Extracorporeal life support (ECLS) is an important device in the management of children with severe refractory cardiac and or pulmonary failure. Actually, two forms of ECLS are available for neonates and children: extracorporeal membrane oxygenation (ECMO) and use of a ventricular assist device (VAD). Both these techniques have their own advantages and disadvantages. The intra-aortic balloon pump is another ECLS device that has been successfully used in larger children, adolescents, and adults, but has found limited applicability in smaller children. In this review, we will present the “state of art” of ECMO in neonate and children with heart failure. ECMO is commonly used in a variety of settings to provide support to critically ill patients with cardiac disease. However, a strict selection of patients and timing of intervention should be performed to avoid the increase in mortality and morbidity of these patients. Therefore, every attempt should be done to start ECLS “urgently” rather than “emergently,” before the presence of dysfunction of end organs or circulatory collapse. Even though exciting progress is being made in the development of VADs for long-term mechanical support in children, ECMO remains the mainstay of mechanical circulatory support in children with complex anatomy, particularly those needing rapid resuscitation and those with a functionally univentricular circulation. With the increase in familiarity with ECMO, new indications have been added, such as extracorporeal cardiopulmonary resuscitation (ECPR). The literature supporting ECPR is increasing in children. Reasonable survival rates have been achieved after initiation of support during active compressions of the chest following in-hospital cardiac arrest. Contraindications to ECLS have reduced in the last 5 years and many centers support patients with functionally univentricular circulations. Improved results have been recently achieved in this complex subset of patients. Mechanical circulatory support is an important tool in the management of children with cardiac failure. Two major forms of mechanical circulatory support are currently available in neonates and children: extracorporeal life support  and ventricular assist device (VAD). Each of these devices has advantages and disadvantages. Another device that has been used in older children is the intra-aortic balloon pump. As mechanical circulatory support in children has evolved, the indications have expanded and outcomes have improved.Survival for children with heart failure on ECLS has progressively improved over the past two decades, and this is occurring despite placing more complex patients on support, including a significant proportion with single-ventricle physiology, end-stage heart failure or in cardiac arrest . SurvivaThe purpose of this review article is to outline current management strategies in the application of ECLS to children with heart disease, review recent evidence about the effect of these strategies, and provide an overview of the topic for clinicians.Indications and contraindications for ECLS in neonates and children with cardiovascular disease have evolved during the past 45 years. Advances in ECLS circuit design and greater recognition that ECLS is beneficial for indications beyond early postoperative support have resulted in steady growth in the number of pediatric patients who receive cardiac ECLS. The indications for ECLS can be divided into two groups: those involving cardiac surgery and not. The indications related to cardiac surgery include preoperative stabilization, failure to wean postcardiotomy, low cardiac output syndrome in the postoperative period, and cardiac arrest. The indications in the absence of cardiac surgery are cardiac arrest, myocarditis and cardiomyopathy, pulmonary hypertension, intractable arrhythmias, and other forms of shock, such as sepsis or Kawasaki’s Disease  is well established. Mechanical circulatory support may be required in the postoperative period, either due to inability to wean from bypass or because of a progressive low cardiac output syndrome. Patients who are subjected to prolonged periods of cardiopulmonary bypass can experience severe postoperative myocardial dysfunction that may prevent successful separation from cardiopulmonary bypass. Children who have preexisting ventricular dysfunction or certain structural cardiac abnormalities [such as hypoplastic left heart syndrome (HLHS)] may be at greatest risk. Once correctable residual anatomic lesions have been excluded, ECLS may be used to provide short-term cardiopulmonary support. In most cases, the cannulation strategy utilized for cardiopulmonary bypass is adequate during and after the transition to ECMO. Veno-arterial ECMO is the most commonly used modality in neonatal and pediatric cardiac patients during the perioperative period although veno-venous ECMO may be used in select circumstances, such as intraoperative pulmonary hemorrhage, which can occasionally occur in lesions, such as Tetralogy of Fallot and its related variants.There is no consensus regarding the optimal timing for ECMO initiation in the perioperative period. Some reports suggest the absence of a clear relationship between timing of ECLS initiation and clinical outcome, whereas others describe improved survival in patients who receive early ECLS support, including those who transition to ECLS in the operating room, compared to those who receive support later in the postoperative period –16. ECMOAlthough the vast majority of neonatal and pediatric cardiac ECLS occurs in patients with structural CHD who are undergoing surgical repair or palliation, patients without structural heart disease may require ECLS to manage severe heart failure.Children with acute fulminant myocarditis can benefit from mechanical circulatory support as a bridge to recovery. Cardiomyopathy (8.3%) and myocarditis (4.5%) are the most common non-structural etiologies of heart failure in neonates and children reported to the ELSO registry . Overallbridge-to-decision and as a bridge-to-bridge (bridge-to-VAD)  . When usExtracorporeal life support can also be used after heart transplantation. Indications for support in this setting include failure to separate from cardiopulmonary bypass and progressive low cardiac output syndrome in the immediate postoperative period, along with circulatory collapse due to rejection or graft vasculopathy later in the course , 31. OveThe list of contraindications to ECLS in children with heart disease is shrinking over time. Absolute contraindications include lethal chromosomal abnormalities, severe irreversible brain injury, and extremely low gestational age and weight (<32 weeks gestation or <1.5 kg). Relative contraindications include moderate intra-ventricular hemorrhage, gestational age <34 weeks, weight <2.0 kg, and certain high-risk congenital heart lesions, such as those that co-exist with congenital diaphragmatic hernia. Data from the ELSO registry indicate that the overall survival rate for premature infants (<37 weeks gestation) who receive cardiac ECLS is 31%, compared to 41% survival in term infants . Weight The approach to cannulation should be flexible and based on the underlying need for ECMO. Transthoracic cannulation of the right atrial appendage and the ascending aorta can be used in cases of failure to wean from cardiopulmonary bypass. In the immediate postoperative period, reopening the sternal wound and direct cardiac cannulation provides the most expeditious route to institute support, especially in patients who suffer cardiac arrest. Adequate venous drainage and excellent arterial perfusion should be assured by chest cannulation, providing the cannulas are placed properly; however, severe hemorrhage and mediastinitis remain disadvantages of chest cannulation, making peripheral cannulation preferable in most other settings.Cannulation of the right internal jugular vein and the common carotid artery provides excellent venous drainage and perfusion and is the preferred cannulation site in neonates and children below 15 kg. Cannulation of the femoral vessels provides adequate venous drainage and perfusion for larger children. A second venous drainage cannula placed in the right internal jugular vein may be added if venous drainage is inadequate through the femoral route. To avoid ischemia of the lower extremities, a perfusion cannula in the distal femoral artery is placed by many groups. Venous congestion of the lower extremities is less frequent and usually does not require treatment.In cases of severe heart dysfunction, inadequate decompression of the left-sided cardiac chambers during ECLS may be seen. A number of strategies can be implemented to address this. Left-sided distension may be managed by increasing ECLS flow to empty the right heart, minimizing pulmonary blood flow, and decreasing pulmonary venous return to the left heart. However, this increases the afterload to the systemic ventricle, which may cause the aortic valve to close, in turn risking thrombus formation in the ventricle as well as hemorrhagic pulmonary edema if the end-diastolic pressure rises too high. In cases of persistent distension of the systemic ventricle, a balloon atrial septostomy can be performed in the cardiac catheterization laboratory. If this is not technically possible, additional surgical options include changing to central cannulation, with or without biatrial cannulation, inserting an atrial vent via a pulmonary vein, or inserting an apical ventricular vent . These vIn selected patients with malignant tachyarrhythmias or bradyarrhythmias, ECLS may be necessary to mitigate cardiogenic shock while medical therapy is being optimized or in order to safely facilitate catheter ablation or pacemaker insertion. Generally, patients with severe myocarditis or dilated cardiomyopathy are supported with ECLS to manage severe arrhythmias. Severe arrhythmias can also present after cardiac surgery or in lesions, such as Ebstein’s malformation –39. CathAs the experience with ECLS has grown, new indications have evolved, including emergent resuscitation. del Nido and colleagues  initiallMany centers previously considered a functionally univentricular circulation a contraindication to ECLS. However, this view is anachronistic and outcomes can be satisfactory in many instances. In addition to the same indications for ECLS as in children with biventricular hearts, shunt thrombosis may also require ECLS and is associated with excellent outcomes when the thrombosis is swiftly identified and corrected. However, while the indications are similar, the cannulation strategies require careful planning.Management of the infant with single ventricular physiology and a systemic-pulmonary shunt can be safely achieved by leaving the shunt patent and increasing the net ECLS circuit flow to compensate for the pulmonary run-off Figure . GeneralThere is limited experience with mechanical support in patients with cavo-pulmonary connections and outcomes are not so good as for other indications, most likely due to a combination of previous surgery, complex physiology, difficult cannulation approach, the need to address venous drainage from both superior and inferior vena cava and the inadequacy of conventional cardiopulmonary resuscitation (CPR) to provide sufficient blood flow if these children suffer of cardiac arrest –54.Patients who have undergone Glenn procedure Figure  have a uAdequate flow drainage is complicated also in Fontan patients , 54. DueMany forms of MCS are available for children with heart failure refractory to conventional management. Actually, ECMO can be used in many settings to support pediatric patients with cardiac disease. However, the use of ECMO in this field requires careful selection of patients and timing of interventions remains challenging. The principles used presently in the application of ECMO to support the failing circulation in children will be used as the foundation for developing innovative circulatory techniques for the future.Dr. MN and Dr. MM wrote 80% of the manuscript. Dr. CC and Dr. PB wrote the remaining 20% of the manuscript. Dr. GM and Dr. AA 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.


