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BACKGROUND: Panic disorder (PD) is a common disabling condition characterized by recurrent panic attacks. Emotional and behavioral impairments are associated with functional connectivity (FC) and network abnormalities. We used the whole brain FC, modular networks, and graph-theory analysis to investigate extensive network profiles in PD. METHOD: The functional MRI data from 82 PD and 97 controls were included. Intrinsic FC between each pair of 160 regions, 6 intra-networks, and 15 inter-networks were analyzed. The topological properties were explored. RESULTS: PD patients showed altered FCs within the right insula, between frontal cortex-posterior cingulate cortex (PCC), frontal cortex-cerebellum, and PCC-occipital cortex (corrected P values < 0.001). Lower connections within the Sensorimotor Network (SMN) and SMN-Occipital Network (OCN) were detected (P values < 0.05). Various decreased global and local network features were found in PD (P values < 0.05). In addition, significant correlations were found between PD symptoms and nodal efficiency (Ne) in the insula (r = -0.273, P = 0.016), and the FC of the intra-insula (r = -0.226, P = 0.041). CONCLUSIONS: PD patients present with abnormal functional brain networks, especially the decreased FC and Ne within insula, suggesting that dysfunction of information integration plays an important role in PD.

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Costa Rica emerged from the seas as a new geological territory during the Miocene as an insular archipelago. It later became part of a continental area once it became a segment of Central America. Two dung beetle genera that colonized this new territory from South and North America, Canthidium and Onthophagus (Coleoptera: Scarabaeidae: Scarabaeinae), are here studied, in the first analysis of a volcanic paleo-archipelago, colonized from its emergence, and then later becoming terra firma. To assess their biodiversity distribution patterns, we analyzed the effect of biogeography, ecosystem origins, and body size on their altitudinal distribution patterns in three geographic basins of Costa Rica. Based on 32 years of collecting representing more than 158,000 specimens from 1017 localities, we undertook Generalized Linear Models of the two dung beetle genera to assess the effects of biodiversity and biogeographical distribution patterns. Canthidium and Onthophagus species ranged from 0 to 3000 m a.s.l., with an abrupt diversity decline at altitudes above 1500 m. Endemic species tended to show a higher altitudinal mean with a narrow altitudinal band distribution than non-endemic dung beetle species. Although there was a trend of decreasing species body size with the increase in altitude, such a trend depended on the distribution pattern of the species group. This possible insular-mediated endemicity mechanism has generated baffling biodiversity levels, considered the highest worldwide per unit area. Costa Rica is an expanse represented by a geographic overlap of two or more temporally disjunct areas and is not part of a natural transition zone. The effect of the insular Miocene origin of Costa Rica still pervades today, reflected by different insular syndromes shown by the dung beetle fauna. The importance of geological origins in generating biodiversity seems to have been an underrated criterion for conservation biology practices and should be considered ex officio.

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Erigeron represents the third largest genus on the Juan Fernández Islands, with six endemic species, five of which occur exclusively on the younger Alejandro Selkirk Island with one species on both islands. While its continental sister species is unknown, Erigeron on the Juan Fernández Islands appears to be monophyletic and most likely evolved from South American progenitor species. We characterized the complete chloroplast genomes of five Erigeron species, including accessions of E. fernandezia and one each from Alejandro Selkirk and Robinson Crusoe Islands, with the purposes of elucidating molecular evolution and phylogenetic relationships. We found highly conserved chloroplast genomes in size, gene order and contents, and further identified several mutation hotspot regions. In addition, we found two positively selected chloroplast genes (ccsA and ndhF) among species in the islands. The complete plastome sequences confirmed the monophyly of Erigeron in the islands and corroborated previous phylogenetic relationships among species. New findings in the current study include (1) two major lineages, E. turricola-E. luteoviridis and E. fernandezia-E. ingae-E. rupicola, (2) the non-monophyly of E. fernandezia occurring on the two islands, and (3) the non-monophyly of the alpine species E. ingae complex.

