Extracellular vesicles (EVs) play an important role in cell-to-cell communication. Although there are different kinds of vesicles, each with their own secretion and capture biology, all of them carry a cargo of proteins, lipids, metabolites and nucleic acids. They act as vehicles for exchange of biological materials and signals and are involved in the regulation of various physiological processes. Liver is an essential organ containing different cell populations fulfilling various functions, which need to be strictly controlled and coordinated. There are a few articles showing the role of liver-derived EVs. On the basis of them, we present here a hypothesis of the implication of such vesicles in the physiology of the liver. Different liver cell types, including hepatocytes, cholangiocytes and stellate cells, secrete and capture EVs and interact with them. Liver injury changes the abundance and cargo of EVs; these changes are likely to be important for the outcome of stress response. Although a substantial effort has been put into the characterization of EVs in isolated populations, it is only recently that some more comprehensive information has begun to appear. In this article, we hypothesize about the role of EVs in liver microenvironment and their possible function using published data from both hepatic and non-hepatic systems. Liver is a multifunctional organ involved in important metabolic functions, synthesis of blood components and storage of lipids and retinol . It is ahttp://www.isevmeeting.org). However, we also provide the type of EVs, if the information is available.All these cellular populations, with their diverse physiological processes, have to be strictly coordinated to perform their functions properly. Recently, the role of secreted extracellular vesicles in the exchange of proteins, nucleic acids and lipids has awakened some considerable interest in the field of intercellular signalling . Two maiThere have been just few studies focusing on the role of circulating liver-derived EVs, or the possible effects that non-hepatic EVs have on liver. Supported by those published articles, and extrapolating information obtained from non-liver systems, we offer here a collection of possible scenarios where EVs may be important for liver functionality.Several reports –8 have iRemarkably, the composition of EVs depends on the particular cell state ; their ePreviously, we have reported that non-tumoral liver-derived cell lines such as MLP29 and primary cultured hepatocytes are able to secrete EVs . A comprOur proteomic analysis of hepatocyte-derived EVs indicates that these vesicles contain several members of the P450, UGT and GST drug-metabolizing protein families. If hepatocyte-derived EVs would be released to the blood stream, it can be hypothesized that EVs might be involved in detoxification of drugs and endogenous toxic substances . SupportHepatocyte-derived EVs contain mRNA , and livIn general, the role of bloodstream mRNAs is still unclear. Since the first descriptions of mRNA associated to EVs, functional transfer of EV-associated RNA to the acceptor cells has been shown . HoweverAlthough it has been suggested that the role of EVs is to protect RNA from RNase activity, the available data are controversial. The first description of RNA in exosomes has shown that mRNA is protected against RNase activity . HoweverHepatitis C virus (HCV) is a major cause of chronic liver disease, with about 170 million people infected worldwide . The HCVLiver is involved in detoxification of xenobiotics and noxious endobiotics, and therefore continuously exposed to injuries; tissue regeneration is an important defensive mechanism maintaining the viability of this organ. In acute liver injury, hepatic regeneration is not unlike a physiological wound-healing process, bringing about transient and reversible changes in the extracellular matrix of the organ and in the proliferative capacity of the hepatocytes , 42.in vivo, EVs derived from a subpopulation of human pluripotent resident liver cells accelerate the morphological and functional recovery of liver in hepatectomized rats (When administered zed rats . This efzed rats . Stem cezed rats , 43, andzed rats .Protein cargo also plays an important role in response to EVs. Cholangiocytes and myofibroblastic hepatic stellate cells release EVs containing active hedgehog ligands in response to platelet-derived growth factor . In the in vivo (Recently, it has been reported that cholangiocytes can secrete EVs . Moreovein vivo .Biliary EVs secreted by cholangiocytes take part in the inactivation of ERK kinase signalling . This paIn developed countries, non-alcoholic fatty liver disease is emerging as a major global liver disorder with the background of an increasing prevalence of obesity and type 2 diabetes . Non-alcThe role of EVs in inflammation is ambiguous because EVs can elicit either inflammation , or immuin vivo is associated with the spontaneous development of hepatocellular carcinoma as a result of aberrant responses to inflammatory and stress signalling (Tumoral cells undergo deregulation of miRNAs expression  and are gnalling , 67. TAKgnalling . The abegnalling .in vivo isolation and characterization of other liver-derived EVs.The appropriate regulation of the components present in any microenvironment is crucial to the stress response. As we have mentioned before, obesity causes leucocyte accumulation . LeucocyMaintaining appropriate signal balance may be crucial for the resolution of liver injuries. In a multicellular environment, EVs from different cell types, carrying different cargos, might interact with parenchymal cells. It is likely that liver dysfunction affects the composition of EVs’ cargo, making those vesicles invaluable in the damage assessment using plasma analysis. From another point of view, the ability of liver cells to uptake circulating EVs makes them a silver bullet in gene and miRNA therapy. The challenge is to decode the EVs’ network and find the tools to control it.


In the last few decades, pediatric medicine has observed a dramatic increase in                    the prevalence of hitherto rare illnesses, among which obesity, diabetes,                    allergies and other autoimmune diseases stand out. In addition, secular trends                    towards earlier onset of puberty and sexual activity contribute to the                    psychological problems of youth and adolescents. All this has occurred in spite                    of the improved health care provision for children, yet traditional concepts of                    medicine have failed to explain these new ?epidemics?.                A recent conference and science school of the European Society of Paediatric                    Endocrinology (ESPE) in Acre, Israel, has taken up this challenge. Experts                    across disciplines including medicine, anthropology and developmental psychology                    discussed potential causes of childhood ill-health from an evolutionary                    point-of-view. Seen from an evolutionary vantage point, the ?epidemics? of                    childhood obesity, diabetes and psychological dysfunction appear, in part, to be                    related to a mismatch between ancestral adaptations and novel environmental                    contingencies. These include changing exposures to pathogens, which impact on                    the function of the immune system, as well as changing patterns of parenting,                    which influence the timing of puberty and the risk for developing                    psychopathology.                             For a few decades, pediatricians have begun to face ?new epidemics? affecting                children?s physical and mental health. Rather than fighting novel infectious                diseases as the leading cause of childhood mortality throughout human history, new                challenges for child healthcare have arisen from overweight, changes in growth                patterns, earlier sexual activity, as well as from other secular transformations                pertaining to the conditions of up-bringing. Ironically, in spite of improved access                to healthcare and treatment options for many childhood diseases, clinicians have                observed a dramatic increase in chronic diseases, including, among many, childhood                obesity, diabetes and autoimmune diseases, particularly since the 1980s .                   In May 2013, a conference of the European Society for Paediatric Endocrinology (ESPE)                on Child Health and Human Evolution in Acre, Israel, took up these open questions                from a scientific angle that has become more and more influential in the recent past                to explain gene-environment interaction with respect to health issues, namely                Evolutionary Medicine.            Homo sapiens?                evolutionary past that brought about physical and psychological adaptations to quite                a diverse spectrum of Environments of Evolutionary Adaptedness (EEAs). Thus, it                offers a great deal of insight into present-day health issues, providing clinical                clues for how the conditions of modern life deviate from ancestral ones, and why                adaptations to past environments can have detrimental effects in contemporary                settings. In this essay, based on the lectures and discussions at the Acre                Conference, we highlight some of the most intriguing aspects of the secular trend                and developmental plasticity with respect to child health and discuss potential                consequences for healthcare and disease prevention.Evolutionary Medicine uses the reconstruction of             Secular trends in childhood obesity represent a growing health issue in the                    developed countries with profound impact on adult health . Over th                Among the most ?obesogenic? factors are increasing energy consumption, mainly                    foods rich in fat and sugar, and decreasing physical activity with the adoption                    of a more sedentary lifestyle - a whole mark of contemporary EEA. Obviously, the                    oversupply of energy and the lack of physical activity and exercise is an                    example of an evolutionary mismatch. Over tens of thousands of years of the                    Paleolithic era, humans lived in conditions where the availability of food could                    change dramatically. Food scarcity and famine probably occurred much more often                    than times of food abundance. According to the ?thrifty gene? hypothesis , humans                 in                        utero. Indeed, the average weight gain during pregnancy has                    increased by 2 kg in 20 years time [Another factor that may profoundly impact the risk of developing overweight and                    obesity is related to the intrauterine environment. ?Fetal programming? suggests                    that intrauterine conditions imprint the fetus? postnatal development with                    long-lasting effects extending into adult life . This maars time .       ars time ,       ars time , cancer ars time , respiraars time , obesityars time  and attears time , to name                Finally, assortative mating, an evolved pattern of how individuals choose their                    sexual partners, which suggests that partner choice is based, in part, on                    physical similarity, may impact the risk of obesity, because of the potential                    accumulation of ?thrifty genes? or genes involved in the regulation of fat                    deposition or satiety. In summary, childhood obesity illustrates at several                    levels how adaptive mechanisms that evolved in the ancestral past can contribute                    to the development of new health hazards. Related to this, diabetes is a                    pressing health issue with some overlapping predispositions with obesity, but                    additional evolutionary factors that might be equally relevant.                                        