The Active Role of Pericytes During Neuroinflammation in the Adult Brain.

Microvessels in the central nervous system (CNS) have one of the highest populations of pericytes, indicating their crucial role in maintaining homeostasis. Pericytes are heterogeneous cells located around brain microvessels; they present three different morphologies along the CNS vascular tree: ensheathing, mesh, and thin-strand pericytes. At the arteriole-capillary transition ensheathing pericytes are found, while mesh and thin-strand pericytes are located at capillary beds. Brain pericytes are essential for the establishment and maintenance of the blood-brain barrier, which restricts the passage of soluble and potentially toxic molecules from the circulatory system to the brain parenchyma. Pericytes play a key role in regulating local inflammation at the CNS. Pericytes can respond differentially, depending on the degree of inflammation, by secreting a set of neurotrophic factors to promote cell survival and regeneration, or by potentiating inflammation through the release of inflammatory mediators (e.g., cytokines and chemokines), and the overexpression of cell adhesion molecules. Under inflammatory conditions, pericytes may regulate immune cell trafficking to the CNS and play a role in perpetuating local inflammation. In this review, we describe pericyte responses during acute and chronic neuroinflammation.

First insights in terrestrial mammals monitoring in the Candelaria and Machay Reserves in the Ecuadorian Tropical Andes.

Habitat disturbance leads to biodiversity decline and modifications in the landscape structure and composition, affecting both dispersal movements and ecological processes at different temporal and spatial scales. The Ecuadorian Tropical Andes harbour suitable habitats for the distribution of a wide variety of species; however, there is a lack of studies focused on mammal diversity and its association with the habitat attributes in the central-eastern slopes. Here, we reported the diversity of terrestrial mammals recorded between 2019 and 2021 in a camera-trap monitoring study in the Candelaria and Machay reserves in the upper basin of the Pastaza River, Ecuador. We performed site-occupancy probability analysis to assess the influence of spatial variables in the species' occurrence and also, based on natural marks, we reported preliminary findings in Andean bear individual identification. We detected 22 species of terrestrial mammals. Alpha diversity was similar between reserves with slightly higher species richness in Machay. Evenness indices showed unequal species distribution, with the Andean bear and domestic dogs exhibiting greater dominance. In addition, species composition was dissimilar between reserves, where the species turnover mostly explained the beta diversity. We observed that Andean bear and puma detections increased according to the natural vegetation cover. Conversely, domestic dogs were frequently detected in cells with an increasing proportion of pastures and crops. Additionally, we identified 26 Andean bears and six individuals recaptured during our study. Our results caution about the disturbance derived from human activities since we recorded unprecedented detections of domestic dogs in wild habitats. Nonetheless, it highlights the importance of private conservation areas (e.g. Candelaria, Machay and others) for supporting the occurrence and dispersal of terrestrial mammal species between larger areas in the upper basin of the Pastaza River.

A review of cardiac troponin I detection by surface enhanced Raman spectroscopy: Under the spotlight of point-of-care testing.

Cardiac troponin I (cTnI) is a biomarker widely related to acute myocardial infarction (AMI), one of the leading causes of death around the world. Point-of-care testing (POCT) of cTnI not only demands a short turnaround time for its detection but the highest accuracy levels to set expeditious and adequate clinical decisions. The analytical technique Surface-enhanced Raman spectroscopy (SERS) possesses several properties that tailor to the POCT format, such as its flexibility to couple with rapid assay platforms like microfluidics and paper-based immunoassays. Here, we analyze the strategies used for the detection of cTnI by SERS considering POCT requirements. From the detection ranges reported in the reviewed literature, we suggest the diseases other than AMI that could be diagnosed with this technique. For this, a section with information about cardiac and non-cardiac diseases with cTnI release, including their release kinetics or cut-off values are presented. Likewise, POCT features, the use of SERS as a POCT technique, and the biochemistry of cTnI are discussed. The information provided in this review allowed the identification of strengths and lacks of the available SERS-based point-of-care tests for cTnI and the disclosing of requirements for future assays design.

HDAC11 activity contributes to MEK inhibitor escape in uveal melanoma.