Elemental homeostasis has been largely characterized using three important elements that were part of the Redfield ratio . These efforts have revealed substantial diversity in homeostasis among taxonomic groups and even within populations. Understanding the evolutionary basis, and ecological consequences of such diversity is a central challenge. Here, we propose that a more complete understanding of homeostasis necessitates the consideration of other elements beyond C, N, and P. Specifically, we posit that physiological complexity underlying maintenance of elemental homeostasis along a single elemental axis impacts processing of other elements, thus altering elemental homeostasis along other axes. Indeed, transcriptomic studies in a wide variety of organisms have found that individuals differentially express significant proportions of the genome in response to variability in supply stoichiometry in order to maintain varying levels of homeostasis. We review the literature from the emergent field of ionomics that has established the consequences of such physiological trade-offs on the content of the entire suite of elements in an individual. Further, we present experimental data on bacteria exhibiting divergent phosphorus homeostasis phenotypes demonstrating the fundamental interconnectedness among elemental quotas. These observations suggest that physiological adjustments can lead to unexpected patterns in biomass stoichiometry, such as correlated changes among suites of non-limiting microelements in response to limitation by macroelements. Including the entire suite of elements that comprise biomass will foster improved quantitative understanding of the links between chemical cycles and the physiology of organisms. Ecological stoichiometry considers individuals as collections of chemical elements akin to a very large molecule. At the most fundamental level, ecological stoichiometry is the study of the sub-organismal mechanisms, and supra-organismal consequences of the principle of mass balance. It operates on the axiom that living entities are not a passive conduit of chemical supply, but rather actively regulating their elemental stoichiometry, referred to as elemental homeostasis . This raises the possibility that genotypes exhibiting differing degrees of elemental homeostasis can vary in other traits. At the simplest level, we can think of elements that share similar properties that can be replaced when supply of one is limiting  model, which included strong positive  and negative  correlations among elements. Considering combinations of elements as phenotypes, as opposed to considering single elements at a time, allows for greater sensitivity in identifying stoichiometric variation because of the fundamental interconnectedness among elements in biomass   with three strains coming from each genus. All three of the Brevundimonas strains were characterized as flexible, whereas Flavobacterium and Sphingomonas each had two strains characterized as inflexible and one strain as flexible. Although we expected strong strain-specific responses, we generally predicted that homoestoichs should exhibit greater changes in other elements  between high and low P supply conditions compared to heterostoichs due to upregulation of compensating mechanisms. Each strain was originally isolated from lakes within Minnesota using either agar plates or dilution isolation as described previously  were present above detection limits in all samples and were used for further analyses. As expected, there was considerable strain-specific variation in biomass P content . Although considerable strain-specific responses preclude identification of any robust patterns, the response of the two homeostoich phenotypes appear to be distinct Figure . Closer It is clear that ionomes are sensitive to both the external environment as well as the genomic composition. Whether such changes in the ionome are ecologically relevant is an important question worthy of attention by both empirical and theoretical practitioners of ecological stoichiometry. Studies with plants indicate that individual growth, even under strong P limitation is better predicted, not by P use efficiency alone, but by the uptake of a few other elements as well (Baxter et al., As our understanding of nutrient limitation shifts from single nutrient models, to more complex, multiple nutrient models predicting co-limitation (e.g., Saito et al., Ionomic data from both the literature and our experiment reveal several correlations, although the functional basis for such correlations appears to be more complex than what can be predicted by linkages based on cellular physiology of element processing. For example, sodium dependent phosphate uptake by cells is a well-established mechanism (discussed in the context of ecological stoichiometry in Jeyasingh and Weider, Genotype-specific effects common in ionomic studies discussed above are similar to discoveries about intraspecific variation in biomass C:N:P stoichiometries (e.g., Bertram et al., All authors made equal, substantial contributions toward the creation of this manuscript, and approve its publication.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.


Cutaneous metaplastic synovial cyst (CMSC), presents as a solitary, tender subcutaneous nodule that usually occurs at the site of previous surgery or trauma. Histologically, the lesion is characterized by a cystic structure with villous-like projections that lined by metaplastic synovial tissue. The main cause remains unclear, but trauma is presumed to be a precipitating factor, as most reported cases have a history of antecedent cutaneous injury. Here we present a case of CMSC in a 51 year old man, presented with a painless deep-seated dermal nodule in the medial aspect of left ankle without history of any trauma or surgery in this site. Immuno-histochemistry study reveals positive reaction for CD68 in the cystic wall and negative reactions for S-100. CMSC is a unique lesion and worthy to attention, and should be included in the differential diagnosis of deep dermal cutaneous cysts. Cutaneous metaplastic synovial cyst (CMSC) is an unusual non-neoplastic and benign subcutaneous cystic lesion that usually occurs as posttraumatic solitary and tender nodule or in the site of previous surgery. This entity is different from synovial cyst described as herniation of synovium through joint capsule . Histologically, CMSC is characterized by cystic space lined by metaplastic synovial tissue accompanied by protrusion of villous structure to cystic spaces and fibrin deposition. The cyst wall lining is composed of alternating foci of cellular area containing polygonal cells with eosinophilic cytoplasm and fibroblast like cells as well as areas without distinct lining only covered by fibrinoid material , 2.Although certain etiology of the disease is not well-known until now, the majority of cases had history of local trauma or surgical manipulation at the same site, therefore mentioned factors proposed as probable etiology of the disease. Therefore, some cases are reported in Ehlers-Danlos syndrome , rheumatHere, we reported an unusual case of CMSC with no history of trauma or surgical procedure.Study populationA 51 year old man who presented with painless subcutaneous mass like lesion in the medial aspect of left ankle was referred to the Imam Khomeini Hospital Complex, Tehran University of Medical Sciences, Tehran, Iran in 2015. He has no history of significant trauma or any surgical procedure in site of lesion. By physical examination, a solitary well-delineated cystic dermal nodule measuring 3.5×1.5 cm was founded. The overlying skin was normal in physical examination without erythema, warmth or tenderness. The lesion was excised by surgery. Through gross examination, we found out a dermal cyst covered by skin with smooth internal surface containing serosal colorless fluid. Microscopic examination shows skin tissue with underlying deep-seated dermal cyst devoid of true epithelial cells with villus-like projections into cystic space . The cysThe first case of CMSC was described by Gonzalez et al. in 1987 .Besides, a child was presented with Ehlers-Danlos syndrome and CMSC without local trauma or previous surgery in site of the lesion. Inappropriate collagen synthesis was offered as probable etiologic factor . OverallA case of CMSC in zygomatic region was analyzed with history of hyaluronic acid filler injection. They proposed immune system inflammatory response to tissue damage due to filler injection as cause of this lesion . SynoviaHistologically, CMSC is a pseudocyst devoid of true epithelial lining characterized by multiple villous structures resembling synovial membrane. The cyst lining has variable cellularity from hypocellular to hypercellular areas contain a mixture of fibroblasts, mononuclear inflammatory cells, epithelioid cells and multinucleated giant cells. The hypercellular area has zonal pattern from the luminal to the outer zone include polymorphic epithelioid and multinucleated giant cells , inflammatory cells (zone 2), vascular tissue (zone 3) and fibroconnective tissues (zone 4). The hypocellular areas are characterized by hyalinized zone of connective tissue and fibrin deposition surrounded by subcutaneous tissue with reactive fibrosis . ImmunohThe diagnosis is cumbersome because CMSC is unfamiliar to dermatologist. It should be differentiated from more common lesions with similar clinical manifestation. Differential diagnosis of similar lesions in this region includes synovial cyst of ankle, which can form a mass like lesion under skin. It may or may not be tender depending on its size and location. Histologically synovial cyst is lined by synovial cells (positive for CD68) with communication to the adjacent joint usually involved by osteoarthrosis or inflammatory process like rheumatoid arthritis .The other differential diagnosis is ganglion cyst that is common in the ankle and is described by cystic lesion containing mucinous fluid and discontinuous layer of pseudosynovial cells flattened and surrounded by connective tissue. Because of histologic findings and clinical sign and symptom, bursitis of the ankle can lie in differential diagnosis of painful mass-like lesions of this region. In comparison with CMSC bursitis is located deeper with more symptoms rather than CMSC.In case of history of surgical intervention, suture granuloma as a granulomatous reaction to foreign materials  can be clinically similar to CMSC or even can be misdiagnosed as recurrence of primary tumor previously excised at the same site . We diagThe diagnosis of CMSC in any dermal or subcutaneous mass-like or cystic lesion in periarticular area should be based on combination of clinical history, careful physical examination, and histologic findings as well as exclusion of clinically and histologically similar lesions of this region. The authors declare that there is no Conflict of Interests.


