, we introduce the key components of the autophagy machinery and signaling pathways that control this process and discuss the evidence that autophagic flux may be impaired and therefore a contributing factor in neurodegenerative disease pathogenesis. Finally, we review the use of autophagy upregulation as a therapeutic strategy to treat neurodegenerative disorders

Human CD34+ cell differentiation toward the endothelial lineage in the zebrafish embryo

Background: The zebrafish embryo (Danio rerio) provides a number of advantages for in vivo study of vertebrate vasculogenesis and angiogenesis, including the possibility to observe in real-time the development of the embryonic vasculature as, for example, by confocal in vivo imaging of endothelial cells sprouting (Lawson et al., 2002). Furthermore, cell transplantation into the zebrafish embryo is a novel and interesting model system for the study of stem cells differentiation and plasticity. (Traver et al., 2003; Traver et al., 2004). Objective: The objective of the present study was to determine whether human CD34+ progenitor (hCD34+) cells differentiate toward the endothelial lineage and contribute to new blood vessels development after transplantation into the zebrafish blastula and embryo. Methods and Results: hCD34+ cells were isolated from cord blood and subsequently labelled with Orange Cell Tracker\u2122 dye (Invitrogen). hCD34+ cells (400-500) were injected into the zebrafish blastula and hCD14- cells were used as controls. Flow cytometry (FACS) analysis of injected embryos dissociated 24- and 44-hours after transplantation (28hpf and 48hpf stage) showed endothelial differentiation of hCD34+ cells, as demonstrated by increased expression levels of endothelial markers (KDR, CD105, CD146), whereas hematopoietic lineage markers (CD3, CD38, CD45 and CD48) were not affected. Further, immunofluorescence analysis showed KDR+ and VE-cadherin+ human cells localized in the dorsal artery and in the cardinal vein. The above changes were not observed in hCD14- cell \u2013 injected zebrafish. In other experiments hCD34+ and hCD14- cells (500-1000) were injected into the sinus venosus of developing Tg(fli1:EGFP) transgenic embryos (48hpf), prior to immune system development. Time-lapse confocal analysis performed 2 hours after transplantation showed circulating hCD34+ cells in developing vessels and no evidence of vascular occlusions. Further, at 12 hours after injection , some hCD34+ cells had incorporated into the vessel wall. One day after transplantation, hCD34+- injected embryos exhibited altered blood vessels sprouting in the growing tail vasculature as well as enhanced and ectopic angiogenesis at the level of the subintestinal vein; these results raised the possibility that hCD34+ cells may synthesize angiogenic factors. In agreement with this hypothesis it was found that hCD34+ cell injection into the zebrafish blastula rescued the vascular phenotype caused by Vegfc knock-down. On the contrary, hCD14- cells injected into the embryo do not seem to affect vascular development. Conclusions: The present study demonstrates the evolutionary conservation of hCD34+ cells differentiation mechanisms toward the endothelial lineage and their angiogenic properties in the zebrafish embryo

Evolutionary conservation of human CD34+ Cell Endothelial Differentiation in the Zebrafish Embryo

Background. We tested the zebrafish embryo as a novel system to study human CD34+ progenitor cells (hCD34+) differentiation toward the endothelial and hematopoietic lineages. Methods and Results. hCD34+ were isolated from cord blood and subsequently labelled either with orange cell trackerTM dye or infected with a lentivirus expressing GFP. Labelled hCD34+ (500\u20131000 cells) were injected into the sinus venosus of developing wt and Tg(fli1:EGFP) transgenic embryos prior to immune system development. Embryos transplanted with labelled hCD14\u2013 cells were used as controls. Time-lapse confocal microscopy at early time points (2 hours) showed hCD34+ circulation in developing vessels of the transgenic embryos and no vascular occlusions. Some hCD34+ integrated into the vessel wall, after their early adhesion to vascular structures. One day after transplantation, hCD34+ injection resulted in severe vascular defects, as increased diameter of dorsal artery (DA) and cardinal vein (CV) (DA=3.8\ub11.4um vs. 8.2\ub12.9um, p<0.05 n=11; CV=6.9\ub10.6um vs. 13\ub12.3um, p<0.01, in control and hCD34+ injected embryos, respectively), altered branching with ectopic sprouts in the growing vasculature (61/72 embryos), and abnormal angiogenesis in the yolk region (14/17 embryos). Further, it was found evidence of hCD34+ differentiation into mature endothelial hVWf - and hCD31 - positive cells. To investigate hCD34+ plasticity in the early events of vascular development, we performed hCD34+ transplantation into the zebrafish blastula. hCD34+ behaved as hemangioblast, homing to both the vascular and the hemato-poietic compartments. Finally, embryos injected with the specific Vegfc morpholino RNA, that display a vascular phenotype, were rescued by hCD34+ cells transplanted 2 hours later at blastula stage (29/32 embryos), suggesting that transplanted hCD34+ show a potent angiogenic activity that affects the developing vasculature of the Vegfc depleted embryos. Conclusions. Our in vivo study demonstrates the evolutionary conservation of hCD34+ differentiation mechanisms in the zebrafish embryo, supported by hCD34+ participation to the zebrafish blood vessels formation, and by hCD34+ production of angiogenic factors acting on resident embryonic vascular cells

Endothelial fate and angiogenic properties of human CD34+ progenitor cells in zebrafish