We previously demonstrated that pan-HDAC inhibitors could limit escape from MEK inhibitor (MEKi) therapy in uveal melanoma (UM) through suppression of AKT and YAP/TAZ signaling. Here, we focused on the role of specific HDACs in therapy adaptation. Class 2 UM displayed higher expression of HDACs 1, 2, and 3 than Class 1, whereas HDACs 6, 8, and 11 were uniformly expressed. Treatment of UM cells with MEKi led to modulation of multiple HDACs, with the strongest increases observed in HDAC11. RNA-seq analysis showed MEKi to decrease the expression of multiple HDAC11 target genes. Silencing of HDAC11 significantly reduced protein deacetylation, enhanced the apoptotic response to MEKi and reduced growth in long-term colony formation assays across multiple UM cell lines. Knockdown of HDAC11 led to decreased expression of TAZ in some UM cell lines, accompanied by decreased YAP/TAZ transcriptional activity and reduced expression of multiple YAP/TAZ target genes. Further studies showed this decrease in TAZ expression to be associated with increased LKB1 activation and modulation of glycolysis. In an in vivo model of uveal melanoma, silencing of HDAC11 limited the escape to MEKi therapy, an effect associated with reduced levels of Ki67 staining and increased cleaved caspase-3. We have demonstrated a novel role for adaptive HDAC11 activity in UM cells, that in some cases modulates YAP/TAZ signaling leading to MEKi escape.

Placental miRNAs Targeting Cellular Stress Response Pathways Are Highly Expressed in Non-Hispanic Black People.

Non-Hispanic Black (NHB) people have a 2.5-fold higher risk of maternal mortality when compared to non-Hispanic White (NHW) people. Neonates of NHB people are more likely to be born preterm and small for gestational age, which may be driven by structural racism. The placenta is very sensitive to the maternal environment and may play a critical role in the translation of environmental stressors to pregnancy outcomes. Our aim was to assess the placental miRNA expression profile in both NHB and NHW people and the association between differentially expressed miRNAs and pregnancy outcomes. Placentas were collected from 50 NHB and 74 NHW people with a normal singleton pregnancy undergoing elective cesarean section at term prior to the onset of labor. Placental miRNA expression was measured via whole-genome small RNA-sequencing in a subset of 77 placentas. Fifteen miRNAs were more highly expressed in the placentas of NHB people. Several of these miRNAs were associated with cellular stress response pathways, suggesting that they may be responding to environmental stressors. Placental miR-192-5p expression was lower among NHB people and was positively associated with neonatal adiposity, suggesting it may be sensitive to structural racism with potential impacts on fetal growth.

Placental miR-3940-3p Is Associated With Maternal Insulin Resistance in Late Pregnancy.

CONTEXT: An increase in maternal insulin resistance (IR) during pregnancy is essential for normal fetal growth. The mechanisms underlying this adaptation are poorly understood. Placental factors are believed to instigate and maintain these changes, as IR decreases shortly after delivery. Methylation of placental gene loci that are common targets for miRNAs are associated with maternal IR. OBJECTIVE: We hypothesized that placental miRNAs targeting methylated loci are associated with maternal IR during late pregnancy. METHODS: We collected placentas from 132 elective cesarean sections and fasting blood samples at delivery to estimate maternal homeostasis model assessment of insulin resistance (HOMA-IR). Placental miRNA expression was measured via whole genome small-RNA sequencing in a subset of 40 placentas selected by maternal pre-gravid body mass index (BMI) and neonatal adiposity. Five miRNAs correlated with maternal HOMA-IR and previously identified as targeting methylated genes were selected for validation in all 132 placenta samples via RT-qPCR. Multiple regression adjusted for relevant clinical variables. RESULTS: Median maternal age was 27.5 years, with median pre-pregnancy BMI of 24.7 kg/m2, and median HOMA-IR of 2.9. Among the 5 selected miRNA, maternal HOMA-IR correlated with the placental expression of miRNA-371b-3p (r = 0.25; P = 0.008) and miRNA-3940-3p (r = 0.32; P = 0.0004) across the 132 individuals. After adjustment for confounding variables, placental miRNA-3940-3p expression remained significantly associated with HOMA-IR (ß = 0.16; P = 0.03). CONCLUSION: Placental miRNA-3940-3p was associated with maternal IR at delivery. This placental miRNA may have an autocrine or paracrine effect-regulating placental genes involved in modulating maternal IR.