Species survival is dependent on successful reproduction. This begins with a desire to mate, followed by selection of a partner, copulation and in monogamous mammals including humans, requires emotions and behaviours necessary to maintain partner bonds for the benefit of rearing young. Hormones are integral to all of these stages and not only mediate physiological and endocrine processes involved in reproduction, but also act as neuromodulators within limbic brain centres to facilitate the expression of innate emotions and behaviours required for reproduction. A significant body of work is unravelling the roles of several key hormones in the modulation of mood states and sexual behaviours; however, a full understanding of the integration of these intrinsic links among sexual and emotional brain circuits still eludes us. This review summarises the evidence to date and postulates future directions to identify potential psycho-neuroendocrine frameworks linking sexual and emotional brain processes with reproduction. Reproduction is an essential physiological process for preservation of the species, which relies on the integration of social and sensory cues with emotional and behavioural outputs that favour the successful production of offspring. Effective mating depends on an appropriate display of sexual behaviours that select for reproductive fitness. The neural command centre for coordinating these complex processes sits within the limbic brain . The limThe amygdala is a small almond-shaped structure lying deep to the antero-inferior temporal lobe. Animal lesion studies have established the amygdala as a central hub for processing emotions and sexual behaviours in rodents and primates , 5, 6. IThe hypothalamus coordinates essential homeostatic mechanisms including the regulation of neuroendocrine axes and autonomic processes. Limbic functions of the hypothalamus have been demonstrated in rats, whereby hypothalamic stimulation leads to increased motivation towards rewarding behaviours . The amyThe hypothalamic–pituitary–gonadal (HPG) axis orchestrates physiological processes required for reproduction. Furthermore, it is known that hypogonadal states in animals and humans are associated with loss of responsiveness to sexual stimuli and loss of appetitive sexual behaviours –14. ReprThis review summarises the literature surrounding key hormones and their roles in sexual and emotional brain processing. We examine common functions and discuss future directions for elucidating the psycho-neuroendocrine links uniting sex, emotion, and reproduction.Kisspeptin refers to a family of peptide hormones cleaved from the product of the Kiss1 gene. They share a common carboxyl terminal sequence obligate for their action on kisspeptin receptors (encoded by Kiss1r) . KisspepKisspeptin-expressing neurones and mRNA have been identified in the bed nucleus of the stria terminalis (BNST), thalamus, and amygdala of rodents . FurtherThe first evidence for kisspeptin’s role in sexual behaviour came from Keller et al., who show that peripheral administration of kisspeptin stimulates the lordosis reflex in female mice . Later, Data also suggest a role for kisspeptin in the display of fear and anxiety. In zebrafish, fear stimuli significantly reduce Kiss1 transcription, while intra-cranial kisspeptin administration attenuates fear responses via effects on the serotonergic system , 39. In Mechanistically, there is observed interplay between kisspeptin signalling and several other neuropeptide systems including serotoninergic , adrenerCollectively, these studies in humans and several other species demonstrate that kisspeptin signalling effectively integrates sensory processing with limbic pathways involved in sexual arousal, positive mood, and anxiety . TogethGnRH is secreted in a pulsatile fashion from the pre-optic area of the hypothalamus. GnRH binds to its receptor on the anterior pituitary to stimulate LH and follicle-stimulating hormone (FSH) release, which induce sex steroid secretion and gametogenesis in the gonads. Once thought to be the master regulator of the HPG axis, it is now known that GnRH release is controlled by two novel neuropeptides; kisspeptin which exerts a stimulatory effect on GnRH  and gonaAnatomical localisation of GnRH-expressing neurones has been studied in many species –51 with In 1973, two independent research groups showed that subcutaneous administration of GnRH to ovariectomised, steroid-replaced female rats potentiates female sexual behaviour, i.e., the lordosis response , 60. FurGnRH agonists are associated with anxiolytic effects in rodent models , whereasGnRH and its receptor are distributed in key brain areas involved in processing olfactory cues and integrating chemosensory information with appropriate sexual behaviours in animals Fig. . By contGnIH belongs to the RF amide family of peptides designated by their C-terminal arginine (R) and amidated phenylalanine (F) residues. First identified in the Japanese quail, GnIH is now known to be a key regulator of reproduction in avian species and has also been found in other vertebrates . Two actGnIH immunoreactive fibres have been identified in brain regions outside the hypothalamus in quail , rats 7, and rheAlongside its inhibitory role on the HPG axis, there is also evidence that GnIH has a negative impact on male sexual behaviours in quail  and ratsThrough its downregulation of the HPG axis and inhibition of sexual behaviours, GnIH may act as a reproductive brake by appropriately inhibiting mating during unfavourable times such as stressful situations Fig. . This isExposure to gonadal steroids such as oestrogen and testosterone during critical developmental windows not only determines sexually dimorphic structural differences, but also influences sex-specific behaviours in many species including humans . OestrogOestrogen receptors (ERs) and testosterone-binding androgen receptors (ARs) are intra-cellular receptors found throughout the brains of rodents , 81. UsiTestosterone was first found to be critical for male sexual behaviour in 1849, when Berthold showed that castrated roosters lost their secondary sexual characteristics and failed to display copulatory behaviours such as vocalisation and aggression. Furthermore, these behaviours were restored by transplantation of functioning testes from another animal . IntracrIn humans, testosterone triggers greater affiliative behaviours in men, e.g., more eye contact, and more self-presentation towards the opposite sex . There iIn ageing men, progressive testosterone decline is associated with low mood, reduced libido, and poor sexual function . TestostIt is known that in women with regular ovulatory cycles, testosterone rises during the follicular phase, reaches maximal levels in the middle third of the cycle, before declining to reach nadir shortly before the next follicular phase . This coAlthough testosterone plays a fundamental part in orchestrating male sexual behaviours, it is interesting to note that the expression of certain male-typical behaviours requires the aromatisation of testosterone into neuroestrogen , 105. ThSexually receptive behaviours in female rodents are mediated by the ventromedial hypothalamus and the medial amygdala, which are major sites of oestrogen action . Both syIn humans, there is evidence that raised oestrogen levels prime females to seek out social signals of masculinity. During peak oestrogen levels, women show highest preference for male-specific odours , and perThus, in humans, both testosterone and oestrogen have effects on female sexual motivation and behaviours with ongoing debate as to which gonadal steroid is most effective. The current guidelines advocate testosterone for the treatment of sexual desire disorders in women  with resTestosterone and oestrogen have well-defined roles in the development and exhibition of sexually driven emotions and behaviours in male and female rodents. This is supported by sex differences in the distribution and density of ERs and ARs within key limbic brain structures. Human studies have found a majority effect for testosterone as a mediator of sexual drive and arousal in men and women; however, questions remain over the role of oestrogen and its interplay with testosterone in the regulation of female sexual behaviours Fig. .Oxytocin and vasopressin are nonapeptide hormones descended from the same ancestral gene; they differ in structure by only two amino acids and are highly conserved among vertebrates . The endOxytocin and vasopressin are synthesised in the magnocellular neurones within the paraventricular nucleus (PVN) and supraoptic nucleus (SON), which extend from the hypothalamus to the posterior pituitary where they are stored prior to peripheral release . CentralIn rodents, oxytocin and vasopressin are co-expressed in several limbic areas including the medial amygdala, nucleus accumbens, the BNST, and olfactory areas . There aAutoradiographical examination in rodents shows high co-localisation of oxytocin and vasopressin-binding sites in limbic brain regions including the amygdala and BNST . HoweverOxytocin enhances sexual behaviours in male and female rodents. Central administration of oxytocin promotes the lordosis response in female rats  facilitaThe monogamous prairie vole provides an excellent model for the study of social bonding. These animals form long-term monogamous relationships and display significantly higher concentrations of oxytocin receptors in their limbic structures  compared to polygamous voles . FurtherIn humans, peripheral oxytocin levels are raised during sexual arousal and copulation in women, and to a lesser degree in men . Recent In keeping with its distribution within the limbic brain, vasopressin’s effects on sexual function are also sexually dimorphic. Vasopressin is essential for male sexual behaviours such as aggression towards competitors and mate guarding. This has been demonstrated in male song sparrows, whereby the use of vasopressin antagonists blocks neural responses related to social stressors . In addiSimilar to oxytocin, vasopressin is also required for the recognition of social cues, as evidenced by distribution of vasopressin receptors in olfactory centres involved in processing social olfactory signals . FurtherIn monogamous species, evidence suggests that vasopressin increases affiliative behaviours. This is demonstrated by administration of vasopressin to male titi monkeys resulting in prolonged time spent with their partner versus an opposite sex stranger . In maleOxytocin and vasopressin are intimately linked in terms of their evolution, distribution within the brain, and their effects on social and sexual behaviours. Both peptides are involved in olfactory recognition in rodents with complementary effects on emotions and prosocial behaviours. Oxytocin is involved in the selection of desirable traits in the opposite sex and promotes intimacy and bonding between