Enabling genome editing in tropical maize lines through an improved, morphogenic regulator-assisted transformation protocol.

The recalcitrance exhibited by many maize (Zea mays) genotypes to traditional genetic transformation protocols poses a significant challenge to the large-scale application of genome editing (GE) in this major crop species. Although a few maize genotypes are widely used for genetic transformation, they prove unsuitable for agronomic tests in field trials or commercial applications. This challenge is exacerbated by the predominance of transformable maize lines adapted to temperate geographies, despite a considerable proportion of maize production occurring in the tropics. Ectopic expression of morphogenic regulators (MRs) stands out as a promising approach to overcome low efficiency and genotype dependency, aiming to achieve 'universal' transformation and GE capabilities in maize. Here, we report the successful GE of agronomically relevant tropical maize lines using a MR-based, Agrobacterium-mediated transformation protocol previously optimized for the B104 temperate inbred line. To this end, we used a CRISPR/Cas9-based construct aiming at the knockout of the VIRESCENT YELLOW-LIKE (VYL) gene, which results in an easily recognizable phenotype. Mutations at VYL were verified in protoplasts prepared from B104 and three tropical lines, regardless of the presence of a single nucleotide polymorphism (SNP) at the seed region of the VYL target site in two of the tropical lines. Three out of five tropical lines were amenable to transformation, with efficiencies reaching up to 6.63%. Remarkably, 97% of the recovered events presented indels at the target site, which were inherited by the next generation. We observed off-target activity of the CRISPR/Cas9-based construct towards the VYL paralog VYL-MODIFIER, which could be partly due to the expression of the WUSCHEL (WUS) MR. Our results demonstrate efficient GE of relevant tropical maize lines, expanding the current availability of GE-amenable genotypes of this major crop.

Impact of cigarette smoke on osteogenic and osteoclast signaling in middle palatal suture.

Considering that smoking is a public health problem that has been growing among adolescents, the aim of this study was to investigate the impact of cigarette smoke on osteogenic and osteoclastogenic signaling in middle palatal suture of rats. Male Wistar rats exposed (n = 30) or not to cigarette smoke (n = 30) were used. Exposure to smoke was carried out for two daily periods of 3 minutes each, with an interval of 12 hours between exposures. After the experimental periods of 3, 7, 14 and 21 days, the animals were euthanized. The collected tissues were analyzed using light microscopy and real-time RT-PCR was performed to investigate gene expression. The data obtained were compared using the Kruskal Wallis and Dunn tests (⍺ = 5%). Morphologically, there were no significant changes in the middle palatal suture of rats exposed or not to cigarette smoke during 3, 7, 14 and 21 days (p> 0.05). On the other hand, osteoclastogenic signaling was increased in animals exposed to smoke and was characterized by a higher production of RANKL at 3 and 14 days (p <0.05), with no change in the synthesis of RANK and osteoprotegerin (p> 0.05). Interestingly, in the exposed animals, an early increase in the synthesis of osteocalcin, bone sialoprotein and osteopontin was also identified at 3 days of exposure (p <0.05), not sustained over time (p> 0.05). Cigarette smoke modulates osteogenic and osteoclastogenic signaling in the middle palatal suture of young rats, although morphological changes have not been evidenced.

Impact of cigarette smoke on osteogenic and osteoclast signaling in middle palatal suture

