INFLUENCE OF ß2-ADRENERGIC SELECTIVE AGONIST FORMOTEROL ON THE MOTOR UNIT OF A MOUSE MODEL OF A CONGENITAL MYASTHENIC SYNDROME WITH COMPLETE VAChT DELETION.

Congenital Myasthenic Syndromes (CMS) are a set of genetic diseases that affect the neuromuscular transmission causing muscular weakness. The standard pharmacological treatment aims at ameliorating the myasthenic symptom by acetylcholinesterase inhibitors. Most patients respond well in the short and medium term, however, over time the beneficial effects rapidly fade, and the efficacy of the treatment diminishes. Increasing evidence shows that ß2-adrenergic agonists can be a suitable choice for the treatment of neuromuscular disorders, including CMS, as they promote beneficial effects in the neuromuscular system. The exact mechanism on which they rely is not completely understood, although patients and animal models respond well to the treatment, especially over extended periods. Here, we report the use of the long-lasting specific ß2-adrenergic agonist formoterol in a myasthenic mouse model (mnVAChT-KD), featuring deletion of VAChT (Vesicular Acetylcholine Transporter) specifically in the α-motoneurons. Our findings demonstrate that formoterol treatment (300 µg/kg/day; sc) for 30 days increased the neuromuscular junction area, induced skeletal muscle hypertrophy and altered fibre type composition in myasthenic mice. Interestingly, ß2-adrenergic agonists have shown efficacy even in the absence of ACh (acetylcholine). Our data provide important evidence supporting the potential of ß2-adrenergic agonists in treating neuromuscular disorders of pre-synaptic origin and characterized by disruptions in nerve-muscle communication, through a direct and beneficial action within the motor unit.

High-Caloric Diets in Adolescence Impair Specific GABAergic Subpopulations, Neurogenesis, and Alter Astrocyte Morphology.

We compared the effects of two different high-caloric diets administered to 4-week-old rats for 12 weeks: a diet rich in sugar (30% sucrose) and a cafeteria diet rich in sugar and high-fat foods. We focused on the hippocampus, particularly on the gamma-aminobutyric acid (GABA)ergic system, including the Ca2+-binding proteins parvalbumin (PV), calretinin (CR), calbindin (CB), and the neuropeptides somatostatin (SST) and neuropeptide Y (NPY). We also analyzed the density of cholinergic varicosities, brain-derived neurotrophic factor (BDNF), reelin (RELN), and cyclin-dependent kinase-5 (CDK-5) mRNA levels, and glial fibrillary acidic protein (GFAP) expression. The cafeteria diet reduced PV-positive neurons in the granular layer, hilus, and CA1, as well as NPY-positive neurons in the hilus, without altering other GABAergic populations or overall GABA levels. The high-sugar diet induced a decrease in the number of PV-positive cells in CA3 and an increase in CB-positive cells in the hilus and CA1. No alterations were observed in the cholinergic varicosities. The cafeteria diet also reduced the relative mRNA expression of RELN without significant changes in BDNF and CDK5 levels. The cafeteria diet increased the number but reduced the length of the astrocyte processes. These data highlight the significance of determining the mechanisms mediating the observed effects of these diets and imply that the cognitive impairments previously found might be related to both the neuroinflammation process and the reduction in PV, NPY, and RELN expression in the hippocampal formation.

Effects of Aging and Nerve Growth Factor on Neuropeptide Expression and Cholinergic Innervation of the Rat Basolateral Amygdala.

The basolateral amygdala (BLA) contains interneurons that express neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP), both of which are involved in the regulation of functions and behaviors that undergo deterioration with aging. There is considerable evidence that, in some brain areas, the expression of NPY and VIP might be modulated by acetylcholine. Importantly, the BLA is one of the brain regions that has one of the densest cholinergic innervations, which arise mainly from the basal forebrain cholinergic neurons. These cholinergic neurons depend on nerve growth factor (NGF) for their survival, connectivity, and function. Thus, in this study, we sought to determine if aging alters the densities of NPY- and VIP-positive neurons and cholinergic varicosities in the BLA and, in the affirmative, if those changes might rely on insufficient trophic support provided by NGF. The number of NPY-positive neurons was significantly reduced in aged rats, whereas the number of VIP-immunoreactive neurons was unaltered. The decreased NPY expression was fully reversed by the infusion of NGF in the lateral ventricle. The density of cholinergic varicosities was similar in adult and old rats. On the other hand, the density of cholinergic varicosities is significantly higher in old rats treated with NGF than in adult and old rats. Our results indicate a dissimilar resistance of different populations of BLA interneurons to aging. Furthermore, the present data also show that the BLA cholinergic innervation is particularly resistant to aging effects. Finally, our results also show that the reduced NPY expression in the BLA of aged rats can be related to changes in the NGF neurotrophic support.