Evaluation of percutaneous renal biopsy complications based on outcomes and indicators of the Nursing Outcomes Classification.

OBJECTIVE: to evaluate the complications of percutaneous renal biopsy based on outcomes and clinical indicators of the Nursing Outcomes Classification. METHOD: a prospective longitudinal study. The sample consisted of 13 patients submitted to percutaneous renal biopsy, with 65 evaluations. The patients were evaluated in five moments in the 24 hours after the procedure, using an instrument developed by the researchers based on five outcomes (Blood coagulation, Circulation status, Blood loss severity, Pain level, Comfort status: Physical) and 11 indicators. The Generalized Estimation Equation Test was used to compare the scores of the indicators. The project was approved by the institutional ethics committee. RESULTS: in the 65 evaluations, a statistically significant difference was identified in the reduction of the scores of the following nursing outcomes: Blood coagulation, "hematuria" indicator; Circulation status, in the "systolic blood pressure and diastolic blood pressure" indicators and Comfort status: physical, in the "physical well-being" indicator. CONCLUSION: the evaluated patients did not show major complications. The clinical indicators signaled changes in circulation status, with reduced blood pressure, as well as in blood clotting observed by hematuria, but without hemodynamic instability. The comfort status was affected by the rest time after the procedure.

A Review on SERS-Based Detection of Human Virus Infections: Influenza and Coronavirus.

The diagnosis of respiratory viruses of zoonotic origin (RVsZO) such as influenza and coronaviruses in humans is crucial, because their spread and pandemic threat are the highest. Surface-enhanced Raman spectroscopy (SERS) is an analytical technique with promising impact for the point-of-care diagnosis of viruses. It has been applied to a variety of influenza A virus subtypes, such as the H1N1 and the novel coronavirus SARS-CoV-2. In this work, a review of the strategies used for the detection of RVsZO by SERS is presented. In addition, relevant information about the SERS technique, anthropozoonosis, and RVsZO is provided for a better understanding of the theme. The direct identification is based on trapping the viruses within the interstices of plasmonic nanoparticles and recording the SERS signal from gene fragments or membrane proteins. Quantitative mono- and multiplexed assays have been achieved following an indirect format through a SERS-based sandwich immunoassay. Based on this review, the development of multiplex assays that incorporate the detection of RVsZO together with their specific biomarkers and/or secondary disease biomarkers resulting from the infection progress would be desirable. These configurations could be used as a double confirmation or to evaluate the health condition of the patient.

Anhydroecgonine methyl ester, a cocaine pyrolysis product, contributes to cocaine-induced rat primary hippocampal neuronal death in a synergistic and time-dependent manner.

Crack cocaine users are simultaneously exposed to volatilized cocaine and to its main pyrolysis product, anhydroecgonine methyl ester (AEME). Although the neurotoxic effects of cocaine have been extensively studied, little is known about AEME or its combination. We investigated cell death processes using rat primary hippocampal cells exposed to cocaine (2 mM), AEME (1 mM) and their combination (C + A), after 1, 3, 6 and 12 h. Cocaine increased LC3 I after 6 h and LC3 II after 12 h, but reduced the percentage of cells with acid vesicles, suggesting failure in the autophagic flux, which activated the extrinsic apoptotic pathway after 12 h. AEME neurotoxicity did not involve the autophagic process; rather, it activated caspase-9 after 6 h and caspase-8 after 12 h leading to a high percentage of cells in early apoptosis. C + A progressively reduced the percentage of undamaged cells, starting after 3 h; it activated both apoptotic pathways after 6 h, and was more neurotoxic than cocaine and AEME alone. Also, C + A increased the phosphorylation of p62 after 12 h, but there was little difference in LC3 I or II, and a small percentage of cells with acid vesicles at all time points investigated. In summary, the present study provides new evidence for the neurotoxic mechanism and timing response of each substance alone and in combination, indicating that AEME is more than just a biological marker for crack cocaine consumption, as it may intensify and hasten cocaine neurotoxicity.

Placental Impact of Dietary Supplements: More Than Micronutrients.