Childhood diabetes type I is another example illustrating secular trends towards                    increased risk for specific metabolic disorders . Type-I                 Helicobacter                        pylori antibodies, a 5-fold higher frequency of Toxoplasma                        gondii antibodies, a 12-fold higher prevalence of hepatitis A                    antibodies, and a 20% higher frequency of Coxsackie virus B4 antibodies as                    compared to Finnish children. In contrast, the figures for autoimmune diseases                    point in the opposite direction, where Karelian children have a six-fold lower                    incidence rate of type-I diabetes, and similar figures have been obtained for                    celiac disease, autoimmune thyreoiditis, and allergen-specific IgE responses                        [One plausible explanation relates to the so-called ?hygiene hypothesis?,                    suggesting that the decrease in diversity of pathogens children are exposed to                    has led to an increase in autoimmune diseases, including type-I diabetes . In supp         . This ex                Another possible factor accounting for the increase of type-I diabetes in                    children resides in changing habits of nurturing and feeding. Early exposure to                    cow milk induces an immune response to bovine insulin, which may later ?spark                    over? to attack endogenous insulin in individuals who lack the tolerance of oral                    ingestion of bovine insulin. In fact, IgG antibodies against bovine insulin are                    elevated in formula-fed infants at three months of age compared to breast-fed                    infants . Again,                                         Secular transitions have not spared family structures and parenting behavior, but                    without evolutionary insights it could not be fully understood why the                    conditions of one?s upbringing have such a great impact on the psychosexual                    development of adolescents . As earl                ESR1) gene who grow up with less sensitive mothers are                    younger at menarche than AA carriers, whereas GG carriers raised by sensitive                    mothers experience menarche later than carriers of the AA genotype.In addition to these environmental factors influencing the psycho-biological                    development in girls, there is evidence for gene-environment interaction,                    suggesting that the quality of maternal care interacts with polymorphic                    variation of estrogen receptor-coding genes in ways that support the assumption                    of ?differential susceptibility? .                       In a psychopathological perspective, individuals with borderline personality                    disorders (BPD) follow a developmental pathway associated with sexual                    risk-taking behavior, associated with frequent disruption of close relationships                    (including partnerships) and reduced investment in their own children . Beyond                                         Over eons, humans have adapted genetically and epigenetically to environmental                    contingencies, which in many ways are dissimilar to contemporary ones.                    Evolutionary Medicine is an emerging field that can help clarify hitherto                    incompletely understood health issues, including the ?new epidemics? of                    childhood that arise from mismatches between genes and environment . Living                 A broad range of diseases that were until recently considered to be rare have now                    exploded in incidence and prevalence rates, at least in the developed countries                    - with the prospect that the situation will be similar in the developing world,                    once the socio-economic status has risen to a similar level. This is, of course,                    not to argue that for these reasons developing countries should be precluded                    from improved health care, an increase in income, education and welfare - quite                    differently, it is rather an opportunity to prevent the developing countries                    from repeating the unfortunate sequence that has inflicted developed                    countries.                Evidence suggests that for European countries the childhood obesity and diabetes                    epidemics have slowed down and might have come to a halt in the last 10 years or                    so . It is                 Homo sapiens has evolved a distinct life-history which includes                    a relatively short infancy, a prolonged childhood and the emergence of                    adolescence as a novel developmental stage not seen in other primates. The                    evolved function of adolescence is most likely associated with the growing need                    of acquiring social skills that are necessary to successfully prepare and adopt                    adult social roles.                Taken together, it seems that our evolved life history patterns have come at a                    cost -?tradeoffs? in evolutionary jargon - which has brought about an increased                    risk for disruption of normal developmental pathways, where the cost becomes                    greater as modern environmental contingencies deviate from ancestral ones.                Insights from Evolutionary Medicine can help us understand, cure and prevent                    physical and psychological disease and disorders in many ways. We believe this                    is the way medicine has to go in the future at all levels, from research to                    practice. Evolutionary Medicine is coming of age.                                                        The authors declare that they have no competing interests.                            Textbook of Evolutionary Psychiatry. The Origins of                    Psychopathology, Oxford University Press, 2008.MB is Professor of Cognitive Neuropsychiatry and Psychiatric Preventive Medicine and                the author of the             Evo-Devo of                    Child Growth, Wiley, 2012. ZH is Professor of Pediatrics and Endocrinology and the author of


Dietary habits influence the risk of cancer of the oesophagus and oesophago-gastric junction, but the role of proportions of the main dietary macronutrients carbohydrates, fats and proteins is uncertain.Data was derived from a nationwide Swedish population-based case-control study conducted in 1995–1997, in which case ascertainment was rapid, and all cases were uniformly classified. Information on the subjects' history of dietary intake was collected in personal interviews. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using logistic regression, with adjustment for potentially confounding factors.Included were 189 oesophageal adenocarcinomas, 262 oesophago-gastric adenocarcinomas, 167 oesophageal squamous cell carcinomas, and 820 control subjects. Regarding oesophageal or oesophago-gastric junctional adenocarcinoma, a high dietary proportion of carbohydrates decreased the risk , and a high portion of fat increased the risk , while a high proportion of protein did not influence the risk . Regarding oesophageal squamous cell carcinoma, the single macronutrients did not influence the risk statistically significantly.A diet with a low proportion of carbohydrates and a high proportion of fat might increase the risk of oesophageal adenocarcinoma. The protocol also prescribed that serial biopsy specimens should be taken every 2 cm from the proximal stomach, through the oesophago-gastric junction, in the oesophagus, until normal squamous-cell epithelium was reached. Additional specimens were to be obtained proximally, distally, and laterally to the tumour. Surgeons and pathologists completed standardised and detailed descriptions of the location of the cancer in operated cases. Moreover, 97% of all biopsies and surgical specimens were re-examined by one pathologist. An adenocarcinoma of the oesophago-gastric junction had to have its centre within 2 cm above, or 3 cm distal to the junction. If Barrett´s oesophagus was detected adjacent to the tumour, Unconditional logistic regression was used to analyse the exposures as continuous variables in relation to risk of the studied tumours and to analyse defined exposure categories as presented above, providing crude and adjusted odds ratios (OR) with 95% confidence intervals (95% CI). The cases in each cancer category were first compared with all controls, and in a second analysis, cases of adenocarcinoma of the oesophagus or oesophago-gastric junction were analysed jointly. In crude models, no adjustments were made, but the frequency matching provided similar distribution of age and sex. In the multivariable model, the results were adjusted for age, sex, reflux symptoms , body mass index (categorised into four groups from high to low by quartiles among the control subjects), tobacco smoking , alcohol use , years of formal education , and total energy intake (categorised in quartiles based on the consumption of the control participants).The study included 189 cases of oesophageal adenocarcinoma (87% participation rate), 262 cases of oesophago-gastric adenocarcinoma (86% participation rate), 167 cases of oesophageal squamous cell carcinoma (73% participation rate), and 820 control subjects (73% participation rate). Characteristics of the participating case and control subjects are presented in High proportions of consumption of carbohydrates were followed by a possibly decreased risk of oesophageal adenocarcinoma . The modA high proportion of carbohydrate intake (highest quartile) was associated with a more than halved risk of oesophago-gastric junctional adenocarcinoma compared to a low proportion  . 10% incSince oesophageal and oesophago-gastric junctional adenocarcinomas are closely located and share many characteristics and aetiological factors, these tumours were combined in an analysis to increase statistical power. A high proportion of carbohydrates decreased the risk of these tumours , while a high proportion of fat increased the risk nearly 2-fold  . A high The proportions of consumption of carbohydrates did not influence the risk of oesophageal squamous cell carcinoma . ParticiGenerally, the crude ORs were similar to the fully adjusted ORs , 3, 4, 5This study indicates that a high proportion of carbohydrate consumption of the total energy intake decreases the risk of oesophageal and oesophago-gastric junctional adenocarcinoma, whereas a high proportion of fat in the diet increases the risk of these tumours, and proportions of protein consumption do not influence this risk. These nutrient proportions did not clearly influence the risk of oesophageal squamous cell carcinoma.Strengths of the study include the population-based design with high participation rates, the thorough and uniform classification of all tumours, the ability to adjust the results for all established risk factors, and the complete and rapid case ascertainment, which enabled personal interviews with all participants. Among weaknesses is the misclassification of the exposure, which might be increased by the fact that we asked about dietary habits 20 years prior to interview. This method of assessment of dietary items 20 years earlier has, however, been validated with good results, showing that such assessment captures a combination of previous and current dietary habits Recent studies have provided further evidence that fruit and vegetable intake is associated with decreased risk of oesophageal and oesophago-gastric junctional adenocarcinoma and oesophageal squamous cell carcinoma, whereas meat and nitrate decreases the risk of these tumours The oesophageal cancer specific mechanisms that might explain the findings of the present study can only be speculated upon. A high fat proportion might be followed by an increased prevalence of obesity, which in turn entails an increased risk of adenocarcinoma of the oesophagus and oesophago-gastric junction, but the results of the present study were thoroughly adjusted for the influence of BMI. Residual confounding by BMI cannot be entirely excluded, but alternative mechanisms should be considered. A high fat proportion diet might cause a slower passage of food through the stomach, i.e. inhibition of gastric emptying by intestinal peptides, In conclusion, this population-based and nationwide Swedish study indicates that a high proportion of consumption of carbohydrates decreases the risk for oesophageal and oesophago-gastric junctional adenocarcinoma, while a high proportion of consumption of fat increases the risk and protein proportions did not influence this risk. Macro nutrient proportions did not much influence the risk of oesophageal squamous cell carcinoma. These differences indicate some separate dietary aetiology for the main histological types of oesophageal cancer.


