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Michelle Miranda Pereira, Debora Maria Befi-Lopes, Aparecido José Couto Soares, Fernanda Chiarion Sassi, Claudia Regina Furquim de Andrade School of Medicine, University of São Paulo, São Paulo, BrazilCorrespondence: Aparecido José Couto SoaresSchool of Medicine, University of São Paulo, Cipotânea Street, 51 – Cidade Universitária – São Paulo, BrazilTel +55 11 3091-7455Email ajcsoares@usp.brBackground: Acute lymphoid leukemia (ALL) is the most prevalent cancer of childhood. Impairment in linguistic and memory skills is a possible late sequela in cancer survivors that can limit their quality of life and the overall performance of the individual in society. There is evidence that survivors of ALL treated exclusively with chemotherapy demonstrate significant differences in long-term linguistic and memory functions and also changes in neuroanatomical integrity. However, most studies described do not count on a speech-language pathologist in their team, which we consider important to discuss. Thus, the objective of the present study was to assess memory and vocabulary skills in the pediatric population diagnosed with acute lymphoid leukemia during chemotherapy treatment.Materials and Methods: An observational cohort study was conducted over a 1.8-year period. Participants of this research were children diagnosed with ALL. All participants were assessed on their linguistic-cognitive skills (ie, vocabulary, short-term memory and lexical access). All data underwent statistical analyses.Results: The results of the current study found no major significant difference in the linguistic-cognitive performance of children with ALL and their healthy controls. Regarding the linguistic variables, we believe that there should be a differentiation between the effects of the drugs and the effects of social communication skills on performance.Conclusion: This first characterization of the linguistic-cognitive abilities of children with ALL did not identify differences between these children and their healthy peers, although we were able to identify variables regarding the multidisciplinary team and social communication that should be considered in future studies.Keywords: interdisciplinary research, leukemia, children, language

Purpose: To continue the validation process of the Decoding Development Monitoring Protocol (PRADE) in software format in the validity evidence stage based on response processes., Methods: 250 individuals participated in this study, 125 individuals from private schools and 125 individuals from public schools. The assessment was carried out in person using the software that hosts the instrument's tasks, which are organized into decoding linguistically balanced words and non-words, respecting the decoding rules of Brazilian Portuguese. The software prepares an individual performance report for each participant, counting the decoding time for each stimulus, as well as the number of words decoded correctly. The data is organized considering the correct decoding time of the stimuli, decoding accuracy and percentage of correct answers. All data underwent statistical analysis using SPSS software., Results: The data indicated an important effect of the length of words and non-words on public and private school students. Furthermore, it was possible to observe the evolution of decoding, depending on the school year, in all the variables studied. In both groups, a strong influence of non-words on student performance throughout Elementary School I was observed., Conclusion: The data indicate validity in the analysis of response processes, since it was possible to adequately characterize the performance of school children public and private throughout Elementary School I, characterizing each group, as well as their differences according to the advancement of schooling.

Cytogenetics has allowed the investigation of chromosomal diversity and repetitive genomic content in wasps. In this study, we characterized the karyotype of the social wasp Mischocyttarus cassununga using conventional cytogenetics and chromosomal mapping of repetitive sequences. This study was undertaken to extend our understanding of the genomic organization of repetitive DNA in social wasps and is the first molecular cytogenetic insight into the genus Mischocyttarus . The karyotype of M. cassununga had a chromosome number of 2 n  = 64 for females and n  = 32 for males. Constitutive heterochromatin exhibited three distribution patterns: centromeric and pericentromeric regions along the smaller arms and extending almost the entire chromosome. The major ribosomal DNA sites were located on chromosome pair in females and one chromosome in males. Positive signals for the microsatellite probes (GA) n and (GAG) n were observed in the euchromatic regions of all chromosomes. The microsatellites, (CGG) n , (TAT) n , (TTAGG) n , and (TCAGG) n were not observed in any region of the chromosomes. Our results contrast with those previously obtained for Polybia fastidiosuscula , which showed that the microsatellites (GAG) n , (CGG) n , (TAT) n , (TTAGG) n , and (TCAGG) n are located predominantly in constitutive heterochromatin. This suggests variations in the diversity and chromosomal organization of repetitive sequences in the genomes of social wasps., Competing Interests: The authors have no conflicts of interest to declare.

Background: The aggregation and spread of misfolded amyloid structured proteins, such as tau and α-synuclein, are key pathological features associated with neurodegenerative disorders, including Alzheimer's and Parkinson's disease. These proteins possess a prion-like property, enabling their transmission from cell to cell leading to propagation throughout the central and peripheral nervous systems. While the mechanisms underlying their intracellular spread are still being elucidated, targeting the extracellular space has emerged as a potential therapeutic approach. The glymphatic system, a brain-wide pathway responsible for clearing extracellular metabolic waste from the central nervous system, has gained attention as a promising target for removing these toxic proteins., Methods: In this study, we investigated the impact of long-term modulation of glymphatic function on tau aggregation and spread by chronically treating a mouse model of tau propagation with a pharmacological inhibitor of AQP4, TGN-020. Thy1-hTau.P301S mice were intracerebrally inoculated with tau into the hippocampus and overlying cortex, and subsequently treated with TGN-020 (3 doses/week, 50 mg/kg TGN-020, i.p.) for 10-weeks. During this time, animal memory was studied using cognitive behavioural tasks, and structural MR images were acquired of the brain in vivo prior to brain extraction for immunohistochemical characterisation., Results: Our findings demonstrate increased tau aggregation in the brain and transhemispheric propagation in the hippocampus following the inhibition of glymphatic clearance. Moreover, disruption of the glymphatic system aggravated recognition memory in tau inoculated mice and exacerbated regional changes in brain volume detected in the model. When initiation of drug treatment was delayed for several weeks post-inoculation, the alterations were attenuated., Conclusions: These results indicate that by modulating AQP4 function and, consequently, glymphatic clearance, it is possible to modify the propagation and pathological impact of tau in the brain, particularly during the initial stages of the disease. These findings highlight the critical role of the glymphatic system in preserving healthy brain homeostasis and offer valuable insights into the therapeutic implications of targeting this system for managing neurodegenerative diseases characterized by protein aggregation and spread., (© 2024. The Author(s).)

Purpose: to verify the performance of children with Developmental Language Disorder in decoding and writing tests in order to better understand their manifestations and the process of acquiring written language skills., Methods: The study subjects were 80 children. The Research Group consisted of 16 children diagnosed with Developmental Language Disorder, 13 males and 3 females, mean age of 7.3. The Control Group counted on 64 subjects paired in gender, age, education and socioeconomic level with the Control Group in a 4:1 ratio. The ability to decode words and pseudowords of both groups was evaluated, measuring the time spent to correctly read words and the percentage of correct answers, also considering the length of the word/pseudoword. The writing evaluation was carried out in the control group, which had its spelling errors analyzed and categorized. All data underwent descriptive and inferential statistical analysis., Results: The data indicated a longer decoding time and a lower percentage of correct answers for the children from the Research Group. Regarding spelling errors, there was a predominance of arbitrary spelling errors., Conclusion: The data showed that children with Developmental Language Disorder tend to have a longer decoding time, greater percentage of errors than their peers and tend to present spelling errors more concentrated in natural orthography.

Objective: To describe the clinical and feeding findings of premature infants with gastroschisis (GTQ) in a neonatal intensive care unit and compare them to preterm infants (NBs) without congenital anomalies., Methods: A retrospective case-control study was conducted with 50 premature NBs (25 with GTQ and 25 without comorbidities - control group). The NBs were compared regarding demographic and clinical data: risk of mortality and speech-language assessment (nonnutritive and nutritive sucking). Subsequently, a multiple logistic regression model was applied to determine the variables associated with the negative speech therapy outcome (speech therapy discharge after more than 7 days considering the first speech therapy evaluation)., Results: The results of the first analysis indicated that there was a difference between the GTQ and the CG for the following variables: total time in days of hospitalization; use of mechanical ventilation (invasive x noninvasive); days of life on the date of the first speech-language assessment; use of alternative feeding route; and the GTQ group had worse results. The results of the multiple logistic regression indicated that the diagnosis of GTQ, the use of invasive mechanical ventilation, and the absence of adequate intraoral pressure during the first speech-language evaluation were risk factors for a negative speech-language outcome., Conclusion: The feeding skills of preterm infants with QTG are related to the severity of the condition (gastrointestinal tract complications), requiring longer hospitalization, use of invasive mechanical ventilation, prolonged use of alternative feeding route and requiring more speech therapy to start oral feeding.