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Memory consolidation is an essential process of long-term memory formation. Neurotrophins have been suggested as key regulators of activity dependent changes in the synaptic efficacy and morphology, which are considered the downstream mechanisms of memory consolidation. The neurotrophin 3 (NT-3), a member of the neurotrophin family, and its high affinity receptor TrkC, are widely expressed in the insular cortex (IC), a region with a critical role in the consolidation of the conditioned taste aversion (CTA) paradigm, in which an animal associates a novel taste with nausea. Nevertheless, the role of this neurotrophin in the cognitive processes that the IC mediates remains unexamined. To answer whether NT-3 is involved in memory consolidation at the IC, adult male Wistar rats were administered with NT-3 or NT-3 in combination with the Trk receptors inhibitor K252a into the IC, immediately after CTA acquisition under two different conditions: a strong-CTA (0.2 M lithium chloride i.p.) or a weak-CTA (0.1 M lithium chloride i.p.). Our results show that NT-3 strengthens the memory trace of CTA, transforming a weak conditioning into a strong one, in a Trk-dependent manner. The present evidence suggests that NT-3 has a key role in the consolidation process of an aversive memory in a neocortical region.

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RATIONALE: Relapse into substance use is often triggered by exposure to drug-related environmental cues. The magnitude of drug seeking depends on the duration of abstinence, a phenomenon known as the incubation of drug craving. Clinical and preclinical research shows that the insular cortex is involved in substance use disorders and cue-induced drug seeking. However, the role of the insula on memory retrieval and motivational integration for cue-elicited drug seeking remains to be determined. OBJECTIVES: We investigated the role of the anterior insular cortex (aIC) and its glutamatergic projection to amygdala nuclei (aIC-AMY) on the expression of conditioned place preference (CPP) during early and late abstinence. METHODS: Male adult C57BL/6J mice underwent amphetamine-induced CPP, and their preference was tested following 1 or 14 days of abstinence. aIC and aIC-AMY functional role in CPP expression was assessed at both abstinence periods by employing optogenetic silencing and behavioral pharmacology. RESULTS: Compared to a single day, an exacerbated preference for the amphetamine-paired context was observed after 14 days of abstinence. Photoinhibition of either aIC or aIC-AMY projection reduced CPP expression following late but not early abstinence. Similarly, the antagonism of aIC NMDA receptors reduced CPP expression after 14 days of abstinence but not 1 day. CONCLUSIONS: These results suggest that aIC and its glutamatergic output to amygdala nuclei constitute critical neurobiological substrates mediating enhanced motivational cue reactivity during the incubation of amphetamine craving rather than contextual memory recall. Moreover, cortical NMDA receptor signaling may become sensitized during abstinence, ultimately modulating disproportioned drug seeking.

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The presentation of novel stimuli induces a reliable dopamine release in the insular cortex (IC) from the ventral tegmental area (VTA). The novel stimuli could be associated with motivational and emotional signals induced by cortical glutamate release from the basolateral amygdala (BLA). Dopamine and glutamate are essential for acquiring and maintaining behavioral tasks, including visual and taste recognition memories. In this study, we hypothesize that the simultaneous activation of dopaminergic and glutamatergic projections to the neocortex can underlie synaptic plasticity. High-frequency stimulation of the BLA-IC circuit has demonstrated a reliable long-term potentiation (LTP), a widely acknowledged synaptic plasticity that underlies memory consolidation. Therefore, the concurrent optogenetic stimulation of the insula's glutamatergic and dopaminergic terminal fibers would induce reliable LTP. Our results confirmed that combined photostimulation of the VTA and BLA projections to the IC induces a slow-onset LTP. We also found that optogenetically-induced LTP in the IC relies on both glutamatergic NMDA receptors and dopaminergic D1/D5 receptors, suggesting that the combined effects of these neurotransmitters can trigger synaptic plasticity in the neocortex. Overall, our findings provide compelling evidence supporting the essential role of both dopaminergic and glutamatergic projections in modulating synaptic plasticity within the IC. Furthermore, our results suggest that the synergistic actions of these projections have a pivotal influence on the formation of motivational memories.

