Previous studies have identified high rates of explosive anger amongst post-conflict populations including Timor-Leste. We sought to test whether explosive anger was integrally associated with symptoms of grief amongst the Timorese, a society that has experienced extensive conflict-related losses. In 2010 and 2011 we recruited adults (n = 2964), 18-years and older, living in an urban and a rural village in Timor-Leste. We applied latent class analysis to identify subpopulations based on symptoms of explosive anger and grief. The best fitting model comprised three classes: grief (24%), grief-anger (25%), and a low symptom group (51%). There were more women and urban dwellers in the grief and grief-anger classes compared to the reference class. Persons in the grief and grief-anger classes experienced higher rates of witnessing murder and atrocities and traumatic losses, ongoing poverty, and preoccupations with injustice for the two historical periods of conflict . Compared to the reference class, only the grief-anger class reported greater exposure to extreme deprivations during the conflict, ongoing family conflict, and preoccupations with injustice for contemporary times; and compared to the grief class, greater exposure to traumatic losses, poverty, family conflict and preoccupations with injustice for both the internal conflict and contemporary times. A substantial number of adults in this post-conflict country experienced a combined constellation of grief and explosive anger associated with extensive traumatic losses, deprivations, and preoccupations with injustice. Importantly, grief-anger may be linked to family conflict in this post-conflict environment. Populations exposed to mass conflict and persecution commonly experience extensive losses , 2, expeAnger as an unwanted and commonly dysfunctional emotional reaction has been associated with feelings of injustice amongst populations that have been exploited and persecuted. Having one’s human rights violated or economic goals systematically undermined can understandably lead to normal anger reactions, however, anger can also be associated with a loss of control, aggression and harm to others, including community members, intimate partners and children –10. AngeStudies amongst post-conflict populations exposed to repeated traumatic losses may shed further light on the role of anger in the grief response. Our past research in Timor-Leste identified what appeared to be a high rate of explosive anger in response to trauma exposure. Explosive anger can express itself as physiological arousal and either verbal or physical aggression, the response characteristically being out of proportion to environmental triggers and experienced as uncontrollable, the subject reacting without immediate thought to the consequences . AlthougA population study in a rural and an urban village of Timor-Leste undertaken in 2004 recorded a prevalence of explosive anger of 38%, based on the international threshold of at least one attack of explosive anger a month (noting that the majority of these persons experienced much more frequent episodes) . In a siDoubts remain, therefore, about the origins and nature of explosive anger and its high prevalence in Timor-Leste, and why it is particularly common amongst women. In our endeavour to understand this phenomenon, we draw on the Adaptation and Development After Persecution and Trauma (ADAPT) model , 29 whicThe history of persecution and conflict in Timor-Leste provided a setting to investigate possible associations between grief and anger amongst a population exposed to extensive traumatic losses. The invasion and occupation of the territory by Indonesia in 1975 provoked a low-grade resistance war waged by members of the indigenous independence movement. During the period of conflict, which culminated in a humanitarian emergency in 1999, an estimated quarter of the indigenous population  died as a consequence of atrocities, warfare, burning of villages, murder, famine and untreated illness. In addition, there was widespread loss of property and livelihoods, and forced displacement of whole communities, with kinship and family groups being dispersed, some as refugees to other countries. In the post-conflict phase, further episodes of violence occurred, particularly in 2006–7, when a period of sustained internal conflict led to extensive injuries, deaths and displacement of communities into makeshift refugee camps. Socio-economic development in the newly independent country has been slow, with many families confronting extreme levels of poverty and deprivation.Our aim was to test whether it is possible to identify a combined pattern of explosive anger and grief symptoms (grief-anger) amongst the Timorese population. We hypothesized that a subpopulation with grief-anger would report high levels of traumatic losses, preoccupations with injustice and ongoing adversity including family conflict in the post-conflict environment. We also examined whether women were more likely than men to experience the putative grief-anger constellation.Between June, 2010 and July, 2011, we conducted a survey of all adults, 18 years and older, living in every household in two administrative villages (sucos), one in Dili, the capital, the other, a rural site an hour’s drive away. Each suco is defined by contiguous hamlets  falling under the administration of one chief (chefe). GPS and aerial mapping produced by the government for census purposes allowed us to identify all households in a setting where there is an absence of street names and many dwellings are located in remote wooded and mountainous areas. Both study sites were extensively affected by mass violence during the Indonesian occupation (1975–1999) and the subsequent internal conflict (2006–7).The team included 18 Timorese field workers with prior survey experience and/or psychology/public health degrees. They received a two-week training course followed by two months of field testing and piloting of survey measures under supervision. Pairs of interviewers were required to achieve a consistent 100 percent level of inter-rater reliability on the core measures. One hour long interviews were conducted in participants’ homes or another location if preferred by respondents, the procedure ensuring maximal privacy and confidentiality. In villages where families live in close proximity to each other, and where overcrowding is a problem, we sought to ensure privacy by taking participants to garden areas or away from the household to somewhere shaded and quite. We also arranged for children to be entertained by one of our colleagues if they were likely to cause a distraction to participants. Households were visited up to five times in order to meet potential participants.The study was approved by the ethics committee of the University of New South Wales, the Ministry of Health of Timor-Leste, and the chiefs of each village. The majority of respondents gave written consent prior to commencement of interviews. Verbal consent was obtained in some cases where respondents were illiterate, trusted witnesses co-signing the forms. The procedure was endorsed by the community and received ethical approval from the University of New South Wales and the Ministry of Health of Timor-Leste.Our selection of constructs and the appropriate measures to assess them was based on theoretical considerations and the empirical findings in our past studies examining explosive anger in Timor-Leste.The protocol, including the grief measure, was iteratively field tested amongst communities geographically adjacent and similar in sociodemographic composition to the sites of the definitive survey. In piloting, we applied an iterative process of feedback in which responses and solicited comments by respondents in the field were analysed and considered by a committee comprising Timorese of diverse backgrounds  and expatriate researchers. Measures were reviewed and revised to ensure that the constructs were understood by the community, items were readily comprehended, both semantically and linguistically, and response options  were appropriately graduated according to the language and culture.The 17 conflict-related traumatic events (TEs) listed in the Harvard Trauma Questionnaire (HTQ)  were modAn inventory of daily adversities was developed based on extensive community consultations and refinement of items during piloting  Table 2Table 2. Respondents were asked to identify and describe the worst human rights violation or other event associated with injustice they had experienced during three defined historical periods: the Indonesian occupation, the period of internal conflict, and contemporary time. Ratings were assigned as 1 for assigning an unjust event; 2 for experiencing preoccupations relating to the event; and 3 for distress related to these preoccupations. The composite index of injustice reflected the addition of scores for each of the three historical time periods (range 0–3).Our community measure of explosive anger was developed, tested and modified serially during piloting to ensure its cultural appropriateness and comprehensibility in the local language, Tetum . The scrWe inquired of all participants whether they had experienced a loss, defined as an event (since 1975) in which someone  close to the individual had died or been killed. Those who identified multiple losses were asked to identify the death that had the most impact on their lives, then recording the cause and time of the death. Almost all of these identified losses were related to traumatic deaths or untreated illness occurring during periods of mass conflict. Based on the identified loss event, participants were then asked to rate each of four grief items on a five-point frequency scale  as experienced in the past four weeks. The initial item pool was derived from the literature and contemporary criteria for assessing prolonged grief , the proPosttraumatic stress disorder (PTSD) symptoms and general symptoms of psychological distress  were assessed using the Harvard Trauma Questionnaire (HTQ) and Kessler-10 respectively, widely used measures applied in our previous studies in Timor-Leste . In our All measures were translated into Tetum, the most widely spoken language in Timor-Leste. Minor inconsistencies were addressed during piloting and the final versions were translated and back-translated into English .We calculated intra-class correlations to assess for possible clustering within households of indices of grief, psychological distress, PTSD and explosive anger. All correlations were low (<0.05) indicating negligible clustering by households We used latent class analysis (LCA) to identify clusters of participants according to their pattern of symptoms of explosive anger and grief (each item scored in a binary manner as present or absent). We tested sequential models  examining a suite of conventional model fit indicators to assess for the best class solution: The Bayesian Information Criterion (BIC), sample size-adjusted Bayesian Information Criterion (SS-BIC), and the Akaike’s Information Criterion (AIC) , 35. LowAfter selecting the best-fitting model, we examined for associations between class membership (with the low symptom class as the reference category) and a range of relevant predictors using multinominal logistic regression analysis. The covariates included: sociodemographic characteristics of gender, residency in urban or rural areas, educational attainment, and employment; traumatic domains comprising conflict-related trauma, witnessing murders and atrocities, traumatic losses, and extreme deprivations; current adversities including indices of poverty, family conflict, and communal conflict; and preoccupations with injustice . Analyses were performed in STATA version 13 and Mplus Version 7.Of the 3368 respondents approached, 2964  completed interviews, a response rate of 83.6% (inability to contact residents was by far the major reason for non-participation).Two hundred and fifty persons (8.4%) met criteria for explosive anger according to IED criteria. A quarter  endorsed one or more symptoms of explosive anger, including sudden anger attacks , loss of control , verbal aggression , destruction of property , and physical aggression .Over half  endorsed one or more symptoms of prolonged grief, including persistent yearnings or longings for the deceased , feelings of bitterness about the death , and feelings of emptiness . A third  reported functional impairment associated with these symptoms.A similar number  met the threshold for PTSD (>2.2) and severe psychological distress  (≥30).Over half of participants (56.1%) reported experiencing one or more conflict-related traumas including political imprisonment, combat, physical assault, torture, and trauma related to involvement in the resistance movement . Four ouDistressing preoccupations with events associated with injustice were reported by 13.1% (n = 388) for the Indonesian occupation (1975–1999), 24.6% (n = 729) for the period surrounding the internal conflict (2002) and 18.5% (n = 549) in contemporary times .Serial model testing concluded after assessing a four class LCA model . Fit indIn comparison to the low symptom class, both the grief and grief-anger classes were associated with PTSD  and severe psychological distress .In comparison to the low symptom reference class, women and urban dwellers were more likely to be assigned to both the grief and grief-anger classes. The two TE domains of witnessing murder and atrocities, and traumatic losses were both associated with the grief and grief-anger classes (relative to the reference class). In addition, however, the grief-anger class reported greater exposure to traumatic losses than the grief class. Also, the grief-anger class alone reported greater exposure to extreme deprivations related to conflict in comparison to the reference class. In relation to ongoing adversities, both the grief and grief-anger classes exceeded the reference class on the index of poverty; the grief-anger class in turn reported higher rates of poverty than the grief class. Only the grief-anger class reported greater levels of family conflict, in comparisons with both the reference low symptom class and the grief class. Compared to the low symptom class, both the grief and grief-anger classes reported greater preoccupations with injustice for the two historical periods of conflict . Only the grief-anger class, however, reported a higher level of preoccupations with injustice for contemporary times compared to the reference class.Our analysis in post-conflict Timor-Leste, identified a typology comprising three subpopulations including those experiencing grief, grief-anger and low symptoms, the first two categories affecting a quarter of adults in the sample. Women and urban-dwellers were more likely to be assigned to both the grief and grief-anger classes. Compared to the low symptom reference class, both the grief and grief-anger classes reported greater exposure to conflict-related murders/atrocities and traumatic losses, more extreme levels of poverty, and distressing preoccupations with injustice related to two successive historical periods of conflict. There were important distinctions between the two morbid classes however, in that the grief-anger class reported greater exposure to traumatic losses (compared to the grief class), greater deprivations during the period of conflict (compared to the reference low symptom class), higher stress levels related to poverty (compared to the grief class), ongoing family conflict (compared to both the reference and grief classes), and preoccupations with injustice for contemporary times (compared to the reference and grief class).Prior to discussing our findings, we consider the strengths and limitations of the study. The sample is one of the largest in the contemporary post-conflict mental health field and we achieved a high response rate. Although sampling was restricted to two localities, the sites were identified initially as being broadly representative of the socio-demographic profile of Timor-Leste as a whole . NeverthCaveats notwithstanding, our findings cast new light on the high prevalence of explosive anger previously identified in community samples in Timor Leste , 9, 20, The grief-anger class reported the greatest exposure to traumatic losses, an important finding given that murder, atrocities and death by untreated illness and famine were widespread during the prolonged period of conflict in Timor-Leste. It may be that in collectivist societies such as Timor-Leste, losses that provoke strong and enduring feelings of injustice are particularly potent in generating the identified combined pattern of grief-anger. To confirm this, replication of our findings will be needed in other conflict-affected settings where traditional family and community values prevail.Importantly, our regression analysis involving relevant covariates added credibility to the distinction we found between the grief-anger and grief classes. Specifically, the grief-anger class stood out in reporting high levels of traumatic loss, extreme deprivations during the period of conflict, severe ongoing poverty and family conflict, and preoccupations with injustice extending over three contiguous historical periods. In relation to the latter finding, we have reported a similar association between the anger component of persistent complex bereavement (PCB) disorder and a sense of injustice amongst refugees from West Papua, a neighbouring territory that has experienced a comparable level of prolonged mass conflict under Indonesian occupation .The finding that a half of the population experienced relatively low levels of grief and anger symptoms offers some insights into the factors that protect post-conflict populations from these adverse psychological outcomes. It is notable that the low symptom group reported a similar level of exposure to the general traumas of conflict, indicating that they had not been sheltered from these events. It was only in the TE domains of witnessing murder/atrocities and traumatic losses that the low symptom group reported lower exposure, suggesting that protection from these salient forms of trauma may act to avert risk of developing the specific grief-anger constellation. Being male, living in a rural environment, experiencing lower levels of poverty and not experiencing family conflict were other factors that appeared protective, noting however, that cause-effect relationships involved remain to be confirmed given the cross-sectional nature of the study.Our findings have potential implications for the individual, the family and the society as a whole, not only in Timor-Leste but in other post-conflict settings worldwide. In particular, confirmation of a grief-anger class and the social factors associated with the pattern, has the potential to add support to a cycles of violence model which postulates that exposure to the traumas of past conflict  may contribute to risk of subsequent family conflict in the aftermath of the violence . We noteIn relation to ongoing adversities, there appeared to be a stepwise relationship between the severity of poverty and the grief-anger, grief and low symptom classes respectively. These observations underscore the interaction between trauma-related mental health problems and socio-economic factors in post-conflict societies. Poverty places stress on individuals, families and communities, compounding past interpersonal and material losses in generating a sense of injustice and anger. In that sense, apart from the immediate hardship incurred by poverty, conditions of extreme material deprivations jeopardise recovery from trauma-related mental health conditions which in turn can impair functioning and reduce the capacity of survivors to engage in gainful employment or other opportunities to improve their economic well-being. .Our study identified a grief-anger constellation comprising a quarter of the study sample in post-conflict Timor-Leste. There were commonalities with the grief group in reporting greater exposure to witnessing murder, traumatic losses and poverty, and experiencing persisting preoccupations with injustice related to two consecutive historical periods of conflict. The grief-anger group was unique however, in reporting extreme levels of traumatic losses, exposure to material deprivations during the period of conflict, preoccupations with injustice in contemporary times and ongoing family conflict. It is a cruel irony that the traumatic rupture of interpersonal bonds during periods of mass conflict can generate a psychological reaction pattern (grief-anger) in survivors which in turn may undermine the survivor’s capacity to achieve a stable family environment in the post-conflict period.S1 Dataset(DTA)Click here for additional data file.


Consumption of a Western Diet (WD) that is high in saturated fat and added sugars negatively impacts cognitive function, particularly mnemonic processes that rely on the integrity of the hippocampus. Emerging evidence suggests that the gut microbiome influences cognitive function via the gut-brain axis, and that WD factors significantly alter the proportions of commensal bacteria in the gastrointestinal tract. Here we review mechanisms through which consuming a WD negatively impacts neurocognitive function, with a particular focus on recent evidence linking the gut microbiome with dietary- and metabolic-associated hippocampal impairment. We highlight evidence linking gut bacteria to altered intestinal permeability and blood brain barrier integrity, thus making the brain more vulnerable to the influx of deleterious substances from the circulation. WD consumption also increases production of endotoxin by commensal bacteria, which may promote neuroinflammation and cognitive dysfunction. Recent findings also show that diet-induced alterations in gut microbiota impair peripheral insulin sensitivity, which is associated with hippocampal neuronal derrangements and associated mnemonic deficits. In some cases treatment with specific probiotics or prebiotics can prevent or reverse some of the deleterious impact of WD consumption on neuropsychological outcomes, indicating that targeting the microbiome may be a successful strategy for combating dietary- and metabolic-associated cognitive impairment. Substantial evidence has linked consumption of a Western Diet (WD), defined here as diets consisting of both high levels of fat  and added sugars, with cognitive dysfunction  and reduced levels of Bacteroidales (Phylum: Bacteroidetes) in rodents, changes that correlated to poor cognitive flexibility  reduced by the WD, whereas Bilophila sp. were elevated in the WD group and barely detectable in the control group  relative to controls that were not given sugar  reduces SCFAs such as acetate, propionate, and butyrate are produced in the gut by microbial-mediated fermentation of indigestible carbohydrates, such as resistant starch and non-starch polysaccharides from cereals, vegetables, and fruits  consists of a structural complex of endothelial cells, pericytes, and glial cells that encompass microvasculature networks within the central nervous system. It serves as a critical regulator for the entry of blood-derived nutrients and compounds required for healthy brain function, while simultaneously precluding the entry of potentially harmful blood-derived toxins. Importantly, WD intake is associated with BBB damage, which may be causally related to WD-induced cognitive dysfunction  impairs insulin receptor function in the hippocampus and reduced hippocampus-dependent spatial memory  receptors, which leads to increased hippocampal long-term potentiation  to treat neurobiological and cognitive dysfunction associated with WD intake and associated metabolic disorders.All three authors contributed to the idea for the manuscript. EN and TH wrote and edited the manuscript. SK edited the manuscript, provided vital input to shape the manuscript, and contributed to the writing. All three authors approved the final manuscript.This work was supported by grant number DK104897 from the National Institute of Diabetes and Digestive and Kidney Diseases and also by the University of Southern California Diabetes and Obesity Research Institute (Awarded to SK).The 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.