Abstract Considering that smoking is a public health problem that has been growing among adolescents, the aim of this study was to investigate the impact of cigarette smoke on osteogenic and osteoclastogenic signaling in middle palatal suture of rats. Male Wistar rats exposed (n = 30) or not to cigarette smoke (n = 30) were used. Exposure to smoke was carried out for two daily periods of 3 minutes each, with an interval of 12 hours between exposures. After the experimental periods of 3, 7, 14 and 21 days, the animals were euthanized. The collected tissues were analyzed using light microscopy and real-time RT-PCR was performed to investigate gene expression. The data obtained were compared using the Kruskal Wallis and Dunn tests (⍺ = 5%). Morphologically, there were no significant changes in the middle palatal suture of rats exposed or not to cigarette smoke during 3, 7, 14 and 21 days (p> 0.05). On the other hand, osteoclastogenic signaling was increased in animals exposed to smoke and was characterized by a higher production of RANKL at 3 and 14 days (p <0.05), with no change in the synthesis of RANK and osteoprotegerin (p> 0.05). Interestingly, in the exposed animals, an early increase in the synthesis of osteocalcin, bone sialoprotein and osteopontin was also identified at 3 days of exposure (p <0.05), not sustained over time (p> 0.05). Cigarette smoke modulates osteogenic and osteoclastogenic signaling in the middle palatal suture of young rats, although morphological changes have not been evidenced.

Resumo Considerando que a fumaça de cigarro é um problema de saúde pública que está crescendo entre os adolescentes, o objetivo deste estudo foi investigar o impacto da fumaça de cigarro na sinalização osteogênica e osteoclastogênica da sutura palatina mediana de ratos. Foram utilizados ratos Wistar machos expostos (n=30) e não expostos à fumaça de cigarro (n=30). A exposição à fumaça de cigarro foi realizada duas vezes ao dia por 3 minutos, com um intervalo de 12 horas entre as exposições. Os animais foram mortos após o período experimental de 3, 7, 14 e 21 dias. Os tecidos coletados foram analisados em microscópico de luz e pelo RT-PCR em tempo real foi realizado para investigar a expressão gênica. s dados obtidos foram comparados usando o testes de Kruskal Wallis e Dunn (⍺ = 5%). Morfoligicamente, não houve mudança significativa na sutura palatina mediana nos ratos expostos ou não à fumaça de cigarro durante os tempo de 3, 7, 14 e 21 dias (p> 0.05). Por outro lado, a sinalização osteogênica esta aumentada nos animais expostos à fumaça e foi caracterizado por um aumento da produção de RANKL aos 3 e 14 dias (p <0.05), sem mudança na síntese da produção de RANK e osteoprotegerina (p> 0.05). Curiosamente, nos animais expostos, também foi observado um aumento precoce da síntese de osteocalcina, sialoproteína óssea e de osteopontina aos 3 dias de exposição, o que não foi mantido ao longo do tempo. A fumaça de cigarro modula a sinalização osteogênica e osteoclastogênica na sutura palatina mediana de ratos jovens, apesar de não tenha sido evidenciado alterações morfológicas.

Prevalence of Dengue, Chikungunya and Zika Viruses in Blood Donors in the State of Pará, Northern Brazil: 2018-2020.

Arboviruses have been reported over the years as constant threats to blood transfusion recipients, given the high occurrence of asymptomatic cases and the fact that the presence of viremia precedes the onset of symptoms, making it possible that infected blood from donors act as a source of dissemination. This work aims to identify the prevalence of dengue virus (DENV), Zika virus (ZIKV) and Chikungunya virus (CHIKV) infection in blood donors during epidemic and non-epidemic periods; classify the donor as symptomatic or asymptomatic; and verify the need to include DENV, CHIKV and ZIKV in the nucleic acid test (NAT) platform in northern Brazil. We investigated 36,133 thousand donations in two years of collection in Northern Brazil. One donor was positive for DENV and one for CHIKV (0.002% prevalence). As the prevalence for arboviruses was low in this study, it would not justify the individual screening of samples from donors in a blood bank. Thus, DENV- and CHIKV-positive samples were simulated in different amounts of sample pools, and both were safely detected by molecular biology even in a pool of 14 samples, which would meet the need to include these three viruses in the routine of blood centers in endemic countries such as Brazil.

In Silico Insights into the Mechanism of Action of Epoxy-α-Lapachone and Epoxymethyl-Lawsone in Leishmania spp.