Biomedical imaging has been undergoing rapid technological advancements over the last several decades and has seen the development of many new applications. Techniques such as functional imaging, spectroscopic imaging, optical imaging, and image-guided interventional techniques (treatment and therapy) have been gaining acceptance in areas ranging from basic research to clinical applications, and from the cellular level to the whole-organ level. Biomedical imaging is an interdisciplinary field that requires collaboration among biologists, chemists, medical physicists, pharmacologists, computer scientists, biomedical engineers, and clinicians of all specialties. Skills and tools provided by radiologists are at the centre of many clinical research programs . BiomediDue to rapid technological developments leading to high spatial and temporal resolution, ultrasound, CT and MRI offer many opportunities for performing functional assessment of different body systems. The emergence and development of functional MR imaging and radionuclide imaging (SPECT/PET) have revolutionised the study of the functions of many body systems. Functional magnetic resonance (MR) encompasses a spectrum of techniques that depict physiological and molecular processes before morphological changes are visible on conventional imaging. Intraoperative MR guidance for neurosurgery improves prevision of tumour resection with use of combined data between functional MRI and SPECT .1HMRS) [31PMRS) [MR spectroscopy is considered to be a method of molecular imaging. The ability to break down the chemical composition of a lesion has contributed significant information to the understanding of the metabolic process of normal and pathological tissues. The techniques most often used in clinical practice are proton spectroscopy (1HMRS) , 5 and p[31PMRS) . The for[31PMRS) . With ra[31PMRS) , 9, and [31PMRS) –14. The [31PMRS) , 16.in vivo in real-time. The use of PET and SPECT lies in monitoring the presence and activity of disease as a function of treatment, particularly in oncology. New functional and metabolic imaging techniques, especially hybrid SPECT/CT, PET/CT and PET/MRI, provide pathological information in addition to morphological information. Combined PET/CT offers superior advantages over each individual imaging modality and has been shown to be valuable in many applications such as cardiac imaging [PET and SPECT are powerful techniques capable of imaging biochemical processes  imaging . It has  imaging –20.in vivo using different imaging modalities and new diagnostic and therapeutic markers. Scintigraphy is in the forefront of molecular imaging because it can directly visualise molecular events occurring at subnanomolar to milimolar levels. Findings in PET and SPECT will continue to direct the progress of molecular imaging [Molecular imaging is a diagnostic method based on the observation of cells and molecular structures  imaging . Molecul imaging . Molecul imaging .Optimal imaging techniques use light emitted through fluorescence or bioluminescence. Biomedical optical imaging is used to investigate tissues from the organelle level to the organ level, to assist in the detection, diagnosis, and treatment of pathological processes in noninvasive or minimally invasive ways to the body. Applications of biomedical optical imaging range from the use of fluorescent and bioluminescent techniques to help identify biomolecular distributions within cells, to micrometre-scale cross-sectional imaging of the retina, to imaging and selective treatment of tumours , 25. Becin vivo imaging at cellular level, to evaluate early neoplastic changes, to enhance molecular imaging performance, to develop novel imaging devices such as image-guided biopsy probes or devices for imaging inside the body, and portable low-cost optical imaging methods for brain function (3).New optical imaging techniques include optical coherence tomography, multiphoton microscopy, total internal reflection fluorescence, and speckle microscopy. Research priorities that are currently being performed or will be targeted include: The classic computer applications are computer-aided detection and computer-aided diagnosis (CAD) in mammography, lung and colon cancer –33.Screening mammography was the first modality to benefit from CAD and clinical studies of CAD system have been conducted successfully to demonstrate the additional value it offers for the detection of breast cancer. The currently accepted model for CAD in mammography is to use it as a “second reader”—that is, the radiologist first searches the images for masses and microcalcifications, and then the computer assists the radiologist to identify additional suspicious regions that may be cancerous, with the final diagnosis and decisions concerning patient care being made by the radiologist , 27. UseThe early detection and removal of colonic polyps can substantially reduce the risk of developing colon cancer. CT colonography, or virtual colonoscopy, has shown promise as a minimally invasive technique to help detect polyps , 30. ManThe detection of subtle focal pulmonary opacities on chest radiograph and CT remains a challenge. While screening and related studies have shown that CT is very sensitive for the detection of lung nodules, it is not very specific for the detection of lung cancers. Hence, there is substantial interest in the development of CAD systems to assist radiologists with the detection of nodules as well as characterisation of detected nodules as benign or malignant. CAD techniques are developed to improve interpretation in chest imaging with the aim of decreasing the intrinsic limitations and variations of human perception by alerting the reader to suspicious areas in chest radiographs and CTs , 33.et al. found that using CAD to detect lesions on images provided less added value to radiologists than CAD diagnosis [Studies have shown that CAD systems have improved the sensitivity in the detection of pulmonary nodules, especially for nodules smaller than 5 mm on CT examinations, which are often overlooked by visual inspection alone , 33. Theiagnosis . Thus, iBiomedical imaging has played an important role in identifying and monitoring the effectiveness of the best treatments for many diseases. The ultimate goal of treatment is to identify the target of treatment and to deliver the maximum therapy to that target with minimal or no damage to the normal cells. Image-guided surgery improves the accuracy of many surgical procedures by eliminating much of the speculation, reducing the risk of human error, and minimising variations among the abilities of individual physicians , 39. ImaReal-time imaging guidance by means of CT, ultrasound or MRI is performed for various treatment options such as minimally invasive image-guided thermal ablation therapies for tumours of the liver, kidney, lung and bone (3). Image-guided minimally invasive therapies of the future may include the delivery of other therapeutics, including proteins, stem cells, gene therapies, chemotherapeutic agents and antibodies (3).biij, available at http://www.biij.org) is an open access multidisciplinary peer-reviewed quarterly online journal, published by the Department of Biomedical Imaging, University of Malaya, Malaysia. biij is a multidisciplinary journal covering all the clinical and technical aspects of biomedical imaging and intervention, radiotherapy and oncology, minimally invasive image-guided therapy, image processing and informatics, molecular medicine, medical physics and radiobiology as well as radiography and bioengineering related to imaging or intervention. Researchers are encouraged to fully utilise the biij as a platform for biomedical imaging research by publishing their original research works and sharing research outcomes with colleagues from the Asia Pacific region and the rest of the world.The Biomedical Imaging and Intervention Journal (


The function of the cerebral cortex requires the coordinated action of two major neuronal subtypes, the glutamatergic projection neurons and the GABAergic interneurons. Although, in terms of numbers, GABAergic interneurons represent a minor cell population compared to glutamatergic neurons in the neocortex, they play an important role in modulating network dynamics of neocortical circuits. Indeed, GABAergic interneurons have been shown to control neuronal excitability and integration, and they have been implicated in the generation of temporal synchrony and oscillatory behavior among networks of pyramidal neurons. Such oscillations within and across neural systems are believed to serve various complex functions, such as perception, movement initiation, and memory. Recently, the development of GABAergic inhibition has been shown to be a key determinant for critical period plasticity of cortical circuits. Critical periods represent heightened epochs of brain plasticity, during which experience can produce permanent, large-scale changes in neuronal circuits. Experience-dependent refinement of neural circuits has been described in many regions within the CNS, suggesting it is a fundamental mechanism for normal vertebrate CNS development. By regulating the onset and closure of critical periods, GABAergic interneurons may influence how experience shapes brain wiring during early life and adolescence. Considering the multifaceted role played by GABAergic cells in the development, function, and plasticity of neural circuits, it is not surprising that alterations in the development of GABAergic circuits per se have been implicated in various neurodevelopmental and psychiatric disorders such as schizophrenia, autism, and epilepsy. However, how modification of GABAergic circuit development contributes to specific pathologies is largely unknown. Furthermore, GABA mimetic drugs, such as benzodiazepines and certain antiepileptic drugs, are widely used in clinical practice, but whether and to what extent these drugs cause deleterious effect on the developing brain is still not clear. A better comprehension of the mechanisms underlying the development and plasticity of GABAergic interneurons will likely indicate which cellular substrates might be affected in neurodevelopmental disorders. At the same time, identifying the genetics variants implicated in these disorders may generate major new insights into the normal and pathological function of GABAergic circuits.Our understanding of GABAergic interneurons function is challenged by their startling heterogeneity; indeed, different subtypes of interneurons display distinct morphology, physiological properties, connectivity patterns, and biochemical constituents. Recent technical advances have significantly accelerated progress in