individuals. Vasopressin has been shown to facilitate male aggression towards competitors and mediate partner preference behaviours in monogamous species. However, its actions can be sexually dimorphic, vary across species, and are likely to be site-specific within the brain. Oxytocin and vasopressin are therefore essential for the formation of successful pair bonds as part of an integrated network of intrinsic links that unite sex, emotion, and reproduction to facilitate species survival Fig. . Future Stress is intimately linked to reproductive function, with both acute and chronic stress being associated with poor reproductive outcomes. The hypothalamic–pituitary–adrenal (HPA) axis governs our innate endocrine response to stressors and in conditions of HPA over- activity such as Cushing’s syndrome; reproductive function is inhibited . CorticoStarting at the apex of the HPG axis, both kisspeptin and GnRH are associated with anxiolytic effects, which are in keeping with their roles in promoting positive emotions and behaviours related to reproduction. There is evidence that GnRH inhibits the stress response through mechanisms that involve blocking the effects of corticotropin-releasing hormone (CRH) . KisspepThe behavioural effects mediated by oxytocin and vasopressin are also closely related to their interaction with the HPA axis. Oxytocin has been shown to suppress sympatho-adrenal activity  and in sStress plays a role in many aspects of sex, emotion, and reproduction. The intrinsic links underlying these interactions are endocrine in nature and involve reciprocal modulation of the HPA and HPG axis at multiple levels.As a relatively young discipline, concepts within endocrinology continue to evolve at a rapid pace with a widening body of research in support of complex and far-reaching interactions between endocrine axes and other essential bodily processes. In particular, there has been intense interest in the interface between neuroendocrinology and limbic brain processing with evidence to suggest that the integral links between sexual, emotional, and reproductive brain processes are endocrine in nature. At the apex of the reproductive axis, kisspeptin represents a key link between sex, emotion, and reproduction. Kisspeptin’s actions within the limbic brain include stimulation of the lordosis reflex  and induThere is also evidence that the HPG axis works in concert with other endocrine mediators including cortisol, oxytocin, and vasopressin to integrate physiological reproductive processes with essential emotions and sexual behaviours required for successful mating.Looking to the future, we are only just beginning to appreciate the outline of these fundamental neuroendocrine frameworks that integrate sex, emotion, and reproduction. A key area of study will be in unravelling the pathways and mechanisms that underlie these interactions between physiology, psychology, and endocrinology. Translational aspects are also important to explore as animal models are limited by the complexity of human sexual behaviour which is more dependent on learning and context rather than stereotyped actions . From a


Microtubule stabilizing drugs such as taxanes have proven fairly effective for treating solid tumors, but the mechanism by which these drugs treat cancer is still unsolved. Intratumor imaging, showing that the cell proliferation rate is low in many chemosensitive human cancers, has challenged the dogma that the cytotoxicity of such compounds is the result of their effect on rapidly dividing cells . MoreoveThe search for next generation microtubule stabilizing drugs with increased efficacy is thus intense. Several strategies have been proposed for the development of potentially more effective and less toxic anticancer drugs. One approach is to find drugs that target nonessential proteins, such as microtubule regulators. If such a target would be more active in cancer cells, or centrally involved in cancer aggressiveness, its inhibition would improve drug selectivity for tumors over normal tissue and contribute to a greater therapeutic window.LIM Kinases (LIMKs) are enzymes whose activity is elevated in cancers compared to normal tissue. LIMKs regulate the architecture of the actin cytoskeleton by phosphorylation and inactivation of actin depolymerization factors of the ADF/cofilin family. Independently of this effect on actin microfilament dynamics, LIMKs also regulate microtubule dynamics, but whether this regulation occurs through a direct binding of LIMK to microtubules or through phosphorylation of an associated protein is still unknown , 4. Whenin vivo growth of different breast cancer derived cell lines in tumor xenograft models [in vivo efficiency of paclitaxel and Pyr1 is microtubule stabilization, reinforcing the notion that targeting microtubule dynamics is central to the therapeutic effects of these drugs.Anticancer drugs are currently classified in two main categories: targeted drugs and cytotoxic drugs. By perturbing a cancer specific pathway that comes down to LIMKs, Pyr1 behaves as a targeted drug. However, because of the induction of cell-cycle arrest and apoptosis, Pyr1 also behaves as a cytotoxic drug, making the mechanism of action of this drug very novel. These properties prompted the investigation of its effect on breast cancer development and the test of the hypothesis that LIMK inhibition could be efficient in paclitaxel resistant cancers. To that aim, Prunier et al. investigated the effect of Pyr1 on the t models . Pyr1's in vivo LIMK inhibition has heterogeneous effect on tumor cell motility and did not prevent metastasis spread. This was unexpected as LIMK inhibition is able, in vitro to impede cell motility and as it has recently been shown that LIMK is required for matrix degradation through phosphorylation of membrane type 1-matrix metalloproteinase [Using fluorescent and luminescent cancer cells, Prunier et al. found that Pyr1 administration has a strong effect on the metastatic load, preventing the growth of metastasis but not their spread. The effect on metastatic load could involve the same mechanisms as those observed in the primary tumor, including an inhibition of integrin-mediated adhesion to the extracellular matrix through an Integrin-Linked-Kinase/β-parvin/LIMK/ cofilin pathway that has been shown to play a central role in tumor initiation and metastatic colonization . Intravioteinase . The com


We found significant differences in floristic diversity and phylogenetic structure across a range of study sites encompassing open vegetation and forest on quartzite (FQ) and on ironstone substrates, commonly termed canga. Substrate and physiognomy were key in structuring floristic diversity in the Espinhaço and physiognomy was more important than substrate in structuring phylogenetic diversity, with neither substrate nor its interaction with physiognomy accounting for significant variation in phylogenetic structure. Phylogenetic clustering was significant in open vegetation on both canga and quartzite, reflecting the potential role of environmental filtering in these exposed montane communities adapted to multiple environmental stressors. In forest communities, phylogenetic clustering was significant only at relatively deep nodes of the phylogeny in FQ while no significant phylogenetic clustering was detected across forest on canga (FC), which may be attributable to proximity to the megadiverse Atlantic forest biome and/or comparatively benign environmental conditions in FC with relatively deep, nutrient-rich soils and access to edaphic water reliable in comparison to those for open vegetation on canga and open or forest communities on quartzite. Clades representing relatively old lineages are significantly over-represented in campos rupestres on quartzite, consistent with the Gondwanan Heritage Hypothesis of Ocbil theory. In contrast, forested sites on canga are recognized as Yodfels. To be effective, conservation measures must take account of the distinct communities which are encompassed within the broad term campos rupestres, and the differing vulnerabilities of Ocbils and Yodfels.Old, climate-buffered infertile landscapes (Ocbils) have attracted increasing levels of interest in recent years because of their exceptionally diverse plant communities. Brazil’s  These hcampos rupestres,’ are an example of a long-standing climate-buffered infertile landscape that contains many endemic species and lineages, which are adapted to very shallow and nutrient-poor soils . Forests and savannas mostly grow at lower altitudes or along the rivers , while patches of campo rupestre grasslands develop at the more exposed sites of mountain tops . Occurring on either quartzitic, arenitic or ironstone substrates, typically at altitudes >900 m above sea level, campos rupestres are estimated to occupy 66,450 km2 and to be home to over five thousand known species  distribution of campo rupestre sites and resulting floristic influences from other habitats, especially savannas  in Minas Gerais, where the iron-rich substrate is known as canga. Comparative studies of the flora of campos rupestres on quartzite and canga have shown strong correlations between plant species abundance, vegetation cover, coverage of individual species and soil properties between these two habitats , and niche conservatism, in which such traits tend to be shared among species that are closely related phylogenetically . Strong campos gerais , in situ diversification and high density of microendemics. On a more local spatial scale, comparing the angiosperm communities on two sites within Itacolomi State Park in Minas Gerais state, campo rupestre vegetation, as predicted by the stress dominance hypothesis  and occurring on contrasting substrates (quartzite and canga) in the Espinhaço range and the Quadrilátero Ferrífero. We construct the first campo rupestre supertree (believed to be the first for any Ocbil) and use it to compare the phylogenetic structure of these contrasting assemblages. Based on earlier studies in campos rupestres, and consistent with Ocbil theory, we predicted that phylogenetic clustering would be prevalent in the assemblages analyzed. Assuming the prevalence of niche conservatism, consistent with Ocbil theory, we anticipated that the high levels of metals in canga might represent an additional environmental filter potentially resulting in accentuated phylogenetic clustering in canga assemblages. We also predicted significant over-representation of clades representing relatively old lineages, consistent with the Gondwanan Heritage Hypothesis of Ocbil theory (campos rupestres (campo rupestre ecosystem as a recently recognized Ocbil and a long-neglected conservation priority (In this study we compare the composition of l theory . We discupestres  and thospriority .brazil AND floristic OR list OR checklist AND “campo rupestre” OR canga) AND PUBYEAR > 1991 AND PUBYEAR < 2016. This search yielded 296 