Epoxy-α-lapachone (Lap) and Epoxymethyl-lawsone (Law) are oxiranes derived from Lapachol and have been shown to be promising drugs for Leishmaniases treatment. Although, it is known the action spectrum of both compounds affect the Leishmania spp. multiplication, there are gaps in the molecular binding details of target enzymes related to the parasite's physiology. Molecular docking assays simulations were performed using DockThor server to predict the preferred orientation of both compounds to form stable complexes with key enzymes of metabolic pathway, electron transport chain, and lipids metabolism of Leishmania spp. This study showed the hit rates of both compounds interacting with lanosterol C-14 demethylase (-8.4 kcal/mol to -7.4 kcal/mol), cytochrome c (-10.2 kcal/mol to -8.8 kcal/mol), and glyceraldehyde-3-phosphate dehydrogenase (-8.5 kcal/mol to -7.5 kcal/mol) according to Leishmania spp. and assessed compounds. The set of molecular evidence reinforces the potential of both compounds as multi-target drugs for interrupt the network interactions between parasite enzymes, which can lead to a better efficacy of drugs for the treatment of leishmaniases.

VEGF and FGF-2 Released In Palatal Suture after Rapid Maxillary Expansion (RME).

Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (FGF-2) have the ability to increase vascular proliferation and permeability. The aim of this study was to quantify the release of two diffusible angiogenic growth factors (VEGF and FGF-2) after rapid maxillary expansion (RME). Thirty animals were randomly assigned to two groups. Control group (5 rats - intact suture) and Experimental groups (25 rats with RME) which were evaluated in different periods of treatment. Five animals were euthanized in different periods of healing at 0, 1, 2, 3, 5 and 7 days after RME. RT-PCR was used to evaluate the gene expression of angiogenic growth factors released on different periods of study. Data were submitted to statistical analysis using ANOVA followed by Tukey test and significance was assumed at a=0.05. RT-PCR showed that mRNAs of VEGF and FGF-2 were expressed in intact palatal suture tissue. mRNAs of VEGF and FGF-2 was upregulated in early periods (24 h) after RME (p<0.001 and p<0.01, respectively). The molecular levels of VEGF never returned to its original baseline values, and FGF-2 expression decreased up to day 5 (p<0.001) and suddenly increased at day 7, returning to its original level. RME increased VEGF secretion, but decreased FGF-2 secretion when compared to intact tissue. The results showed that these angiogenic growth factors are released and regulated in the palatal suture tissue after RME and could make an important contribution to the knowledge of overall reparative response of the suture tissue during the bone remodeling process.

Effects of experimental conditions on solubility measurements for BCS classification in order to improve the biowaiver guidelines

Among the methods described for determining the solubility, shake-flask is suitable to evaluate the equilibrium solubility according to the BCS. Nevertheless, experimental conditions related to the shake-flask method are not well described. Evaluating the effects of experimental conditions on solubility measurements by shake-flask method is important and contributes in biowaiver decision. For this work, propranolol hydrochloride and nimesulide were used as model compound of high and low solubility, respectively. Equilibrium solubility was evaluated at 37 ºC, 100 rpm during 48 hours in buffer media. Effects of the rotation speed, temperature, substance in excess and aliquot withdrawn were evaluated. Small variations of temperature caused significant differences in the solubility and then this parameter must be controlled. Excess of raw material influenced the results of the nimesulide, then, little excess is recommended. Rotation speed did not cause differences in the equilibrium solubilities, but at 150 rpm the equilibrium was reached faster. Aliquot did not present significant differences, but excessive withdrawn should be avoided. Therefore, the evaluation of equilibrium solubility using shake-flask method must be performed in physiological pH conditions, 37 ± 1 ºC, substance in excess 10% above saturation, 50, 100 or 150 rpm and aliquot withdrawn not more than 10% of the media volume.