PURPOSE: Maternal nutrition is a key modifier of fetal growth and development. However, many maternal diets in the United States do not meet nutritional recommendations. Dietary supplementation is therefore necessary to meet nutritional goals. The effects of many supplements on placental development and function are poorly understood. In this review, we address the therapeutic potential of maternal dietary supplementation on placental development and function in both healthy and complicated pregnancies. METHODS: This is a narrative review of original research articles published between February 1970 and July 2020 on dietary supplements consumed during pregnancy and placental outcomes (including nutrient uptake, metabolism and delivery, as well as growth and efficiency). Impacts of placental changes on fetal outcomes were also reviewed. Both human and animal studies were included. FINDINGS: We found evidence of a potential therapeutic benefit of several supplements on maternal and fetal outcomes via their placental impacts. Our review supports a role for probiotics as a placental therapeutic, with effects that include improved inflammation and lipid metabolism, which may prevent preterm birth and poor placental efficiency. Supplementation with omega-3 fatty acids (as found in fish oil) during pregnancy tempers the negative effects of maternal obesity but may have little placental impact in healthy lean women. The beneficial effects of choline supplementation on maternal health and fetal growth are largely attributable to its placental impacts. l-arginine supplementation has a potent provascularization effect on the placenta, which may underlie its fetal growth-promoting properties. IMPLICATIONS: The placenta is exquisitely sensitive to dietary supplements. Pregnant women should consult their health care practitioner before continuing or initiating use of a dietary supplement. Because little is known about impacts of many supplements on placental and long-term offspring health, more research is required before robust clinical recommendations can be made.

Selective Regional Isolation of Brain Microvessels.

The study of the regionalized function of the blood-brain barrier at the level of brain endothelial cells and pericytes is essential to understand the biological properties and molecular mechanisms regulating this biological barrier. The isolation of blood vessels from specific brain regions will allow to understand regional differences in susceptibility to pathological phenomena such as ischemia, traumatic brain injury, and neurodegenerative diseases, such as Alzheimer disease. Here, we propose an efficient and fast method to isolate brain endothelial cells and pericytes from a specific cerebral region. The isolated brain endothelial cells and pericytes are viable to perform conventional molecular and histological techniques such as Western blots, immunocytofluorescence, and scanning electron microscopy.

Complementarity of pain assessment instruments in children with central nervous system impairment / Complementariedad de instrumentos de medición del dolor en niños con compromiso del sistema nervioso central / Complementaridade de instrumentos de mensuração de dor em crianças com comprometimento do sistema nervoso central

ABSTRACT Objective: To evaluate the complementarity of the revised Face, Legs, Activity, Cry, Consolability scale and of the Inventory of Pain Behavior in Neurological Disability for the assessment of pain in children with severe neurological impairment. Method: Cross-sectional study conducted in pediatric units of a university hospital in the southern region of Brazil. The sample consisted of 26 children with severe neurological impairment, hospitalized from January to August 2019, and their caregivers. The data were analyzed by descriptive statistics; Kappa Coefficient, Fisher's Exact Test and Spearman's Coefficient were used (p≤0,05). Results: Most children primary diagnosis was cerebral palsy (80.8%). Pain was present in 50.0% of children with the application of the scale and in 34.6% with that of the inventory. Considering the two instruments, there was good agreement (84.6%) between respondents (k=0.692; 95% CI 0.437-0.967; p=0.000). Conclusion: The instruments can be used complementarily to assess pain in children with this profile.

RESUMEN Objetivo: Evaluar la complementariedad de la Escala revised Faces, Legs, Activity, Cry and Consolability y del Inventario de Comportamientos del Dolor en la Deficiencia Neurológica para la medición del dolor en los niños con severo compromiso neurológico. Método: Estudio transversal, realizado en las unidades pediátricas de un hospital universitario de la región sur de Brasil. Muestra de 26 niños con severo compromiso neurológico, internados desde enero a agosto de 2019, y sus cuidadores. Datos analizados por estadística descriptiva; se utilizó el coeficiente Kappa, la Prueba Exacta de Fisher y la Coeficiente de Spearman (p≤0,05). Resultados: La mayoría de los niños tenía un diagnóstico primario de parálisis cerebral (80,8%). La puntuación indicó presencia de dolor en 50% de los niños por la aplicación de la escala, y en 34,6%, por el inventario. Considerando los dos instrumentos, hubo una buena concordancia (84,6%) entre los encuestados (k=0, 692; IC 95% 0,437-0,967; p=0,000). Conclusión: Los instrumentos se puede utilizar de forma complementaria para evaluar el dolor en este perfil de niños.