Preterm infants present an immature neurobehavioral profile at birth, even in absence of severe brain injuries and perinatal complications. As such, they require a long-lasting hospitalization in the Neonatal Intensive Care Unit (NICU), which is thought to grant at-risk newborns’ survival, but still entails a number of physical, painful, and socio-emotional stressors. Hence, preterm birth and NICU stay represent an early adverse experience, which has been linked to detrimental consequences for neurological, neuro-endocrinal, behavioral, and socio-emotional development, as well as to disease later in life. Recent advances in the behavioral epigenetic field are helping us to unveil the potential mechanisms through which early NICU-related stress may lead to negative developmental outcomes. From this perspective, telomere regulation might be a key programming mechanism. Telomeres are the terminal portion of chromosomes and are known to get shorter with age. Moreover, telomere length (TL) is affected by the exposure to stress during early development. As such, TL might be an innovative biomarker of early adverse exposures in young infants and children. Unfortunately, there is paucity of studies investigating TL in populations of preterm infants and its association with known NICU-related stressors remains unexplored. In the present paper, the potential relevance of TL for research and clinical work with preterm infants will be underlined in the light of recent contributions linking progressive telomere shortening and early exposure to adverse experiences and stressful environments in humans. Finally, insights will be provided to guide clinically relevant translational research on TL in the field of VPT birth and NICU stay. Preterm birth  is estimated to account for about 11% of all live births worldwide . NotablyTelomeres are repeat sequences at the ends of chromosome arms, known to shorten because of cellular aging and in relation to disease conditions . Recent Preterm infants need long-term hospitalization and specialized multi-disciplinary interventions in NICU. During this time, they are exposed to painful procedures , 8 and hThe NICU environment is far from being a surrogate of the maternal womb and represents a source of distress in newborns due to three main aspects: (1) sensorial adverse stimulation, (2) invasive and painful procedures, and (3) limited opportunities for parental caregiving and alteration of parent–infant bonding.First, infants are exposed to physical and sensorial stimulations  characterized by levels of intensity and length of exposure which easily exceed the average recommended exposure standard for newborns . As suchAdditionally, invasive and painful clinical procedures  are needed to improve the health status of preterm infants. Due to their immature neurobehavioral profile, preterm infants present lower threshold and higher sensitization to external perturbations, so that even routine handling  might be responded to with a heightened physiological response . InterveNotably, preterm newborns are suddenly separated from the mother after birth, which may disrupt the biological routes to caregiving and parental bonding  and alteDuring the last decades, DC strategies have been developed to minimize the negative effects of preterm birth, optimize infants’ neurobehavioral development, and improve interactions between parents and infants , 27. DC The quality of DC is critical to counterbalance the effect of NICU-related stressors on preterm infants’ developmental trajectories. Preterm infants hospitalized in NICUs characterized by high quality of DC practices exhibit better neurobehavioral development during the first month of life , less inNR3C1, encoding for cortisol receptors and SLC6A4, encoding for the serotonin transporter  current evidence pertaining to TL analysis and prematurity; (2) methodological issues; and (3) implications for research and clinical practice.To best of our knowledge, only few studies have investigated TL in the preterm infants. One study reported no significant differences between mean leukocyte TLs of preterm infants compared with their full-term counterparts at birth . HoweverN = 5). Vasu and colleagues , socio-demographic , and contextual factors . This implies that researchers need to pay attention to different conditions that interact in a complex manner in affecting TL in preterm infants. For instance, preterm infants born at lower gestational age might have longer telomeres at birth but greater stress-related erosion due to cumulative NICU-related stress and pain. Consistently, careful selections of preterm infants in terms of inclusion criteria should be pursued to avoid mixed samples of subjects, which might affect the quality of life, developmental trajectories, and number of stressful intervention to which they are exposed during the NICU stay.Differences in DC practices among diverse NICUs and countries represent another source of variation . DespiteFinally, the context and timing of early adversity exposure might be critical in setting a TL-related biomarker of later-in-life health and disease , 66. As Figure Future investigation could also consider the protective effects of DC practices in reducing or slowing the rate of telomere erosion. Studies in adult human subjects have demonstrated that positive changes in life-style have the possibility to result in significantly increased telomerase activity and improved telomere maintenance capacity in human immune-system cells . SimilarThe study of TL in preterm infants is an intriguing area of research and a potential source of scientifically sound beneficial information for NICU staff. Although now it remains a hypothetical scenario, it is possible to speculate that TL measure might have some clinical implications for NICU-related stress and infant DC. For example, given the evidence for association between TL and pain , one couLP conceived the work, wrote the first draft of the manuscript, and approved the final version of the paper. GSM contributed to literature review, revised English language, and approved the final version of the paper. RG supervised the work, by providing unique expertise in relation to biological sections, and he approved the final version of the paper. RM contributed to the conception of the work, supervised the writing, and approved the final version of the paper. All authors agree to be accountable for all aspects of the work.The 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.


Mycobacterium leprae infection. The findings described here highlight an important area of investigation, as leprosy is one of the main causes of infection in the peripheral nervous system.Neurotrophins are a family of proteins that regulate different aspects of biological development and neural function and are of great importance in neuroplasticity. This group of proteins has multiple functions in neuronal cells, as well as in other cellular populations. Nerve growth factor (NGF) is a neurotrophin that is endogenously produced during development and maturation by multiple cell types, including neurons, Schwann cells, oligodendrocytes, lymphocytes, mast cells, macrophages, keratinocytes, and fibroblasts. These cells produce proNGF, which is transformed by proteolytic cleavage into the biologically active NGF in the endoplasmic reticulum. The present review describes the role of NGF in the pathogenesis of leprosy and its correlations with different clinical forms of the disease and with the phenomena of regeneration and neural injury observed during infection. We discuss the involvement of NGF in the induction of neural damage and the pathophysiology of pain associated with peripheral neuropathy in leprosy. We also discuss the roles of immune factors in the evolution of this pathological process. Finally, we highlight avenues of investigation for future research to broaden our understanding of the role of NGF in the pathogenesis of leprosy. Our analysis of the literature indicates that NGF plays an important role in the evolution and outcome of  The most extensively studied neurotrophin is nerve growth factor (NGF). This growth factor consists of 118 amino acids and has a molecular weight of 130 kDa. NGF was discovered as a growth factor that participates in the responses of sympathetic and sensory neurons regulating differentiation, neuronal regeneration, and the perception of pain Figure . In sensNerve growth factor was first described by Rita Levi-Montalcini , who shoNerve growth factor is endogenously produced as proNGF during development and into maturity by immune and nerve cells, as well as peripheral effector cells, such as keratinocytes, melanocytes, smooth muscle cells, fibroblasts, and Schwann cells , 17, 18.Following its synthesis, NGF enters nerve endings by endocytosis, a process that permits NGF transport to the nerve cell body through contact with NGF receptors (NGF-R) located on the plasma membrane , 26. SevNerve growth factor has been suggested to be involved in apoptosis of sympathetic and sensory neurons , 21. TheThus, in addition to its primary functions in the nervous system, NGF participates in inflammatory processes and immune responses. NGF concentrations are increased during tissue inflammatory processes. Increased NGF levels can produce hyperalgesia through direct activation of nociceptors, which leads to CNS activation and neurogenic inflammation. Moreover, this process leads to local events comprising activation-induced release of histamine and increased numbers of mast cells and other immune system cells , 19.Labouyrie et al. described the relationship between NGF and cells in normal and pathologic human lymphoid tissues and showed that NGF is involved in inflammatory or lymphoproliferative disorders . ImmunorDifferent inflammatory and autoimmune diseases lead to altered expression of NGF. Increased anti-NGF antibody levels have been detected in patients with rheumatoid arthritis, systemic lupus erythematosus, and