Irony Understanding in Children With Attention-Deficit/Hyperactivity Disorder: A Comparative Analysis.

INTRODUCTION: Attention-deficit/hyperactivity disorder (ADHD) is a neurodevelopmental disorder characterized by persistent patterns of inattention, hyperactivity, and impulsivity that can significantly impact a child's daily life and academic performance. Some studies have noted challenges in social cognition among children with ADHD, specifically in aspects like emotion perception and processing, empathy, Theory of Mind, and pragmatics. One of the lesser-explored aspects of ADHD is its potential impact on higher-level language skills, such as irony understanding. Our goal in this study was to compare irony comprehension in two groups: schoolchildren with normal development and schoolchildren with ADHD. METHODS: We performed a comparative cross-sectional study on a total of 35 children aged between 6 and 12 years: 17 with ADHD and 18 not neurodevelopmentally impaired. Irony comprehension was evaluated using an assessment method validated for the Portuguese population: Turtle on the Island - Battery of Assessment of Executive Functions in Children (TI-BAFEC). We also applied the EACE (Emotional Awareness and Coping in Children) scale. It is the Portuguese adaptation of the Assessment of Children's Emotion Skills and assesses children's emotional knowledge. RESULTS: We obtained two comparable groups concerning sociodemographic data. The analysis of the data, using the Mann-Whitney U Test, showed that in all parameters, the control group consistently exhibited superior performance compared to the ADHD group. However, only TI-BAFEC (p = 0.005) and emotional behavior (p = 0.007) showed a significant statistical variance between the two groups. CONCLUSION: This article shows that children with ADHD encounter challenges in grasping irony. However, the underlying cognitive processes contributing to these difficulties warrant further investigation.

Assessing the impact of post-mortem damage and contamination on imputation performance in ancient DNA.

Low-coverage imputation is becoming ever more present in ancient DNA (aDNA) studies. Imputation pipelines commonly used for present-day genomes have been shown to yield accurate results when applied to ancient genomes. However, post-mortem damage (PMD), in the form of C-to-T substitutions at the reads termini, and contamination with DNA from closely related species can potentially affect imputation performance in aDNA. In this study, we evaluated imputation performance (i) when using a genotype caller designed for aDNA, ATLAS, compared to bcftools, and (ii) when contamination is present. We evaluated imputation performance with principal component analyses and by calculating imputation error rates. With a particular focus on differently imputed sites, we found that using ATLAS prior to imputation substantially improved imputed genotypes for a very damaged ancient genome (42% PMD). Trimming the ends of the sequencing reads led to similar improvements in imputation accuracy. For the remaining genomes, ATLAS brought limited gains. Finally, to examine the effect of contamination on imputation, we added various amounts of reads from two present-day genomes to a previously downsampled high-coverage ancient genome. We observed that imputation accuracy drastically decreased for contamination rates above 5%. In conclusion, we recommend (i) accounting for PMD by either trimming sequencing reads or using a genotype caller such as ATLAS before imputing highly damaged genomes and (ii) only imputing genomes containing up to 5% of contamination.

100 ancient genomes show repeated population turnovers in Neolithic Denmark.