this field. In particular, the development of genetic strategies based on interneuron cell type-specific promoters and fluorescent protein reporters has allowed efficient high-resolution labelling of specific GABAergic interneuron classes in intact or semi-intact tissues, such as organotypic brain cultures.Contributions to this special issue of provide an overview of recent discoveries in the field of GABAergic circuit development and related brain disorders. The genetic program for the construction of cortical GABAergic network is initiated early during brain development, and it orchestrates cell type specification, migration, and some aspects of synaptic connectivity. On the other hand, the establishment of mature patterns of GABAergic innervation and inhibitory transmission is not achieved until adolescence and is profoundly influenced by neuronal activity and experience. E. Rossignol describes the tightly controlled genetic cascades that determine the great diversity of cortical GABAergic interneurons and how dysfunctions in genes important for their generation, specification, and maturation might contribute to various neurodevelopmental disorders. B. Chattopadhyaya describes the molecular mechanisms underlying the activity-dependent maturation of GABAergic innervation in the postnatal brain. Several articles in the special issue have investigated the evidence linking dysfunction in GABAergic signaling and plasticity to specific neurodevelopment disorders, such as autism , schizophrenia , and epilepsy . The developmental role of GABAergic circuits is not limited either to the brain or to the developmental phase. A. E. Allain et al. discuss the role of GABA and GABAergic receptors in motoneuron development and in immature hypoglossal motoneurons of the spastic mouse, a model of human hyperekplexic syndrome. B. Imbrosci and T. Mittman describe the response of the GABAergic system to cortical injuries in the adult and how this response could be manipulated to help the functional recovery of patients.In the last decades, cell-based therapies using GABAergic neuronal grafts have emerged as a promising treatment, since they may restore the lost equilibrium by cellular replacement of the missing/altered inhibitory neurons or modulating the hyperactive excitatory system. Advances in this field are reviewed by V. Broccoli and M. Dolado. It is becoming increasingly clear that the strength of GABAergic synaptic transmission is dynamic. R. Wright et al. review some of the sophisticated ways in which GABA-A receptor driving force can vary within neuronal circuits. P. Méndez and A. Bacci discuss the plasticity and modulation of adult cortical and hippocampal GABAergic synaptic transmission, while P. Wenner describes new insight into the mechanisms of GABAergic homeostatis in developing motor networks. Finally, A. Ludwig et al. provide evidence that the trophin nurturin is implicated in the developmental regulation of the cotransporter KCC2, a key molecular player in the establishment of the chloride-gradient, which in turn regulates the strength of GABAergic transmission.We hope that this special issue will serve to emphasize the new technical and conceptual advances in the field of GABAergic circuits development and to highlight the importance of this network for neurological disorders.Graziella Di CristoGraziella Di CristoTommaso PizzorussoTommaso PizzorussoLaura CanceddaLaura CanceddaEvelyne SernagorEvelyne Sernagor


Osseous metaplasia in the gastrointestinal tract is a rare phenomenon.We present the case of a 62-year-old Hispanic man with two colonic polypoid lesions, one of which, upon resection and histopathological examination, was found to be a traditional serrated adenoma with a focus of stromal osseous metaplasia.Our patient’s case is the third report of stromal osseous metaplasia in a traditional serrated adenoma of the sigmoid colon. Epithelial metaplasia is a common occurrence in the gastrointestinal tract (GIT), especially in the upper GIT (excluding the liver and pancreas). However, mesenchymal metaplasia, particularly osseous metaplasia (OM), is a rare and incidental finding in the GIT and is composed of histologically benign bone tissue -3. FewerIn 1939, Dukes described four cases of OM in colonic adenocarcinomas; since then, it has been described in both benign and malignant GIT lesions. The rectal region is particularly affected (adenocarcinoma), although the spectrum of presentation includes Barrett’s esophagus; tubular, tubulovillous and serrated adenomas; juvenile and hyperplastic polyps; and appendicular tumors  of colonic mucosa (sigmoid) with exuberant, villiform growth pattern and complex serration. High power examination disclosed eosinophilic pencillate cells, numerous ectopic crypts and low-grade epithelial dysplasia. Figures  and 2 ThMesenchymal metaplasia, particularly osseous metaplasia, is an unusual finding in GIT, composed of histologically benign bone tissue. This is an unexpected variation of the usual histological features encountered in colorectal TSA. In 1939, Dukes described four cases of OM in colonic adenocarcinoma; since then, it has been described in both benign and malignant GIT lesions, but the rectal region is particularly affected .Despite the absence of a conclusive explanation of the induction mechanism of ossification, the hypothesis proposed by Rhone and Horowitz in 1976 postulated the metaplasia of a pluripotent cell into an osteoblast under the influence of factors generated by epithelial cells. One possible mechanism includes the production of bone morphogenetic proteins (BMP)-5 and BMP-6 by epithelial cells and BMP-2 and BMP-4 by adjacent fibroblasts ,13,15,16An extensive review of the literature reveals that OM is mostly described in association with colorectal carcinomas and adenomas (both tubular and villous); however, no risk association has been determined for sex, age or polyp size ,8,9,11. According to our bibliographic review, our patient’s case is only the third report of osseous metaplasia in the stroma of a traditional serrated adenoma located in the sigmoid colon. No characteristic has been identified that would suggest a traumatic or ischemic process, nor any foci of calcification, either proximal or distal.Written informed consent was obtained from the patient for publication of this case report and accompanying images. A copy of the written consent is available for review by the Editor-in-Chief of this journal.The authors declare that they have no competing interests.NM performed the histological examination and diagnosis of the colonic polyp. JB and LR conducted a thorough literature review on osseous metaplasia in the gastrointestinal tract and were the major contributors in writing the manuscript. All authors read and approved the final manuscript.


The model facultative aerobe Escherichia coli, the microbial cell factory par excellence, has elaborate sensing and signal transduction mechanisms that respond to the availability of electron acceptors and alternative carbon sources in the surrounding environment. In particular, the ArcBA and CreBC two-component signal transduction systems are largely responsible for the metabolic regulation of redox control in response to O2 availability and carbon source utilization, respectively. Significant advances in the understanding of the biochemical, genetic, and physiological duties of these regulatory systems have been achieved in recent years. This situation allowed to rationally-design novel engineering approaches that ensure optimal carbon and energy flows within central metabolism, as well as to manipulate redox homeostasis, in order to optimize the production of industrially-relevant metabolites. In particular, metabolic flux analysis provided new clues to understand the metabolic regulation mediated by the ArcBA and CreBC systems. Genetic manipulation of these regulators proved useful for designing microbial cells factories tailored for the synthesis of reduced biochemicals with added value, such as poly(3-hydroxybutyrate), under conditions with restricted O2 supply. This network-wide strategy is in contrast with traditional metabolic engineering approaches, that entail direct modification of the pathway(s) at stake, and opens new avenues for the targeted modulation of central catabolic pathways at the transcriptional level.Bioprocesses conducted under conditions with restricted O E. coli remains as the microbial cell factory (MCF) par excellence, mainly because this well- characterized enterobacterium can easily and rapidly grow on cheap substrates and can be modified as desired through a broad variety of molecular tools. As fermentation technologies designed for the production of bulk bioproducts need further innovation to became economically and environmentally sound , the environmental performance of PHAs equals that of petrochemical polymers. This situation clearly calls for alternative bioprocesses and biocatalysts to meet the technical and environmental requirements of a sound production process.The design and development of efficient MCFs for PHA production received much attention in the last few decades. Although different microorganisms have been exploited for polymer production, catalyst  as it docatalyst . The maiE. coli arc mutants are unregulated for respiration under micro-oxic conditions. Some enzymes of the TCA cycle are not repressed and the pool of reducing equivalents is elevated, and thus available to be funneled into reduced biochemicals. The PHB biosynthetic genes from Azotobacter sp. strain FA8  circumventing overflow metabolism by re-directing central metabolic fluxes and preserving sufficient precursors for amino acids synthesis, the actual building blocks of proteins.Acetate formation in organism , 97, 98.organism . This, iorganism . These aE. coli. Vemuri et al.  enhancement in the rate of substrate uptake, [ii] reduction of flux(es) to undesirable by-products, and enhancement of precursors and cofactor availability, [iii] introduction of relevant heterologous pathway(s) and optimization of the activity of its constituent enzymes, and [iv] export of the product to the extracellular medium in order to shift the concentration equilibrium towards product formation.In the first place, genome-scale and specific metabolic models are starting to play an important role in deciphering the cellular and metabolic characteristics of microbial cells n silico , 110. ThThis exciting and extremely dynamic scenario will certainly lead to the development of better strategies to manipulate central and peripheral pathways to enhance the production of reduced biochemicals and other molecules of industrial interest.Click here for additional data file.