thyroiditis and are thought to contribute to the immune dysfunction and nerve damage observed in these diseases , 35, 36.via the TrkA receptor. This process has previously been described for sensory neurons  that prevents the proliferation of the bacillus. In the lepromatous form, the cell-mediated immune response is characterized by an anti-inflammatory cytokine profile (Th2) that contribute to multiplication of the bacillus in macrophage phagosomes. In the borderline form, the patients exhibit immunological and histopathological characteristics that vary between those of the tuberculoid and lepromatous forms .Nerve injury-associated tissue damage is arguably the most important clinical consequence of leprosy . In the Leprosy is the most common cause of non-traumatic neuropathy and is a classic example of an infectious neurodegenerative disease of the peripheral nervous system. It is estimated that more than one-fourth of patients with leprosy have some degree of disability and that about half of these patients have grade 2 disability corresponding to permanent neurological damage –59.Studies have shown that different levels of NGF are associated with lepromatous and tuberculoid leprosy such that higher levels of NGF are associated with lepromatous forms and low levels of NGF are associated with tuberculoid forms of the disease . SpecifiOther studies have suggested that NGF may restore pain sensitivity. Thus, NGF could play an important role in the prevention of ulcerations resulting from nociception loss, as observed in leprosy and other peripheral neuropathies , 61. SomImmunostaining of damaged tissues using anti-NGF and anti-NGF-R antibodies revealed higher expression of NGF in patients with lepromatous leprosy and lower expression of NGF in those with the indeterminate form of the disease . In leprSeveral important findings have elucidated the role of NGF as an immune response mediator . In addiImbalance in the proNGF/NGF ratio, increased expression levels of TNF-α and p75 neurotrophin receptor, and impaired TrkA phosphorylation have been reported in microvascular preparations from Cre-proNGF transgenic mice when compared to normal control animals . TNF-α cM. leprae is the Schwann cell, which is the predominant cell type in the peripheral nervous system. Degradation of Schwann cells by M. leprae favors the development of peripheral neuropathy, which is the leading cause of morbidity in patients with leprosy. NGF may act as a protective factor for Schwann cells, and low levels of NGF may contribute to the development of neuropathy  have revealed an interaction between NGF and TGF-β , 63. Neur = 0.8722 and r = 0.7257, respectively), with highly significant p-values for the two correlations   , 49. Thectively) , 64. In ctively) , 66.During healing, NGF activates processes involved in the restoration of innervation , 67. In Cell-mediated immune responses may be beneficial against bacterial infections; however, inflammation can lead to irreversible tissue damage. Nerve injury occurs in approximately 10% of patients with paucibacillary leprosy and 40% of patients with multibacillary leprosy and is particularly acute in patients with reverse reactions , 70.TGF-β participates in the tissue repair process as an anti-inflammatory agent during intense inflammation by inducing nerve and tissue regeneration , 71. HigIn lepromatous leprosy, neurological manifestations progress slowly over many years and involve small nerve branches and multiple nerve trunks within the skin , 74. PerThe expression of p75 receptor in peripheral nerve cells is induced by loss of contact between the target organ and the axon . Some stProspective cohort studies and intervention studies evaluating patients with different clinical forms of the disease and different reactive states (erythema nodosum and reverse reaction types) may help better elucidate the relationship between NGF and the immune response and the factors contributing to the protection and regeneration of nerves affected during infection. Further analysis of the relationship among tissue levels of NGF and a large panel of pro- and anti-inflammatory cytokines, blood levels of NGF, and the immune response is important for a better understanding of the involvement of NGF in the pathophysiology of chronic granulomatous peripheral nerve infections, especially leprosy.This review demonstrates that, within lesions associated with leprosy, NGF and TGF-β respond to inflammatory processes and tissue damage while triggering tissue remodeling. Further studies are needed to elucidate the broad role of NGF in the pathogenesis of leprosy. Although our understanding of the effects of NGF on nerve damage has increased, further insights into the functional roles of NGF (and other neurotrophins) in normal skin and during disease progression are needed.TA, JS, AF, LF, and JQ conceived and wrote the manuscript.The 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.


Ronald Ma and Barry Popkin discuss the urgent need and challenges to reduce intergenerational transmission of obesity and diabetes. Globally, efforts to prevent diabetes and obesity have attained mixed success.Although prevalence of adult-onset diabetes is declining in some countries, the prevalence of childhood obesity and young-onset diabetes, with risk of associated complications, continues to increase in many high- and lower-income countries . More thOngoing efforts to promote lifestyle intervention have had limited impact on the obesity and diabetes epidemic because of several critical factors. One factor comprises early life effects, a new field that has emerged surrounding developmental origins of many noncommunicable diseases (NCDs). This is the impact of nutritional and other insults during pregnancy and the first 2 years of life on the long-term risk of diabetes. Beginning with pathbreaking work by Barker on the relationship between low birth weight infants and subsequent risk of many adverse cardiometabolic outcomes ,6, this A second critical factor involves risks affecting populations of non-European origin, including Hispanic, African, Middle Eastern, or Asian groups –18. ThesAnother aspect of developmental influences relates to maternal hyperglycemia, gestational diabetes mellitus (GDM), maternal obesity, and the resulting in utero overnutrition. Maternal obesity, GDM, and excessive gestational weight gain lead to macrosomia and excess adiposity in offspring and increase the risk of childhood obesity ,21. FurtThere is 1 last set of changes we are seeing across many countries. As overweight and obesity has increased globally , researcThis intergenerational transmission of risk poses particular challenges to our current approach to obesity and diabetes prevention. Recent trials have highlighted that interventions initiated in pregnancy, such as increasing physical activity in overweight mothers, are likely to be too late to have an impact on pregnancy outcome –25. HencSeveral strategies have been explored over recent years, including restricting advertisement of unhealthy foods to children, front-of-pack labelling, improving school meals, using taxation to reduce the consumption of unhealthy food items, and providing different incentives to encourage intake of healthy foods or to encourage the production of healthier foods . HoweverA keen sense of urgency is therefore needed in approaching the current conundrum. Not only is the global burden of diabetes giving rise to escalating healthcare expenditure that is threatening economic development of emerging economies, but the worsening epidemic of childhood obesity and young-onset diabetes and the increasing prevalence of GDM and diabetes affecting women of reproductive age are fuelling an evolving cycle of intergenerational risk transmission. Emerging data linking GDM with adverse neurocognitive development in offspring of mothers with GDM is particularly alarming , with ma


Thegray literature, ie, thesis, dissertations, books, guidelines and dictionary wasalso explored to ensure the robustness needed to clarify the topics not covered bythe studies. the definitions aim to facilitate the identification of the nursing diagnosis fornonverbal critically ill patients and to support future teaching and research onthe nursing diagnosis of Acute Pain (00132). However, the inability tocommunicate verbally, present in many patients in intensive care units, does not negatethe possibility that they are experiencing pain and need to have treatmentDue to this problem, statistics for pain prevalence are practically non-existent fornonverbal patients and patients suffering from severe illnesses or with cognitiveimpairment are often excluded from pain prevalence studiesIn their daily practice, ICU nurses must have quick decision-making skills based on theobservation of physiological and behavioral changes in patients. The direction of thenursing care depends on the nurses’ rapid observation, since complications that affectthe recovery of their patients can be preventedThis rapid assessment is expressed through the identification of the nursing diagnosis(ND). For this, nursing professionals can rely on their own taxonomies, such as theNANDA International classification (NANDA-I)These studies aim to improve the DCs commonly identified for the NDsThese validation studies provide refinement and improvement of classifications, whichfavors the exercise of critical thinking and decision-making and improves communicationand standardized nursing recordsTherefore, the objective of this study is to develop conceptual and operationaldefinitions of each DC and conceptual definitions of the related factors present inNANDA-IThe methodological process of this integrative review was in agreement with therecommendations of the Preferred Reporting Items for Systematic Reviews andMeta-Analyzes - PRISMAThe selected studies were thoroughly read and data were extracted using an instrumentvalidated in Brazilth and 16th, 2017, an integrative review wasconducted with the