RESUMO Objetivo: Avaliar a complementaridade da Escala revised Faces, Legs, Activity, Cry and Consolability e do Inventário de Comportamentos da Dor na Deficiência Neurológica para mensuração da dor em crianças com comprometimento neurológico severo. Método: Estudo transversal, realizado em unidades pediátricas de um hospital universitário da região Sul do Brasil. Amostra de 26 crianças com comprometimento neurológico severo, internadas de janeiro a agosto de 2019, e seus cuidadores. Dados analisados por estatística descritiva; Coeficiente de Kappa, Teste Exato de Fisher e Coeficiente de Spearman foram utilizados (p≤0,05). Resultados: Maioria das crianças apresentou diagnóstico primário de paralisia cerebral (80,8%). Presença de dor foi pontuada em 50% das crianças com a aplicação da escala e, em 34,6%, com inventário. Considerando os dois instrumentos, houve boa concordância (84,6%) entre os respondentes (k=0,692; IC 95% 0,437-0,967;p=0,000). Conclusão: Os instrumentos podem ser utilizados de forma complementar na avaliação da dor neste perfil de crianças.

Evaluation of percutaneous renal biopsy complications based on outcomes and indicators of the Nursing Outcomes Classification / Evaluación de las complicaciones de la biopsia renal percutánea con base en los resultados e indicadores de la Nursing Outcomes Classification / Avaliação de complicações da biópsia renal percutânea com base nos resultados e indicadores da Nursing Outcomes Classification

Abstract Objective: to evaluate the complications of percutaneous renal biopsy based on outcomes and clinical indicators of the Nursing Outcomes Classification. Method: a prospective longitudinal study. The sample consisted of 13 patients submitted to percutaneous renal biopsy, with 65 evaluations. The patients were evaluated in five moments in the 24 hours after the procedure, using an instrument developed by the researchers based on five outcomes (Blood coagulation, Circulation status, Blood loss severity, Pain level, Comfort status: Physical) and 11 indicators. The Generalized Estimation Equation Test was used to compare the scores of the indicators. The project was approved by the institutional ethics committee. Results: in the 65 evaluations, a statistically significant difference was identified in the reduction of the scores of the following nursing outcomes: Blood coagulation, "hematuria" indicator; Circulation status, in the "systolic blood pressure and diastolic blood pressure" indicators and Comfort status: physical, in the "physical well-being" indicator. Conclusion: the evaluated patients did not show major complications. The clinical indicators signaled changes in circulation status, with reduced blood pressure, as well as in blood clotting observed by hematuria, but without hemodynamic instability. The comfort status was affected by the rest time after the procedure.

Objetivo: evaluar las complicaciones de la biopsia renal percutánea con base en los resultados e indicadores clínicos de la Nursing Outcomes Classification. Método: estudio longitudinal prospectivo. La muestra fue de 13 pacientes a los que se les realizó biopsia renal percutánea, con 65 evaluaciones. Los pacientes fueron evaluados en cinco momentos en las 24 horas posteriores al procedimiento, utilizando un instrumento desarrollado por los investigadores con base en cinco resultados (Coagulación sanguínea, Estado circulatorio, Severidad de la pérdida de sangre, Nivel de dolor, Estado de comodidad: física) y 11 indicadores. Se utilizó la Prueba de Ecuación de Estimación Generalizada para comparar los puntajes de los indicadores. El proyecto fue aprobado por el comité ético institucional. Resultados: en las 65 evaluaciones, se identificó una diferencia estadísticamente significativa en la reducción de los puntajes de los resultados de enfermería Coagulación sanguínea, indicador "hematuria"; Estado circulatorio, en los indicadores "presión arterial sistólica y presión arterial diastólica" y en el Estado de comodidad: física, en el indicador de "bienestar físico". Conclusión: los pacientes evaluados no presentaron mayores complicaciones. Los indicadores clínicos apuntaban a cambios en el estado circulatorio, con reducción presión arterial, así como en la coagulación sanguínea verificada por hematuria, pero sin inestabilidad hemodinámica. El estado de comodidad se vio afectado por el tiempo de descanso posterior al procedimiento.