A unidade afetivo-cognitiva: aspectos conceituais e metodológicos a partir da psicologia histórico-cultural / The affective-cognitive unity: theoretical and methodological aspects from the perspective of cultural-historical psychology / La unidad afectivo-cognitiva: aspectos conceptuales y metodológicos a partir de la psicología histórico-cultural

Este estudo teórico-metodológico teve por objetivo sistematizar o conceito de unidade afetivo-cognitiva a partir da Psicologia Histórico-Cultural. Esse objetivo se justifica pela escassez de estudos acerca do tema na Psicologia e pela observância de que há uma predominância de visões dualistas acerca da razão e da emoção nos estudos psicológicos. Nesse sentido, o estudo da unidade afetivo-cognitiva pode trazer acréscimos à Psicologia por discutir os aspectos metodológicos da cisão razão/emoção e por evidenciar a união entre esses processos como parte essencial do processo humano de apreensão da realidade. Sendo assim, buscou-se discutir a constituição da consciência humana e da estrutura da atividade como unidades de análise da unidade afetivo-cognitiva. A partir de Leontiev e Vigotski, analisou-se a estrutura da atividade humana e sua expressão pelos significados sociais e sentidos pessoais como unidade afetivo-cognitiva. Essa investigação resultou na constatação de que a estrutura da atividade e a constituição da consciência humana demandam funções afetivo-cognitivas para formar a imagem subjetiva da realidade objetiva no psiquismo humano. Por isso, indicou-se, conforme afirma Vigotski, que entender a unidade afetivo-cognitiva como sistema semântico da consciência demanda o destrinchamento da relação entre a atividade humana e a forma como o ser torna essa atividade consciente.

This theoretical-methodological study aimed to systematize the concept of affective-cognitive unity in Historical-Cultural Psychology. This objective is justified by the scarcity of studies on the subject in Psychology and based on the observation that there is a predominance of dualistic views on reason and emotion in psychological studies. In this sense, the study of the affective--cognitive unit can present improvements to Psychology by discussing the methodological aspects of the reason/emotion split and for making evident the union between these processes as an essential part of the human process of apprehension of reality. Thus, we sought to discuss the constitution of human consciousness and the structure of activity as units of analysis of the affective--cognitive unity. Based on Leontiev and Vygotsky, we analyzed the structure of human activity and its expression by social meanings and personal senses as an affective-cognitive unit. This investigation resulted in the realization that the structure of activity and the constitution of human consciousness demand affective-cognitive functions to form the subjective image of objective reality in the human psyche. Therefore, it was pointed out, as Vygotsky states, that understanding the affective-cognitive unity as a semantic system of consciou-sness requires the study of how human activity relates to the way that the human being is aware and conscious of this activity.

Este estudio, teórico-metodológico, tuvo como objetivo sistematizar el concepto de unidad afectivo-cognitiva a partir de la Psicología Histórico-Cultural. Este objetivo se justifica por la escasez de estudios acerca del tema en la Psicología y por la observancia de que hay un predominio de visiones dualistas acerca de la razón y la emoción en los estudios psicológicos. En este sentido, el estudio de la unidad afectivo-cognitiva puede incrementar a la Psicología por discutir los aspectos metodológicos de la escisión razón / emoción y por evidenciar, la unión entre estos procesos como parte esencial del proceso humano de aprehensión de la realidad. Siendo así, se buscó discutir la constitución de la conciencia humana y de la estructura de la actividad como unidades de análisis de la unidad afectivo-cognitiva. A partir de Leontiev y Vigotski, se analizó la estructura de la actividad humana y su expresión por los significados sociales y sentidos personales como unidad afectivo-cognitiva. Esta investigación resultó en la constatación de que la estructura de la actividad y la constitución de la conciencia humana demandan funciones afectivo-cognitivas para formar la imagen subjetiva de la realidad objetiva en el psiquismo humano. Por eso, se indica, según afirma Vigotski, que entender la unidad afectivo-cognitiva como sistema semántico de la conciencia, demanda desmenuzar la relación entre la actividad humana y la forma como el ser hace esa actividad consciente.

Human pegivirus (HPgV, GBV-C) RNA in volunteer blood donors from a public hemotherapy service in Northern Brazil.