Introduction: Computed tomography angiography (CTA) and ventilation/perfusion (V/Q) single photon emission computed tomography/CT (SPECT/CT) images have been widely used to detect PE, but few studies have performed a direct comparison between them. We aimed to evaluate the performance of these tests in the same group of patients, selected from the routine practice of a general hospital., Methods: Patients with suspected acute PE were prospectively submitted to CTA and V/Q SPECT/CT. General radiologists and nuclear physicians, respectively, interpreted the images. Data regarding age, sex, time between examinations, symptoms, and Wells score were also recorded. The final diagnosis was decided through a consensus among the clinicians, taking into account clinical, laboratory, follow-up, and all imaging procedures data., Results: Twenty-eight patients (15 male, 13 female, and median age of 51.5 years) were studied. Median duration of the onset of symptoms was 4 (1-14) days, and the median Wells score was 3.5 (1.5-6). Sensitivity, specificity, positive and negative predictive values, and accuracy were 84.6%, 80.0%, 78.6%, 85.7%, and 82.1% for V/Q SPECT/CT, and 46.1%, 100%, 100%, 68.2%, and 75.0% for CTA. The overall agreement between the methods was 57.1%. Of the 22 patients with negative CTA, 10 (45.4%) had positives V/Q SPECT/CT and seven of them classified as true positives., Conclusions: Our results suggest that V/Q SPECT/CT is more sensitive and accurate than CTA when interpreted by general radiologists and nuclear medicine physicians.

Among Meliponini species, c -heterochromatin can occupy large portions of chromosomes. This characteristic could be useful for understanding evolutionary patterns of satellite DNAs (satDNAs), although few sequences have been characterized in these bees. In Trigona , phylogenetically represented by clades A and B, the c -heterochromatin is mostly located in one chromosome arm. Here we used different techniques, including restriction endonucleases and genome sequencing followed by chromosomal analysis, to identify satDNAs that may be contributing to the evolution of c -heterochromatin in Trigona . Our results revealed a highly abundant ThyaSat01-301 satDNA, corresponding to about 13.77% of the Trigona hyalinata genome. Another seven satDNAs were identified, one corresponding to 2.24%, and the other six corresponding to 0.545% of the genome. The satDNA ThyaSat01-301 was shown to be one of the main constituents of the c -heterochromatin of this species, as well as of other species belonging to clade B of Trigona . However, this satDNA was not observed on the chromosomes of species from clade A, demonstrating that the c -heterochromatin is evolving divergently between species of clade A and B, as a consequence of the evolution of repetitive DNA sequences. Finally, our data suggest the molecular diversification of the karyotypes, despite a conservated macrochromosomal structure on the genus.

Decoding skills are crucial for literacy development and they tend to be acquired early in transparent languages, such as Brazilian Portuguese. It is essential to better understand which variables may affect the decoding process. In this study, we investigated the processes of decoding as a function of age of children who are exposed to a transparent language. To this end, we examined the effects of grade, stimulus type and stimulus extension on the decoding accuracy of children between the ages of six and 10 years who are monolingual speakers of Brazilian Portuguese. The study included 250 children, enrolled from the first to the fifth grade. A list of words and pseudowords of variable length was created, based on Brazilian Portuguese structure. Children assessment was conducted using the computer program E-prime ® which was used to present the stimuli. The stimuli were programmed to appear on the screen in a random order and children were instructed to read them. The results indicate two important moments for decoding: the acquisition and the mastery of decoding skills. Additionally, the results highlight an important effect of the extent and type of stimuli and how it interacts with the school progress. Moreover, data indicate the multifactorial nature of decoding acquisition and the different interactions between variables that can influence this process. We discuss medium- and long-term implications of it, and possible individual and collective actions which can improve this process.

Objectives: The aims of this study were to evaluate and compare the effects that dexmedetomidine and methadone, either alone or in combination, have on the ocular variables of healthy adult cats when administered intramuscularly, as well as their reversal with atipamezole., Methods: A randomized crossover blinded study of 10 healthy cats was used to assess the effect of 0.2 mg/kg methadone (MET), 7.5 μg/kg dexmedetomidine (D7), 10 μg/kg dexmedetomidine (D10), 7.5 μg/kg dexmedetomidine and 0.2 mg/kg methadone (DM7) and 10 μg/kg dexmedetomidine and 0.2 mg/kg methadone (DM10) on intraocular pressure (IOP), tear production and pupil diameter (PD). The animals were evaluated for 30 mins. Afterwards, atipamezole was administered and ocular variables were evaluated for 30 mins., Results: D10, DM7 and DM10 significantly decreased mean IOP but MET or D7 did not. Tear production decreased significantly in all treatments, corresponding to 18%, 59%, 63%, 86% and 98% in MET, D7, D10, DM7 and DM10, respectively. PD increased in all treatments, but MET showed the highest PD. Thirty minutes after atipamezole (RT30), IOP returned to baseline with no difference between groups, and there was a significant increase in tear production, but the means were still different from baseline., Conclusions and Relevance: Dexmedetomidine decreases IOP and tear production but increases PD in healthy cats. Atipamezole can partly reverse those alterations. Low-dose dexmedetomidine (7.5 µg/kg) promotes sedation without changing the IOP. All protocols significantly decrease tear production, and Schirmer tear test after sedation is not representative of non-sedated values. Methadone induces quick onset mydriasis without changing the IOP.

Leaf-cutting ants are among the New World's most conspicuous and studied ant species due to their notable ecological and economic roles. Cytogenetic studies carried out in Atta show remarkable karyotype conservation among the species. We performed classical cytogenetics and physical mapping of repetitive sequences in the leaf-cutting ant Atta cephalotes (Linnaeus, 1758), the type species of the genus. Our goal was to test the karyotype conservation in Atta and to understand the genomic organization and diversity regarding repetitive sequences in leaf-cutting ants. Atta cephalotes showed 2 n  = 22 (18m + 2sm + 2st) chromosomes. The heterochromatin followed a centromeric pattern, and the GC-rich regions and 18S rDNA clusters were co-located interstitially in the 4th metacentric pair. These cytogenetic characteristics were observed in other Atta species that had previously been studied, confirming the karyotype conservation in Atta . Evolutionary implications regarding the conservation of the chromosome number in leaf-cutting ants are discussed. Telomeric motif (TTAGG) n was detected in A. cephalotes as observed in other ants. Five out of the 11 microsatellites showed a scattered distribution exclusively on euchromatic areas of the chromosomes. Repetitive sequences mapped on the chromosomes of A. cephalotes are the first insights into genomic organization and diversity in leaf-cutting ants, useful in further comparative studies.

A substantial portion of the eukaryotic genome includes repetitive DNA, which is important for its stability, regulation, and architecture. Fungus-farming ant genomes show remarkable structural rearrangement rates that were necessary for the establishment of their agriculture-based lifestyle, highlighting the relevance of this peculiar group in understanding the repetitive portion of ant genome. Chromosomal banding studies are in accordance with genomic data because they show that repetitive heterochromatic sequences of basal and derivative Attina species are GC-rich, an uncommon trait in Formicidae. To understand the evolutionary dynamics of heterochromatin in Attina, we compared GC-rich heterochromatin patterns between the Paleoattina and Neoattina clades of this subtribe. To this end, we hybridized the Mrel-C 0 t probe (highly and moderately repetitive DNA) obtained from Mycetomoellerius relictus, Neoattina with GC-rich heterochromatin, in karyotypes of Paleoattina and Neoattina species. Additionally, we mapped the repetitive sequences (GA) 15 and (TTAGG) 6 in species of the two clades to investigate their organization and evolutionary patterns in the genome of Attina. The Mrel-C 0 t probe marked the heterochromatin in M. relictus, in other Mycetomoellerius spp., and in species of Mycetarotes, Cyphomyrmex, and Sericomyrmex (Neoattina). In Mycetomoellerius urichii, only pericentromeric heterochromatin was marked with Mrel-C 0 t. No marking was observed in Paleoattina species or in Atta and Acromyrmex (Neoattina). These results indicated that different evolutionary events led to heterochromatin differentiation in Attina. The most likely hypothesis is that GC-rich heterochromatin arose in the common ancestor of the two clades and accumulated various changes throughout evolution. The sequences (GA) 15 and (TTAGG) 6 located in euchromatin and telomeres, respectively, showed more homogeneous results among the species., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Many neurodegenerative diseases, including Alzheimer's disease and Parkinson's disease, are characterised by the accumulation of misfolded protein deposits in the brain, leading to a progressive destabilisation of the neuronal network and neuronal death. Among the proteins that can abnormally accumulate are tau and α-synuclein, which can propagate in a prion-like manner and which upon aggregation, represent the most common intracellular proteinaceous lesions associated with neurodegeneration. For years it was thought that these intracellular proteins and their accumulation had no immediate relationship with extracellular homeostasis pathways such as the glymphatic clearance system; however, mounting evidence has now suggested that this is not the case. The involvement of the glymphatic system in neurodegenerative disease is yet to be fully defined; however, it is becoming increasingly clear that this pathway contributes to parenchymal solute clearance. Importantly, recent data show that proteins prone to intracellular accumulation are subject to glymphatic clearance, suggesting that this system plays a key role in many neurological disorders. In this review, we provide a background on the biology of tau and α-synuclein and discuss the latest findings on the cell-to-cell propagation mechanisms of these proteins. Importantly, we discuss recent data demonstrating that manipulation of the glymphatic system may have the potential to alleviate and reduce pathogenic accumulation of propagation-prone intracellular cytotoxic proteins. Furthermore, we will allude to the latest potential therapeutic opportunities targeting the glymphatic system that might have an impact as disease modifiers in neurodegenerative diseases., (© 2022. The Author(s).)