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Environmental enrichment (EE) is known to improve memory and cognition and modulate the impact of aversive stimuli in animals, promoting the development of resilience to stressful situations. Likewise, it is known that EE can modulate synaptic plasticity as is the case of long-term potentiation (LTP). These findings have been described initially in ex vivo preparations, suggesting that the effects of EE are the result of an early modification of the synaptic excitability and transmission. In this regard, it is known that metaplasticity refers to the persistent modification, by previous activity, in the ability to induce synaptic plasticity. Our previous studies have shown that prior training in conditioned taste aversion (CTA) prevents the subsequent induction of LTP in the projection from the basolateral nucleus of the amygdala (Bla) to the insular cortex (IC) in vivo. In addition, we have shown that CTA extinction allows the induction but not the maintenance of IC-LTP of the Bla-IC pathway. Recently, we also showed that prior exposure to environmental enrichment for three weeks reduces the strength of CTA, restoring the brain-derived neurotrophic factor (BDNF) levels in the IC. The present study aimed to analyze the effects of brief exposure to an enriched environment on the strength of aversive memory, as well as on the in vivo IC-LTP. To do so, adult rats were exposed for seven days to an EE, either before CTA training or LTP induction in the Bla-IC pathway. Our results demonstrate that a seven-day exposure to an enriched environment attenuates the aversive response to a strong CTA and allows the induction but not the maintenance of LTP in the insular cortex. These findings provide evidence that metaplastic regulation in a neocortical region takes part in the mechanisms through which brief exposure to enriched environments attenuates an aversive response.

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RATIONALE: In a social context, individuals are able to detect external information from others and coordinate behavioral responses according to the situation, a phenomenon called social decision-making. Social decision-making is multifaceted, influenced by emotional and motivational factors like stress, sickness, and hunger. However, the neurobiological basis for motivational state competition and interaction is not well known. OBJECTIVE: We investigated possible neural mechanisms through which internal states could shape social behavior in a social affective preference (SAP) test. In the SAP test, experimental rats given a choice to interact with naïve or stressed conspecifics exhibit an age-dependent preference to interact with stressed juvenile conspecifics, but avoid stressed adult conspecifics. First, we assessed the effect of food and water deprivation on SAP behavior. Behavior in the SAP test requires the insular cortex, which receives input from the ingestion-related peptides melanin-concentrating hormone (MCH) and orexin neurons of the lateral hypothalamus (LH). This study aimed to evaluate the role of LH and insular MCH and orexin in SAP test. METHODS: SAP tests were conducted in rats that were sated, food and water deprived or allowed 1 h of access to food and water after 14 h of deprivation (relieved condition). Separate cohorts of sated rats received cannula implants for microinjection of drugs to inhibit the LH or to block or stimulate MCH or orexin receptors in the insula prior to SAP tests or social interaction tests. RESULTS: Food and water deprivation prior to SAP tests with juvenile rats caused a shift in preference away from the stressed rat toward the naïve juveniles. Pharmacological inhibition of LH with muscimol (100 ng/side) abolished the preference for the juvenile-stressed conspecific, as well as the preference for the adult naïve conspecific. The blockade of MCH receptor 1or orexin receptors in the insular cortex with SNAP94847 (50 µM) or TCS1102 (1 µM), respectively, also abolished the preference for the stressed juvenile conspecific, but only the antagonism of orexin receptors was able to abolish the preference for the adult naïve conspecific. Microinjection of increasing doses (50 or 500 nM) of MCH or orexin-A in the insular cortex increased the interaction time in the one-on-one social interaction test with juvenile conspecifics; however, only the microinjection of orexin-A increased the interaction time with adult naïve conspecifics. CONCLUSIONS: Taken together, these results suggest that lateral hypothalamus peptides shape the direction of social approach or avoidance via actions MCH and orexin neurotransmission in the insular cortex.