literature available at the databases Medical Literature Analysis andRetrieval System Online (MEDLINE via Pubmed), Cochrane Library, The Cumulative Index toNursing and Allied Health Literature (CINAHL), and in the Latin American & CaribbeanHealth Sciences Literature (LILACS). These databases were chosen due to their broadcoverage in the areas of health and nursing.Between February 8Medical Subject Headings Terms, Health Science Descriptors and key words from 17 DCs and3 RFs were used in the literature search along with term “dor aguda/acute pain” or“dor”, according to the result obtained. For the DCs “expressive behavior” and “narrowedfoccus” in the MEDLINE database (via Pubmed) the time criteria was withdrawn, sincethere were relevant results from the year 2011. The term “dor” was used only in theLILACS database, since many DCs and RFs did not have any result with the term “doraguda/acute pain”, so this strategy was used to increase the number of results. Theinclusion criteria were: results addressing pain in humans, published between the years2013-2017, in English, Spanish or Portuguese.After reading the titles and abstracts, 51 studies were selected for full text readingin Lilacs, 42 in MEDLINE, 19 in CINAHL and none in Cochrane. After the exclusion of theduplicates (n=30) and the unavailable studies (n=2), 80 studies proceed for full textreading. Of these, 16 had relevant content for the development of the conceptual andoperational definitions of the DCs and RFs presented in NANDA-I for the ND Acute Pain(00132), according to --As it was not possible to elaborate all the conceptual and operational definitions usingonly the studies of the systematic review, gray literature was also explored. Thus, adictionaryThe studies selected supported the development of this study. None of them had level ofevidence I or II, due to the characteristics of the subject; however, they providedvaluable contributions that supported the development of the definitions.The journals in which the studies were published are worth highlighting: British Journalof Anaesthesia, Canadian Journal of Emergency Medicine, Escola Anna Nery Revista deEnfermagem, International Journal of Behavioral Medicine, International Journal ofNursing Terminologies and Classifications, Pain Management Nursing (four studies),Revista Colombiana de Psiquiatría, Revista Dor (two studies), Revista Latino-Americanade Enfermagem, Revista de Neuro-psiquiatría e The Journal of Pain (two studies). The studies included in the review, classified according to title, country, year andobjective, are presented in -,,-,-,-,-,,Regarding language, ten studies were in EnglishThere were two reviews and 14 clinical studies, of which eleven were conducted withadults and three with children. Only two were conducted in critical care units. Sixstudies excluded the patients who could not communicate, which highlights the need topromote further research for this specific population.,,,-,,Regarding pain assessment, the verbal report of the patient, without the use of anappropriate instrumentRegarding the method, there were descriptive and qualitative studies, systematic reviewsof the literature and of patient records, observational studies and controlled studieswithout randomization.-,,,-,) and level VI -,,,,-The evidence levelsThe operational and conceptual definitions of the DCs for the ND Acute Pain (00132),developed after integrative review of the literature and additions from the grayliterature, are presented next:Self-report of intensity using standardized pain scale  - Conceptual definition:It is a subjective evaluation) makes a verbal reportSelf-report of pain characteristics using standardized pain instrument  - Conceptual definition: Itis a subjective evaluation) makes a verbal report) characterizing aspects of their pain other than intensity: location,,,-Distraction Behavior - Conceptual definition: It is the lack ofconcentration in the surroundings) to recognize the individual oblivious to the environment.Expressive behavior  -Conceptual definition: Expressive behavior refers to behavioral reactions tostimuli,,,Protective Behavior - Conceptual definition: Reaction to externalaggression  in an attempt to protect or defendHopelessness - Conceptual definition: It’s an emotion before, during and after the painful event. Record size immediately afterelevation of upper eyelids.Evidence of pain using standardized pain behavior checklist for those unable tocommunicate verbally  - Conceptual definition: It isthe use of a scale or classification previously validated to identify the existence ofpain in a specific population ; Operational definition:Pain assessment through the identification of the presence or absence of the items onthe scales. E.g. Behavioural Pain Scale (BPS) for adults in mechanical ventilation,validated in Brazil.Facial expression of pain  - Conceptual definition: It is the way theface,,Self-focused - Conceptual definition: Focus is the point to whichsomething convergesNarrowed focus - Conceptual definition: Focus is the point to whichsomething convergesProtective Behavior - Conceptual definition: Voluntary or involuntarybody movementAppetite change - Conceptual definition: Change in the desire toeatChange in physiological parameter  - Conceptual definition: Changes in vital signs caused by the release ofcatecholaminesPositioning to ease pain - Conceptual definition: Unusual bodypostureProxy report of pain behavior/activity change  - Conceptual definition: Exposition, description ornarrationThe conceptual definitions of the RFs for the ND acute pain (00132), constructed afteran integrative review of the literature and additions from the gray literature, arepresented next:Biological injury agent  -Conceptual definition: Something that causes an injury Physical injury agent  - Conceptual definition:Something that causes an injury ) related to a physical event of an accidental or surgical damage on organictissues.Chemical injury agent  - Conceptual Definition: Something that causes an injury (a traumatictissue change)The symptom of pain is a pressing issue among hospitalized patients, since millions ofpeople suffer from pain as a result of trauma, surgery or diseasesThis study aims to help nurses in clinical practice to correctly name and identify thesigns and symptoms (DCs) presented by the patient and the related factors (RFs) for thecorrect indication of the ND Acute Pain (00132). Thus, with the indication of an ND, thecare plan can be correctly implemented and the patient can receive the best availabletreatment to alleviate their suffering and prevent complications from inadequate painmanagement. -Therefore, a validation study addressing the components of the ND to improve the alreadyexisting taxonomies can be essential, since it can give the necessary strength to theNursing praxis, increasing the visibility of the profession. In addition, studiesindicate gaps in the knowledge about pain of the nursing team on critical careunitsMoreover, since the conceptual and operational definitions clarify and reinforce theitems in an ND, this study contributes to the development of specific language, whichallows for future discussions on the best available evidence on these components andtheir applicability to specific populations.Validation studies provide refinement and improvement of classifications, which favorsthe exercise of critical thinking and decision-making and improves communication andstandardized nursing recordsThe use of standardized language favors universality and dissemination of information.Therefore, it also benefits teaching activities, since students can better understandthe phenomenon studied.The present study aimed to elaborate the conceptual and operational definitions of theDCs and the conceptual definitions of the RFs of the ND Acute Pain (00132), addressingnonverbal patients in the ICU. The importance of describing this phenomenon is evident,since it can provide the scientific support for nurses to identify an ND thatrepresents, in fact, the patient’s responseIn addition to the frequency of the ND, it is relevant to determine the accuracy of thedefining characteristics for specific populations. This knowledge can help understandinghow the human response is presented, with its antecedents and consequences, and can alsosupport a quality careAnother clinical validation studyThese different data show the variation between nations and populations, but also showsome similarities. Therefore, these data indicate the importance of replicatingvalidation studies for different populations and of evaluating the clinical validity ofNDs after the content validation.And, finally, nursing practice is favored by the uniformity in the evaluation andindication of the diagnosis. This way, the patient also benefits, since the nurse willhave consistent evidence to make an ND and determine the next stages of the nursingprocessPain is a frequent event in ICU patients and this study considers particularly those whocan not communicate. The main characteristic of this population hinders theidentification of clinical evidences that lead to the ND Acute Pain.Despite the scarcity of studies addressing this issue for this population, there aremethods to describe and evaluate data that allow studying this phenomenon with the useof procedures that are commonly present in critical care.This way, the use of proper procedures and the identification of the definingcharacteristics and related factors of this ND must be in the methodology that will leadto the accuracy of these elements for this population.For this to occur, the development of the conceptual and operational definitions of theDCs and the conceptual definitions of the RFs of the ND Acute Pain (00132) of theNANDA-I, supported by the literature review and additions from gray literature,completes the first stage of a validation study of the ND. The next stages to bedeveloped in this study are the content analysis and clinical validation of this ND fornonverbal critically ill patients.