Major migration events in Holocene Eurasia have been characterized genetically at broad regional scales1-4. However, insights into the population dynamics in the contact zones are hampered by a lack of ancient genomic data sampled at high spatiotemporal resolution5-7. Here, to address this, we analysed shotgun-sequenced genomes from 100 skeletons spanning 7,300 years of the Mesolithic period, Neolithic period and Early Bronze Age in Denmark and integrated these with proxies for diet (13C and 15N content), mobility (87Sr/86Sr ratio) and vegetation cover (pollen). We observe that Danish Mesolithic individuals of the Maglemose, Kongemose and Ertebølle cultures form a distinct genetic cluster related to other Western European hunter-gatherers. Despite shifts in material culture they displayed genetic homogeneity from around 10,500 to 5,900 calibrated years before present, when Neolithic farmers with Anatolian-derived ancestry arrived. Although the Neolithic transition was delayed by more than a millennium relative to Central Europe, it was very abrupt and resulted in a population turnover with limited genetic contribution from local hunter-gatherers. The succeeding Neolithic population, associated with the Funnel Beaker culture, persisted for only about 1,000 years before immigrants with eastern Steppe-derived ancestry arrived. This second and equally rapid population replacement gave rise to the Single Grave culture with an ancestry profile more similar to present-day Danes. In our multiproxy dataset, these major demographic events are manifested as parallel shifts in genotype, phenotype, diet and land use.

Population genomics of post-glacial western Eurasia.

Western Eurasia witnessed several large-scale human migrations during the Holocene1-5. Here, to investigate the cross-continental effects of these migrations, we shotgun-sequenced 317 genomes-mainly from the Mesolithic and Neolithic periods-from across northern and western Eurasia. These were imputed alongside published data to obtain diploid genotypes from more than 1,600 ancient humans. Our analyses revealed a 'great divide' genomic boundary extending from the Black Sea to the Baltic. Mesolithic hunter-gatherers were highly genetically differentiated east and west of this zone, and the effect of the neolithization was equally disparate. Large-scale ancestry shifts occurred in the west as farming was introduced, including near-total replacement of hunter-gatherers in many areas, whereas no substantial ancestry shifts happened east of the zone during the same period. Similarly, relatedness decreased in the west from the Neolithic transition onwards, whereas, east of the Urals, relatedness remained high until around 4,000 BP, consistent with the persistence of localized groups of hunter-gatherers. The boundary dissolved when Yamnaya-related ancestry spread across western Eurasia around 5,000 BP, resulting in a second major turnover that reached most parts of Europe within a 1,000-year span. The genetic origin and fate of the Yamnaya have remained elusive, but we show that hunter-gatherers from the Middle Don region contributed ancestry to them. Yamnaya groups later admixed with individuals associated with the Globular Amphora culture before expanding into Europe. Similar turnovers occurred in western Siberia, where we report new genomic data from a 'Neolithic steppe' cline spanning the Siberian forest steppe to Lake Baikal. These prehistoric migrations had profound and lasting effects on the genetic diversity of Eurasian populations.

Luxación intrusiva en dentición primaria: reporte de caso / Luxação intrusiva na dentição decídua: relato de caso clínico / Intrusive luxation in primary dentition: clinical report

La luxación intrusiva es el tipo más común de lesión traumática en la dentición primaria, caracterizada por el desplazamiento del diente hacia el interior del alvéolo. Requieren minucioso examen clínico, buena radiografía y seguimiento del caso, siendo el prognóstico desfavorable, principalmente por la probabilidad de daño al germen del diente permanente. Este trabajo tiene como objetivo reportar un caso clínico de luxación intrusiva en dentición primaria, desde el diagnóstico hasta el seguimiento. Se trata de un paciente masculino, de 3 años, que acudió a la Emergencia de la Clínica de Especialidades Infantiles ­ Universidad Estatal de Londrina (UEL) por traumatismo dentoalveolar en la región antero-superior ocurrido hace tres días. En el examen físico, se observó que el diente 61 presentaba intrusión de grado II con un tercio de la corona visible y el diente 62 intrusión de grado I, con más de un tercio de la corona visible. Radiográficamente se observa inclinación vestibular, sin fractura de la placa ósea, por lo que se optó por un tratamiento conservador, a la espera de la reerupción espontánea de los dientes afectados. Se informó a los padres sobre posibles consecuencias de este tipo de traumatismo y la necesidad de consultas de seguimiento clínico y radiológico. Después de ocho meses de seguimiento, los dientes erupcionaron satisfactoriamente. Es de suma importancia establecer un protocolo de observación y control del diente en el cual se ha identificado el trauma, observando signos y síntomas de los tejidos involucrados en el trauma para reducir secuelas en ambas denticiones