BACKGROUND: Human pegivirus (HPgV)-formerly known as GBV-C-is a member of the Flaviviridae family and belongs to the species Pegivirus C. It is a non-pathogenic virus and is transmitted among humans mainly through the exposure to contaminated blood and is often associated with human immunodeficiency virus (HIV) infection, among other viruses. This study aimed to determine the prevalence of HPgV viremia, its association with HIV and clinical epidemiological factors, as well as the full-length sequencing and genome characterization of HPgV recovered from blood donors of the HEMOPA Foundation in Belém-PA-Brazil. METHODS: Plasma samples were obtained from 459 donors, tested for the presence of HPgV RNA by the RT-qPCR. From these, a total of 26 RT-qPCR positive samples were submitted to the NGS sequencing approach in order to obtain the full genome. Genome characterization and phylogenetic analysis were conducted. RESULTS: The prevalence of HPgV was 12.42%. We observed the highest prevalences among donors aged between 18 and 30 years old (16.5%), with brown skin color (13.2%) and men (15.8%). The newly diagnosed HIV-1 prevalence was 26.67%. The HPgV genotype 2 (2a and 2b) was identified. No data on viral load value was found to corroborate the protective effect of HPgV on HIV evolution. CONCLUSIONS: This study provided information regarding the HPgV infection among blood donors from HEMOPA Foundation. Furthermore, we genetically characterized the HPgV circulating strains and described by the first time nearly complete genomes of genotype 2 in Brazilian Amazon.

The Effect of Cigarette Smoking And Low-Level Laser Irradiation in RANK/RANKL/OPG Expression.

The objective of this study was to investigate the effects of low-level laser therapy (LLLT) and cigarette smoke on alveolar socket osteoclastogenesis signaling after tooth extraction, in rats. Sixty male Wistar rats were randomly assigned to four groups with 15 animals each: Control Group (with right maxillary molar extraction - ME), Experimental I (with ME and LLLT), Experimental II (with ME and cigarette smoke) and Experimental III group (with ME, LLLT and cigarette smoke). Euthanasia was performed at 3, 7 and 14 days postoperative. qRT-PCR was used to evaluate expression of Tnfrsf11a (RANK), Tnfsf11 (Rankl) and Tnfrsf11b (OPG). Data were submitted to statistical analysis using two-way ANOVA followed by Bonferroni test (α=0.05). There was an upregulation of RANK, RANKL and OPG genes over all the time of healing in Exp I group compared to control group. Exp II group showed a decreased expression of all genes over time, whereas Exp III genes expression were higher than Exp II values but lower than Control and Exp I values over time. The results of this study concluded that the LLLT had a positive effect, whereas cigarette smoke had a negative effect on RANK, RANKL and OPG gene expression in bone remodeling process.

Dental tooth decay profile in an institutionalized elder population of Northern Portugal / Perfil de cárie dentária numa população idosa institucionalizada do Norte de Portugal

Objectives: the purpose of this study was to assess the dental tooth decay profile of an elderly population from Northern Portugal. Material and Methods: this is a descriptive, cross-sectional study where a questionnaire, intended to assess the institutionalized elders oral health condition, was applied with the purpose of quantifying certain oral conditions. Tooth decay was assessed using the DMFT Index for the crown's tooth and the individual radicular caries index (RCIi) for the root's condition assessment. Univariate analysis and multivariable logistical regressions were performed (p < 0.05) using the IBM© SPSS© Statistics vs.19.0. Results: three hundred and seventy-two elderly participants were assessed with an average age of 78.8 years, of which 260 were women (69.9%). Of the sample 30.4% had no literacy and 18.3% were dependent on their daily oral hygiene care, 30.9% (95%CI:26.2%-35.6%) were edentulous and 84.9% (95%CI:81.3%-88.5%) had less than 20 teeth. The average number of teeth was 8.9 (± 8.7) and the DMFT was 25.6 (± 7.3). The RCIi for men was 44.3% (± 30.4%) and 39.4% (± 31.2%) for women. Using multivariate logistical regression model the outcome RCIi≥20% was shown to be significantly and positively associated with the use of removable partial dentures (metal partial dentures: OR = 5.348 (95% CI: 1.176-24,329; p = 0.030) as well as with daily tooth brushing (no toothbrushing: OR = 2.802 (95% CI: 1.445-5.433); p = 0.002). Conclusion: the prevalence of tooth decay in this elder institutionalized population of Northern Portugal is high, mostly due to bad oral hygiene habits associated to the use of metal removable partial dentures. Institutionalized elderly need protocols tailored to their oral health needs and integrated efforts in the institutions are needed to effectively answer the demands related to the elderly cognitive and motor skills (ageing deterioration) decline of physiological functions.(AU)