Intracortical microstimulation (ICMS) is one of the most widely employed techniques for providing causal evidence of the relationship between neuronal activity and specific motor, perceptual, or even cognitive functions. In recent years, several new types of linear multielectrode silicon probes have been developed, allowing researchers to sample neuronal activity at different depths along the same cortical site simultaneously and with high spatial precision. Nevertheless, silicon multielectrode probes have been rarely employed for ICMS studies and, more importantly, it is unknown whether and to what extent they can be used for combined recording and stimulation experiments. Here, we addressed these issues during both acute and chronic conditions. First, we compared the behavioral outcomes of ICMS delivered to the hand region of a monkey's motor cortex with multielectrode silicon probes, commercially available multisite stainless-steel probes and single-tip glass-coated tungsten microelectrodes. The results for all three of the probes were reliable and similar. Furthermore, we tested the impact of long-train ICMS delivered through chronically implanted silicon probes at different time intervals, from 1 to 198 days after ICMS sessions, showing that although the number of recorded neurons decreased over time, in line with previous studies, ICMS did not alter silicon probes' recording capabilities. These findings indicate that in ICMS experiments, the performance of linear multielectrode silicon probes is comparable to that of both single-tip and multielectrode stainless-steel probes, suggesting that the silicon probes can be successfully used for combined recording and stimulation studies in chronic conditions. The electrical stimulation of brain tissue marked the beginning of modern electrophysiology and strongly contributed to neuroscientific progress by allowing researchers to study brain organization and function in many animal species, including humans. Initially, electrical stimulation could be applied only to the cortical surface and induced gross activation that involved large neuronal populations , previously used for other neurophysiological experiments (see below). Before recordings, the monkey was habituated to sitting in a primate chair and to interacting with the experimenters. It was then trained to perform the visuomotor tasks described in previous studies . All experimental protocols were approved by the Veterinarian Animal Care and Use Committee of the University of Parma  and authorized by the Italian Ministry of Health .We performed both acute experiments, in which a single electrode/probe was temporarily inserted into the brain and then retracted at the end of the experimental session, and chronic experiments, in which probes were permanently implanted in the brain tissue for several months.After acute single-cell recording experiments with different types of linear multielectrode probes were conducted . Dexamethasone and prophylactic broad-spectrum antibiotics were administered pre- and postoperatively. Furthermore, analgesics were administered intra- and postoperatively. Dexamethasone administration was continued for 1 week after the surgery. During all surgeries, hydration of the monkey was maintained with continuous infusion of saline solution. A heating pad stabilized the monkey's body temperature throughout the surgical procedure. Heart rate, respiratory depth, and body temperature were continuously monitored. Upon recovery from anesthesia, the animal was returned to its home cage and closely monitored until complete recovery.The neuronal recordings described in this paper were performed by means of two 3D arrays of 64 sputter-deposited platinum electrodes arranged into 2 parallel lines of 4 shafts, each of which had eight recording channels. Probes were implanted with a vertical approach, approximately 1 mm laterally to the mesial wall. Previous reports provide more details on the methodology of probe fabrication , and recorded in sets of 16 channels. All final quantitative analyses were performed offline, as described in the subsequent sections. From the same chronically implanted probes, ICMS was performed at the end of the recording sessions. Monopolar, biphasic trains of cathodic square-wave pulses were delivered through a constant current stimulator , with the following parameters, which are based on previous ICMS studies on area F6  was detected by applying to the filtered  wide-band activity a negative threshold corresponding to 2.5 standard deviations from the mean peak height. Spikes were then sorted using standard principal component and template matching techniques, provided by dedicated offline sorting software (Plexon Inc.), as described elsewhere  relative to those that exhibited the opposite effect (n = 1). Notably, most of the sites that initially showed SUA still did that after ICMS as well (14/16). Figure Figure as well 1/16. FiguFigure In this study, we compared the performance of linear multielectrode silicon probes in acute ICMS experiments in the motor cortex of a monkey with the performance of linear multielectrode stainless-steel probes and classical tip electrodes. In addition, we verified the long-term single-neuron recording capabilities of chronically implanted silicon probes previously used for both ICMS and single-neuron recordings.The results of the acute ICMS experiments showed that all three probes could trigger similar behavioral reactions at comparable current-intensity thresholds. We used a single tip-electrode for comparison because most extant ICMS studies on both motor and nonmotor brain regions have been carried out with this type of electrode. However, because we sequentially replicated cortical penetrations in the same position with different probes , it could be argued that we failed to capture subtle differences among probes, thus producing false-negative results. Nevertheless, it is worth noting that, with all probes, the minimum and average current-intensity thresholds reflect the well-established properties of the motor cortex, that is, greater excitability of the cortical sites located closer to or within the anterior bank of the central sulcus , to deliver ICMS at all the available sites  of microelectrode arrays chronically implanted in 27 monkeys over a period of about 15 years showed that the recording duration ranged from 0 to 2,104 days, with a considerable case-by-case variability and with a number of possible different failure modes (Barrese et al., An important limitation of the present study consists in the low number of observations carried out on a single animal with a single set of probes. A comprehensive retrospective report on a large set (CF performed the experiments, analyzed the data, prepared figures and wrote and revised the manuscript; MM performed the experiments, analyzed the data and revised the manuscript; AL performed the experiments, analyzed the data and revised the manuscript; ML performed the experiments, analyzed the data and revised the manuscript; LB designed the research, supervised the experiments, wrote and revised the manuscript.The 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.


Lactoferrin is a multifunctional glycoprotein found in the milk of most mammals. In addition to its well-known role of binding iron, lactoferrin carries many important biological functions, including the promotion of cell proliferation and differentiation, and as an anti-bacterial, anti-viral, and anti-parasitic protein. These functions differ among lactoferrin homologs in mammals. Although considerable attention has been given to the many functions of lactoferrin, its primary nutritional contribution is presumed to be related to its iron-binding characteristics, whereas the role of glycosylation has been neglected. Given the critical role of glycan binding in many biological processes, the glycan moieties in lactoferrin are likely to contribute significantly to the biological roles of lactoferrin. Despite the high amino acid sequence homology in different lactoferrins (up to 99%), each exhibits a unique glycosylation pattern that may be responsible for heterogeneity of the biological properties of lactoferrins. An important task for the production of biotherapeutics and medical foods containing bioactive glycoproteins is the assessment of the contributions of individual glycans to the observed bioactivities. This review examines how the study of lactoferrin glycosylation patterns can increase our understanding of lactoferrin functionality. Lactoferrin is a highly glycosylated protein that was first isolated from bovine milk in 1939 by Sorensen and Sorensen . It has Milk lactoferrin possesses varying numbers of potential glycosylation sites, depending on the species of origin . Althoug3+ ions +1, 204.09 m/z [HexNAc + H]+1, and 366.14 m/z [HexNAc − Hex + H]+1. Each spectrum can be screened for the presence of fragment ions. After N-glycan identification, molecules can be entered into digital data libraries. These libraries typically include mass, retention time, and monosaccharide compositions. The availability of these libraries has driven the development of software programs capable of interrogating these libraries in real time to identify molecules in chromatograms based on the accurate mass and retention time. Hua and Kronewitter [N-linked glycans, illustrating the enabling utility of this approach.For particularly complex mixtures, measuring the accurate mass is not sufficient to identify structures. Therefore the MS/MS mode is used, where precursor ions are fragmented to generate product ions that are analyzed by QTOF. Fragmentation is performed by collision-induced dissociation (CID). Precursor ions strike collision gas molecules (nitrogen), leading to their fragmentation . These pnewitter ,70 estabN-glycans for which very limited information is currently available. The combination of these novel approaches with advanced mass spectrophotometric tools and bioinfomatic libraries, will enable structure-activity studies to be made of the naked protein backbone and the glycosylated form, and these technologies will enable the identification of key specific glycan compositions for functional activities. Numerous products are already on the market, or under development.Research in multiple disciplines interfacing glycobiology continues to discover novel biological roles for the multi-functional protein lactoferrin. It is, therefore expected that the diverse structures and functions of lactoferrin will remain a popular target for investigations, and that this research will reveal additional functions and health benefits in the near future. While the backbone sequence of milk proteins has been extensively elucidated via proteomics studies, the roles of glycosylation in contributing to, or altering these protein bioactivities, have largely been ignored. Advancements in cataloguing the complexity and unique glycosylation patterns of lactoferrin in different biological sources are necessary to for next phase of research: illuminating the exceptional contribution of glycans to biological and physicochemical properties of a wide range of proteins. The application of novel deglycosylating enzymes will play important roles for the determination of the contribution of these glycans to the function of lactoferrin. Especially, considering EndoBI-1’s ability to release specific glycan structures at different conditions, this will enable the discrimination of the activity of multiple glycans pools. Moreover, the application of novel enzymes will help to determine the actual sites of