papers. Analysis of their abstracts identified papers with comprehensive floristic lists (for angiosperms) for sites in Espinhaço range or the Quadrilátero Ferrífero as candidates for inclusion in our study. Details of all selected studies and all Angiosperm species reported therein were entered in a database developed in the ‘plotsamples’ module of Brahms software  in the original lists, and where no further reliable identification in the virtual herbaria consulted was found, were assigned to these taxa. Names that were qualified with ‘aff.’ in the original lists  were removed from this study because the use of affinis in this context indicates that the specimen resembles the species mentioned but is not referable to it.A literature search was conducted to locate published botanical studies that include species lists for locations in the Espinhaço range and the Quadrilátero Ferrífero. The bibliographic search was carried out in Scopus in October 2015, using the following search terms: ALL reported in the publication. Where a single study encompassed multiple vegetation types, information contained within the publication was used to subdivide the list into sampling sites which were more homogeneous with respect to vegetation type. We categorized the physiognomy of each site as “forest” for vegetation with a canopy or as “open vegetation” in the case of grassland or savanna. The Espinhaço is very complex in geology  and substrate  and specialists suggest campo rupestre sensu stricto does not occur on alkaline soils . From the initial database (Supplementary Data Table 1 and Supplementary Data canga (FC), 9 as forest on quartzite (FQ), 11 as open vegetation on canga (OC) and 22 as open vegetation on quartzite (OQ).For the analyses reported here, we excluded from our initial database all sites for which we could not determine the physiognomy of the vegetation or the substrate of the site, as well as all sites where anthropogenic disturbance was reported to be prominent. Reports of Figure 2), we extracted the list of 2920 species , we manually replaced “-” with “.”  in our dated megatree  and in different vegetation physiognomies (open vegetation or forest or both).We used iTOL  to visuaTo evaluate the floristic differences between the plant communities in different habitats  we calculated a metric of beta diversity and two metrics of phylogenetic alpha diversity using Vegan  and PicaWe used the 47 site matrix and the Espinhaço megatree to calculate two phylogenetic metrics: the Mean Pairwise Distance (MPD) and the Mean Nearest Taxon Distance (MNTD). To interpret the values of MPD and MNTD we compared the observed values of MPD and MNTD with those obtained using bootstrap analysis of a null model, where we shuffled the tips of the phylogenetic tree 999 times and recalculated values. This generates the Standard Effect Size (SES) for each metric, Net Relatedness Index (NRI) and Nearest Taxon Index (NTI), respectively. At the level of individual sites, values of NRI and NTI larger than 1.96 or smaller than -1.96 are significantly greater than the mean values obtained from the null model and indicate sites with significant phylogenetic clustering or overdispersion, respectively.t-tests on the subsets of interest. To evaluate which factors best explain variation in the degree of phylogenetic clustering between communities we used ANOVA, with post hoc Tukey tests to determine which means differed significantly from each other  and a given presence-absence matrix (showing where each species occurs) whether a particular evolutionary lineage is over- or under-represented in each site. Using the iTOL tree viewer we plotted our nodesig outputs for individual sites  suggested strong associations between certain clades and particular substrate-physiognomy combinations and a relatively few and scattered species found on both substrates and/or in both vegetation physiognomies. For example, Xyridaceae and Eriocaulaceae were very strongly associated with OQ while Poaceae, also found mainly in open habitats, had several species reported from both canga and quartzite. Groups of species confined to forest habitats on canga were infrequent but examples were seen in the early branching angiosperms, e.g., some Annonaceae and Lauraceae. Among the families represented by numerous species, Myrtaceae were notable for having many species reported from both open and forest habitats and/or from both canga and quartzite.Merged species lists for the 47 sites included in our analysis yielded 2920 species representing 789 genera and 135 families, with species recorded per site ranging from 20 to 503 and averaging 148. Our initial visualization  shows a primary division driven by substrate, with all but three of the canga sites forming a distinct group that includes just one site on quartzite at ITA. The remaining three canga sites  form a cluster which is nested within a major grouping comprising all the quartzitic sites (except the ITA site mentioned above). The next level of grouping appears to be influenced more by physiognomy: within the main canga grouping, three forest sites form a sub-group distinct from the remainder, which are from open vegetation. Similarly, the main quartzitic group is divided into two subgroups: one dominated by sites with OQ but including two forest sites on quartzite while the second subgroup is more heterogeneous. The first dichotomy in the heterogeneous subgroup separates a group of quartzite sites with open vegetation from the remaining sites which in turn are divided into four groupings. The composition of the smallest groupings reflects geographical locations, with sites frequently appearing least dissimilar to another site at the same location. The major groupings detected in the Ward similarity analysis were also reflected in the NMDS analysis , which classified each site with the corresponding habitat type.The Ward similarity analysis illustrated the influence of substrate, physiognomy and location on floristic similarity values. The dendrogram . Significant phylogenetic clustering as measured by NRI and NTI was evident in mean values both for open sites on canga and for open sites on quartzite. However, for forest sites on canga, neither NRI nor NTI showed means significantly greater than zero while forest sites on quartzite showed significant phylogenetic clustering as measured by mean NRI but not by NTI.Analysis of phylogenetic structure revealed the general prevalence of phylogenetic clustering in the open p = 0.02) while neither substrate nor the interaction of substrate and physiognomy were significant (Table 3). Tukey tests on the results of the ANOVA showed that neither NRI nor NTI values differed significantly between communities on quartzite and those on canga (p > 0.05 in each case). In contrast, NTI values for communities with open physiognomies were significantly greater than those for forest formations (p = 0.0165) while NRI values did not differ significantly between open and forest formations (p > 0.05).Analysis of variance results suggest that substrate and physiognomy account for some variation in NRI  while the interaction of substrate and physiognomy was clearly not significant. In contrast, for NTI physiognomy had significant explanatory power  were over-represented in forest sites on canga while the cyperid and xyrid monocots and consecutive monocot clades above them tended to be under-represented in these sites. In contrast, FQ bore assemblages in which Eudicots were over-represented, specifically Pentapetalids, while Narthecidae and several other monocot clades were under-represented. Open vegetation on iron showed an over-representation of the genus Eremanthus (Asteraceae) and under-representation of the xyrid monocots. Commelinids and cyperids + xyrids were over-represented in OQ while Rosids and Subrosids were under-represented. Tabulation of the estimated crown ages for these clades (Table 4) showed that clades which are significantly over-represented in assemblages on canga generally represent younger lineages than those which are significantly over-represented in assemblages on quartzite. The oldest over-represented clades (>100 Mya) are all over-represented in FQ while the three youngest are all over-represented on FC sites (≤37 Mya).Clades indicated by nodesig as being significantly over- or under-represented in at least half of the sites from a particular habitat type are shown in campo rupestre to date and the only one to include sites on canga, a substrate that is a target for extractive industry and thus one of the most threatened habitats in Brazil . FC sites were also notable for an over-representation of clades representing relatively young lineages consistent with expectations for young, often disturbed, fertile landscapes  provides a useful framework for investigation of the impact of environment on patterns of biodiversity. Moreover, our study is the most comprehensive study of community phylogenetic structure in n Brazil . Our aimcanga having a flora more similar to that of other sites on canga than to the majority of sites on quartzite. The NMDS ordination also showed the importance of substrate, but physiognomy appeared even more important, with only one site crossing an otherwise clear separation between the floras of forest and open vegetation. Considered in combination, these results based on floristic composition provide strong support for our a priori recognition of four habitat types based on substrate and physiognomy: FC, FQ, OC and OQ. This is of great importance from a biodiversity perspective. The Espinhaço has substantial environmental heterogeneity related to both substrate, altitude, topographic position and microrelief. While the windward face of a mountain can harbor forests, the top of the same mountain bears campo rupestre, with forests restricted to small depressions where deeper soils accumulate. At a landscape scale, we can see sharp transitions between vegetation types and, as we see in our analysis, each habitat type has its own flora with its own environmental processes shaping its composition, in terms not only of individual species, but also of lineages.Similarity analysis clearly showed the importance of substrate to floristic composition, with most communities on campo rupestre communities on both canga and quartzite and in both open and forest physiognomies, but the degree of phylogenetic clustering detected differed between the habitat types and with the metric used. Phylogenetic clustering was most evident in open assemblages, with both OC and OQ having mean NRI and NTI significantly different from zero, indicating significant phylogenetic clustering both near the tips of the phylogeny (NTI) and also involving the deeper nodes in the tree (NRI). Phylogenetic clustering was particularly pronounced in open assemblages on quartzite (OQ) with mean NRI and NTI values exceeding those reported by campo rupestre vegetation