The mechanisms involved in the development of resistance to infection/reinfection by Schistosoma mansoni still arouse great interest and controversy. Some authors demonstrate that resistance to infection is attributed to a mixed Th1 and Th2 response and resistance to reinfection after repeated treatments through mechanisms associated with the Th2 response. Through flow cytometry, the phenotypic characterization of B and T lymphocytes in individuals residing in endemic areas with low parasite loads over 10 years was evaluated for the first time in humans. In this study, individuals with low parasite loads for Schistosoma mansoni had a higher proportion of Th1 and Th2 cells. In addition, lymphocytes from these individuals showed a higher degree of expression of costimulatory molecules CD28 and CTLA-4 and regulatory molecules FoxP3 and IL-10, when compared to individuals with high parasite loads. Our data indicate that the control of the parasite load of S. mansoni must be associated with a Th1, Th2, and regulatory response, and that further studies are needed to elucidate the possibility of mechanisms associated with the hyporesponsiveness of lymphocytes from individuals with high parasite loads., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The fungus-farming ant genus Cyphomyrmex (subtribe Attina, clade Neoattina) comprises 23 described species that are widely distributed throughout the Neotropics. Species within Cyphomyrmex have taxonomic issues such as Cyphomyrmex rimosus (Spinola, 1851) which is likely a species complex. Cytogenetics is a useful tool for evolutionary studies and understanding species with dubious taxonomy. In this study, we characterized the karyotype of C. rimosus from Viçosa, Minas Gerais State, southeastern Brazil using classical and molecular cytogenetic techniques to enrich the chromosomal information about Cyphomyrmex. The karyotype of C. rimosus from the rainforest of southeastern Brazil (2n = 22, 18m + 4sm) notably contrasts with that previously described for this species in Panama (2n = 32). This intraspecific chromosomal variation suggests the existence of a species complex within this taxon according to the previous hypothesis derived from morphological analysis. We detected GC-rich heterochromatic regions in C. rimosus and, using repetitive DNA probes, showed that this heterochromatin shares repetitive sequences with other Neoattina species already studied, enhancing the importance of this specific genome region in the understanding of Attina evolution. Mapping of microsatellite (GA)15 on C. rimosus was restricted to the euchromatic regions of all chromosomes. The single intrachromosomal rDNA sites observed in C. rimosus follow the general genomic organization trend of ribosomal genes in Formicidae. Our study extends the data of chromosome mapping on Cyphomyrmex and reinforces the importance of cytogenetic studies in different localities to better understand taxonomic issues in widely distributed taxa such as C. rimosus., (© 2023 S. Karger AG, Basel.)

Satellite DNAs (satDNAs) and transposable elements (TEs) are among the main components of constitutive heterochromatin (c-heterochromatin) and are related to their functionality, dynamics, and evolution. A peculiar case regarding the quantity and distribution of c-heterochromatin is observed in the genus of bees, Melipona, with species having a low amount of heterochromatin and species with high amount occupying almost all chromosomes. By combining low-pass genome sequencing and chromosomal analysis, we characterized the satDNAs and TEs of Melipona quadrifasciata (low c-heterochromatin) and Melipona scutellaris (high low c-heterochromatin) to understand c-heterochromatin composition and evolution. We identified 15 satDNA families and 20 TEs for both species. Significant variations in the repeat landscapes were observed between the species. In M. quadrifasciata, the repetitive fraction corresponded to only 3.78% of the genome library studied, whereas in M. scutellaris, it represented 54.95%. Massive quantitative and qualitative changes contributed to the differential amplification of c-heterochromatin, mainly due to the amplification of exclusive repetitions in M. scutellaris, as the satDNA MscuSat01-195 and the TE LTR/Gypsy_1 that represent 38.20 and 14.4% of its genome, respectively. The amplification of these two repeats is evident at the chromosomal level, with observation of their occurrence on most c-heterochromatin. Moreover, we detected repeats shared between species, revealing that they experienced mainly quantitative variations and varied in the organization on chromosomes and evolutionary patterns. Together, our data allow the discussion of patterns of evolution of repetitive DNAs and c-heterochromatin that occurred in a short period of time, after separation of the Michmelia and Melipona subgenera., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Cytogenetic studies on fungus-farming ants have shown remarkable karyotype diversity, suggesting different chromosomal rearrangements involved in karyotype evolution in some genera. A notable cytogenetic characteristic in this ant group is the presence of GC-rich heterochromatin in the karyotypes of some ancient and derivative species. It was hypothesized that this GC-rich heterochromatin may have a common origin in fungus-farming ants, and the increase in species studied is important for understanding this question. In addition, many genera within the subtribe Attina have few or no cytogenetically studied species; therefore, the processes that shaped their chromosomal evolution remain obscure. Thus, in this study, we karyotyped, through classical and molecular cytogenetic techniques, the fungus-farming ants Cyphomyrmextransversus Emery, 1894, Sericomyrmexmaravalhas Ješovnik et Schultz, 2017, and Mycetomoelleriusrelictus (Borgmeier, 1934), to provide insights into the chromosomal evolution in these genera and to investigate the presence the GC-rich heterochromatin in these species. Cyphomyrmextransversus (2n = 18, 10m + 2sm + 6a) and S.maravalhas (2n = 48, 28m + 20sm) showed karyotypes distinct from other species from their genera. Mycetomoelleriusrelictus (2n = 20, 20m) presented the same karyotype as the colonies previously studied. Notably, C.transversus presented the lowest chromosomal number for the genus and a distinct karyotype from the other two previously observed for this species, showing the existence of a possible species complex and the need for its taxonomic revision. Chromosomal banding data revealed GC-rich heterochromatin in all three species, which increased the number of genera with this characteristic, supporting the hypothesis of a common origin of GC-rich heterochromatin in Attina. Although a single chromosomal pair carries rDNA genes in all studied species, the positions of these rDNA clusters varied. The rDNA genes were located in the intrachromosomal region in C.transversus and M.relictus , and in the terminal region of S.maravalhas . The combination of our molecular cytogenetic data and observations from previous studies corroborates that a single rDNA site located in the intrachromosomal region is a plesiomorphic condition in Attina. In addition, cytogenetic data obtained suggest centric fission events in Sericomyrmex Mayr, 1865, and the occurrence of inversions as the origin of the location of the ribosomal genes in M.relictus and S.maravalhas . This study provides new insights into the chromosomal evolution of fungus-farming ants., (Gisele Amaro Teixeira, Luísa Antônia Campos Barros, Hilton Jeferson Alves Cardoso de Aguiar, Denilce Meneses Lopes.)