Resumo Objetivo: avaliar as complicações da biópsia renal percutânea com base nos resultados e indicadores clínicos da Nursing Outcomes Classification. Método: estudo longitudinal prospectivo. A amostra foi de 13 pacientes submetidos à biópsia renal percutânea, com 65 avaliações. Os pacientes foram avaliados em cinco momentos nas 24 horas após o procedimento, por meio de um instrumento desenvolvido pelos pesquisadores com base em cinco resultados (Coagulação sanguínea, Estado circulatório, Gravidade da perda de sangue, Nível de dor, Estado de conforto: físico) e 11 indicadores. Utilizou-se o Teste de Equações de Estimação Generalizadas para comparação entre os escores dos indicadores. O projeto foi aprovado pelo comitê de ética institucional. Resultados: nas 65 avaliações foi identificada diferença estatisticamente significativa na redução dos escores dos resultados de enfermagem Coagulação sanguínea, indicador "hematúria"; Estado circulatório, nos indicadores "pressão arterial sistólica e pressão arterial diastólica" e no Estado de conforto: físico, no indicador "bem-estar físico". Conclusão: os pacientes avaliados não apresentaram complicações maiores. Os indicadores clínicos apontaram alterações no estado circulatório, com redução da pressão arterial, bem como na coagulação sanguínea constatada pela hematúria, porém sem instabilidade hemodinâmica. O estado de conforto foi afetado pelo tempo de repouso após o procedimento.

Reflexões sobre a disciplina de história da psicologia no estado do Mato Grosso do Sul / Reflections on History of Psychology's course in Mato Grosso do Sul / Reflexiones acerca de la asignatura de Historia de la Psicología en Mato Grosso do Sul

Este artigo objetivou analisar a organização do ensino da disciplina de História da Psicologia, baseando-nos em cursos de graduação em Psicologia, no estado do Mato Grosso do Sul. Inicialmente, inventariaram-se as instituições que possuem cursos de Psicologia, acessando os seus sites e aplicando questionários aos seus coordenadores. Em seguida, organizaram-se algumas características gerais dos cursos e das instituições anuentes. Posteriormente, discutiu-se como o ensino da História da Psicologia auxilia na formação de estudantes, introduzindo-os ao estudo da Psicologia e às suas contendas científicas e profissionais. Ele contribui, com críticas, ao que se empreende no presente, embora apresente dificuldades de proporcionar algo introdutório mesclado a reflexões avançadas em uma disciplina. Por fim, observou-se que ele pode ser alocado e articulado com outras disciplinas, fazendo interface com os domínios da Ontologia, Epistemologia, Lógica e Ética.

This article aimed to analyze the History of Psychology subject teaching organization, based on the Psychology undergraduate courses in Mato Grosso do Sul. Initially, institutions with Psychology courses were inventoried; their websites were accessed and surveys were applied to their coordinators. Ten, some general characteristics of these courses and institutions were organized. Afterwards, it was discussed how the teaching of History of Psychology helps the students training, introducing them to Psychology and its scientific and professional strife. It also contributes with criticism on what have been done in the present, although it has some boundaries providing introductory issues mixed to advanced reflections on Psychology. Finally, it was observed that this teaching could be placed and articulated with other subjects, interfacing with Ontology, Epistemology, Logics and Ethics domains.

Este artículo tuvo como objetivo analizar la organización de la enseñanza de la asignatura de Historia de la Psicología, basándose en cursos de grado en Psicología, en el estado de Mato Grosso do Sul. Al principio, se inventariaron las instituciones que poseen cursos de Psicología, accediendo a sus sitios web y aplicando cuestionarios a sus coordinadores. A continuación, se organizaron algunas características generales de los cursos y de las instituciones anuentes. Después, se discutió cómo la enseñanza de la Historia de la Psicología auxilia en la formación de estudiantes, introduciéndolos al estudio de la Psicología y a sus contiendas profesionales y científicas; contribuye, con críticas, a lo que se emprende en el presente; presenta dificultades de proporcionar algo introductorio y múltiple, mezclado con reflexiones avanzadas en una disciplina; puede ser asignado y articulado con otras disciplinas; puede hacer interfaz con los dominios de la Ontología, Epistemología, Lógica y Ética.