on quartzite at two sites in Minas Gerais and by campo rupestre formations associated with quartzitic outcrops. .We found phylogenetic clustering to be common in campos rupestres where a clear pre-dominance of stress-tolerant strategies was reported [and the few species with a greater level of competitiveness had relatively low abundance]. However, recent reviews of the application of phylostructure metrics to community ecology have cautioned against the use of phylogenetic patterns as proxies for community assembly mechanisms  in explaining variation in NRI. Surprisingly, substrate x physiognomy interactions did not explain significant variation in either metric. The significance of physiognomy in explaining variation in NTI in our study is consistent with the observations that across campo rupestre the development of forest tends to occur in areas that are more favorable in terms of water and/or nutrient availability  together with the xyrid clade (Eriocaulaceae + Xyridaceae). These latter families include ‘everlasting’ flowers from the iconic megadiverse genera Paepalanthus and Xyris, long considered emblematic of open campo rupestre communities and an important source of income for rural, low-income families  a wide range of root specializations to survive P limitation. The ages of the two monocot clades which are over-represented in OQ sites are 67 and 96 Mya, respectively (Table 4), lending further support to the interpretation of open campo rupestre systems on quartzite as Ocbils. In contrast to the monocot over-representation in OQ sites, the only clade that our study showed to be over-represented in most OC sites is the genus Eremanthus (estimated age c. 20 Mya). These tree-like, woody Asteraceae are characteristic of the transition between forest and open habitats on canga, often forming homogenous stands known as ‘candeias.’Phylogenetic clustering was most evident in OQ, and the clade-by-clade results generated by nodesig revealed over-representation in at least half of the OQ sites in our study of two major clades of monocots: the larger being the commelinid monocot clade, which includes the Poales and three other orders of monocots, a result consistent with the wide diversity of different types of open vegetation encompassed within our OQ study sites. The smaller over-represented clade in OQ is nested within Poales, and comprises the cyperid clade , a result consistent with the inclusion of forest assemblages on quartzite within the campo rupestre Ocbil. While potentially attributable to niche conservatism of lifeform, this result is also consistent with our tree-wide phylostructure metrics for which phyloclustering was detected by mean NRI (reflecting relatively deep nodes) but not at the level of NTI and may reflect the relative isolation of most FQ sites from the humid forest biome. In a study of forest islands on quartzite in the Serra do Cipó, The cyperid + xyrid clade, so characteristic of OQ, is significantly under-represented in most of the study sites for FC and for FQ. FQ, was revealed by nodesig to have significant over-representation of the early branching clades in our Espinhaço megatree: specifically the Eudicot clade, the clade comprising Sabiales to Asterales and the pentapetalids. These three clades represent the oldest lineages which are significantly over-represented in any habitat in our study ; the clade formed by Myrtaceae together with Vochysiaceae, and nested within, the family Myrtaceae and the genus Myrcia. The prominence of Solanum and Solanaceae in FC is consistent with qualitative and quantitative descriptions of canga vegetation by earlier authors  are represented by multiple species in FC, but not elsewhere in our study. Notably, the six clades reported above as significantly over-represented in the majority of FC sites are among the seven youngest over-represented lineages in our study (see Table 4).Our tree-wide metrics detected no significant phylogenetic structure across study sites for FC, with neither mean NRI nor mean NTI differing significantly from zero, but clade-by-clade analysis indicated significant over-representation of several dicot clades in FC. These included: the family Solanaceae and a subclade within it encompassing  authors . The impst biome  in which diverse . Furthercampo rupestre soils  and FC with open campo rupestre vegetation on quartzite (OQ) and FQ. Their results confirm the observations of many earlier authors  and on quartzite (OQ) showed greater variation in soil moisture than did forest sites, with recorded moisture levels frequently indicating severe water deficits in grassland on canga (OC) and even more frequently in grassland on quartzite (OQ), a difference which canga soils resulting in greater water retention and availability. No water deficits were recorded for FQ while interpretation of results for FC was complicated by technical/methodological issues. These results suggest that soil moisture factors may represent particularly strong filters in OQ.Two very recent, synthetic studies of re soils  have bee authors ,b, that canga (OC) and soil temperature usually exceeds air temperature to a greater extent than in grassland on quartzite (OQ), a difference which the authors attribute to substrate albedo: dark, red, dense canga absorbing more energy than light, pale quartzite. Furthermore, the frequency and maximum duration of events in which soil temperature reached >35°C was highest in grassland on canga (OC). Surprisingly, events where the soil temperature exceeded 35°C were also recorded in FQ, albeit with lower frequency and duration than in grassland on canga (OC). Thus soil temperature factors may represent particularly strong filters in OC but also, secondarily in FQ.Parallel monitoring of air and soil temperatures showed different patterns across the four sites compared by campo rupestre soils differing more in physical than in chemical attributes: sandy soils with greater levels of exchangeable Al3+, associated with granite/gneiss and quartzite substrates as compared to soils with greater accumulation of organic matter and clayey/silty textures found on canga and itabirite. These authors conclude that the low biomass of campo rupestre vegetation in general is attributable to low nutrient levels, especially to low phosphorus (P) rather than to metal toxicity. They report P amounts to be particularly critical for vegetation on canga from mining areas including those in the Iron Quadrangle of Minas Gerais though, puzzlingly, the lowest P levels they report for any site within our study area are actually for a quartzite site within the Quadrilátero Ferrífero.A cluster analysis of soil profiles from high altitude rocky complexes across Brazil , distingcampo rupestre, particularly acute in quartzitic soils, relative to those on canga, and a more significant constraint to growth than metal toxicity. Other factors such as soil moisture, water deficits and very high temperatures vary in frequency and intensity across the four habitat types studied and are variously reported to be more severe in grassland (on OQ and/or OC in the case of water deficits and soil temperatures) or FQ (soil temperatures). In these studies no single potential environmental filter was reported to be most severe for FC, a result congruent with the lack of phylogenetic clustering detected in this habitat. Together these considerations may suggest that this assemblage is less likely to be a result of relatively severe environmental filtering than others in our study, though of course in practice environmental factors need to be considered in combination rather than individually in order for their filtering effects to be understood.In summary, low nutrient levels are prevalent across campo rupestre flora and soils on quartzite and canga substrates, our results on phylogenetic structure, ages of over-represented clades in each habitat and the identities of these clades, lend support to the recognition of both open and forest assemblages on quartzite as Ocbils, with their significant over-representation of clades from several ancient lineages consistent with Gondwanan Heritage Hypothesis predictions  in protected area networks and sustainable management plans. Our conclusion that the forest communities on canga are quite different in phylogenetic structure from other campo rupestre assemblages and should in fact be interpreted as Yodfels requires further exploration before it can be considered sufficiently robust to inform appropriate conservation measures, which differ greatly between Ocbils and Yodfels (canga may harbor diverse subsets of Atlantic Forest diversity could lead to the identification of win–win approaches, whereby the vegetation restoration projects which are a legal requirement following mine closure (Our study provides insights for those concerned with the conservation and sustainable management of areas of  Yodfels . However closure  could alcangas of Carajás in the Brazilian Amazon. Increased investment in floristic inventory in these areas over the past decade is resulting in the publication of site-based lists suitable for inclusion in such studies (To our knowledge, this study is one of the most inclusive to date to focus primarily on non-forest ecosystems. However, increasing adoption of a very broad definition of ‘Rupestrian Grasslands’ to include all high altitude rocky complexes in Brazil highlights the opportunity for an even more inclusive analysis including other rupestrian environments associated with highlands, such as the tepuis of Roraima and the  studies .As more comprehensive data on species distributions becomes better understood and accessible, covering other rupestrian environments on highlands in different parts of the continent, studies such as the present one will elucidate processes underlying biodiversity dynamics at ever-increasing spatial scales and ecological heterogeneity.DZ co-designed and initiated this study, collated the plot data used in the analysis, contributed to integration and interpretation of the data, and contributed to the final manuscript. MM contributed to collation of the plot data used in the analysis, conducted and documented the bulk of the similarity and phylogenetic analyses, contributed to integration and interpretation of the results, and contributed to the final manuscript. TM conducted statistical analysis, contributed to integration, visualization and interpretation of the results, also contributing toward the final manuscript. EN co-designed this study, contributed to statistical analysis and to integration, and interpretation of the results, and played a lead role in drafting the final manuscript. All authors have approved the final version to be published and agree to be accountable for all aspects of the work, ensuring questions related to its accuracy and integrity are appropriately investigated and resolved.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.