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Increased activity in the insula has been consistently reported to be associated with anxiety and anxiety-related disorders. However, little is known on how the insula regulates anxiety. The present study aims at determining the role of the insula on the effects of glucocorticoids in anxiety. A combination of pharmacological manipulations, including blockade of adrenal GC synthesis by metyrapone and intra-insular microinjections of corticosterone, corticosterone-BSA, mineralocorticoid receptor (MR) antagonist spironolactone and glucocorticoid receptor (GR) antagonist mifepristone, were used to assess the short-term (5 min) effects of intra-insular corticosterone in two anxiety-like behaviors in male Sprague-Dawley rats. The elevated plus maze (EPM) and Novelty Suppressed Feeding (hyponeophagia) were utilized. We found that corticosterone in the insula is sufficient to prevent the anxiolytic effects corticosterone synthesis blockade in anxiety, and that intra-insular corticosterone has anxiolytic or anxiogenic effects depending on the amount of corticosterone microinjected and the arousal associated to the test, without affecting the HPA axis. Glucocorticoid anxiolytic effects in the insula are mediated by MRs, while its anxiogenic effects are dependent on a mifepristone-sensitive membrane-bound mechanism. Anxiety appears to be modulated at the insula through a competition between fast MR-dependent anxiolytic and membrane-associated anxiogenic signaling pathways that orchestrate the behavioral response to stress and determines the resulting level of anxiety.

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Resumen Objetivo: Explorar si voluntarios sanos presentarían correlación entre su puntaje en un test psicológico emocional y las activaciones de áreas cerebrales relacionadas con las emociones medidas con resonancia magnética funcional (RMf). Material y métodos: Estudio exploratorio de prueba diagnóstica, prospectivo, con diseño del propio individuo como control, con muestra de 12 participantes. Se categorizó a cada participante con un puntaje de estabilidad emocional derivado de un test psicológico y se utilizaron estímulos emocionales audiovisuales durante las adquisiciones de RMf. Resultados: La sumatoria de clusters de activación medidos en cantidad total de vóxeles durante los estímulos negativos en áreas cerebrales relacionadas con las emociones mostró una correlación negativa estadísticamente significativa para nuestro tamaño de muestra respecto de los puntajes en el test emocional, con rho de Spearman de −0,623 y p = 0,0428. Conclusiones: Los paradigmas de RMf utilizados permitieron cuantificar las activaciones cerebrales ante estímulos emocionales de valencia positiva y negativa, y los resultados obtenidos abren una perspectiva hacia la posibilidad de utilizar test psicológicos y secuencias de RMf para predecir la posibilidad de aparición de síntomas de patologías psicológicas o psiquiátricas ante factores desencadenantes en población sana que presente en estos test valores cercanos al límite de la normalidad.

Abstract Objective: To explore if healthy volunteers would present a correlation between their score in emotional psychological test and the activations of brain areas related to emotions measured with functional magnetic resonance imaging (fMRI). Material and methods: Exploratory study of a prospective diagnostic test, with the individual’s own design as a control, with a sample of 12 participants. Each participant was categorized with an emotional stability score derived from a psychological test and audiovisual emotional stimuli were used during fMRI acquisitions. Results: The sum of activation clusters measured in total number of voxels during negative stimuli in brain areas related to emotions showed a statistically significant negative correlation for our sample size with respect to the scores in the emotional test, with Spearman’s rho of −0.623 and p = 0.0428. Conclusions: The fMRI paradigms used made it possible to quantify brain activations in response to emotional stimuli of positive and negative valence, and the results obtained open a perspective towards the possibility of using psychological tests and fMRI sequences to predict the possibility of the appearance of symptoms of psychological or psychiatric pathologies in response to triggering factors in a healthy population that present values close to the normal limit in these tests.