The Effects of Enhanced Enforcement at Mexico's Southern Border: Evidence From Central American Deportees.

Immigration enforcement cooperation between final-destination and transit countries has increased in the last decades. I examine whether the Southern Border Plan, an immigration enforcement program implemented by the Mexican government in 2014, has curbed intentions of unauthorized migrants from El Salvador, Guatemala, and Honduras to migrate to the United States. I use the announcement of the Southern Border Plan to implement a difference-in-differences approach and compare the evolution of short-run intentions to engage in additional unauthorized crossings of Central American (treatment group) relative to Mexican deportees (comparison group). The findings suggest that increased enforcement in Mexico decreases the likelihood of attempting repeated unauthorized crossings.

Sleep loss disrupts pericyte-brain endothelial cell interactions impairing blood-brain barrier function.

Sleep loss in the rat increases blood-brain barrier permeability to circulating molecules by disrupting interendothelial tight junctions. Despite the description of the ultrastructure of cerebral microvessels and the evidence of an apparent pericyte detachment from capillary wall in sleep restricted rats the effect of sleep loss on pericytes is unknown. Here we characterized the interactions between pericytes and brain endothelial cells after sleep loss using male Wistar rats. Animals were sleep-restricted 20 h daily with 4 h sleep recovery for 10 days. At the end of the sleep restriction, brain microvessels (MVs) were isolated from cerebral cortex and hippocampus and processed for Western blot and immunocytochemistry to evaluate markers of pericyte-endothelial cell interaction (connexin 43, PDGFR-ß), tight junction proteins, and proinflammatory mediator proteins (MMP9, A2A adenosine receptor, CD73, NFκB). Sleep restriction reduced PDGFR-ß and connexin 43 expression in MVs; in addition, scanning electron microscopy micrographs showed that pericytes were detached from capillary walls, but did not undergo apoptosis (as depicted by a reduced active caspase-3 expression). Sleep restriction also decreased tight junction protein expression in MVs and increased BBB permeability to low- and high-molecular weight tracers in in vivo permeability assays. Those alterations seemed to depend on a low-grade inflammatory status as reflected by the increased expression of phosphorylated NFκB and A2A adenosine receptor in brain endothelial cells from the sleep-restricted rats. Our data show that pericyte-brain endothelial cell interaction is altered by sleep restriction; this evidence is essential to understand the role of sleep in regulating blood-brain barrier function.

Indoleamine 2,3-dioxygenase in melanoma progression and BRAF inhibitor resistance.

Indoleamine 2,3-dioxygenase (IDO) is associated with the progression of many types of tumors, including melanoma. However, there is limited information about IDO modulation on tumor cell itself and the effect of BRAF inhibitor (BRAFi) treatment and resistance. Herein, IDO expression was analyzed in different stages of melanoma development and progression linked to BRAFi resistance. IDO expression was increased in primary and metastatic melanomas from patients' biopsies, especially in the immune cells infiltrate. Using a bioinformatics approach, we also identified an increase in the IDO mRNA in the vertical growth and metastatic phases of melanoma. Using in silico analyses, we found that IDO mRNA was increased in BRAFi resistance. In an in vitro model, IDO expression and activity induced by interferon-gamma (IFNγ) in sensitive melanoma cells was decreased by BRAFi treatment. However, cells that became resistant to BRAFi presented random IDO expression levels. Also, we identified that treatment with the IDO inhibitor, 1-methyltryptophan (1-MT), was able to reduce clonogenicity for parental and BRAFi-resistant cells. In conclusion, our results support the hypothesis that the decreased IDO expression in tumor cells is one of the many additional outcomes contributing to the therapeutic effects of BRAFi. Still, the IDO production changeability by the BRAFi-resistant cells reiterates the complexity of the response arising from resistance, making it not possible, at this stage, to associate IDO expression in tumor cells with resistance. On the other hand, the maintenance of 1-MT off-target effect endorses its use as an adjuvant treatment of melanoma that has become BRAFi-resistant.