Many databases have been constructed to provide computationally predicted miRNA targets. However, they cannot provide the miRNA targets expressed in a specific tissue and related to a specific disease at the same time. Moreover, they cannot provide the common targets of multiple miRNAs and the common miRNAs of multiple genes at the same time. To solve these two problems, we construct a database called CSmiRTar (Condition-Specific miRNA Targets). CSmiRTar collects computationally predicted targets of 2588 human miRNAs and 1945 mouse miRNAs from four most widely used miRNA target prediction databases  and implements functional filters which allows users to search (i) a miRNA’s targets expressed in a specific tissue or/and related to a specific disease, (ii) multiple miRNAs’ common targets expressed in a specific tissue or/and related to a specific disease, (iii) a gene’s miRNAs related to a specific disease, and (iv) multiple genes’ common miRNAs related to a specific disease. We believe that CSmiRTar will be a useful database for biologists to study the molecular mechanisms of post-transcriptional regulation in human or mouse. CSmiRTar is available at  MicroRNAs (miRNAs), 20–25 nucleotides non-coding RNAs, play important roles in the post-transcriptional regulation of gene expression –3. Via bUnderstanding the miRNA-target interactions is the crucial step to discern the roles of miRNAs in different biological processes . Many daThe complex circuitry of miRNA-mRNA interactions show the dynamic regulation of gene expression. Recent study showed that overexpressed MRE (miRNA response element)-containing transcripts can soak up the miRNA and upregulate its target genes . MoreoveTo meet these two needs, we develop a database called CSmiRTar (Condition-Specific miRNA Targets). CSmiRTar collects computationally predicted targets of 2588 human miRNAs and 1945 mouse miRNAs from four most widely used miRNA target prediction databases . CSmiRTar implements (i) a tissue filter for users to search the miRNA targets expressed in a specific tissue, (ii) a disease filter for users to search the miRNA targets related to a specific disease, and (iii) a database filter for users to search the miRNA targets supported by multiple existing miRNA target prediction databases. Moreover, CSmiRTar allows users to search the common targets of a set of input miRNAs under a specific physiological condition and the common miRNAs of a set of input genes under a specific physiological condition. We believe that CSmiRTar will be a useful database for biologists to study the molecular mechanisms of post-transcriptional regulation in human or mouse.Five data sources were used to construct CSmiRTar. First, the experimentally validated human and mouse miRNA targets were retrieved from miRTarBase , which mCSmiRTar was built using the scripting language PHP and Codelgniter framework. Crawler was used to retrieve raw data from other databases and Python was used to process the raw data. The processed data was stored in MySQL. The Interactive bar chart was generated by Highcharts.CSmiRTar provides both a search mode and a browse mode. In the search mode, users have four possible ways to search CSmiRTar. First, users can input a miRNA and search its targets which are (i) expressed in a specific tissue, (ii) related to a specific disease, or/and (iii) supported by multiple existing miRNA target prediction databases. After submission, users will see the search results sorted by the number of supported databases or the average normalized score see . Second,In the browse mode, users have two possible ways to browse CSmiRTar. First, users can click on a human/mouse miRNA name and get the miRNA’s targets supported by one or multiple existing miRNA target prediction databases see . Second,Identifying the functional targets is a crucial step to dissect the function of miRNAs. Using existing miRNA target prediction databases usually returns thousands of predicted targets per miRNA. Therefore, it is very hard for researchers to choose the biologically plausible candidates for further experimental validation. Besides, it can be expected that many of the predicted targets are non-functional since miRNAs only regulate their target genes in specific tissues, cell types and disease states.To solve this problem, we implement three different kinds of filters  to efficiently reduce the number of predicted miRNA targets but still keep the functional ones. To show the effectiveness of our filters, we prepare a benchmark set by randomly selecting several experimentally validated miRNA-target pairs in specific tissues and cancers from OncomiRDB . As showAs shown in An important step to reconstruct the ceRNA network is to identify the shared miRNAs of ceRNAs. CSmiRTar allows users to input a set of genes (e.g. ceRNAs) to search the shared miRNAs which regulate these genes. As shown in In CSmiRTar, users can input a set of miRNAs to search their common target genes. As shown in In this article, we present CSmiRTar which provide computationally predicted targets of 2588 human miRNAs and 1945 mouse miRNAs. CSmiRTar implements (i) a tissue filter for users to search the miRNA targets expressed in a specific tissue, (ii) a disease filter for users to search the miRNA targets related to a specific disease, and (iii) a database filter for users to search the predicted miRNA targets supported by multiple existing databases,. Moreover, CSmiRTar allows users to search the common targets of a set of input miRNAs under a specific physiological condition and the common miRNAs of a set of input genes under a specific physiological condition. We provide many case studies to show the effectiveness of our filters in reducing the number of predicted miRNA targets but still keep the functional ones. However, users should note that some functional miRNA targets may not be kept when applying both the tissue and disease filters if they are not expressed in normal tissues but are abnormally expressed in disease states. Nevertheless, we believe that CSmiRTar will be a useful database for biologists to study the molecular mechanisms of post-transcriptional regulation in human and mouse.


PGDT protocols help to titrate fluids, vasopressors, or inotropes to hemodynamic target values. There is considerable evidence that PGDT can improve patient outcome in high-risk patients if both fluids and inotropes are administered to target hemodynamic variables reflecting blood flow. Despite this evidence, PGDT strategies aiming at an optimization of blood flow seem to be not well implemented in routine clinical care. The analysis of the arterial blood pressure waveform using invasive uncalibrated pulse contour analysis can be used to assess hemodynamic variables used in PGDT protocols. Pulse contour analysis allows the assessment of stroke volume (SV)/cardiac output (CO) and pulse pressure variation (PPV)/stroke volume variation (SVV) and thus helps to titrate fluids and vasoactive agents based on principles of “functional hemodynamic monitoring.” Pulse contour analysis-based PGDT treatment algorithms can be classified according to the hemodynamic variables they use as targets: PPV/SVV, SV/CO, or a combination of these variables. From a physiologic point of view, algorithms using both dynamic cardiac preload and blood flow variables as hemodynamic targets might be most effective in improving patient outcome. Future research should focus on the improvement of hemodynamic treatment algorithms and on the identification of patient subgroups in which PGDT based on uncalibrated pulse contour analysis can improve patient outcome. PGDT aims at an optimization of basic and advanced global hemodynamic variables to maintain adequate oxygen delivery to the end-organs. PGDT protocols help to titrate fluids, vasopressors, or inotropes to hemodynamic target values that can be assessed with different hemodynamic monitoring technologies.The analysis of the arterial pressure waveform using uncalibrated pulse contour analysis can be used to estimate stroke volume (SV), cardiac output (CO), and dynamic variables of cardiac preload .In this article, we will describe the physiological background and the clinical application of PGDT using invasive uncalibrated pulse contour analysis for the assessment of hemodynamic values.Numerous randomized-controlled trials and meta-analyses demonstrate that there is an increasing body of evidence that PGDT can contribute to an improvement in patient outcome –9 and guA meta-analysis including 29 studies demonstrated that preemptive PGDT strategies targeting cardiac index (CI) or oxygen delivery improve patient outcome in terms of mortality and postoperative complications in moderate- and high-risk surgical patients .In accordance, a meta-analysis including 32 randomized-controlled trials showed that protocol-based optimization of tissue perfusion in terms of optimization of hemodynamics decreases postoperative mortality and organ dysfunction in high-risk surgical patients, particularly when CI, oxygen delivery, and oxygen consumption are used to guide therapy  allows not only the monitoring of arterial blood pressure but also the estimation of SV, CO, and PPV/SVV Figure . An arteThere are a variety of different algorithms for pulse contour analysis that enable SV to be estimated from the arterial blood pressure waveform , 17. TheBesides the estimation of SV, pulse contour analysis allows the assessment of dynamic cardiac preload variables  that—based on heart-lung interactions during mechanical ventilation—can be used to predict fluid responsiveness .Although pulse contour analysis can be easily used in patients with an arterial catheter, the method has several limitations that are crucial to know to avoid erroneous measurements. First, pulse contour analysis depends on an optimal arterial pressure signal. Therefore, to assure impeccable arterial blood pressure waveform recording, one has to meticulously avoid clotting of the arterial catheter, over- or underdamping of the tubing system, or incorrect zeroing of the pressure transducer and monitoring system. In addition, the clinical usefulness of pulse contour analysis is limited in patients with high-grade cardiac arrhythmias and rapid changes or profound abnormalities in vasomotor tone  . The useThe cardiac function curve  describes the relation of ventricular preload or left-ventricular end-diastolic pressure and SV. A left ventricle functioning on the steep part of the cardiac function curve will increase SV after an increase in cardiac preload . This state of “preload reserve” is clinically referred to as “fluid responsiveness,” i.e., an increase in blood flow following fluid administration. Because ventricular function is a major determinant of the shape of the cardiac function curve, fluid administration must be performed cautiously to avoid fluid overload and circulatory failure, especially in patients with poor ventricular function in whom the heart is already working on the flat part of the curve.Based on these basic physiologic principles, pulse contour analysis provides crucial hemodynamic variables reflecting fluid responsiveness  and blood flow  that can be used in PGDT protocols to titrate fluids and vasoactive agents based on principles of “functional hemodynamic monitoring” . FunctioInvasive uncalibrated pulse contour analysis is frequently used for the assessment of hemodynamic variables within PGDT protocols , 26, 27.These treatment algorithms can be classified according to the hemodynamic variables they use as targets: some algorithms are solely based on either dynamic cardiac preload variables  or blood flow variables ; other algorithms combine these dynamic cardiac preload and blood flow variables .The OPTIMISE trial is an example for a study using pulse contour analysis solely to optimize blood flow . In thisCompared with the approach of maximizing SV by using the full cardiac preload reserve, PGDT algorithms targeting both dynamic cardiac preload parameters and SV/CO may help to better tailor the hemodynamic management to the individual patient , 29.In the ongoing follow-up study of the OPTIMISE trial  as a member of the medical advisory board and received honoraria for giving lectures and refunds of travel expenses from Pulsion Medical Systems SE. BS received institutional research grants, unrestricted research grants, and refunds of travel expenses from Tensys Medical Inc. . BS received honoraria for giving lectures and refunds of travel expenses from CNSystems Medizintechnik AG . BS received research support from Edwards Lifesciences . DR collaborates with Pulsion Medical Systems SE as a member of the medical advisory board and received honoraria for giving lectures and refunds of travel expenses from Pulsion Medical Systems SE. DR provided advising services for and received honoraria for giving lectures from Masimo  and Fresenius Kabi .