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In this study the authors use the Fishers ecological knowledge (FEK) from the south coast of the São Paulo State, in southeastern Brazil, to recover records of Atlantic nurse shark (Ginglymostoma cirratum), a threatened with extinction species, in a mosaic of marine-protected areas (MPAs) of the south coast of São Paulo, in southeastern Brazil, in the southernmost distribution of the species in the Southwest Atlantic Ocean. For half a century, in the study area only one male of G. cirratum was officially recorded, in 1967. Retrieving records from artisanal fishers, recreational anglers, fishing guides and spearfishers added 30 more records of G. cirratum, including 2 recent captures (February/2022 and May/2022), especially around coastal islands that are covered by MPAs. Two uncommon fishing techniques, but which provided the highest number of records for the species, were documented. These are fishing nets and small longlines adapted for fishing close to the rocks, being an apparently traditional activity of a few natives of the region. As Brazil has a long history of intense catches of threatened elasmobranchs and problems with fisheries monitoring, the consideration of the FEK in the fisheries monitoring carried out by the authorities can be useful to promote improvements in data collection, especially of rare and endangered species such as G. cirratum. The strengthening of the protection of the MPAs and the articulation of research and management institutions with native people and tourists who use these areas should also be considered.

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Previous studies have shown that dopaminergic activity modulates the salience of novel stimuli enabling the formation of recognition memories. In this work, we hypothesize that dopamine released into the insular cortex (IC) from the ventral tegmental area (VTA) inputs enables the acquisition to consolidate object recognition memory. It has been reported that short training produces weak recognition memories; on the contrary, longer training produces lasting and robust recognition memories. Using a Cre-recombinase under the tyrosine hydroxylase (TH+) promoter mouse model, we photostimulated the VTA-IC dopaminergic pathway during short training or photoinhibited the same pathway during long training while mice explored objects. Our results showed that the photostimulation of the VTA-IC pathway during a short training enables the acquisition of recognition memory. Conversely, photoinhibition of the same pathway during a long training prevents the acquisition of recognition memory. Interestingly, the exploration time of the objects under photoinhibition or photostimulation of the dopaminergic VTA-IC pathway was not altered. Significantly, this enhancement of acquisition of the object recognition memory through the photostimulation of the VTA dopaminergic neurons could be impaired by the blockage of the D1-like receptors into the IC, either before or after the photostimulation. Altogether, our results suggest that dopamine released by the VTA is required during the acquisition to consolidate the object recognition memory through D1-like receptors into the IC without affecting the activity or the motivation to explore objects.

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Activation of the insula is found in all anxiety-related disorders and increased insular-prefrontal cortex (PFC) functional connectivity is associated with reduced anxiety. In this study, the combined stimulation of the insula and PFC using the dTMS H4 (insula+LPFC) and H2 (PFC) coils were used to reduce anxiety in 13 subjects experiencing occupational stress, and 55 participants suffering from generalized anxiety disorder (GAD). The combined HF stimulation of the insula and PFC significantly decreased anxiety scores according to the HARS, CAS, and STAI anxiety scales, leading to a reduction in anxiety according to HARS of 88.7% and 70.7% in participants with occupational stress and the clinical sample of participants diagnosed with GAD, respectively. The findings suggest that the prefrontal-insular axis is critical for the regulation of anxiety and its stimulation can be used for the treatment of anxiety in people suffering from occupational stress and GAD.

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Homeostatic challenges may alter the drive for social interaction. The neural activity that prompts this motivation remains poorly understood. In the present study, we identify direct projections from the hypothalamic supraoptic nucleus to the cortico-amygdalar nucleus of the lateral olfactory tract (NLOT). Dual in situ hybridization with probes for pituitary adenylate cyclase-activating polypeptide (PACAP), as well as vesicular glutamate transporter (VGLUT)1, VGLUT2, V1a and V1b, revealed a population of vasopressin-receptive PACAPergic neurons in NLOT layer 2 (NLOT2). Water deprivation (48 h, WD48) increased sociability compared to euhydrated subjects, as assessed with the three-chamber social interaction test (3CST). Fos expression immunohistochemistry showed NLOT and its main efferent regions had further increases in rats subjected to WD48 + 3CST. These regions strongly expressed PAC1 mRNA. Microinjections of arginine vasopressin (AVP) into the NLOT produced similar changes in sociability to water deprivation, and these were reduced by co-injection of V1a or V1b antagonists along with AVP. We conclude that, during challenge to water homeostasis, there is a recruitment of a glutamatergic-multi-peptidergic cooperative circuit that promotes social behavior.