Objetivos: O objetivo deste estudo foi avaliar o perfil de cárie dentária numa população idosa do norte de Portugal. Material e Métodos: Trata-se de um estudo descritivo, transversal, onde foi aplicado um questionário destinado a avaliar a condição de saúde oral dos idosos institucionalizados, com o objetivo de quantificar determinadas condições orais. A cárie dentária foi avaliada usando o índice CPOD para a coroa do dente e o índice de cárie radicular individual (IRCi) para a avaliação da condição da raiz. Foi realizada análise univariada e regressão logística multivariável (p <0,05) usando o IBM© SPSS© Statistics vs.19.0. Resultados: avaliaram-se 372 idosos, com idade média de 78,8 anos, dos quais 260 eram mulheres (69,9%). Da amostra, 30,4% não tinham escolaridade e 18,3% dependiam de outros para realizar os seus cuidados diários de higiene oral; 30,9% (IC95%: 26,2%-35,6%) eram desdentados e 84,9% (IC95%: 81,3%-88,5%) tinham menos de 20 dentes. O número médio de dentes foi de 8,9 (± 8,7) e o CPOD foi de 25,6 (± 7,3). O ICRi para o género masculino foi de 44,3% (± 30,4%) e de 39,4% (± 31,2%) para o feminino. Usando o modelo de regressão logística multivariada, o resultado ICRi≥20% mostrou-se significativa e positivamente associado ao uso de próteses parciais removíveis (próteses parciais esqueléticas: OR= 5,348 (IC95%: 1,176-24,329; p = 0,030), bem como com escovagem diária (sem escovagem: OR= 2,802 (IC95%: 1,445-5,433); p = 0,002). Conclusão: a prevalência de cárie dentária nesta população idosa institucionalizada do Norte de Portugal é alta, principalmente devido à má higiene oral e hábitos associados ao uso de próteses parciais removíveis esqueléticas. Idosos institucionalizados precisam de protocolos adaptados às suas necessidades de saúde oral e esforços integrados nas instituições para atender efetivamente às necessidades relacionadas com o declínio das funções fisiológicas do idoso, cognitivas e motoras (deterioração do envelhecimento). (AU)

Neoechinorhynchus buttnerae parasitic infection in tambaqui (Colossoma macropomum) on fish farms in the state of Amazonas / Parasitismo por Neoechinorhynchus buttnerae em tambaqui (Colossoma macropomum) de pisciculturas do Estado do Amazonas

The record of occurrence of acanthocephalan parasites on tambaqui farms in Northern Brazil has increased in recent years along with reports of associated economic losses, justifying further epidemiological studies of this parasite. As such, this study evaluated the occurrence of the acanthocephalan Neoechinorhynchus buttnerae and the parasite-host relationship in tambaqui (Colossoma macropomum) from three fish farms in the Amazonian town of Rio Preto da Eva, Brazil. Fish from the farms (N = 18 per farm) were collected for parasitological examination, determination of the relative condition factor and correlation tests between the parameters assessed. Of the fish examined, 37 were parasitized with N. buttnerae (68.5% prevalence), with intensity of infection variation ranging from 1 to 1,219 specimens per fish (12,279 specimens counted). In two farms was detected the high mean intensity of infection, 230.8 and 451.3 parasites per fish, with a total count of 4,155 and 8,123 parasites, respectively, whereas the third farm had a single fish infected with only one specimen. The weight-length ratio equation indicated that the fish were growing at the same rate in weight and length. It was observed significant correlation between parasite intensity and fish length, as well as significant negative correlation with parasite intensity and condition factor. The gastrointestinal tract contained ostracod invertebrates, which are intermediate N. buttnerae hosts. The high N. buttnerae intensity affect their physiological condition as suggested by the negative correlation between parasitic intensity and condition factor, emphasizing the need to implement suitable prophylactic measures to prevent infection by the parasite.