ADRA2b polymorphism as a tool to investigate naturally occurring differences in NA availability. We next review associative learning processes that—modulated by the LC-NA system—result in such implicit attentional biases. Further, we demonstrate how NA may influence aversive and appetitive conditioning linked to anxiety disorders as well as addiction and depression.Emotionally arousing events are typically better remembered than mundane ones, in part because emotionally relevant aspects of our environment are prioritized in attention. Such biased attentional tuning is itself the result of associative processes through which we learn affective and motivational relevance of cues. We propose that the locus coeruleus-noradrenaline (LC-NA) system plays an important role in the genesis of attentional biases through associative learning processes as well as their maintenance. We further propose that individual differences in and disruptions of the LC-NA system underlie the development of maladaptive biases linked to psychopathology. We provide support for the proposed role of the LC-NA system by first reviewing work on attentional biases in development and its link to psychopathology in relation to alterations and individual differences in NA availability. We focus on pharmacological manipulations to demonstrate the effect of a disrupted system as well as the  Indeed, whereas attentional prioritization measures preexisting filters that inform what we will see of the world before we ever encounter it, many clinical studies have focused on another form of attentional bias: difficulty with attentional disengagement from salient stimuli once they have already been observed [Emotional salience enhances both attention and memory. For example, we typically remember emotionally arousing events better than mundane ones, reliving the birth of a child or a teenage humiliation with a high degree of vividness decades later –3. We ree retina , the inftization , or seletization , and biatization . Accorditization . Moreovetization , 15 and tization –18. In atization . It shouobserved . In thisAttentional biases appear early in development and specific biases predict later emergence of a range of maladaptive outcomes. A body of recent research has focused on the etiology of maladaptive attentional biases in childhood and adolescence and has suggested a causal role for such biases in the development of anxiety disorders . Researcrelevant either due to explicit task-related demands, visual salience, or motivational/affective salience acquired through life experience [Although the role of NA in guidance of attention to salient aspects of the environment has been thoroughly reviewed elsewhere , 27–29),–29,27–29perience , 29, 34,perience , improviperience . In theiperience . Importaperience . Moreoveperience .ADRA2b gene, which codes for alpha2b NA receptors and is thought to be associated with higher levels of intercellular NA [ADRA2b in emotional enhancement of memory, we used genotyping to examine the role of NA in affectively biased attentional prioritization, which might partly account for emotional enhancement of memory effects. As enhanced encoding of emotionally salient stimuli has been found to predict both subsequent recall and recognition memory . Yet, when T2 is emotionally salient, the attentional blink is somewhat reduced, in a robust finding we refer to as emotional sparing. Following on the work of Di Lollo and colleagues [ADRA2b deletion variant show emotional sparing for both positive and negative stimuli, deletion carriers show an even greater sparing effect for negative stimuli, indicating a role for naturally occurring NA differences in biases in attentional prioritization [ADRA2b deletion carriers subjectively perceive emotionally salient stimuli to be more perceptually vivid  relative to neutral stimuli than noncarriers [ADRA2b deletion variant makes it a tractable tool for examining naturally occurring NA variation-related activity of alpha2b receptors in humans. However, other receptor subtypes also play an important role in modulating NA's effects on cognition. A substantial amount of animal research has demonstrated the importance of high affinity alpha2 and lower affinity alpha1 receptors for optimal functioning of the prefrontal cortex (PFC). More specifically, it has been shown that moderate levels of NA promote PFC functions such as working memory and top-down attention mechanisms as well as decision-making and emotion regulation , we hypy . Our owcarriers  . ThAs mentioned above, implicit biases in attentional prioritization not only influence what we encode and remember but they are also themselves the product of learning and memory . Our resclassical conditioning paradigm, an animal learns to associate an initially neutral stimulus (CS+) with an aversive stimulus or event (US) that elicits an innate response [operant conditioning or reinforcement learning, an animal learns that performing a certain action  is followed by a specific outcome . Similar paradigms have been developed to study associative learning in humans. In the following paragraphs, we will review research on aversive and appetitive conditioning in both human and nonhuman animals, focusing on the role of NA and its relation to psychopathology.Associative learning is used as an umbrella term to refer to different types of learning that are characterized by the development of conscious or unconscious associations between a certain cue or action and the occurrence of a specific stimulus. For example, in an aversive response . After lThe study of aversive conditioning in nonhuman animals has a long history of employing mild electric shocks as US and tones or lights as typical CS+ stimuli. Robust conditioning can be achieved after only a few continuous pairings of the CS with the US. Aversive conditioning in humans can employ a wide range of possible CS and US , and theThe brain circuitry underlying aversive conditioning is also quite well mapped. Research in nonhuman animals as well as lesion and neuroimaging studies in humans has identified the amygdala, the hippocampus, and the ventromedial prefrontal cortex as key nodes in brain systems underlying aversive conditioning. The amygdala plays a role in integration of information about CS and US and controlling fear responses via projections to autonomic and endocrine control systems in the brainstem . LesionsADRA2b deletion variant showed greater susceptibility to intrusive traumatic memory than noncarriers, suggesting a role for these receptors in the intrusive memory component of PTSD [Alterations in this circuit mediated by the LC-NA system are thought to underlie maladaptive patterns of fear learning expressed as fear and anxiety disorders such as PTSD , 71. Fea of PTSD .While one long-prevalent idea has been that PTSD results from disturbances in memory consolidation —a procesAppetitive conditioning is an associative learning process by which initially neutral stimuli or events become associated with a reward and hence gain motivational salience . In appeConverging evidence from human and nonhuman studies suggests that the amygdala plays a key role in appetitive conditioning. The amygdala has been shown to be critical for outcome evaluation and cost estimation , 102 as Increasing evidence suggests that the LC-NA system not only is important for aversive conditioning but also plays a role in reward processing related to addiction. Decades of research have established that dopamine (DA) is essential for the reinforcing effects of various rewards such as drugs –119. A sPutative neuronal mechanisms underlying the role of LC-NA in attentional mechanisms related to reward have been further elucidated in a recent study suggesting a major role of the LC-NA system in modulating neural gain . Under sADRA2b polymorphism) may explain greater predisposition to maladaptive habit formation observed in some individuals. In fact, both human and nonhuman studies have revealed that chronic or acute stress can bias behaviour towards the habitual [A prevailing view in the addiction literature is to characterize addiction as a disorder of appetitive learning : On the habitual , 136 addhabitual , 138. Pahabitual . As deschabitual , can in habitual  and thushabitual .While addiction is characterized by attentional biases associated with increased approach motivation, the opposite picture is present in patients with major depressive disorder (MDD). Anhedonia—the inability to experience pleasure—is a cardinal symptom of depression , 142. ImIn summary, a large body of literature suggests that NA-mediated alterations and individual differences in the appetitive associative learning system give rise to specific patterns of biased attention. Attentional biases can both be strengthened  and weakened  through reward learning and can develop into deeply habitual patterns of orienting to the world that underlie the etiology and maintenance of psychopathology.In summary, we have argued that NA plays an important role in the genesis and maintenance of biased attention patterns that are established via associative learning processes. Here, we first reviewed evidence for the emergence of attentional biases linked to psychopathology in development and the role of putative individual differences in NA availability in such biases. We next reviewed associative learning processes that can give rise to such biases, as well as evidence suggesting a role for NA in specific patterns of fear learning linked to PTSD and appetitive learning linked to both addiction and depression. Based on convergent evidence, we propose that attentional biases play a key role in creating and maintaining prioritization of relevant cues as well as the transfer of reward learning to habitual behaviours associated with addiction. We hypothesize that after attentional biases for reward-related cues are formed through associative learning processes, they are themselves used to inform and prompt behaviours. More specifically, they may facilitate the formation of habitual behaviours by redirecting attention from the outcome to the cue. This is a possible mechanism that could explain why habitual behaviours are performed even if the outcome changes towards the negative. In addition, such biases themselves form deeply habitual patterns of orienting to the world, which can play an important role in etiology and maintenance of psychopathology.ADRA2b genotype as a source of naturally occurring differences in NA availability. It will be important to delineate how both operant and Pavlovian conditioning are affected by these manipulations and whether it is actual learning rate or reward sensitivity that is affected. Future research should aim to disentangle these two components of reward learning. If stress is used as a means to activate the LC-NA system, the intensity and type of stressor need to be considered [A number of outstanding questions remain. First and foremost, little is known about the role of NA in appetitive learning in humans. While previous research in humans demonstrated a role of stress in habit formation and Pavlovian-instrumental transfer, it remains to be investigated whether initial reward learning is affected by NA availability. Future research can examine this by manipulating NA availability, for example, through acute stress induction or by using the nsidered . Effectsnsidered , 155. Thnsidered , 157. ThMoreover, the proposed link between associative learning and attentional biases needs to be tested directly in humans. That is, once the role of NA in associative learning is fully established, one should examine whether newly learned associations result in attentional biases for cue- or outcome-related stimuli.In addition, the directionality of the proposed link needs to be investigated further. While converging evidence suggests that associative learning processes form attentional biases, attentional biases are likely to influence later instances of emotional learning. It is unclear whether activity of the LC-NA would further reinforce existing biases by influencing subsequent learning processes or whether one of main roles of this neurotransmitter system is to facilitate learning processes that give rise to attentional biases. It is likely that the process can be mediated at both ends; however, this problem needs to be investigated in more detail.