Background: Williams-Beuren syndrome is a multisystemic disorder caused by a microdeletion of the 7q11.23 region. Although familial cases with autosomal dominant inheritance have been reported, the vast majority are sporadic., Objective: To investigate the main complaints and clinical findings of patients with Williams-Beuren syndrome., Methods: A total of 757 parents of patients registered in the Brazilian Association of Williams-Beuren Syndrome (ABSW) received a questionnaire via WhatsApp from March to July 2017., Results: In total, 229 parents answered the survey. Age of diagnosis ranged from 2 days to 34 years (median: 3 years). The main clinical findings reported by the parents were abdominal colic (83.3%), failure to thrive (71.5%), feeding difficulty in the first year (68.9%), otitis (56.6%), urinary tract infections (31.9%), precocious puberty (27.1%) and scoliosis (15.9%). Cardiac defects were present in 66% of patients, and the most frequent defect was supravalvular aortic stenosis (36%). Arterial hypertension was reported in 23%. Hypercalcemia was reported in 10.5% of patients, mainly during the first year of life. Hyperacusis and hypersociability were common complaints (both present in 89%). Other behavioral and neuropsychiatric symptoms reported by the parents included attention deficit (89%), anger crises (83%), excessive fear (66%), depression (64%), anxiety (67%) and hypersexuality (33%). The most common complaints were hypersensitivity to sounds, talkative personality, emotional dependence and learning difficulties. In 98.3%, the parents denied family history., Conclusions: Williams-Beuren syndrome requires close follow-up with different medical specialties due to their variable clinical comorbidities, including language and school learning difficulties, behavioral and psychiatric problems.

Genome changes, evidenced through karyotype or nuclear genome size data, can result in reproductive isolation, diversification and speciation. The aim of this study was to understand how changes in the karyotype such as chromosome number and nuclear genome size accompanied the evolution of neotropical stingless bees, and to discuss these data in a phylogenetic context focusing on the karyotype evolution of this clade. We sampled 38 species representing the three Neotropical Meliponini groups; 35 for karyotype analyses and 16 for 1C value measurement. The chromosome number varied from 2n = 16 to 2n = 34, with distinct karyotypic formulae and the presence of a few polymorphisms, such as B chromosomes in one species and arm size differences between homologous chromosomes in two species. The mean 1C value varied from 0.31 pg to 0.92 pg. We associated empirical data on chromosome number and mean 1C value to highlight the importance of Robertsonian fusion rearrangements, leading to a decrease in chromosome number during the Neotropical Meliponini evolution. These data also allowed us to infer the independent heterochromatin amplification in several genera. Although less frequent, Melipona species with 2n = 22 represent evidence of Robertsonian fissions. We also pointed out the importance of chromosomal rearrangements that did not alter chromosome number, such as inversions and heterochromatin amplification., (© 2021 Royal Entomological Society.)

The major rDNA genes are composed of tandem repeats and are part of the nucleolus organizing regions (NORs). They are highly conserved and therefore useful in understanding the evolutionary patterns of chromosomal locations. The evolutionary dynamics of the karyotype may affect the organization of rDNA genes within chromosomes. In this study, we physically mapped 18S rDNA genes in 13 Neotropical ant species from four subfamilies using fluorescence in situ hybridization. Furthermore, a survey of published rDNA cytogenetic data for 50 additional species was performed, which allowed us to detect the evolutionary patterns of these genes in ant chromosomes. Species from the Neotropical, Palearctic, and Australian regions, comprising a total of 63 species from 19 genera within six subfamilies, were analysed. Most of the species (48 out of 63) had rDNA genes restricted to a single chromosome pair in their intrachromosomal regions. The position of rDNA genes within the chromosomes appears to hinder their dispersal throughout the genome, as translocations and ectopic recombination are uncommon in intrachromosomal regions because they can generate meiotic abnormalities. Therefore, rDNA genes restricted to a single chromosome pair seem to be a plesiomorphic feature in ants, while multiple rDNA sites, observed in distinct subfamilies, may have independent origins in different genera., (© 2021 Royal Entomological Society.)

Background: Acute lymphoid leukemia (ALL) is the most prevalent cancer of childhood. Impairment in linguistic and memory skills is a possible late sequela in cancer survivors that can limit their quality of life and the overall performance of the individual in society. There is evidence that survivors of ALL treated exclusively with chemotherapy demonstrate significant differences in long-term linguistic and memory functions and also changes in neuroanatomical integrity. However, most studies described do not count on a speech-language pathologist in their team, which we consider important to discuss. Thus, the objective of the present study was to assess memory and vocabulary skills in the pediatric population diagnosed with acute lymphoid leukemia during chemotherapy treatment., Materials and Methods: An observational cohort study was conducted over a 1.8-year period. Participants of this research were children diagnosed with ALL. All participants were assessed on their linguistic-cognitive skills (ie, vocabulary, short-term memory and lexical access). All data underwent statistical analyses., Results: The results of the current study found no major significant difference in the linguistic-cognitive performance of children with ALL and their healthy controls. Regarding the linguistic variables, we believe that there should be a differentiation between the effects of the drugs and the effects of social communication skills on performance., Conclusion: This first characterization of the linguistic-cognitive abilities of children with ALL did not identify differences between these children and their healthy peers, although we were able to identify variables regarding the multidisciplinary team and social communication that should be considered in future studies., Competing Interests: The authors have no conflicts of interest to declare. This study was conducted in accordance with the Declaration of Helsinki., (© 2021 Miranda Pereira et al.)

Purpose: To verify whether the performance in the first assessment of pretend play of children with diagnostic hypothesis of developmental language disorder, correlates with the performance in formal tests of non-verbal intellectual function and both receptive and expressive vocabulary after 5 years old., Methods: The research comprised 26 subjects, 19 of whom were male, and 7 were female, with an mean age of 4:10. All participants presented a diagnostic hypothesis of developmental language disorder based on inclusion and exclusion criteria described internationally. Data were analyzed in relation to pretend play, vocabulary and performance in standardized intellectual assessment test; we also investigated correlation between them. All data underwent statistical analysis and the confidence intervals were 95%., Results: As for pretend play, it was found that most children with suspected developmental language disorder present more primitive symbolic development. Regarding vocabulary, the data show greater performance in receptive vocabulary than in expressive. In the assessment of the intelligence quotient, the children obtained, in their majority, classifications in the mean score and superior score to their age. Regarding the interaction between studied variables, no correlation was found., Conclusion: No relationship was found between the studied variables. The data are discussed in the light of international literature and promote important reflections on the symbolic-linguistic development of this population.

Manganese porphyrins are well-known protectors against the deleterious effects of pro-oxidant species such as superoxide ions and hydrogen peroxide. The present study investigated the antioxidant cytochrome c-like activities of Mn(III)TMPyP [meso-tetrakis (4-N-methyl pyridinium) porphyrin] against superoxide ion and hydrogen peroxide that remained unexplored for this porphyrin. The association of TMPyP with a model of the inner mitochondrial membrane, cardiolipin (CL)-containing liposomes, shifted +30 mV vs. NHE (normal hydrogen electrode) redox potential of the Mn (II) /Mn (III) redox couple. In CL-containing liposomes, Mn (III) TMPyP was reduced by superoxide ions and recycled by Fe (III) cytochrome c to the oxidized form. Similarly, isolated rat liver mitoplasts added to a sample of Mn (II) TMPyP promoted immediate porphyrin reoxidation by electron transfer to the respiratory chain. These results show that Mn (III) TMPyP can act as an additional pool of Fe (III) cytochrome c capable of transferring electrons that escape from the IV complex back into the respiratory chain. Unlike Fe (II) cytochrome c, Mn (II) TMPyP was not efficient for hydrogen peroxide clearance. Therefore, by reducing cytochrome c, Mn (II) TMPyP can indirectly contribute to hydrogen peroxide elimination., Competing Interests: Declaration of competing interest Authors declare no conflicts of interest and no competing interests., (Copyright © 2021 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.)

Schistosomiasis is a parasitic disease that affects about 166 million people around the world. It is estimated that 5%-10% of individuals with schistosomiasis develop severe forms of the disease, which are characterized by pulmonary hypertension, ascites, periportal fibrosis, and other significant complications. The chronic phase of the disease is associated with a Th2 type immune response, but evidence also suggests there are roles for Th1 and Th17 in the development of severe disease. The aim of this study was to evaluate the CD4 + T lymphocyte profile of patients with different degrees of periportal fibrosis secondary to schistosomiasis. These individuals had been treated for schistosomiasis, but since they live in a S. mansoni endemic area, they are at risk of reinfection. They were evaluated in relation to the degree of periportal fibrosis and classified into three groups: without fibrosis or with incipient fibrosis (WF/IFNE), n=12, possible periportal fibrosis/periportal fibrosis, n=13, and advanced periportal fibrosis/advanced periportal fibrosis with portal hypertension, n=4. We observed in the group without fibrosis a balance between the low expression of Th2 cytokines and high expression of T reg cells. As has already been described in the literature, we found an increase of the Th2 cytokines IL-4, IL-5, and IL-13 in the group with periportal fibrosis. In addition, this group showed higher expression of IL-17 and IL-10 but lower IL-10/IL-13 ratio than patients in the WF/IFNE group. Cells from individuals who present any level of fibrosis expressed more TGF-β compared to the WF/IFNE group and a positive correlation with left lobe enlargement and portal vein wall thickness. There was a negative correlation between IL-17 and the thickness of the portal vein wall, but more studies are necessary in order to explore the possible protective role of this cytokine. Despite the fibrosis group having presented a higher expression of pro-fibrotic molecules compared to WF/IFNE patients, it seems there is a regulation through IL-10 and T reg cells that is able to maintain the low morbidity of this group., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Santana, de Almeida, Lopes, Page, Oliveira, Souza, Ribeiro, Gutiérrez, Carvalho and Cardoso.)