A luxação intrusiva é o tipo de lesão traumática mais frequente na dentição decídua, caracterizada pelo deslocamento do dente para o alvéolo. As intrusões requerem exame físico minucioso, radiografias de valor diagnóstico e acompanhamento do caso, pois o prognóstico é desfavorável, principalmente pela probabilidade de danos ao germe dentário permanente. Este trabalho tem como objetivo relatar um caso clínico de luxação intrusiva na dentição decídua, desde o diagnóstico até o acompanhamento. Trata-se de paciente do sexo masculino, 3 anos de idade, atendido no Pronto Socorro do Ambulatório de Especialidades Infantis da Universidade Estadual de Londrina (UEL) devido a trauma dento-alveolar em região ântero-superior ocorrido há três dias. Ao exame físico intraoral, observou-se que o dente 61 apresentava intrusão grau II (um terço da coroa visível) e o dente 62 apresentava intrusão grau I, com mais de um terço da coroa visível. Radiograficamente foi observada inclinação vestibular, sem fratura da tábua óssea. Assim, optou-se pelo tratamento conservador, aguardando a reerupção espontânea dos dentes envolvidos. Além disso, os pais foram informados sobre as possíveis consequências deste tipo de trauma e a necessidade de consultas de proservação clínica e radiográfica. Após oito meses de acompanhamento, os dentes erupcionaram satisfatoriamente. É de extrema importância estabelecer um protocolo de observação e controle do dente traumatizado, observando sinais e sintomas dos tecidos envolvidos no trauma para reduzir sequelas em ambas as dentições.

Intrusive dislocation is the most frequent type of traumatic injury in the primary dentition, characterized by displacement of the tooth into the alveolus. Intrusions require a thorough physical examination, radiographs of diagnostic value, and follow-up of the case, since the prognosis is unfavorable, mainly due to the probability of damage to the permanent tooth germ. This work aims to report a clinical case of intrusive dislocation in the primary dentition, from diagnosis to follow-up. This is a male patient, 3 years old, who attended in the Emergency of the Children's Specialty Clinic ­ State University of Londrina (UEL) due to dento-alveolar trauma in the antero-superior region that occurred three days ago. On clinical examination, tooth 61 had grade II intrusion (one-third of the crown visible), and tooth 62 with grade I intrusion, with more than one-third of the crown visible. Radiographically, a buccal inclination was observed, with no fracture of the bone plate. Thus, we opted for conservative treatment, waiting for the spontaneous re-eruption of the teeth involved. Furthermore, parents were informed about the possible consequences of this type of trauma and the need for clinical and radiographic follow-up consultations. After eight months of follow-up, the teeth erupted satisfactorily. It is extremely important to establish a protocol for observation and control of the traumatized tooth, observing signs and symptoms of the tissues involved in the trauma to reduce sequelae in both dentitions

Retrospective analysis of surgical and oncological results of laparoscopic surgeries performed by residents of coloproctology / Análise retrospectiva dos resultados cirúrgicos e oncológicos em cirurgias videolaparoscópicas realizadas por residentes de coloproctologia

ABSTRACT Introduction: with the improvement and wide acceptance of laparoscopy in colorectal operations, there was a need for specific training of surgeons in training. There are few studies evaluating the postoperative results of laparoscopic colectomies performed by resident physicians and their impact on patient safety. Purpose: to analyze the surgical and oncological results of laparoscopic colectomies performed by coloproctology residents and compare them with data in the literature. Methods: this is a retrospective analysis of patients undergoing laparoscopic colorectal surgery performed by resident physicians at the Hospital das Clínicas de Ribeirão Preto, between 2014 and 2018. The clinical characteristics of the patients were studied, as well as the main surgical and oncological aspects in a period of one year. Results: we analyzed 191 operations, whose main surgical indication was adenocarcinoma, most of them stage III. The mean duration of surgeries was 210±58 minutes. There was a need for a stoma in 21.5% of the patients, mainly loop colostomy. The conversion rate was 23%, with 79.5% due to technical difficulties, and the main predictors of conversion were obesity and intraoperative accidents. The median length of stay was 6 days. Preoperative anemia was associated with a higher rate of complications (11.5%) and reoperations (12%). Surgical resection margins were compromised in 8.6% of cases. The one-year recurrence rate was 3.2% and the mortality rate was 6.3%. Conclusions: videolaparoscopic colorectal surgery performed by residents showed efficacy and safety similar to data found in the literature.