In vivo antitumoral effect of 4-nerolidylcatechol (4-NC) in NRAS-mutant human melanoma.

NRAS-mutations arise in 15-20% of all melanomas and are associated with aggressive disease and poor prognosis. Besides, the treatment for NRAS-mutant melanoma are not very efficient and is currently limited to immune checkpoints inhibitors or aggressive chemotherapy. 4-nerolidylcathecol (4-NC), a natural product extracted from Pothomorphe umbellata, induces apoptosis in melanoma cells by ROS production, DNA damage and increased p53 expression, in addition to inhibiting invasion in reconstructed skin. Moreover, 4-NC showed cytotoxicity in BRAF/MEKi-resistant and naive melanoma cells by Endoplasmic Reticulum (ER) stress induction in vitro. We evaluated the in vivo efficacy and the systemic toxicity of 4-NC in a NRAS-mutant melanoma model. 4-NC was able to significantly suppress tumor growth 4-fold compared to controls. Cleaved PARP and p53 expression were increased indicating cell death. As a proof of concept, MMP-2 and MMP-14 gene expression were decreased, demonstrating a possible role of 4-NC in melanoma invasion inhibition. Toxicological analysis indicated minor changes in the liver and bone marrow, but this toxicity was very mild when compared to other proteasome inhibitors and ER stress inductors already described. Our data indicate that 4-NC can counteract melanoma growth in vivo with minor adverse effects, suggesting further investigation as a potential NRAS-mutant melanoma treatment.

Metalloproteinases Suppression Driven by the Curcumin Analog DM-1 Modulates Invasion in BRAF-Resistant Melanomas.

BACKGROUND: Melanoma is the most aggressive skin cancer, and BRAF (V600E) is the most frequent mutation that led to the development of BRAF inhibitors (BRAFi). However, patients treated with BRAFi usually present recidivism after 6-9 months. Curcumin is a turmeric substance, and it has been deeply investigated due to its anti-inflammatory and antitumoral effects. Still, the low bioavailability and biodisponibility encouraged the investigation of different analogs. DM-1 is a curcumin analog and has shown an antitumoral impact in previous studies. METHODS: Evaluated DM-1 stability and cytotoxic effects for BRAFi-sensitive and resistant melanomas, as well as the role in the metalloproteinases modulation. RESULTS: DM-1 showed growth inhibitory potential for melanoma cells, demonstrated by reduction of colony formation, migration and endothelial tube formation, and cell cycle arrest. Subtoxic doses were able to downregulate important Metalloproteinases (MMPs) related to invasiveness, such as MMP-1, -2 and -9. Negative modulations of TIMP-2 and MMP-14 reduced MMP-2 and -9 activity; however, the reverse effect is seen when increased TIMP-2 and MMP-14 resulted in raised MMP-2. CONCLUSION: These findings provide essential details into the functional role of DM-1 in melanomas, encouraging further studies in the development of combinatorial treatments for melanomas.

Decitabine limits escape from MEK inhibition in uveal melanoma.

MEK inhibitors (MEKi) demonstrate anti-proliferative activity in patients with metastatic uveal melanoma, but responses are short-lived. In the present study, we evaluated the MEKi trametinib alone and in combination with drugs targeting epigenetic regulators, including DOT1L, EZH2, LSD1, DNA methyltransferases, and histone acetyltransferases. The DNA methyltransferase inhibitor (DNMTi) decitabine effectively enhanced the anti-proliferative activity of trametinib in cell viability, colony formation, and 3D organoid assays. RNA-Seq analysis showed the MEKi-DNMTi combination primarily affected the expression of genes involved in G1 and G2/2M checkpoints, cell survival, chromosome segregation and mitotic spindle. The DNMTi-MEKi combination did not appear to induce a DNA damage response (as measured by γH2AX foci) or senescence (as measured by ß-galactosidase staining) compared to either MEKi or DNMTi alone. Instead, the combination increased expression of the CDK inhibitor p21 and the pro-apoptotic protein BIM. In vivo, the DNMTi-MEKi combination was more effective at suppressing growth of MP41 uveal melanoma xenografts than either drug alone. Our studies indicate that DNMTi may enhance the activity of MEKi in uveal melanoma.