The eukaryote genome is enriched by different types of repetitive DNA sequences and is most abundant in heterochromatin regions. Historically, no function has been assigned to these sequences, which makes them the target of studies that have demonstrated their structural and functional importance in the genome. Despite having a constant chromosome number, the genus Melipona has species with wide variation in heterochromatin content, from 8 to 73%, which is an important feature to be investigated regarding its origin and evolution. In the present study, a repetitive DNA sequence of Melipona mondury was isolated by restriction enzyme digestion. This sequence was used to hybridize chromosomes of eight Melipona species that include representatives of the four subgenera and present divergent characteristics in relation to the heterochromatin content. Considering that rDNA localization has shown differences in Melipona, 16 species of this genus were analyzed with 18S rDNA probe. Our data suggest that heterochromatin growth occurred independently in the Michmelia and Melikerria subgenera, considering that the isolated repetitive DNA sequence was shared only by the Michmelia species. Amplification possibly occurred from the centromeric region, causing the displacement of the rDNA sites to the ends of the chromosomes. The repetitive DNA sequence used is a constituent of Michmelia heterochromatin, which that arose from the common ancestor of the species of this subgenus.

The genus Partamona includes 33 species of stingless bees, of which 11 were studied cytogenetically. The main goal of this study was to propose a hypothesis about chromosomal evolution in Partamona by combining molecular and cytogenetic data. Cytogenetic analyses were performed on 3 Partamona species. In addition, the molecular phylogeny included mitochondrial sequences of 11 species. Although the diploid number was constant within the genus, 2n = 34, B chromosomes were reported in 7 species. Cytogenetic data showed karyotypic variations related to chromosome morphology and the amount and distribution of heterochromatin and repetitive DNA. The molecular phylogenetic reconstruction corroborated the monophyly of the genus and separated the 2 clades (A and B). This separation was also observed in the cytogenetic data, in which species within each clade shared most of the cytogenetic characteristics. Furthermore, our data suggested that the B chromosome in the genus Partamona likely originated from a common ancestor of the species that have it in clade B and, through interspecific hybridization, it appeared only in Partamona rustica from clade A. Based on the above, Partamona is an interesting genus for further investigations using molecular mapping of B chromosomes as well as for broadening phylogenetic data., (© 2021 S. Karger AG, Basel.)

Gnamptogenys includes 138 described species that are widely distributed, with high diversity, in the Neotropics. Some Neotropical species have taxonomic issues, as is the case with Gnamptogenys striatula, for which morphological variations have been observed between different populations. For the ant species with taxonomic issues, classical and molecular cytogenetic studies have assisted in the resolution of these issues. Cytogenetic studies of Gnamptogenys are scarce and have only been reported for 14 taxa. These reports have rarely presented chromosomal morphology. Considering the importance of the taxonomic revision of some species, such as G. striatula, the present study cytogenetically characterized four species of Gnamptogenys: G. striatula, G. moelleri, G. regularis, and G. triangularis, discussing their phylogenetic and biogeographic characteristics. The number of chromosomes ranged from 2n = 26 to 2n = 44, with distinct karyotypes at both species and population levels. All four species presented a pair of 18S rDNA gene markers that coincided with GC-rich regions. In the case of G. striatula from the Atlantic rainforest, a chromosomal polymorphism was observed, with chromosomal translocations being the likely origin of this polymorphism. Two populations of G. striatula showed karyotype differences, thus corroborating previous morphological data indicating the existence of a species complex in this taxon. In addition, G. regularis showed a polymorphism involving a chromosome pair bearing ribosomal genes, possibly caused by unequal crossing-over. Although G. moelleri has a well-defined taxonomy, a population from the eastern Amazon rainforest presented a divergent karyotype from the Atlantic rainforest populations, suggesting the existence of a cryptic species in this taxon.

Purpose: To investigate lexical performance and verbal short-term memory ability in premature infants at preschool age and compare the results with their full-term peers., Methods: Forty preschool children aged between 4 and 5 years and 11 months participated on the study and were equally divided into two groups considering their gestational age: preterm and full-term. Groups did not differ on age, family income and maternal education. Participants were paired according to their chronological age and their socioeconomic status. Their performance was assessed using expressive vocabulary and verbal short-term memory tests. Inferential statistical analysis was performed using the Mann-Whitney and the Fisher exact test., Results: Group performance did not differ on vocabulary, but premature children showed an inferior performance on nonword repetition tasks., Conclusion: These data indicates that preschoolers born premature performed statistically lower than their peers born full-term on nonword repetition task. Thus, premature birth was associated to vocabulary development on typically developing range, but also to verbal short-term memory impairments.

Studies in several organisms have contributed to the understanding of heterochromatin and its biological importance. In bees of the tribe Meliponini, the presence of chromosomes with totally heterochromatic arms has been attributed to the mechanism of karyotype evolution in which this group accumulated heterochromatin to maintain telomere stability after centric fission events. In the present study, the use of classical and molecular cytogenetic techniques as well as automated image analysis software for the description of the karyotypes of Partamonachapadicola and P. nhambiquara bee species revealed variability in the compaction and patterns of chromatin structure. Although both species have the same chromosome number as other species in the genus Partamona (2n = 34), C-banding and image analyses indicated the existence of chromosomes with 3 regions of different staining intensities, suggesting a chromatin structure with distinct patterns and characteristics. Repetitive DNA probes hybridized only in the euchromatic regions, whereas the regions with intermediate staining intensity did not show any hybridization signals. This suggests that these regions present features more similar to heterochromatin. Evidence of the existence of a chromatin class with intermediate condensation compared to euchromatin and heterochromatin indicates a potential mechanism for heterochromatin amplification and demonstrates the need for further studies on this topic. This previously unrecognized class of chromatin should be taken into account in the study of all Meliponini chromosomes., (© 2020 S. Karger AG, Basel.)

The characterization of karyotypes is an important aspect in understanding the structure and evolution of genomes. Polybia is a genus of social wasps of the family Vespidae. This genus has 58 species, but for only 8 of these chromosome number and morphology have been reported in the literature. The aim of this study was to describe and characterize the Polybia fastidiosuscula Saussure karyotype, presenting the first case of a B chromosome in Vespidae. In addition, we investigated the chromatin composition of this species through C-banding, base-specific fluorochrome staining, and physical mapping of 7 microsatellites and 18S rDNA. Four colonies of P. fastidiosuscula from Minas Gerais and Paraná states, Brazil, were analyzed. The chromosome number identified was 2n = 34, and 2 colonies presented a B chromosome. We characterized the chromatin composition of this species, analyzing the existence of different microsatellite-rich heterochromatic regions which are also enriched with AT or GC base pairs. We suggest an intraspecific origin of the B chromosome based on the homology of the heterochromatic composition with A chromosomes and also verify that the TTAGG and TCAGG sequences are not telomeric, but only microsatellites that occur in the centromeres of most chromosomes, as well as GAG and CGG., (© 2021 S. Karger AG, Basel.)