In mammals several cell adhesion molecules are involved during the pre- and postnatal development of all organ systems. A very prominent member of this family is the neural cell adhesion molecule (NCAM). Interestingly, NCAM can be a target for a special form of posttranslational modification: polysialylation. Whereas nearly all extracellular proteins bear mono-sialic acid residues, only a very small group can be polysialylated. Polysialic acid is a highly negatively-charged sugar polymer and can comprise more than 90 sialic acid residues in postnatal mouse brains increasing dramatically the hydrodynamic radius of their carriers. Thus, adhesion and communication processes on cell surfaces are strongly influenced allowing, e.g., the migration of neuronal progenitor cells. In the developing brain the essential role of polysialylated NCAM has been demonstrated in many studies. In comparison to the neuronal system, however, during the formation of other organs the impact of the polysialylated form of NCAM is not well characterized and the number of studies is limited so far. This review summarizes these observations and discusses possible roles of polysialylated NCAM during the development of organs other than the brain. Escherichia coli (E. coli) K1, are also able to generate different polysialic acid (polySia) species . Ho. Ho22]. trans mode, increasing amounts of polySia abolish the interaction between NCAM molecules expanding the area between cells. Due to the dramatic increase of the hydrodynamic radius also cell-cell interactions mediated by other adhesion molecules like cadherins can also be negatively affected [Nevertheless, the most prominent example is still the tuning of cell-cell adhesion mechanism by the inhibition of the homophilic NCAM-NCAM interaction. More than twenty-five years ago Rutishauser and co-workers proposed that the cell-cell interaction is triggered depending on the polySia ratio of NCAM ,54. Whilaffected . Thus, paffected ,57.As already mentioned, polySia can regulate physiological processes independently of its carrier. Especially Sato’s and Kitajima’s groups identified several biomolecules, which bind polySia inducing or inhibiting distinct signaling pathways ,58. InteMoreover, fibroblast growth factor 2 (FGF2) was shown to be an interaction partner of polySia . The comIn addition to the presented modulation of NCAM functions as well as interaction partners, some others functionalities of polySia were described . In addition, interstitial cells are located between the tubules. In addition to interstitial macrophages, Leydig cells are primarily present in the interstitium. Leydig cells belong to the endocrine system and they release, inter alia, testosterone and other androgens, such as androstenedione and dehydroepiandrosterone (DHEA). Thus, Leydig cells are important regulators for sexual development and spermatogenesis. Mayerhofer et al. detected NCAM on Leydig cells of adult testes in the beginning of the 1990s . ShortlyNot only interstitial cells, but also Sertoli cells and germ cells, seem to express the polysialylated form of NCAM during fetal development. In feline  and murine  testes, membranes of Sertoli cells and/or spermatogonia are polySia-positive .Since neurotrophins like BDNF, NGF, and NT-3 are discussed to support the forming of the seminiferous cord, as well as the persistence of germ cells , and polThis possibility is supported by findings using the roe deer as a model for seasonal initiation and termination of spermatogenesis . In wildThe initiation of polysialylation during these key points of seasonal spermatogenesis and the ability of polySia to modulate cell-cell interaction and communication, as well as the functionality of growth factor, may contribute to the regulatory system of spermatogenesis. A study showing that spermatogonia differentiation is reversible and stem cell potential is regained when their connections are detached ,79 let uDuring epididymal transit sperm matures. This maturation step includes an exchange of several surface components between epithelial cells and sperm cells, which is essential for them to become fertile . InteresNevertheless, also during the postnatal development of murine epididymis polysialylated NCAM is expressed . In addiRemarkably, the reduction of the polySia levels comes along with increasing quantities of extracellular collagen during the formation of these contractile areas. A comparable interrelation was also observed in the postnatal tunica albuginea , which eIn mammals the placenta is essential for the development of new life. Interestingly, during pregnancy in humans the placenta consists of embryonic, as well as maternal, cells. This is possible, since embryonic cells invade the uterus leading to the crucial formation of a nutrient/waste exchange system . TherebyRemarkably, at term nearly no polysialylation occurs. Fisher and co-workers performed in addition functional assays to get an idea, which processes may depend on the polysialylation status of invasive cytotrophoblasts. By an enzymatic removal of polySia they demonstrated in cell culture models that the migration, as well as invasion capacity of invasive cytotrophoblasts, are significantly impaired .Furthermore, it seems that cancer cells of gestational trophoblastic disease tumors overexpress polySia . The occEspecially due to pioneering work of the lab of Jürgen Roth and cooperation partners, we have known for many years that the polysialylated form of NCAM is not only restricted to the neuronal system during pre- and postnatal development in mammals. In addition to the outlined organs, the development of other physiological systems seems to be supported by polySia-NCAM. For instance, during hair follicle formation, as well as during the development of the digestive and the respiratory elements, the polysialylated form of NCAM is present, as shown in Lackie et al. . FurtherTaken together, the described localization, as well as the abilities of polySia to influence cell-cell interaction/communication and proliferation, in addition to differentiation processes, seems to trigger the maturation of organs throughout the body. Nevertheless, organ and/or cell specific ablation of polySia in murine systems will be necessary to determine exactly the various functions of polySia as a posttranslational modification of NCAM in all different physiological systems.


The biosynthesis, structural diversity, and functionality of collagens of sponge origin are still paradigms and causes of scientific controversy. This review has the ambitious goal of providing thorough and comprehensive coverage of poriferan collagens as a multifaceted topic with intriguing hypotheses and numerous challenging open questions. The structural diversity, chemistry, and biochemistry of collagens in sponges are analyzed and discussed here. Special attention is paid to spongins, collagen IV-related proteins, fibrillar collagens from demosponges, and collagens from glass sponge skeletal structures. The review also focuses on prospects and trends in applications of sponge collagens for technology, materials science and biomedicine. Collagens constitute a superfamily of long-lived extracellular matrix structural proteins of fundamental evolutionary significance, found in both invertebrate and vertebrate taxa. They are among the most studied proteins due to their important functions in mammals, including humans. In addition to their structural function in cartilage and skin formation ,2, as weMarine vertebrate collagens have attracted scientific attention, mostly as products of fisheries . In partIn contrast to marine vertebrate collagens, similar structural proteins found in marine invertebrates represent one of the most ancient protein families within Metazoa. Marine invertebrate collagens arose earlier than their vertebrate analogs, and possess diverse unique structural features, including very special structure–function interrelations. Collagens from poriferans, coelenterates, annelids, mollusks, echinoderms, and crustaceans have been discussed in detail in several review papers  are the most simple and ancient multicellular organisms on our planet, and mostly live attached to a suitable substratum  on the seabed. Poriferans diverged from other Metazoans earlier in evolutionary history than any other known animal phylum, extant or extinct , with thDifferent systems of terminology relating to poriferan collagens are found in the literature, as sponges also display considerable polymorphism with respect to their collagenous structures. The insolubility of most poriferan collagens has been the main obstacle to carrying out any detailed biochemical analysis. Studies on the morphology and nanotopography of the collagenous fibrils have shown that they are dispersed throughout the intracellular matrix within the skeletons of sponges. Cuticular structures have been found in some sponges, but their molecular composition has not been determined .Spongia graminea, which they called spongin A and spongin B  motif predisposed for silica precipitation, and provides a novel template for biosilicification in nature [Hyp–4Hyp Monorhaphis sp. [H. sieboldi, collagen isolated and identified from Monorhaphis sp. was matched only to the type I collagen pre-pro-alpha (I) chain (COL1A1) from dog (AAD34619)  [Collagen has also been reported as the main organic component of the spicules of the glass sponge phis sp.   .C. reniformis has been successfully used as an alternative by the Ehrlich research group [The existence of naturally occurring collagen–silica-based composites in the form of spicules of glass sponges stimulated material scientists to develop analogous hybrid materials. Due to the limited available amounts of glass sponge collagen for the development of silica-based composite materials, fibrillar collagen from the demosponge ch group ,129. Morch group ,137,138.ch group . In workch group , collagech group . It was ch group . Such noch group .We have presented here only a brief discussion of selected examples, which nonetheless provide novel data concerning poriferan collagens. In spite of the progress made in this research field, numerous open questions remain. For example, additional investigations are necessary to obtain understanding of the nature and origin of halogenated spongins. It is still not clear how many collagen and/or keratin domains they contain. Additionally, the unique resistance of these biopolymers against diverse chemicals and enzymes remains poorly investigated. The possible role of collagens in the spiculogenesis of demosponges and formation of axial filaments must also be researched. The discovery of crystalline proteins within amorphous biosilica-based structures in sponges is ground-breaking in the understanding of biomineralization. What can be discovered about the crystallinity of collagen within biosilica-related structures in sponges? The existence of collagen-based crystals within siliceous biominerals could revolutionize our understanding of the origin and evolution of collagens, from the point of view of biomineralization in sponges as the first multicellular organisms on Earth. Further, the relationship of collagen and chitin in the skeletal structures of diverse sponge classes and orders is entirely unknown. Consequently, we believe that the use of modern X-ray imaging techniques based on the “diffraction before destruction” principle is the best way forward to gain understanding of the principles of the unique organization of collagen within both fossil and recent collagen-based biomineralized constructs.Novel approaches must be proposed which will bring together modern bioanalytical and molecular biology methods for better understanding of the fundamental principles of collagen fibrillogenesis and the mechanisms of its cross-linking in sponges, as well as details of the structural organization of poriferan collagens at the molecular and atomic levels. The best way to address this challenging task on these levels is by coherent synergetic collaboration using explicit reasoning and well-tested explanatory principles of multidisciplinary knowledge, experience, and new technologies. Finally, we suggest that studying the processes of marine farming of the collagen-producing demosponges has implications for a variety of practical large-scale applications, ranging from the design of highly effective extraction techniques to the development of novel collagen-containing composites for biomedicine and technology.