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Birds that inhabit white-sand ecosystems (WSE) in the Amazon are adapted to habitats distributed as isolated patches. These environments occur in sandy soils that are extremely poor in nutrients, have low floristic diversity and support bird assemblages restricted to WSE. We investigated whether bird species specialized in WSE have morphological or ecological traits that distinguish them from generalist birds that share the same habitat but are not restricted to WSE. We collected morphological and ecological data from 22 specialist and 102 generalist bird species from WSE and described their ecomorphological diversity using multivariate analyses and measures of functional diversity. Understory insectivorous species that move alone or in pairs, were the group with the most species among specialist birds from WSE. In contrast, canopy frugivorous species that form mono- or heterospecific groups were more frequent among generalist species. Specialist and generalist birds overlapped extensively in ecomorphological space, with specialists occupying a narrower space compared to generalists. Functional diversity, however, was not different between specialist and generalist species when controlling for the number of species within the communities. Further studies comparing the ecomorphological diversity of bird assemblages of other environments in the Amazon may highlight the ecological pressures leading to the functional diversity of specialist species in WSE observed in this study.(AU)

As aves que habitam os ecossistemas de areia branca (EAB) na Amazônia estão adaptadas a habitats distribuídos em manchas isoladas. Esses ambientes ocorrem em solos arenosos extremamente pobres em nutrientes, possuem baixa diversidade florística e suportam assembleias de aves restritas a EAB. Investigamos se espécies de aves especializadas em EAB possuem características morfológicas ou ecológicas que as distinguem de aves generalistas que compartilham o mesmo habitat, mas não estão restritas a EAB. Coletamos dados morfológicos e ecológicos de 22 espécies de aves especialistas e 102 generalistas de EAB, e descrevemos sua diversidade ecomorfológica usando análises multivariadas e medidas de diversidade funcional. Espécies insetívoras do sub-bosque que se movem solitárias ou em pares foram o grupo com maior número de espécies dentro das aves especialistas da EAB. Em contraste, espécies frugívoras de dossel que formam grupos mono ou heteroespecíficos foram mais frequentes entre as espécies generalistas. Aves especialistas e generalistas sobrepuseram-se extensivamente no espaço ecomorfológico, com especialistas ocupando um espaço mais estreito em comparação com generalistas. A diversidade funcional, no entanto, não foi diferente entre espécies especialistas e generalistas ao controlar o número de espécies dentro das comunidades. Estudos futuros comparando a diversidade ecomorfológica de assembleias de aves de outros ambientes na Amazônia podem destacar as pressões ecológicas que levam à diversidade funcional de espécies especialistas em EAB observadas neste estudo.(AU)

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Introduction: Vascular lesions in insular glioma surgery can severely impact patients' quality of life. This study aims to present the results of our dissections and authors' reflections on the insular vascular anatomy. Matherials and Methods: The insular vascularization was examined using ×3 to ×40 magnification in 20 cadaveric cerebral hemispheres in which the arteries and veins had been perfused with colored silicone. Results: In insular gliomas, this individualization of the anatomical structures is rarely possible, as the gyri are swollen by the tumor and lose their individuality. In the transsylvian approaches, the anatomical parameters for delimiting the insula in tumors are best provided by the superior and inferior circular sulci. The branches of the MCA are easily identified in the transcortical approach, but only at the end of the surgery after the tumor is resected.). One of the factors under-discussed in the literature is the involvement of the lenticulostriate arteries by the medial part of the tumor. In our experience of 52 patients (article submitted to publishing), LSTa were founded to be involved by the tumor in 13 cases. In 39 patients, there was no involvement of the LSTa, which allowed a more aggressive resection. Early preoperative identification of the anterior perforated substance on the MRI and its proximity to the tumor may help determine the route of the LSTa over the medial tumor boundaries. Discussion: Our reflections introduced our imaging and anatomical concept regarding LSTa in insular glioma surgery. Accurate identification of origin, route, and distribution of the LSTa is pivotal to surgical success, especially in the lateral group. The anatomical knowledge of their path directly impacts the extent of tumor resection and functional preservation. Conclusion: Knowledge of microsurgical anatomy, brain mapping, and surgical experience counts a lot in this type of surgery, creating a reasonable procedure flowchart to be taken intraoperatively.