We compare the sets of experimentally validated long intergenic non-coding (linc)RNAs from human and mouse and apply a maximum likelihood approach to estimate the total number of lincRNA genes as well as the size of the conserved part of the lincRNome. Under the assumption that the sets of experimentally validated lincRNAs are random samples of the lincRNomes of the corresponding species, we estimate the total lincRNome size at approximately 40,000 to 50,000 species, at least twice the number of protein-coding genes. We further estimate that the fraction of the human and mouse euchromatic genomes encoding lincRNAs is more than twofold greater than the fraction of protein-coding sequences. Although the sequences of most lincRNAs are much less strongly conserved than protein sequences, the extent of orthology between the lincRNomes is unexpectedly high, with 60 to 70% of the lincRNA genes shared between human and mouse. The orthologous mammalian lincRNAs can be predicted to perform equivalent functions; accordingly, it appears likely that thousands of evolutionarily conserved functional roles of lincRNAs remain to be characterized. Genome analysis of humans and other mammals reveals a surprisingly small number of protein-coding genes, only slightly over 20,000 . Recent analysis of the mammalian genomes and transcriptomes, in particular, using the RNAseq technology, shows that, in addition to protein-coding genes, mammalian genomes encode many long non-coding RNAs. For some of these transcripts, various regulatory functions have been demonstrated, but on the whole the repertoire of long non-coding RNAs remains poorly characterized. We compared the identified long intergenic non-coding (linc)RNAs from human and mouse, and employed a specially developed statistical technique to estimate the size and evolutionary conservation of the human and mouse lincRNomes. The estimates show that there are at least twice as many human and mouse lincRNAs than there are protein-coding genes. Moreover, about two third of the lincRNA genes appear to be conserved between human and mouse, implying thousands of conserved but still uncharacterized functions. The great majority of mammalian genome sequences are transcribed, at least occasionally, a phenomenon known as pervasive transcription A popular view that the vast majority of lincRNAs are by-products of background transcription, “simply the noise emitted by a busy machine” Given the general lack of strong sequence conservation, identification of lincRNAs on genome scale relies on expression analysis which makes comprehensive characterization of the mammalian lincRNome an elusive goal. The combination of different experimental approaches applied to transcriptomes of several species has resulted in continuous discovery of new transcripts Here we re-analyze the most reliable available sets of human and mouse lincRNAs using the latest next generation sequencing (RNAseq) data and apply a maximum likelihood approach to obtain a robust estimate of the size of the mammalian lincRNome. The results suggest that mammalian genomes are likely to encode at least twice as many lincRNAs as proteins.We performed comparative analysis of the recently reported validated sets of 4662 human lincRNAs A substantial fraction of the vast mammalian transcriptome, most likely the lower expressed transcripts, is expected to be non-functional. Therefore, to minimize the contribution of transcriptional noise, cut-off values were imposed on expression levels of lincRNA genes and their putative orthologs that were used for the lincRNome size estimation. Similarly, a series of cut-off values was applied for the fraction of indels in pairwise genomic alignments .The sets of experimentally validated lincRNAs are random samples from complete sets of lincRNAs (lincRNomes) for the corresponding species.The experimentally validated lincRNA sets for human and mouse are uncorrelated with each other.Lh and Lm be the sizes of the experimentally validated sets of lincRNAs for human and mouse, respectively. Also let Kh be the number of confirmed human lincRNAs that have an expressed orthologous sequence in mouse and Km be the corresponding number of mouse lincRNAs. Finally, Kb is the number of confirmed, expressed human lincRNAs whose orthologs in mouse are also confirmed lincRNAs. If the orthology relations between the human and mouse lincRNAs are strictly one-to-one, the number of confirmed mouse lincRNAs for which the human ortholog is also a confirmed lincRNA should be Kb as well. This is indeed the case in practice, with a few exceptions.Let Nb, ii) unique to human, Nh-Nb, and ii) unique to mouse, Nm-Nb; here Nh and Nm are the total sizes of the complete human and mouse lincRNomes, respectively. Assumption (2) allows us to compute the probability of observing a particular set of Kh, Km and Kb simply by counting the number of possible samples of Lh and Lm lincRNAs drawn at random from the respective pools of Nh and Nm that result in the given set of Kh, Km and Kb values:Given assumption (1), the lincRNAs can be partitioned into three pools: i) those present in both species, pool size To assess the robustness of the estimates, ranges of open reading frame size thresholds used to eliminate putative protein-coding genes and RPKM  .The relatively poor sequence conservation and often low expression of lincRNAs hamper robust estimation of the size of the lincRNome from expression data alone and render comparative-genomic estimation an essential complementary approach. Strikingly, the estimates obtained here by combining comparative genomic and expression analysis suggest that the mammalian lincRNome is at least twice the size of the proteome In order to assess the reliability and robustness of the model with respect to parameters, we produced series of estimates of the total size of the human and mouse lincRNomes and their conserved subset with varying thresholds on expression level, extent of sequence similarity and the maximum allowed open reading frame size. Nevertheless, it is impossible to rule out some sources of bias that might have affected the estimates. For example, some orthologous lincRNA genes might remain undetected because they were not included in the UCSC genome alignments due to high divergence or synteny breaks in . Such under-detection of orthologs could cause an underestimate of evolutionary conserved lincRNA genes although it has been reported that the of breakpoints is not large <250) for the human/mouse genomic comparison 50 for thcis to regulate protein expression but also intrinsically and independently in trans, likely as non-coding RNAs Some of the transcripts identified as lincRNAs potentially might represent fragments generated from long  5′UTRs or 3′UTRs of protein-coding genes. Such transcripts could results from utilization of alternative poly(A) addition signals and/or could represent alternative splice forms separated by long introns de novo gene birth The possibility that some lincRNA genes encode short peptides that are translated, perhaps in a tissue-specific manner, is the subject of an ongoing debate Taking all these problems into account, here we adopted a simple, conservative approach by excluding from the analysis lincRNAs containing relatively long ORFs, under a series of ORF length thresholds. However, it should be noted that human and mouse lincRNAs used in this study had been previously filtered for the presence of evolutionary conserved ORFs and the presence of protein domains, and the most questionable transcripts were removed at this stage Questions about functional roles of lincRNAs and the fraction of the lincRNAs that are functional loom large. For a long time, the prevailing view appeared to be that, apart from a few molecular fossils such as rRNA, tRNA and snRNAs, RNAs did not play an important role in extant cells. More recently, the opposite position has become popular, namely that  every detectable RNA molecule is functional. It has been repeatedly pointed out that this view is likely to be too extreme The results of this work predict that, despite the fact that on average sequence conservation between orthologous lincRNAs is much lower than the conservation between protein-coding genes As the human lincRNA data set, the ‘stringent set’ of 4662 lincRNAs, which is a subset of the over 8000 human lincRNAs described in a recent comprehensive study http://www.ncbi.nlm.nih.gov/Traces/sra; Study ERP000546; runs ERR030888 to ERR030903). For mouse, RNAseq data of the ENCODE project ftp://hgdownload.cse.ucsc.edu/goldenPath/mm9/encodeDCC/wgEncodeLicrRNAseq/).Expression of the lincRNAs was assessed by analysis of the available RNAseq data. For human, the run files of the Illumina Human Body Map 2.0 project for adipose, adrenal, brain, breast, colon, heart, kidney, liver, lung, lymph node, ovary, prostate, skeletal muscle, testis, thyroid, white blood cells, were downloaded from The NCBI Sequence Read Archive . The reads were aligned with the cognate genomic sequences using TopHat Pre-built Bowtie indices of human and mouse, based on UCSC hg19 and mm9, were downloaded from Bowtie FTP site (http://www-huber.embl.de/users/anders/HTSeq) to calculate the number of aligned reads (“counts”). The RPKM  and ending with a stop codon. The ORFs were identified by using the ATG_EVALUATOR program Kh), human lincRNA gene coordinates of assembly hg19 were converted to mouse mm9 using the liftOver tool of the UCSC Genome Browser Lh), for 3529 putative orthologous regions were identified in the mouse genome. These sequences were checked for the evidence of expression in mouse tissues using the RNAseq data. Exon coordinates of putative lincRNAs were obtained by mapping their coordinates onto exons of all known genes of mm9 assembly of UCSC Genome Browser. The sums of exons were then used in expression level calculation to normalize for sequence length. Out of the 3369 putative lincRNAs for which the exon models could be determined, 2872 had expression level greater than zero. Similarly, the subset of mouse lincRNAs with expressed putative orthologs in human (Km) was found by converting the coordinates of initial 4156 mouse lincRNAs (Lm) from mm9 to hg19 and searching for the evidence of expression in human tissues. The exon models could be determined for 3656 of the 3677 putative lincRNAs, out of which 3157 had expression level greater than zero.To obtain the subset of human lincRNAs with expressed orthologs in mouse (Kb) was obtained by selecting those lincRNAs whose putative orthologs in another species overlap with the validated lincRNAs of that species. That is, we searched for the overlap of putative orthologs of human lincRNAs (in hg19 coordinates) with the mouse lincRNAs . The overlap was determined using intersectBed from BEDtools package with the command line option -s (“force strandedness”). This resulted in 196 pairs of unique human and mouse lincRNAs. Approximate indel values were estimated from the sequence length differences between the lincRNAs and their orthologs, i.e. the following formula was used:llincRNA is the total length of lincRNA exons, and Lortholog is the total length of the exons of lincRNA ortholog. Manual examination of orthologous lincRNA alignments and putative orthologs suggested that approximately 5% of the alignments with the largest INDEL values were unreliable. Thus, all lincRNA alignments with INDEL >95% were removed from further analysis. Similarly, a cut-off was imposed on expression level of putative human and mouse orthologs of lincRNA. This cut-off was set at the lowest 5% of the expression levels of the 196 orthologous validated lincRNA genes  levels, and all ORF thresholds. The results of bootstrap analysis are given in the Supporting Table S1Comprehensive information on the human and mouse lincRNA sets.(XLS)Click here for additional data file.