Background: Stroke is a leading cause of death and disability globally, with an increasing incidence in low- and middle-income countries (LMICs). The successful treatment of acute stroke requires an organized, efficient and well-resourced emergency care system. However, debate exists surrounding the prioritization of stroke treatment programs given the high costs of treatment and the increased incidence of hemorrhagic stroke in LMICs. Economic data is helpful to guide evidence-based priority setting in health systems development, particularly in low-resource settings where scarcity requires careful stewardship of resources. This systematic review surveys the existing evidence surrounding the cost-effectiveness of interventions to address acute stroke in LMIC settings., Methods: The authors conducted a PRISMA style systematic review of economic evaluations of interventions to address acute stroke in LMICs. Five databases were systematically searched for articles, which were then reviewed for inclusion., Results: Of the 153 unique articles identified, 11 met the inclusion criteria. Four studies demonstrate the heavy economic burden on patients and households due to stroke. Two studies estimate that preventive measures are more cost-effective than acute treatments. Four studies directly examine the cost-effectiveness of thrombolysis and thrombectomy in three middle-income countries (Iran, China, and Brazil) with results ranging from roughly $2578 to $34,052 (2019 USD) per quality adjusted life-year saved. These results are similar to the cost-effectiveness ratios estimated in high-income settings. Finally, one study examined a care bundle that included acute treatment elements., Conclusions: The findings reinforce the need for additional research support informed decision-making. The available evidence suggests that preventive measures should be prioritized over emergency treatment for acute stroke, particularly in settings of resource scarcity. Cost-effectiveness ratios do not compare favorably to estimates for other emergency care interventions in LMICs, such as basic emergency care training, implementation of triage systems, and basic trauma care. Cost-effectiveness is also likely to vary depending on local epidemiology. Overall, decision-makers should balance the economic evidence alongside social, political and cultural priorities when making resource allocation choices., Competing Interests: The authors declared no conflicts of interest., (© 2020 African Federation for Emergency Medicine. Publishing services provided by Elsevier.)

Fungus-farming ants of the genus Mycetophylax exhibit intra and interspecific chromosome variability, which makes them suitable for testing hypotheses about possible chromosomal rearrangements that endure lineage diversification. We combined cytogenetic and molecular data from Mycetophylax populations from coastal environments to trace the evolutionary history of the clade in light of chromosomal changes under a historical and geographic context. Our cytogenetic analyses revealed chromosomal differences within and among species. M. morschi exhibited three distinct karyotypes and considerable variability in the localization of 45S rDNA clusters. The molecular phylogeny was congruent with our cytogenetic findings. Biogeographical and divergence time dating analyses estimated that the most recent common ancestor of Mycetophylax would have originated at about 30 Ma in an area including the Amazon and Southern Grasslands, and several dispersion and vicariance events may have occurred before the colonization of the Brazilian Atlantic coast. Diversification of the psammophilous Mycetophylax first took place in the Middle Miocene (ca. 18-10 Ma) in the South Atlantic coast, while "M. morschi" lineages diversified during the Pliocene-Pleistocene transition (ca. 3-2 Ma) through founder-event dispersal for the Northern coastal regions. Psammophilous Mycetophylax diversification fits into the major global climatic events that have had a direct impact on the changes in sea level as well as deep ecological impact throughout South America. We assume therefore that putative chromosomal rearrangements correlated with increased ecological stress during the past climatic transitions could have intensified and/or accompanied the divergence of the psammophilous Mycetophylax. We further reiterate that "M. morschi" comprises a complex of at least three well-defined lineages, and we emphasize the role of this integrative approach for the identification and delimitation of evolutionary lineages.

Schistosomiasis affects about 240 million people worldwide and is estimated that about 700 million people live in areas at risk of infection. In the context of immune response associated with infection by Schistosoma mansoni, the role of memory T cells is not well understood., Aim: To evaluate the frequency of memory CD4 + and CD8 + T cells from individuals resistant and susceptible to Schistosoma mansoni infection., Methods and Results: We selected individuals with low (resistant) and high (susceptible) parasite burden using databases generated during previous studies carried out in the same endemic area. The cell surface markers were performed using flow cytometry. In this study, the resistant individuals showed an increase in the CD4 + memory T-cell pool associated with an increase in the central memory cell (TCM) and a decrease in the effector memory cell (T EM ). Individuals susceptible to infection had higher frequencies of effector memory cells compared to resistant individuals., Conclusions: These data suggest that resistance to S mansoni infection may be associated with an increase in the number of CD4 + memory T cells and susceptibility to infection is associated with a decrease in the central memory cell as well as high proportions of effector memory cells., (© 2019 John Wiley & Sons Ltd.)

It is thought that two evolutionary mechanisms gave rise to chromosomal variation in bees: the first one points to polyploidy as the main cause of chromosomal evolution, while the second, Minimum Interaction Theory (MIT), is more frequently used to explain chromosomal changes in Meliponini and suggests that centric fission is responsible for variations in karyotype. However, differences in chromosome number between Meliponini and its sister taxa and in the karyotype patterns of the Melipona genus cannot be explained by MIT, suggesting that other events were involved in chromosomal evolution. Thus, we assembled cytogenetical and molecular information to reconstruct an ancestral chromosome number for Meliponini and its sister group, Bombini, and propose a hypothesis to explain the evolutionary pathways underpinning chromosomal changes in Meliponini. We hypothesize that the common ancestor shared by the Meliponini and Bombini tribes possessed a chromosome number of n = 18. The karyotype with n = 17 chromosomes was maintained in Meliponini, and variations of haploid numbers possibly originated through additional Robertsonian fissions and fusions. Thus, the low chromosome number would not be an ancestral condition, as predicted by MIT. We then conclude that Robertsonian fission and fusions are unlikely to be the cause of chromosomal rearrangements that originated the current karyotypes in Meliponini., Competing Interests: The authors have declared that no competing interests exist.

Increasing evidence suggests that nerve fibers responding to noxious stimuli (nociceptors) modulate immunity in a variety of tissues, including the skin. Yet, the role of nociceptors in regulating sterile cutaneous inflammation remains unexplored. To address this question, we have developed a detailed description of the sterile inflammation caused by overexposure to UVB irradiation (i.e., sunburn) in the mouse plantar skin. Using this model, we observed that chemical depletion of nociceptor terminals did not alter the early phase of the inflammatory response to UVB, but it caused a significant increase in the number of dendritic cells and αβ + T cells as well as enhanced extravasation during the later stages of inflammation. Finally, we showed that such regulation was driven by the nociceptive neuropeptide calcitonin gene-related peptide. In conclusion, we propose that nociceptors not only play a crucial role in inflammation through avoidance reflexes and behaviors, but can also regulate sterile cutaneous immunity in vivo., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

The constant refinement of tests used in animal research is crucial for the scientific community. This is particularly true for the field of pain research, where ethical standards are notably sensitive. The formalin test is widely used in pain research and some of its mechanisms resemble those underlying clinical pain in humans. Immediately upon injection, formalin triggers two waves (an early and a late phase) of strong, nociceptive behaviour, characterised by licking, biting, lifting and shaking the injected paw of the animal. Although well characterised at the behaviour level, since its proposal over four decades ago, there has not been any significant refinement to the formalin test, especially those combining minimisation of animal distress and preservation of behavioural outcomes of the test.  Here, we propose a modified and improved method for the formalin test. We show that anaesthetising the animal with the inhalable anaesthetic sevoflurane at the time of the injection can produce reliable, robust and reproducible results whilst animal distress during the initial phase is reduced. Importantly, our results were validated by pharmacological suppression of the behaviour during the late phase of the test with gabapentin, the anaesthetic showing no interference with the drug. In addition, we demonstrate that this is also a useful method to screen for changes in pain behaviour in response to formalin in transgenic lines., Competing Interests: No competing interests were disclosed.

Background: The immunopathogenesis of severe asthma has been associated with an inefficient regulatory response. There are a few studies about the CD4 T cells profile among individuals with severe asthma refractory to treatment., Objective: To evaluate the CD4 T lymphocyte profile from individuals with severe asthma according to their response to treatment, relating to their atopy status and age of asthma onset., Methods: We evaluated nineteen individuals with severe asthma refractory to treatment (SAR), 21 with well-controlled or partly controlled severe asthma (CSA) and 23 with mild-to-moderate asthma (MMA). Lymphocytes were obtained from PBMC, and the frequency of expression of different molecules in this population was assessed using the flow cytometry., Results: We observed the frequency of CD4 + IFN-γ + T cells was higher in atopic individuals with SAR than with CSA. In addition, among the atopic and early-onset asthma (EOA), the frequency of CD4 + CTLA-4 + T cells was lower in the SAR group than the CSA group. In relation to non-atopic and late-onset asthma (LOA) phenotypes, we noted the frequency of CD4 + FoxP3 + T cells was lower in individuals with SAR than with CSA. We also observed among the LOA patients, the frequency of CD4 + TGF-β + T cells was decreased in SAR group than the in CSA group., Conclusion and Clinical Relevance: Our data suggest that refractoriness to treatment in asthma is associated with a lower expression of distinct regulatory molecules by CD4 T cells between those who are atopic and have EOA and those who are non-atopic and have LOA. Thus, these results may contribute to the identification of new regulatory strategies to treat asthma according to their phenotypes., (© 2019 John Wiley & Sons Ltd.)