RESUMO Introdução: com o aperfeiçoamento e a ampla aceitação da laparoscopia nas operações colorretais, houve necessidade de treinamento específico dos cirurgiões em formação. Existem poucos estudos avaliando os resultados pós operatórios das colectomias videolaparoscópicas realizadas por médicos residentes e seu impacto na segurança do paciente. Objetivo: analisar os resultados cirúrgicos e oncológicos das colectomias videolaparoscópicas realizadas por residentes de coloproctologia e comparar com dados da literatura. Métodos: trata-se de uma análise retrospectiva de pacientes submetidos a cirurgias colorretais laparoscópicas, realizadas por médicos residentes do Hospital das Clínicas de Ribeirão Preto, entre 2014 e 2018. Foram estudadas as características clínicas dos pacientes bem como os principais aspectos cirúrgicos e oncológicos em um período de um ano. Resultados: analisou-se 191 operações, cuja principal indicação cirúrgica foi adenocarcinoma, a maioria estadio III. A duração média das cirurgias foi 210±58 minutos. Houve necessidade de estoma em 21,5% dos pacientes, principalmente colostomia em alça. A taxa de conversão foi 23%, sendo 79,5% por dificuldades técnicas, e os principais fatores preditores de conversão foram obesidade e acidentes intra-operatórios. A mediana do tempo de internação foi 6 dias. Anemia pré operatória associou-se a uma maior taxa de complicações (11,5%) e reoperações (12%). Houve comprometimento das margens de ressecção cirúrgica em 8,6% dos casos. A taxa de recidiva em um ano foi de 3,2%, e a taxa de mortalidade, 6,3%. Conclusões: a cirurgia colorretal videolaparoscópica realizada por residentes apresentou eficácia e segurança semelhante aos dados encontrados na literatura.

Atividade antimicrobiana in vitro das folhas de araçá (Psidium cattleianum Sabine) contra micro-organismos da mucosa oral / In vitro antimicrobial activity of leaves of araçá (Psidium cattleianum Sabine) against oral micro-organisms

Introdução: A espécie Psidium cattleianum Sabine tem despertado o interesse dos pesquisadores por apresentar, além de atividade cicatrizante, analgésica e antioxidante, propriedades antimicrobianas frente a micro-organismos da mucosa oral que podem atuar como agentes cariogênicos. Objetivo: Foi avaliada, neste trabalho, a atividade antimicrobiana de extratos bruto e fracionados das folhas do araçá, além do seu perfil cromatográfico. Material e método: Para avaliação da atividade antimicrobiana, foi utilizada a técnica de microdiluição, para determinação da Concentração Inibitória Mínima (CIM), e repique do inóculo em Ágar Muller Hinton (Himedia), para averiguação da Concentração Bactericida Mínima (CBM). O extrato foi testado nas concentrações entre 10 e 500 µg/mL. O perfil cromatográfico foi realizado pelo método de Cromatografia Líquida de Alta Eficiência (CLAE). Resultado: Os resultados obtidos foram submetidos a uma análise descritiva e foi possível observar a atividade inibitória dos extratos do P. cattleianum contra S. mutans e S. oralis. Conclusão: A atividade antimicrobiana dos extratos de P. cattleianum contra micro-organismos orais justifica maiores estudos para a utilização medicinal dessa espécie, como, por exemplo, sua utilização em enxaguantes bucais.

Introduction: The specie Psidium cattleianum Sabine has aroused the interest of researchers to exhibit, healing activity, analgesic, antioxidant, antimicrobial properties against the micro-organisms of the oral mucosa that can act as cariogenic agents. Objective: It was analyzed in this study the antimicrobial activity of crude extracts and fractions and chromatographic profile of araçá. Material and method: For antimicrobial activity, we used the technique of on broth microdilution for determining the Minimum Inhibitory Concentration (MIC) and replating of inoculum in Muller Hinton (Himedia) agar for finding the Minimum Bactericidal Concentration (MBC). The extract was tested at concentrations between 10 and 500 µg/mL. The chromatographic profile was performed by the method of High Performance Liquid Chromatography. Result: The results were submitted to a descriptive analysis was possible to observe the inhibitory activity of the extracts of P. cattleianum against S. oralis and S. mutans. Conclusion: The antimicrobial activity of P. cattleianum extracts against oral microorganisms justifies the use of this species as a medicinal plant.