Nevertheless, the knowledge of the mechanism(s) underlying this fundamental step in embryogenesis is still scarce, especially if compared to other model organisms. We will review here the current knowledge on the post-transcriptional control of gene expression in mouse early embryos and discuss some of the unanswered questions concerning this fascinating field of biology.Fertilization is a very complex biological process that requires the perfect cooperation between two highly specialized cells: the male and female gametes. The oocyte provides the physical space where this process takes place, most of the energetic need, and half of the genetic contribution. The spermatozoon mostly contributes the other half of the chromosomes and it is specialized to reach and to penetrate the oocyte. Notably, the mouse oocyte and early embryo are transcriptionally inactive. Hence, they fully depend on the maternal mRNAs and proteins stored during oocyte maturation to drive the onset of development. The new embryo develops autonomously around the four-cell stage, when maternal supplies are exhausted and the zygotic genome is activated in mice. This oocyte-to-embryo transition needs an efficient and tightly regulated translation of the maternally-inherited mRNAs, which likely contributes to embryonic genome activation. Full understanding of post-transcriptional regulation of gene expression in early embryos is crucial to understand the reprogramming of the embryonic genome, it might help driving reprogramming of stem cells  Both gametes are shaped and programmed to fuse with each other as they touch. In just few hours after fertilization, the haploid parental pronuclei become clearly visible inside the mouse zygote (one-cell embryo) . Before Inside the environment provided by the oocyte, a complex series of remarkable tasks has to be accomplished rapidly and precisely. First, fertilization triggers the completion of the II meiotic division (MII) of the oocyte. Then, the zygote must resume the mitotic cell cycle, remodel the chromatin, activate transcription, and initiate the embryonic developmental program ,3 Figur. NoteworThe two-cell embryo stage marks the transition from maternal-to-zygotic gene dependence ,5. In faFertilization creates totipotent cells capable of generating all tissues of the new organism. In this respect, the one-cell embryo can be considered a totipotent cell. However, since the mouse zygote is essentially a transient state in which a diploid nucleus is never formed, the first totipotent cells originated by fertilization are the blastomeres of the two-cell embryo. As embryonic development proceeds, this capability will be progressively reduced as a consequence of the increasing differentiation of the daughter cells into specific cell types. At the blastocyst stage, totipotency will be restricted to pluripotency, but the exact time when this boundary arises is still controversial –10. For In this review we will focus our attention on one peculiar aspect of early embryonic development: the post-transcriptional control of gene expression. This step in gene expression is essential for the zygote and it is likely to be relevant for the correct onset of the embryonic developmental program. In particular, we will underline the strict connection between a finely tuned translation regulation, chromatin remodeling and zygote genome activation in the mouse embryo. We will also discuss some intriguing new findings about the intracellular localization of maternal mRNAs in mouse.2.Chromatin is thought to be the principal controller of the access of the transcription machinery to the mammalian genome. Chromatin structure is organized in nucleosomes, wherein double-stranded DNA is wrapped around histones, and higher-ordered structures obtained by folding of the chromosomes . Genome Chromatin modifications play an essential role during embryonic development, permitting different cells and tissues to progressively acquire specific patterns of gene expression . ExtensiUpon fertilization, the highly condensed paternal chromatin is rapidly reorganized in order to acquire the appropriate epigenetic state. First, protamines are replaced by histones, followed by active DNA demethylation of the chromatin in the paternal pronucleus that will be completed before DNA replication begins ,23. Thus3.Smarca4) is required for histone methylation and for ZGA in the two-cell embryo [Recently, a novel role for sperm epigenetic marks has been proposed in the establishment of embryonic totipotency . Cairns l embryo . Importal embryo . Chromatl embryo .Dppa3, developmental pluripotency-associated 3) is an oocyte-specific maternal-effect protein. STELLA is required to maintain methylation marks on most of the maternal genome [de novo establishment of germline methylation, and also plays a broader role in maintaining maternal and paternal methylation patterns in early embryos [Zar1 knockout zygotes pronuclei form and DNA replication starts, but the maternal and paternal genomes remain separate for unknown reasons [Maternally-inherited proteins have also been linked to maintenance of the parental imprinting in the early embryo. STELLA .Dicer and RISC activity and are likely to be responsible for maternal RNA degradation in zygotes. Since the field of non-coding RNAs is rapidly expanding, it is likely that new insights into their contribution to gene expression regulation in early mammalian development will be unveiled soon.The total mRNA levels decrease by 60% from MII-arrested oocytes to late 1-cell embryos , and deg6.Maternal mRNAs are stored during oogenesis in an inactive state until they are recruited for translation. Their unmasking is accompanied by the elongation of the poly(A) tail, followed by complex changes in protein synthesis patterns ,52–54. TD. melanogaster embryo. A remarkable example is represented by the Bicoid (bcd) mRNA, which is pre-localized at the anterior pole of the oocyte through interaction with sequence-specific RNA-binding proteins (RBPs), and whose translation after fertilization produces a Bcd protein gradient that determines the anterior cell fate [In a wide range of vertebrate and invertebrate embryos, the intracellular localization of mRNAs directs protein synthesis to subcellular domains, thereby establishing embryonic polarity . Translaell fate ,62.C. elegans gonads and embryos, RBPs control the spatial and temporal expression of specific genes [Also in ic genes ,64. For ic genes ,66. Furtic genes ,68.in vivo and to provide clear-cut answers to these issues.When mRNAs are not engaged in translation, they must be saved from the default pathway of degradation. For this reason, RNAs are always bound to proteins in complexes called ribonucleoproteins (RNPs) within the cell. RNPs, in turn, can be organized in multi-molecular structures named RNA granules, which change in shape, components and size in response to environmental cues. In somatic cells two types of granules, the P bodies and the stress granules, have been extensively studied ,70. The 7.It has been recently proposed that the cortex of growing oocytes serves as an mRNA storage compartment, and that it contains a new kind of transient RNA granules related to P-bodies, named sub-cortical RNP domain (SCRD) . Althouget al. [et al. [Padi6 is a maternal effect gene required for development beyond the two-cell stage and for CPL formation. Padi6 null embryos showed impaired transcription and dysregulation of mRNA translation, supporting the hypothesis that the two-cell arrest was due to defective activation of the embryonic genome. A similar role in CPL formation has been described for the maternal-factors MATER [Some recent papers hypothesize that a structure named cytoplasmic lattice (CPL) could be the site from which maternal factors are released during early embryogenesis, thus playing a role in translational control of maternal transcripts. CPL is a fibrillar matrix that was described many years ago in the oocyte as composed of RNAs and proteinaceous components. This structure persists until the blastocyst stage but its function has remained elusive, even though it was proposed to represent a ribosomal storage site –79. Espoet al.  and Yurt [et al.  demonstrrs MATER  and FLOPrs MATER . This lars MATER . Interesrs MATER . This cors MATER ) and TLErs MATER . Altoget8.C. elegans GLD-1 protein, Sam68, SLM-1 and SLM-2, are also potential candidates for a role in early embryogenesis [The events leading to the development of an organism must be strictly and timely controlled. At the onset of embryogenesis, post-transcriptional mechanisms are the main regulators of gene expression. Although in lower multicellular organisms, these mechanisms rely on the function of RBPs –63, muchogenesis . In partogenesis  and sperogenesis , and it ogenesis  and femaogenesis . Neverthogenesis , possiblogenesis . Altoget9.in vitro culturing of mammalian oocytes and embryos to be used for assisted reproduction. In mammals, the paucity of the starting material has strongly limited classical biochemical approaches used to elucidate the molecular mechanisms of post-transcriptional control of gene expression. Nevertheless, the amazing improvement of in vivo techniques for the analysis of protein and RNA functions, coupled with the development of high-sensitivity molecular tools, is rapidly filling the gap of our knowledge on the regulation of post-transcriptional control of gene expression also in mammalian oocytes. Thus, although much still remains to be discovered in order to build up a comprehensive vision of the mechanisms driving the onset of mammalian development, it is foreseen that the next years will witness a bloom of studies in this fascinating field of biology.Fertilization is the most important step for conservation of the species. The fusion of two gametes is required to generate a new organism in higher eukaryotes, and assortment of maternal and paternal chromosomes ensures diversity of the offspring. Importantly, the first steps of embryo development occur in the absence of transcription of the embryonic genome, and rely on post-transcriptional regulation of maternally-inherited mRNAs. Thus, appropriate control of degradation or translation of maternal transcripts is likely required to drive the onset of embryonic genome activation. It is predictable that full understanding of post-transcriptional regulation of gene expression in early embryos will shed light on the molecular mechanisms that regulate reprogramming of the embryonic genome. This knowledge might open new avenues also for the reprogramming of stem cells to be used in regenerative medicine or for the treatment of genetic diseases. On the other hand, such studies might also aid in improving protocols for


Anterior cervical spine fusion and stabilization with plating are well-established surgical procedures for the treatment of myelopathy, cervical spine traumas, and spinal infectious diseases. Various complications have been described in the literature, more frequently, intraoperative bleeding, peri- or postoperative hypopharyngeal, and/or esophageal ruptures with mediastinal deep infection and loosening and extrusion of the screws from the plating. Screw migration has also been observed as a complication of the procedure, either early in the postoperative period or delayed, even after many years. In some instances, the esophageal perforation can recover spontaneously with absence of complications, even if a case of plate failure and graft migration with lethal sudden airways obstruction has been reported. We describe a case of hypopharyngeal screw migration after cervical spine stabilization with plating never described before in the literature. Due to the improved safety and outcomes, anterior surgical approaches to the cervical spine have gained popularity in the last decade. More commonly, the indications for this procedure are represented by cervical degenerative, neoplastic, and traumatic lesions.Several complications are reported, mostly due to bleeding and rupture of hypopharyngeal and/or esophageal walls with mediastinal deep infection and loosening and extrusion of the screws from the plating. Though rare, migration of the screw after plating has been reported –12, the A 65-years-old lady was referred to us from the Emergency Department, due to odynophagia, and sense of foreign body in her throat suddenly occurred 7 days before without any other symptom. Three years before, the patient underwent surgery with cervical spine stabilization with plating for C5-C6 disc herniation with a good outcome and absence of complications, apart from moderate dysphagia in the early postoperative period. A metallic foreign body (screw) was easily visible during laryngoscopy  in the pA microlaryngoscopy was then performed, the screw  being eaRadiographic  and endoIn the last decade, there have been a growing development and refinement of surgical procedures indicated for spinal stabilization and fusion for cervical degenerative, neoplastic, and traumatic lesions. These procedures contemplate an anterior cervical access and the placement of plates and screws to guarantee an immediate stabilization and to improve the rate of bone fusion, avoiding the use of prolonged bracing with shorter hospital stay and early mobilization. Nonetheless, different complications can occur, such as cervical soft tissue swelling, hematomas, bleeding, breaches of the pharyngeal or esophageal walls, and, less often, screw displacement or migration. The latter, though rare, has been reported, the esophagus being the most frequent site of migration –12.Esophageal injury after anterior cervical spine procedures is a rare complication. Sun described in a cohort of 2348 patients who underwent anterior cervical surgery only 5 esophageal perforations (0.2%) . Screw mMigration can also be observed further on. Gazzeri et al.  reportedIn some instances, the esophageal perforation can spontaneously recover with absence of late complications. Yee and Terry  and PompFailure at the screw-bone interface occurs by either screw backout or screw cutout, the main predisposing factor that exposes to screw pullout appearing to be the initial malposition or suboptimal position of the screw.Coe and Vaccaro , in theiAs a matter of fact, in the most updated devices, the screws can be locked to avoid the risk of implant loosening and/or dislodgement with possible upper digestive tract damages.Anterior cervical spine fusion and stabilization with plating are well-established surgical procedures for the treatment of degenerative, traumatic, or infective nature. Various complications have been described in the literature, either in the early postoperative period or delayed. Among the latter, hypopharyngeal and/or esophageal ruptures with mediastinal deep infection and loosening and extrusion of the screws from the plating have been reported. In case of screw migration, dysphagia can often represent the only symptom. Considering the long time required for the screw to migrate , an accurate endoscopy and radiographic study of upper aerodigestive tract are in our opinion strongly advisable to promptly rule out a hypopharyngeal or esophageal foreign body when the patient's history reveals cervical spine previous surgery.