Objective: Pain is the main reason patients report Osteoarthritis (OA), yet current analgesics remain relatively ineffective. This study investigated both peripheral and central mechanisms that lead to the development of OA associated chronic pain., Design: The monoiodoacetate (MIA) model of OA was investigated at early (2-6 days post injection) and late (>14 days post injection) time points. Pain-like behaviour and knee histology were assessed to understand the extent of pain due to cartilage degradation. Electrophysiological single-unit recordings were taken from spinal wide dynamic range (WDR) neurons to investigate Diffuse Noxious Inhibitory Controls (DNIC) as a marker of potential changes in descending controls. Immunohistochemistry was performed on dorsal root ganglion (DRG) neurons to assess any MIA induced neuronal damage. Furthermore, qPCR was used to measure levels of glia cells and cytokines in the dorsal horn., Results: Both MIA groups develop pain-like behaviour but only late phase (LP) animals display extensive cartilage degradation. Early phase animals have a normally functioning DNIC system but there is a loss of DNIC in LP animals. We found no evidence for neuronal damage caused by MIA in either group, yet an increase in IL-1β mRNA in the dorsal horn of LP animals., Conclusion: The loss of DNIC in LP MIA animals suggests an imbalance in inhibitory and facilitatory descending controls, and a rise in the mRNA expression of IL-1β mRNA suggest the development of central sensitisation. Therefore, the pain associated with OA in LP animals may not be attributed to purely peripheral mechanisms., (Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The immune response induced by Schistosma mansoni antigens is able to prevent immune-mediated diseases. Conversely, the inflammatory response in cutaneous leishmaniasis (CL), although responsible for controlling the infection, is also associated with the pathogenesis of disease. The aim of this study was to evaluate the potential of the S. mansoni Sm29 antigen to change certain aspects of the profiles of monocyte derived dendritic cells (MoDCs) and lymphocytes from subjects with CL in vitro . Expression of surface molecules and intracellular cytokines in the MoDCs and lymphocytes as well as the proliferation of Leishmania braziliensis were evaluated by flow cytometry. Levels of cytokines were evaluated in culture supernatants by ELISA. It was observed that stimulation by rSm29 increased the frequency of expression of CD83, CD80, CD86, and IL-10R in MoDCs compared to non-stimulated cultures. Additionally rSm29 decreased the frequency CD4 + and CD8 + T cells expressing CD28 and increased the frequency of CD4 + CD25 hi and CD4 + CTLA-4 + T lymphocytes. Addition of rSm29 to cultures increased IL-10 levels and decreased levels of IL-12p40 and IFN-γ, while not altering TNF levels compared to non-stimulated cultures. This study showed that rSm29 induced a regulatory profile in MoDCs and lymphocytes and thereby regulated the exaggerated inflammation observed in CL. Considering that there are few therapeutic options for leishmaniasis, the use of rSm29 may be an alternative to current treatment and may be an important strategy to reduce the healing time of lesions in patients with CL.

Stingless bees of the genus Melipona are subdivided into 4 subgenera called Eomelipona, Melikerria, Melipona sensu stricto, and Michmelia according to species morphology. Cytogenetically, the species of the genus Melipona show variation in the amount and distribution of heterochromatin along their chromosomes and can be separated into 2 groups: the first with low content of heterochromatin and the second with high content of heterochromatin. These heterochromatin patterns and the number of chromosomes are characteristics exclusive to Melipona karyotypes that distinguish them from the other genera of the Meliponini. To better understand the karyotype organization in Melipona and the relationship among the subgenera, we mapped repetitive sequences and analyzed previously reported cytogenetic data with the aim to identify cytogenetic markers to be used for investigating the phylogenetic relationships and chromosome evolution in the genus. In general, Melipona species have 2n = 18 chromosomes, and the species of each subgenus share the same characteristics in relation to heterochromatin regions, DAPI/CMA3 fluorophores, and the number and distribution of 18S rDNA sites. Microsatellites were observed only in euchromatin regions, whereas the (TTAGG)6 repeats were found at telomeric sites in both groups. Our data indicate that in addition to the chromosome number, the karyotypes in Melipona could be separated into 2 groups that are characterized by conserved cytogenetic features and patterns that generally are shared by species within each subgenus, which may reflect evolutionary constraints. Our results agree with the morphological separation of the Melipona into 4 subgenera, suggesting that they must be independent evolutionary lineages., (© 2019 S. Karger AG, Basel.)

The genus Melipona is subdivided into four subgenera based on morphological characteristics, and two groups based on cytogenetic patterns. The cytogenetic information on this genus is still scarce, therefore, the goal of this study was to characterize Melipona paraensis, Melipona puncticollis, and Melipona seminigra pernigra using the following techniques: C-banding, DAPI/CMA3 fluorochromes, and FISH with an 18S rDNA probe. Melipona paraensis (2n=18) and M. seminigra pernigra (2n=22) were classified as high heterochromatin content species (Group II). Their euchromatin is restricted to the ends of the chromosomes and is CMA3+; the 18S rDNA probe marked chromosome pair number 4. Melipona puncticollis (2n=18) is a low heterochromatin content species (Group I) with chromosome pair number 1 marked with CMA3 and 18S rDNA. Low heterochromatin content is a putative ancestral karyotype in this genus and high content is not a monophyletic trait (Melikerria presents species with both patterns). Differences concerning the karyotypic characteristics can be observed among Melipona species, revealing cytogenetic rearrangements that occurred during the evolution of this genus. Studies in other species will allow us to better understand the processes that shaped the chromatin evolution in Melipona.

Promoter-based genetic recombination (via, e.g., Cre-lox) is most useful when all cells of interest express a particular gene. The discovery that the actin-binding protein advillin is expressed in all somatic sensory neurons has been exploited repeatedly to drive DNA recombination therein, yet specificity of expression has not been well demonstrated. Here, we characterize advillin expression amongst sensory neurons and in several other neural and non-neural tissues. We first validate an advillin antibody against advillin knock-out tissue, advillin promoter-driven EGFP, and advillin mRNA expression. In the dorsal root ganglion (DRG), advillin is enriched in non-peptidergic nociceptors. We also show that advillin expression, and advillin promotor-driven EGFP and Cre-recombinase expression, occurs in multiple tissues including the dorsal habenula of the epithalamus, endocrine cells of the gut, Merkel cells in the skin, and most strikingly, throughout the autonomic nervous system (sympathetic, parasympathetic, and enteric neurons) in mice, rats, and non-human primates. In the mouse pelvic ganglion, advillin immunoreactivity is most intense in pairs of small neurons, and concentrated in spine-like structures on the axon initial segment contacted by sympathetic preganglionic axons. In autonomic targets (iris and blood vessels), advillin is distributed along cholinergic parasympathetic axons and in sympathetic varicosities. Developmentally, advillin expression is absent from sympathetics at postnatal day 4 but begins to emerge by day 7, accounting for previous reports (based on embryonic expression) of advillin's specificity to sensory neurons. These results indicate that caution is warranted in interpreting previous studies in which advillin-driven genomic editing is either constitutive or performed after postnatal day 4.

Background: Cri du Chat syndrome (CdCS) is a genetic syndrome caused by deletions in the short arm of chromosome 5. Although the main clinical features of CdCS are well known, the neurocognitive and behavioural characteristics of the phenotype are rarely described in detail in the literature. In this study, we analysed the main phenotypic features of CdCS from a parental perspective., Method: A questionnaire was sent to 700 Brazilian families that were registered in the Brazilian Association of CdCS. The questions involved specific domains of CdCS, such as pregnancy and birth conditions, recurrence of the disease in the family, current major health problems, and aspects of cognitive development., Results: In total, 73 questionnaires were completed: 44 females and 29 males, ranging from 9.5 months old to 40 years old (mean = 13.8 years; median = 12 years). Most of the parents noticed the typical cat-like cry at birth (94.4%). The age at diagnosis of CdCS ranged from the time of birth to 180 months (mean = 14 months; median = 6 months), while one case was diagnosed during pregnancy. In all of the cases, the diagnosis of CdCS was made by G-banding karyotype analysis. In 66.2% of the cases, the parents underwent cytogenetic investigation. A total of 52.1% of the parents answered that they did not remember what the recurrence risk of CdCS was in their family. The main health problems that were reported were as follows: swallowing problems (80.3%), feeding problems (80.3%), congenital heart disease (31.5%), spine abnormalities (28.8%), and neurological symptoms (20.5%), including seizures (11%). The behavioural problems that were reported were as follows: aggressive behaviour, stereotypies, anxiety, phobias, and genital manipulation/masturbation. Neurodevelopmental delay was reported in all of the cases. Independent walking was achieved in 72.2% of the patients. Approximately 50% of the patients never presented expressive language, and most of the patients are dependent on others for their daily activities., Conclusions: The questionnaire was a pioneer initiative in the CdCS support group, and the answers used in this study can improve the health care assistance to these patients because they focus attention on the demands from a parental perspective. In addition, nearly half of the families stated that they did not remember information regarding recurrence risk, which reinforces the importance of genetic counselling follow-up and the need for the expansion of genetic services in Brazil., (© 2018 MENCAP and International Association of the Scientific Study of Intellectual and Developmental Disabilities and John Wiley & Sons Ltd.)