Objective-: The vascular competence of human-derived hematopoietic progenitors for postnatal vascularization is still poorly characterized. It is unclear whether, in the absence of ischemia, hematopoietic progenitors participate in neovascularization and whether they play a role in new blood vessel formation by incorporating into developing vessels or by a paracrine action. Methods and Results-: In the present study, human cord blood-derived CD34 + (hCD34 +) cells were transplanted into pre- and postgastrulation zebrafish embryos and in an adult vascular regeneration model induced by caudal fin amputation. When injected before gastrulation, hCD34 + cells cosegregated with the presumptive zebrafish hemangioblasts, characterized by Scl and Gata2 expression, in the anterior and posterior lateral mesoderm and were involved in early development of the embryonic vasculature. These morphogenetic events occurred without apparent lineage reprogramming, as shown by CD45 expression. When transplanted postgastrulation, hCD34 + cells were recruited into developing vessels, where they exhibited a potent paracrine proangiogenic action. Finally, hCD34 + cells rescued vascular defects induced by Vegf-c in vivo targeting and enhanced vascular repair in the zebrafish fin amputation model. Conclusion-: These results indicate an unexpected developmental ability of human-derived hematopoietic progenitors and support the hypothesis of an evolutionary conservation of molecular pathways involved in endothelial progenitor differentiation in vivo

Role of E2F1\u2013Cyclin E1-Cyclin E2 Circuit in Human Coronary Smooth Muscle Cell Proliferation and Therapeutic Potential of Its Downregulation by siRNAs

Aberrant coronary vascular smooth muscle cell (CSMC) proliferation is a pivotal event underlying intimal hyperplasia, a phenomenon impairing the long-term efficacy of bypass surgery and angioplasty procedures. Consequently research has become focused on efforts to identify molecules that are able to control CSMC proliferation. We investigated downregulation of CSMC growth by small interfering RNAs (siRNAs) targeted against E2F1, cyclin E1, and cyclin E2 genes, whose contribution to CSMC proliferation is only now being recognized. Chemically synthesized siRNAs were delivered by two different transfection reagents to asynchronous and synchronous growing human CSMCs cultivated either in normo- or hyperglycemic conditions. The depletion of each of the three target genes affected the expression of the other two genes, demonstrating a close regulatory control. The clearest effects associated with the inhibition of the E2F1\u2013cyclin E1/E2 circuit were the reduction in the phosphorylation levels of the retinoblastoma protein pRB and a decrease in the amount of cyclin A2. At the phenotypic level the downmodulation of CSMC proliferation resulted in a decrease of S phase matched by an increase of G1-G0 phase cell amounts. The antiproliferative effect was cell\u2013donor and transfectant independent, reversible, and effective in asynchronous and synchronous growing CSMCs. Importantly, it was also evident in hyperglycemia, a condition that underlies diabetes. No significant aspecific cytotoxicity was observed. Our data demonstrate the interrelation among E2F1\u2013cyclin E1-cyclin E2 and the pivotal role this circuit exerts in CSMC proliferation. Additionally, our work validates the concept of utilizing anti\u2013E2F1\u2013cyclin E1-cyclin E2 siRNAs to develop a potential novel therapy to control intimal hyperplasia

Tau isoforms imbalance impairs the axonal transport of the amyloid precursor protein in human neurons

Tau, as a microtubule (MT)-associated protein, participates in key neuronal functions such as the regulation of MT dynamics, axonal transport, and neurite outgrowth. Alternative splicing of exon 10 in the tau primary transcript gives rise to protein isoforms with three (3R) or four (4R) MT binding repeats. Although tau isoforms are balanced in the normal adult human brain, imbalances in 3R:4R ratio have been tightly associated with the pathogenesis of several neurodegenerative disorders, yet the underlying molecular mechanisms remain elusive. Several studies exploiting tau overexpression and/or mutations suggested that perturbations in tau metabolism impair axonal transport. Nevertheless, no physiological model has yet demonstrated the consequences of altering the endogenous relative content of tau isoforms over axonal transport regulation. Here, we addressed this issue using a trans-splicing strategy that allows modulating tau exon 10 inclusion/exclusion in differentiated human-derived neurons. Upon changes in 3R:4R tau relative content, neurons showed no morphological changes, but live imaging studies revealed that the dynamics of the amyloid precursor protein (APP) were significantly impaired. Single trajectory analyses of the moving vesicles showed that predominance of 3R tau favored the anterograde movement of APP vesicles, increasing anterograde run lengths and reducing retrograde runs and segmental velocities. Conversely, the imbalance toward the 4R isoform promoted a retrograde bias by a significant reduction of anterograde velocities. These findings suggest that changes in 3R:4R tau ratio has an impact on the regulation of axonal transport and specifically inAPPdynamics, which might link tau isoform imbalances with APP abnormal metabolism in neurodegenerative processes.Fil: Lacovich, Valentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Biología Celular y Neurociencia ; Argentina. International Clinical Research Center; Estados UnidosFil: Espindola, Sonia Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular ; ArgentinaFil: Alloatti, Matías. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Biología Celular y Neurociencia ; ArgentinaFil: Pozo Devoto, Victorio Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Biología Celular y Neurociencia ; ArgentinaFil: Cromberg, Lucas Eneas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Biología Celular y Neurociencia ; ArgentinaFil: Carna, Mária E.. International Clinical Research Center; Estados UnidosFil: Forte, Giancarlo. International Clinical Research Center; Estados UnidosFil: Gallo, Jean Marc. King's College London; Reino UnidoFil: Bruno, Luciana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Física de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Física de Buenos Aires; ArgentinaFil: Stokin, Xgorazd B.. Anne's University Hospital; República ChecaFil: Avale, Maria Elena. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular ; ArgentinaFil: Falzone, Tomas Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Biología Celular y Neurociencia ; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentin

Additional file 1 of Unraveling axonal mechanisms of traumatic brain injury

Additional file 1: Fig. S1. Characterization of injury system, Related to Fig. 1

Additional file 20 of Unraveling axonal mechanisms of traumatic brain injury

Additional file 20. Key Resources Table: Table including antibodies, resources, kits, cells, and software with catalog numbers