Glioblastoma Multiforme (GBM) is the most common and lethal of human primary central nervous system (CNS) tumors, with a median survival of 14-16 months despite optimal surgery, radiation and chemotherapy. A reason for this dismal prognosis is insufficient understanding of the ontogeny of GBMs, which are highly heterogeneous at a pathological level. This pathological diversity, between and within GBMs as well as varying grades of gliomas, has not been fully explained solely on the grounds of oncogenic stimulus. Interaction with the tumor microenvironment is likely a source of this pathological heterogeneity, as well as the inherent characteristics of the tumor cell of origin. Currently, controversy exists on whether the initial transformed cell is a differentiated astrocyte, progenitor or neural stem cell. Putative Cancer Stem Cells (CSCs), which have features of normal stem cell plus the ability to recapitulate the tumor phenotype in vivo in small numbers, have been identified from a variety of solid human cancers, including GBMs. Evidence suggesting that regions harboring normal stem cells in the adult CNS, such as the subventricular zone and the dentate gyrus, are more prone to viral and chemical oncogenesis, is supportive of the hypothesis that brain tumors arise from stem cells. However, it is still to be determined whether the appearance of brain tumor stem cells (BTSC) is the cause or consequence of tumor initiation and progression. This review discusses emerging evidence highlighting the relevance of the state of differentiation and regional heterogeneity in the ontogeny of GBM. This is an area of high interest in cancer in general, with potential significant therapeutic and prognostic implications. Gliomas compose almost 60% of total human primary CNS malignancies , 2. ThesFor decades, it was widely assumed that differentiated glia were the only cells capable of transformation as the adult brain was thought to be mitotically inactive . The demConditional mouse modeling combined with lineage tracing techniques makes it possible to target these distinct neurogenic regions in adults, as well as cell populations at different stages of their differentiation. These and other reagents have facilitated experiments towards a better understanding of the ontogeny of GBMs, which have underscored the relationship between gliomagenesis and brain development. A relationship further highlighted by the The Cancer Genome Atlas (TCGA) which sub-classified GBMs to four gene signature profiles , which are characteristic of distinct stages, prognosis and patterns of differentiation , 20. ThiCNS development proceeds in a hierarchical manner; whereby, mature differentiated cells such as neurons and the varying glial subtypes  arise from CNS stem cells. These CNS stem cells arise from the pseudostratified neuroepithelium that lines the cerebral ventricles early in embryonic development and stains positive for Nestin . By the The subventricular zone (SVZ) is the largest germinal region, and it exhibits a high degree of organization, Figure Similarly, neural stem cells within the hippocampus seem to have a radial glial origin, Figure A third population of CNS precursors in the adult brain, is a subpopulation of NG2 expressing cells shown to play a glial precursor role. The NG2 proteoglycan is believed to be an in vivo marker for oligodendrocytes progenitors found in the developing brain , 29, 30.All of the neural stem and progenitor populations described above have different proliferative capacities and varying ability to generate astrocytes, oligodendrocytes and neurons that may reflect distinct patterns of gene expression. Thus regional differences in neuroglial precursor populations may dictate a significant component of CNS tumor gene expression and biology.All forms of cancer arise from the acquisition of genetic alteration, leading to several aberrant biological properties required for transformation. These biological properties include, sustained proliferation, evading growth suppression, resisting cell death, enabling replicative immortality, inducing angiogenesis and activating invasion and metastasis . In the PDGFR, IDH1/IDH2, EGFR or NF1 genes respectively [Comparative gene expression analysis in leukemias show that the different forms of leukemias appear to arise from distinct points in the hematopoietic lineage, that are susceptible to specific translocations. Furthermore, the biology behind this translocations-lineage stage mechanism might lie in the capacity of translocations to activate key leukemogenic programs , 34, reiectively , 20, 39.There exists debate about the Cancer Stem Cell (CSC) hypothesis and the existence of BTIC in brain tumors. Unlike leukemias discussed above, there is no singular cell surface or genetic signature of the BTIC. Initial enthusiasm with CD133 has recently been questioned by studies demonstrating that alternations in mitochondrial function among glioma cells induce CD133 expression. ConversAnother but not mutually exclusive hypothesis is that some GBMs can arise from the transformation of terminally differentiated glia. Retroviral transduction of INK4a/Arf-/- mature astrocytes with a constitutively active mutant EGF receptor (EGFRvIII), prevalent in human GBMs, induces astrocyte de-differentiation and GBM formation. A phenoTo elucidate the ontogeny of gliomas conditional and somatic strategies coupled to lineage tracing techniques have come to the forefront, as summarized on Table The first germline derived mouse models for brain tumors, were models that over expressed viral oncogenes . However12Ras) under the control of a human hGFAP promoter was developed [To study the role of Ras signaling in gliomas, a mouse model carrying a constitutively active Ras , but not of neural stem cells or other lineages was observed. Further transcriptome and marker staining analysis confirmed upon tumor formation the OPC nature of the tumor cells. This demonstrates that in p53/Nf1 mutant driven gliomas, mutations may initially occur in either NSC or OPC, but only OPC provides the susceptible cellular context needed for transformation [Despite all the insights from the models discussed above, the true cell of origin is still controversial. This mainly due to the fact that the cell of origin is difficult to identify by analyzing cells within terminal stage tumors, whose genetic identity could be concealed by the acquired plasticity. A recent study aimed at addressing this issue analyzed proliferative abnormalities in distinct lineages prior to malignancy . Using aormation . This hiGerminal regions such as the SVZ have long been proposed as more gliomagenic, as this region is more susceptible to viral and chemical oncogenesis -61. In cNf1/p53 flox background induced astrocytoma formation with 100% penetrance [Work by Alcantarallaguno et al. demonstrates that viral mediated Cre-recombinase targeting of the SVZ in an netrance . Conversnetrance .Rb, p53 and Pten Jacks et al showed that the initial combination of genetic mutations in the SVZ dictates the pathogenesis and phenotype of the tumors that develop. In these mice, recombination of Pten/p53 gave rise to gliomas, whereas deletion of Rb/p53 generated primitive neuroectodermal tumors (PNET) [Using mice bearing conditional alleles of s (PNET) . This moOther retroviral-based approaches such as the RCAS system have allowed lineage tracing of specific cell populations through the process of tumorigenesis . The virIn contrast to targeting the neural-glial progenitor population, a study used the RCAS system, with injection of the RCAS-PDGF + RCAS-Cre virus into various regions of a Nestin-tva; Ink4aARF null; Pten flox adult mice. These regions included the SVZ, left and right cerebral hemispheres and cerebellum, with development of gliomas in all of the areas targeted . SimilarMouse models of gliomagenesis have been pivotal in determining how the process of differentiation is integrally related to the process of gliomagenesis and the maintenance of the neoplastic phenotype. They indicate that not only the nature of transforming genetic alteration, but also the differentiation status of the neuro-glial cell, modulates gliomagenesis. Of interest, the oncogenic stimuli can themselves alter the state of neuro-glial differentiation, to make them more susceptible to transformation.However, the question of ontogeny of gliomas remains in its infancy despite these mouse models. One of the problems is that the models to date target, using broadly expressed promoters such as Nestin and GFAP. For example, targeting the SVZ with CMV, GFAP or Nestin promoters target the stem and progenitor cells, but also differentiated glia that resides in this region. Use of cell specific promoters to drive expression of transforming alterations in neuro-glial cells at different stage of the normal lineage hierarchies is crucial, towards our understanding of initiating events in gliomagenesis within a defined cellular compartment. Identifying the cell of origin of gliomas and the region in the brain where these cells reside would permit a more systematic analysis of the genetic lesions involved in tumor initiation and progression. Moreover, the gene signature of the cell of origin may elucidate key molecular pathways and driver mutations that could lead to the development of new therapeutic approaches to prevent and target early stage disease. Finally, the integration of developmental biology, genomics and cancer biology will lead to the production of more accurate models, where single cell genetic manipulations in unambiguously labeled cells, will allow a better understanding of the ontogeny of gliomas.