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It has been shown that exposure to an enriched environment (EE) can modulate the physiological impact of aversive stimuli in animals, promoting adaptive attitudes, as well as the development of resilience to stressful situations. These changes are known to be related to increased levels of some trophic factors, such as brain-derived neurotrophic factor (BDNF), which has been considered a regulatory protein for synaptic plasticity in the adult brain. Our previous studies have demonstrated that in the insular cortex (IC), a brain region of the temporal lobe implicated in the acquisition, consolidation, and retention of conditioned taste aversion (CTA) task, BDNF can reverse the CTA memory deficit caused by a protein synthesis inhibitor. Likewise, our research group have also shown that BDNF is required for the maintenance of CTA long-term memory. Here we evaluate the effects of the exposure to an enriched environment on the CTA memory strength, using a weak and strong version of this paradigm. The exposure to an EE for 21 days was able to attenuate the strong-CTA response through the restoration of BDNF levels in the IC of adult rats. These results provide evidence that environmental enrichment is capable of reducing the strength of an aversive memory trace, restoring the BDNF levels in a neocortical region of the adult brain.

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The ability to retrieve contextual fear memories depends on the coordinated activation of a brain-wide circuitry. Transition from recent to remote memories seems to involve the reorganization of this circuitry, a process called systems consolidation that has been associated with time-dependent fear generalization. However, it is unknown whether emotional memories acquired under different stress levels can undergo different systems consolidation processes. Here, we explored the activation pattern and functional connectivity of key brain regions associated with contextual fear conditioning (CFC) retrieval after recent (2 days) or remote (28 days) memory tests performed in rats submitted to strong (1.0 mA footshock) or mild (0.3 mA footshock) training. We used brain tissue from Wistar rats from a previous study, where we observed that increasing training intensity promotes fear memory generalization over time, possibly due to an increase in corticosterone (CORT) levels during memory consolidation. Analysis of Fos expression across 8 regions of interest (ROIs) allowed us to identify coactivation between them at both timepoints following memory recall. Our results showed that strong CFC elicits higher Fos activation in the anterior insular and prelimbic cortices during remote retrieval, which was positively correlated with freezing along with the basolateral amygdala. Rats trained either with mild or strong CFC showed broad functional connectivity at the recent timepoint whereas only animals submitted to the strong CFC showed a widespread loss of coactivation during remote retrieval. Post-training plasma CORT levels are positively correlated with FOS expression during recent retrieval in strong CFC, but negatively correlated with FOS expression during remote retrieval in mild CFC. Our findings suggest that increasing training intensity results in differential processes of systems consolidation, possibly associated with increased post-training CORT release, and that strong CFC engages activity from the aIC, BLA and PrL - areas associated with the Salience Network in rats - during remote retrieval.

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Island ecosystems differ in several elements from mainland ecosystems and may induce variations related to natural selection and patterns of adaptation in most aspects of the biology of an organism. Thylamys elegans (Waterhouse, 1839) is a marsupial endemic to Chile, distributed from Loa River to Concepción. Historically, three subspecies have been described: Thylamys elegans elegans, Thylamys elegans coquimbensis and Thylamys elegans soricinus. For this research, two morphometric approaches and a biomechanical model were used to compare the mandible shapes and biomechanics between two Chilean mouse opossum populations belonging to different subspecies: one from the coastal desert of Chile (T. e. coquimbensis) and the other from the central inland region (T. e. elegans). Additionally, mandibles of insular populations found in the Reserva Nacional Pinguino de Humboldt (RNPH)), from which the subspecies association is unknown, were also included. The results showed that insular populations have differences in mandibular shapes, sizes and biomechanical characteristics compared to continental populations, which may be related to environmental variables like aridity and vegetation cover, prey type, insularity effects and/or the founder effect on micromammals, apart from vicariance hypotheses and other selective pressures.