Nos últimos anos tem crescido o registro de ocorrências de parasitos acantocéfalos em pisciculturas de tambaqui da região Norte do Brasil, com relatos de perdas econômicas, justificando assim a necessidade de aprofundar dados epidemiológicos desta doença parasitária. Assim, o objetivo deste estudo foi avaliar a ocorrência do acantocéfalo Neoechinorhynchus buttnerae e a relação parasito-hospedeiro em tambaqui (Colossoma macropomum) de três pisciculturas do município de Rio Preto da Eva (AM), Brasil. Foram amostradas três pisciculturas no município alvo, coletando 18 peixes em cada uma, para análises parasitológicas e cálculo do fator de condição relativo e correlações associadas. Foram quantificados 12.279 espécimes de N. buttnerae em 37 peixes parasitados (prevalência 68,5%), com intensidade de infecção variando de 1 a 1.219 parasitos por peixe. Duas propriedades apresentaram as maiores intensidades médias de infecção, 230,8 e 451,3 parasitas por peixe; com 4.155 e 8.123 parasitos quantificados, respectivamente, enquanto na terceira foi encontrado apenas 1 parasito em 1 peixe infectado. A equação da relação peso-comprimento indicou que os peixes estavam crescendo na mesma proporção em peso e comprimento. Foi observada correlação significativa entre intensidade parasitária e comprimento dos peixes, assim como correlação negativa significativa entre intensidade parasitária e fator de condição. No trato gastrointestinal dos tambaquis foi observada a presença de invertebrados ostracodas; hospedeiros intermediários desta espécie de acantocéfalo. Portanto, a alta carga parasitária de N. buttnerae pode ter comprometido o desempenho dos peixes e afetado a sua condição fisiológica como sugerido pela correlação negativa entre intensidade parasitária e fator de condição, e enfatiza-se a importância do emprego de medidas adequadas de profilaxia para prevenção de infecções por este parasito.

Neoechinorhynchus buttnerae parasitic infection in tambaqui (Colossoma macropomum) on fish farms in the state of Amazonas / Parasitismo por Neoechinorhynchus buttnerae em tambaqui (Colossoma macropomum) de pisciculturas do Estado do Amazonas

The record of occurrence of acanthocephalan parasites on tambaqui farms in Northern Brazil has increased in recent years along with reports of associated economic losses, justifying further epidemiological studies of this parasite. As such, this study evaluated the occurrence of the acanthocephalan Neoechinorhynchus buttnerae and the parasite-host relationship in tambaqui (Colossoma macropomum) from three fish farms in the Amazonian town of Rio Preto da Eva, Brazil. Fish from the farms (N = 18 per farm) were collected for parasitological examination, determination of the relative condition factor and correlation tests between the parameters assessed. Of the fish examined, 37 were parasitized with N. buttnerae (68.5% prevalence), with intensity of infection variation ranging from 1 to 1,219 specimens per fish (12,279 specimens counted). In two farms was detected the high mean intensity of infection, 230.8 and 451.3 parasites per fish, with a total count of 4,155 and 8,123 parasites, respectively, whereas the third farm had a single fish infected with only one specimen. The weight-length ratio equation indicated that the fish were growing at the same rate in weight and length. It was observed significant correlation between parasite intensity and fish length, as well as significant negative correlation with parasite intensity and condition factor. The gastrointestinal tract contained ostracod invertebrates, which are intermediate N. buttnerae hosts. The high N. buttnerae intensity affect their physiological condition as suggested by the negative correlation between parasitic intensity and condition factor, emphasizing the need to implement suitable prophylactic measures to prevent infection by the parasite.(AU)