Energy balance plays an important role in the control of reproduction. However, the cellular and molecular mechanisms connecting the two systems are not well understood especially in teleosts. The hypothalamus plays a crucial role in the regulation of both energy balance and reproduction, and contains a number of neuropeptides, including gonadotropin-releasing hormone (GnRH), orexin, neuropeptide-Y, ghrelin, pituitary adenylate cyclase-activating polypeptide, α-melanocyte stimulating hormone, melanin-concentrating hormone, cholecystokinin, 26RFamide, nesfatin, kisspeptin, and gonadotropin-inhibitory hormone. These neuropeptides are involved in the control of energy balance and reproduction either directly or indirectly. On the other hand, synthesis and release of these hypothalamic neuropeptides are regulated by metabolic signals from the gut and the adipose tissue. Furthermore, neurons producing these neuropeptides interact with each other, providing neuronal basis of the link between energy balance and reproduction. This review summarizes the advances made in our understanding of the physiological roles of the hypothalamic neuropeptides in energy balance and reproduction in teleosts, and discusses how they interact with GnRH, kisspeptin, and pituitary gonadotropins to control reproduction in teleosts. A close connection between energy balance and reproduction has been well documented in mammals . Energy Initiation of reproduction is affected by the amount of body energy reserves and is responsive to diverse metabolic factors. The neuroendocrine mechanisms responsible for the association between energy balance and fertility are represented by metabolic hormones and neuropeptides that affect the hypothalamic center controlling the expression and release of gonadotropin-releasing hormone (GnRH) , 3. TherMost feeding-related neuropeptides in mammals have also been identified in fish species , suggestThis review focuses on the role of the neuropeptides that regulate feeding and energy balance on reproduction in teleosts, and discusses if the metabolic control of reproduction is conserved from fish to mammals.In teleosts, as in other vertebrates, reproduction is coordinated by the HPG axis. The hypothalamus produces GnRH, which regulates the synthesis and release of gonadotropins (GTHs), follicle-stimulating hormone (FSH), and luteinizing hormone (LH), from the pituitary. The GTHs act on the gonads to stimulate gonadal development through the secretion of sex steroid hormones. These steroids, in turn, feedback to the brain and the pituitary to complete the HPG axis and to regulate the reproductive cycle , 11. ThuIn the early 1970s, two research groups simultaneously reported the isolation of a LH-releasing factor from the hypothalamus of pigs and sheep , 13, andDistribution of three different forms of GnRH in the brain was first reported in a perciform fish, the sea bream . GnRH1 nThe function of GnRH in the central regulation of LH release has been recognized in all orders of teleosts. Although the assay for FSH peptide is lacking for most fish species, studies in the rainbow trout –48 and tperciformes  and the nucleus posterioris periventricularis (NPPv)  , 84, whiss2 mRNA . A recenss2 mRNA . The facss2 mRNA  suggestsebrafish , and senebrafish , 87. Thekiss1 and kiss2 mRNA levels are increased significantly at the start of the pubertal phase together with GnRH2 and GnRH3 mRNAs  neurons in the zebrafish , suggestGnIH or RFamide-related peptide (RFRP), which has a characteristic C-terminal LPXRFa motif (X = L or Q), is a hypothalamic neuropeptide that was originally identified from the quail as a neuropeptide that inhibits gonadotropin release from the pituitary . ExtensiIn vivo studies using the goldfish show that GnIH decreases plasma LH levels as in avian and mammalian species  peptide decreases the firing frequency of non-hypophsiotropic terminal nerve GnRH3 neurons in the dwarf gourami . Since GA number of hypothalamic neuropeptides have been identified in fish species , and fouOrexin has two well conserved molecular forms, a 33-amino acid peptide known as orexin A (OXA) and a 28-amino acid peptide known as orexin B (OXB) derived from the same precursor [see review in Ref. ]. OrexinIn mammals, orexin is known to stimulate the HPG axis via GnRH secretion –155. In NPY which is composed of 36 amino acid residues, was first identified in the porcine brain , and wasCentrally or peripherally injected NPY increases plasma LH levels in the goldfish, common carp, rainbow trout, and in the sea bass , 111, inPlecoglossus altivelis) (in vitro in the goldfish (In the brown trout  and the tivelis)  and the tivelis) , suggestgoldfish  and in tgoldfish .In vitro treatment with NPY stimulates LH release from pituitary cells in the goldfish   . In teleculture) . On the GAL is a 29-amino acid peptide, expressed in the central nervous system and in the intestine. GAL stimulates feeding in the goldfish  and the in vitro GnRH release in rats  are seen in the sea bass . SimilarAs mentioned before, among the different forms of GnRH, neuronal fibers of GnRH2  are widely distributed in the vertebrate brain. In an insectivore, the musk shrew, GnRH2 stimulates sexual behavior and seduces food intake , 187, inIn fish species, the suppressive effect of GnRH2 on feeding has also been confirmed. Food consumption is significantly decreased by intracerebroventricular injections of GnRH2 but not GnRH3 in a dose dependent manner in the goldfish , 118 andin vitro  encoded unmodified peptide, was first characterized in rats , and wasThe number of studies about the function of nesfatin-1 in the control of reproduction is still limited. However, recent studies showed that nesfatin-1 acts as an inhibitory signal in the control of fish reproduction. Although nesfatin-1 plays a stimulatory role in LH secretion in rats , an intrLeptin is primarily produced by adipocytes of the white adipose tissue , and secin vitro LH release from the pituitary culture in the sea bass   , suggestIn addition to its primary role in reproduction, GnIH stimulates food intake in chickens  and in rTo monitor the amount of energy stock, central metabolic neuropeptide neurons receive peripheral signals including leptin and ghrelin. For example, in mammals leptin receptor is expressed in many metabolic neuropeptide neurons including orexin, NPY, GAL, MCH, POMC, CART, and CCK neurons . IDouble immunostaining revealed interactions among orexigenic/anorexigenic neuropeptide neurons in teleosts, in particular in the goldfish Figure . Among aAmong orexigenic neuropeptide neurons, NPY and orexin neurons make reciprocal connections in fish as in mammals. NPY neuronal fibers make close appositions with orexin neurons, whereas orexin neuronal fibers make close appositions with NPY neurons in the NPPv in the goldfish . FurtherMoreover, the orexigenic and the anorexigenic circuits are also connected with each other. α-MSH neuronal fibers make close appositions with NPY neurons, whereas NPY neuronal fibers project to α-MSH neurons in the goldfish . In addivice versa (Studies using antagonists against of the receptors for metabolic neuropeptides further provided possible interaction among central metabolic neuropeptides in the goldfish. For example, GAL mediates the orexigenic action of orexin, and orexin mediates the orexigenic action of GAL . GAL alsce versa . Besidesce versa . Furtherce versa . These rIn addition to the evident neuronal interactions in the goldfish, more combinations of neuronal interactions were reported in other fish species. In the masu salmon  and the Significant amount of information about the relationship among orexigenic and anorexigenic neuropeptides have been accumulated, particularly in the goldfish. However, the knowledge of direct interactions among these neurons is still not enough to draw a complete diagram of the neuronal circuit to control food intake and reproduction in fish. In particular, metabolic regulation of kisspeptin and GnRH neurons are still unknown, while it is suggested from mammalian studies. Further fiber projection studies using double immunostaining and localization of the neuropeptide receptors in certain neuronal cell bodies need to be performed.There are many differences in the regulatory mechanism of food intake and reproduction not only between mammals and fish but also between fish species. The significant difference between fish species might be the result of the adaptation to a wide range of feeding habits and reproductive strategies. Therefore we have to be careful to combine data obtained from different species.Several studies reported that the responses of the reproductive system to metabolic signals differ depending on the sex and the stage of sexual maturation. In fact, gonadal steroids modulate the effect of NPY on GnRH and LH release in the goldfish . Each stAnimals might change the metabolic control of reproduction according to the available energy stock. For example, short term food limitation attenuates sexual motivation, while remaining energy stock still maintains activity of the HPG axis. On the other hand, long term food limitation depletes the energy stock and stops the HPG axis to prioritize the energy supply to the survival. Thus, feeding conditions and the timing of experiment might be important to obtain comparable data.The endocannabinoid system is involved in a variety of physiology including pain-sensation, mood, and memory. Importantly, both energy balance and reproduction are modulated by the endocannabinoid system. Endocannabinoids modulate several hypothalamic metabolic neuropeptides in mammals . TIn summary, the cellular and molecular basis for the integration of feeding and reproduction involves a complex interaction of the reproductive system with metabolic neuropeptides and peripheral fuels. The metabolic neuropeptides, particularly orexin, NPY, PACAP, MCH, nesfatin, GnRH2, and CCK play an important role in the reproduction by either regulating GnRH neurons in the hypothalamus or by stimulating gonadotropes in the pituitary. Peripheral metabolic signals such as ghrelin and leptin also act on the pituitary to stimulate LH secretion. It should be, however, noted that compared to mammals, fishes show a great variety of feeding and reproductive habits. The variations of metabolic control of reproduction in different teleost species may reflect different requirement of energy status for reproduction in different species. Compared to mammals, fish represent a vast phylogenetic group, which shows a significant level of diversity with regards to morphology, ecology, behavior, and genomes . Thus, sThe authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.