Greater emphasis on the study of intact cellular networks in their physiological environment has led to rapid advances in intravital imaging of the central nervous system (CNS), while the peripheral system remains largely unexplored. To assess large networks of sensory neurons, we selectively label primary afferents with GCaMP6s in male and female C57bl/6 mice and visualize their functional responses to peripheral stimulation in vivo . We show that we are able to monitor the activity of hundreds of sensory neurons simultaneously, with sufficient sensitivity to detect, in most cases, single action potentials with a typical rise time of around 200 ms, and an exponential decay with a time constant of approximately 700 ms. With this technique we are able to characterize the responses of large populations of sensory neurons to innocuous and noxious mechanical and thermal stimuli under normal and inflammatory conditions. We demonstrate that the majority of primary afferents are polymodal with between 50-80% of thermally sensitive DRG neurons responding also to noxious mechanical stimulation. We also specifically assess the small population of peripheral cold neurons and demonstrate significant sensitization to cooling after a model of sterile and persistent inflammation, with significantly increased sensitivity already at decreases of 5°C when compared to uninflamed responses. This not only reveals interesting new insights into the (patho)physiology of the peripheral nervous system but also demonstrates the sensitivity of this imaging technique to physiological changes in primary afferents.

Objective: The identification of oropharyngeal aspiration is paramount since it can have negative consequences on a compromised respiratory status. Our hypothesis was that dysphagia in neurologically intact children with respiratory disease is associated to specific clinical markers., Study Design: Using the medical files we conducted a retrospective, observational cohort study on children admitted to the pediatric hospital unit due to respiratory disease. We collected data on specific parameters of a clinical swallowing assessment and dysphagia was classified according to the Dysphagia Management Staging Scale. We also included the following clinical markers: age, days of hospitalization, need for orotracheal intubation (OTI), duration of orotracheal intubation (in hours), number of previous hospital admissions due to respiratory disease, number of previous hospital admissions due to other causes, and previous orotracheal intubations., Results: The final study sample consisted of 102 patients (mean age of 5.88 months). For the purposes of statistical analysis, the patients were grouped according to the classification of dysphagia (ie, no dysphagia, mild dysphagia, and moderate-severe dysphagia). Data analysis indicated that the clinical markers of orotracheal intubation (P = 0.042), duration of orotracheal intubation (P = 0.025), and days of hospitalization (P = 0.037) were significant in children with moderate-severe dysphagia., Conclusions: Our data indicate that neurologically intact children with respiratory disease who were submitted to prolonged OTI (ie, over 48 h) should be prioritized for receiving a detailed swallowing assessment., (© 2018 Wiley Periodicals, Inc.)

Objective to investigate the performance of European Portuguese children in a Brazilian test of expressive vocabulary, seeking to identify differences between age groups and gender, and to verify its applicability in this population. Methods the sample consisted of 150 typical developed children, of both genders, between the ages of 5 and 6. All children attended public schools in the north area of Portugal. To assess the semantic performance, the expressive vocabulary sub-test of the language test (ABFW) was used, considering the percentage of usual verbal assignments and the classification (adequate/inadequate) according to the Brazilian reference values. Results the performance of the European Portuguese children indicated that at age 6 they have a higher percentage of correct answers in expressive vocabulary. As for the gender, there were only occasional differences: the girls showed a greater dominance in the semantic fields of clothing (both ages) and furniture and utensils (at age 5), whereas the boys showed more dominance in the semantic field means of transportation (6 years). Regarding classification, there was no difference between age groups in overall performance. Only the semantic field shapes and colors had more individuals of 6 years with inadequate performance. Conclusion the reference values adopted in the Brazilian population for semantic performance indicated that more than 80% of the children of each age group could have their performance classified as adequate. Such evidence suggests that this tool shows potential as an instrument of quantitative vocabulary's assessment of 5 and 6-years old children in European Portuguese.

Coinfection with leishmaniasis and schistosomiasis has been associated with increased time to healing of cutaneous lesions of leishmaniasis. The objective of this study was to evaluate the effect of Leishmania braziliensis infection on co-cultures of monocyte-derived dendritic cells (MoDCs) with autologous lymphocytes from patients with schistosomiasis and patients with cutaneous leishmaniasis. MoDCs were differentiated from peripheral blood monocytes, isolated by magnetic beads, infected with L. braziliensis, and co-cultured with autologous lymphocytes. Expression of HLA-DR, CD1a, CD83, CD80, CD86, CD40, and the IL-10 receptor (IL-10R) on MoDCs as well as CD28, CD40L, CD25, and CTLA-4 on lymphocytes were evaluated by flow cytometry. The production of the cytokines IL-10, TNF, IL-12p40, and IFN-γ were evaluated by sandwich ELISA of the culture supernatant. The infectivity evaluation was performed by light microscopy after concentration of cells by cytospin and Giemsa staining. It was observed that the frequency of MoDCs expressing CD83, CD80, and CD86 as well as the MFI of HLA-DR were smaller in the group of patients with schistosomiasis compared to the group of patients with leishmaniasis. On the other hand, the frequency of IL-10R on MoDCs was higher in patients with schistosomiasis than in patients with leishmaniasis. CD4 + and CD8 + T lymphocytes from patients with schistosomiasis presented a lower frequency of CD28 and a higher frequency of CTLA-4 compared to lymphocytes from patients with leishmaniasis. Levels of IL-10 were higher in the supernatants of co-cultures from individuals with schistosomiasis compared to those with leishmaniasis. However, levels of TNF, IL-12p40, and IFN-γ were lower in the group of individuals with schistosomiasis. Regarding the frequency of MoDCs infected by L. braziliensis after 72h in culture, it was observed that higher frequencies of cells from patients with schistosomiasis were infected compared to cells from patients with leishmaniasis. It was concluded that MoDCs from patients with schistosomiasis are more likely to be infected by L. braziliensis, possibly due to a lower degree of activation and a regulatory profile., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Women suffer chronic pain more frequently than men. It is not clear whether this is due to differences in higher level cognitive processes or basic nociceptive responses. In this study we used a mouse model of neuropathic pain to dissociate these factors. We performed RNA-seq on purified peripheral afferent neurons, but found no striking differences in gene expression between male and female mice, neither before nor after nerve injury. Similarly, spinal cord immune responses between the sexes appeared to be indistinguishable when studied by flow cytometry or qRT-PCR. Differences emerged only upon studying peripheral immune cell infiltration into the dorsal root ganglion, suggesting that adaptive immune responses in neuropathic pain could be sexually dimorphic.

The dorsal root ganglia (DRG) and trigeminal ganglia (TG) are clusters of cell bodies of highly specialized sensory neurons which are responsible for relaying information about our environment to the central nervous system. Despite previous efforts to characterize sensory neurons at the molecular level, it is still unknown whether those present in DRG and TG have distinct expression profiles and therefore a unique molecular fingerprint. To address this question, we isolated lumbar DRG and TG neurons using fluorescence-activated cell sorting from Advillin-GFP transgenic mice and performed RNA sequencing. Our transcriptome analyses showed that, despite being overwhelmingly similar, a number of genes are differentially expressed in DRG and TG neurons. Importantly, we identified 24 genes which were uniquely expressed in either ganglia, including an arginine vasopressin receptor and several homeobox genes, giving each population a distinct molecular fingerprint. We compared our findings with published studies to reveal that many genes previously reported to be present in neurons are in fact likely to originate from other cell types in the ganglia. Additionally, our neuron-specific results aligned well with a dataset examining whole human TG and DRG. We propose that the data can both improve our understanding of primary afferent biology and help contribute to the development of drug treatments and gene therapies which seek targets with unique or restricted expression patterns.