Drosophila . ThWolbachia are potent regulators of viral infection and are used to control the vector capacity of some mosquitoes [Drosophila midgut [Acetobacter pomorum activates the NF-κB cascade, leading to the transcriptional induction of the Platelet-Derived-Growth Factor/Vascular Endothelial Growth Factor (PDGF-VEGF) homologue, Pvf2 (Drosophila in this era of (re)emerging arboviruses.In insects, endosymbionts such as squitoes –56. In msquitoes . Recentla midgut . This stue, Pvf2 . In paraue, Pvf2  and bindDrosophila gut microbiota, and the ease of genetically manipulating Drosophila, the fruit fly will continue to thrive as a workhorse for biological discoveries in this area.The gut is a major interface between a host and microbes. Whether we consider the emerging notion that the gut microbiota influences host physiology, the relation of GI inflammation to health and disease, or that the gut of insect vectors is a first point of contact with human parasites, it is increasingly important to elucidate the complexity and plethora of interactions between the GI tract and microbes. Due to the strong conservation of both structure and function of the gut epithelium, the less complex microbial community that composes the


To report the case of a 39-year-old man with bilateral Eales disease.  The clinical, angiographic, and ultrasonographic evaluations of the patient were thoroughly and specifically carried out. The treatment consisted of pars plana vitrectomy and endolaser treatment at the time of surgery to the left eye and laser scatter retinal photocoagulation to the right eye. Postoperatively, the visual acuity improved at 20/ 20 to the left eye. Eales disease is a clinical diagnosis of exclusion as many other retinal disorders can mimic it, especially conditions of retinal inflammation or neovascularization. The condition, with a characteristic clinical picture, fluorescein angiographic finding, and natural course, is considered a specific disease entity, easily distinguished from other vasculitides and retinopathies. The condition usually affects healthy young adult males in their second decade and is observed in men of all age groups being bilateral in up to 90% of the patients although symptoms often present unilaterally. Peak age of onset for Eales disease is 20-35 years, with a reported range of 13-63 years [2]. Eales disease is a primary idiopathic peripheral obstructive retinal vasculopathy of unclear etiology. The disease, also known as angiopathia retinae juvenilis, periphlebitis retinae, primary perivasculitis of the retina, is characterized by three overlapping stages of venous inflammation , ischemia, and retinal neovascularization, being hallmarked by recurrent retinal and vitreal hemorrhages .The etiopathogenic diagnosis is one of presumption. The cause of this disease is not known. However, the Polymerase Chain Reaction detected the deoxyribonucleic acid of the bacterium Mycobacterium tuberculosis in a significant number of patients [3-5], and anti-tubercular therapy (in patients with exposure to tuberculosis) are recommended in the early stages. For patients with a later stage of the disease, the laser scatter retinal photocoagulation  in the areas of retinal non-perfusion and direct photocoagulation of new and underlying feeder vessels were suggested. For nonresolving vitreous hemorrhages and/ or retinal detachment, the pars plana vitrectomy (no later than 6 months following the onset of hemorrhage) with or without other vitreoretinal surgical procedures is necessary.Therapeutically, the treatment has to be adapted to the evolution stage of the disease. The steroidal and non-steroidal agents, intravitreal anti-vascular endothelial growth factor (VEGF) therapy [As many other retinal disorders can mimic it, especially conditions of retinal inflammation or neovascularization, Eales disease is a clinical diagnosis of exclusion. With a characteristic clinical picture, fluorescein angiographic finding, and natural course, the disease is considered a specific disease entity, easily distinguished from other vasculitides and retinopathies.AcknowledgmentsAll the authors have made substantial contributions to the manuscript ; they have provided the corresponding author with a written permission to be named in the manuscript. The authors do not have a financial relationship. No organization sponsored the research. DisclosuresThe authors have no proprietary of commercial interest in any of the materials discussed in this article; they declare no conflict of interest. The authors have nothing to disclose.


Mammalian genomes are scattered with thousands of copies of endogenous retroviruses (ERVs), mobile genetic elements that are relics of ancient retroviral infections. After inserting copies into the germ line of a host, most ERVs accumulate mutations that prevent the normal assembly of infectious viral particles, becoming trapped in host genomes and unable to leave to infect other cells. While most copies of ERVs are inactive, some are transcribed and encode the proteins needed to generate new insertions at novel loci. In some cases, old copies are removed via recombination and other mechanisms. This creates a shifting landscape of ERV copies within host genomes. New insertions can disrupt normal expression of nearby genes via directly inserting into key regulatory elements or by containing regulatory motifs within their sequences. Further, the transcriptional silencing of ERVs via epigenetic modification may result in changes to the epigenetic regulation of adjacent genes. In these ways, ERVs can be potent sources of regulatory disruption as well as genetic innovation. Here, we provide a brief review of the association between ERVs and gene expression, especially as observed in pre-implantation development and placentation. Moreover, we will describe how disruption of the regulated mechanisms of ERVs may impact somatic tissues, mostly in the context of human disease, including cancer, neurodegenerative disorders, and schizophrenia. Lastly, we discuss the recent discovery that some ERVs may have been pressed into the service of their host genomes to aid in the innate immune response to exogenous viral infections. A retroviral genome exists in different forms during its replication cycle. A viral particle, or virion, protects the RNA genome of the retrovirus during escape from the host cell and infection of new cells. A virion that enters a new host cell deploys its genomic payload, using its own reverse transcriptase to convert the RNA viral genome into a DNA copy which is integrated into the host genome, referred to as a provirus Figure . SubsequThese ancient genomic residents represent a potent source of genomic and regulatory variability. The high degree of homology between these ERV copies, and the presence of the long terminal repeats (LTRs) at either end of each copy Figure , provideCertain stages of mammalian pre-implantation embryo and germ cell development characterized by multiple waves of epigenetic reprogramming pose a unique challenge for the control of endogenous retroviral activity. During the two waves of epigenetic reprogramming that occur in primordial germ cells (PGCs) and fertilized oocytes, a considerable amount of DNA demethylation occurs. Examination of global DNA methylation at these stages have shown that levels within human and mouse pre-implantation embryos decrease beginning at the 1- to 2-cell stage, depending on the species, and up to or soon after the blastocyst stage . For example, elevated expression of the ERV-H family has been observed within both naïve-like and primed hESC sub-populations  layer of blastocysts, and plays a critical role in the gas, nutrient, and waste exchange required for normal embryonic growth. It is well established that both mouse and human placentas are hypomethylated compared to other somatic cells derived from either env gene from the ERV-fb1 element and is transcribed within the placenta. Within normal human placentas, suppressyn is co-expressed with Syncytin-1 in the syncytiotrophoblast layer  and a corresponding immunosuppressive domain. Additionally, both ERV-V and ERV-K expression has been observed within placental trophoblast cells at the maternal-fetal interface, although corresponding in vitro functional assays have not yet been completed to directly support in vivo findings  represents the best-preserved and most recently active ERV, containing a substantial number of loci that have predicted coding potential throughout different primate genomes. It has also been observed that ERV-K encodes a small accessory protein, Rec, in naïve ES cells and human blastocysts. Overexpression of Rec protein within human pluripotent cells increases the innate antiviral response and can inhibit exogenous viral infections, suggesting an immunoprotective role of the ERV-K Rec protein during early embryonic development -inducible enhancers have been dispersed via ERV insertions adjacent to IFN-inducible genes independently over mammalian evolution. This has resulted in regulatory networks of genes able to work in concert due to the presence of these ERV sequences. Further, CRISPR-Cas9 deletion of a MER41 insertion upstream of The relationship between ERVs and the human genome is a diverse and complicated one, resulting from millions of years of co-evolution. ERVs are known to be involved in disease through insertional mutagenesis, as targets of epigenetic repression, and via recombination of sequences between the homologous copies of these elements scattered across the genome. Throughout mammalian evolution, the deleterious effects of ERVs have been balanced by the benefits gained from innovative co-option of their sequences and proteins by their host genomes. These innovations include the intimate relationship between ERV activity with embryonic and placental development, as well as a number of ERV-associated regulatory networks that have become important components of the normal function of our genome. An innate immune response to exogenous retroviral infection is likely only one of several ERV functional roles. Once thought to have been quiescent, dead residents of the human genome, we are only beginning to uncover the scope of how actively intertwined our biology is with these long-time genomic partners.TM and JR drafted the manuscript and figures. SC and LC edited the manuscript.TM was supported by the National Library of Medicine of the National Institutes of Health under Award Number T15LM007088. JR was supported by the Collins Medical Trust Foundation and Glenn/AFAR Scholarship for Research in the Biology of Aging. LC and SC were supported by NIH/NICHD R01HD086073-A1, National Centers for Translational Research in Reproduction and Infertility (NCTRI). Additional funding for SC came from the Georgeanna Jones Foundation for Reproductive Medicine, Medical Research Foundation of Oregon, and the Collins Medical Trust. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health, the Collins Medical Trust Foundation, or Medical Research Foundation of Oregon.The 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.