Nos últimos anos tem crescido o registro de ocorrências de parasitos acantocéfalos em pisciculturas de tambaqui da região Norte do Brasil, com relatos de perdas econômicas, justificando assim a necessidade de aprofundar dados epidemiológicos desta doença parasitária. Assim, o objetivo deste estudo foi avaliar a ocorrência do acantocéfalo Neoechinorhynchus buttnerae e a relação parasito-hospedeiro em tambaqui (Colossoma macropomum) de três pisciculturas do município de Rio Preto da Eva (AM), Brasil. Foram amostradas três pisciculturas no município alvo, coletando 18 peixes em cada uma, para análises parasitológicas e cálculo do fator de condição relativo e correlações associadas. Foram quantificados 12.279 espécimes de N. buttnerae em 37 peixes parasitados (prevalência 68,5%), com intensidade de infecção variando de 1 a 1.219 parasitos por peixe. Duas propriedades apresentaram as maiores intensidades médias de infecção, 230,8 e 451,3 parasitas por peixe; com 4.155 e 8.123 parasitos quantificados, respectivamente, enquanto na terceira foi encontrado apenas 1 parasito em 1 peixe infectado. A equação da relação peso-comprimento indicou que os peixes estavam crescendo na mesma proporção em peso e comprimento. Foi observada correlação significativa entre intensidade parasitária e comprimento dos peixes, assim como correlação negativa significativa entre intensidade parasitária e fator de condição. No trato gastrointestinal dos tambaquis foi observada a presença de invertebrados ostracodas; hospedeiros intermediários desta espécie de acantocéfalo. Portanto, a alta carga parasitária de N. buttnerae pode ter comprometido o desempenho dos peixes e afetado a sua condição fisiológica como sugerido pela correlação negativa entre intensidade parasitária e fator de condição, e enfatiza-se a importância do emprego de medidas adequadas de profilaxia para prevenção de infecções por este parasito.(AU)

Co-exposure to titanium dioxide nanoparticles (NpTiO2) and lead at environmentally relevant concentrations in the Neotropical fish species Hoplias intermedius.

Growing production and utilization of titanium dioxide nanoparticles (NpTiO2) invariably lead to their accumulation in oceans, rivers and other water bodies, thus increasing the risk to the welfare of this ecosystem. The progressive launch of these nanoparticles in the environment has been accompanied by concern in understanding the dynamics and the toxic effect of these xenobiotic in different ecosystems, either on their own or in tandem with different contaminants (such as organic compounds and heavy metals), possibly altering their toxicity. Nevertheless, it remains unknown if these combined effects may induce damage in freshwater organisms. Therefore, this study aimed to analyze the consequences caused by NpTiO2, after a waterborne exposure of 96 h to a Neotropical fish species Hoplias intermedius, as well as after a co-exposure with lead, whose effects for fish have already been well described in the literature. The characterization of NpTiO2 stock suspension was carried out in order to provide additional information and revealed a stable colloidal suspension. As a result, NpTiO2 showed some genotoxic effects which were observed by comet assay in gill, kidney and brain cells. Also, the activity of brain acetylcholinesterase (AChE) has not changed, but the activity of muscle AChE decreased in the group exposed only to PbII. Regarding the hepatic antioxidant system, catalase (CAT) did not show any change in its activity, whereas that of superoxide dismutase (SOD) intensified in the groups submitted only to PbII and NpTiO2 alone. As for lipid peroxidation, there was a decrease in the group exposed to the NpTiO2 alone and to the co-exposed group (NpTiO2+PbII). As far as metallothionein is concerned, its concentration rose for the co-exposed group (NpTiO2+PbII) and for the group exposed to PbII alone. Overall, we may conclude that NpTiO2 alone caused DNA damage to vital tissues. Also, some impairment related to the antioxidant mechanism was described but it is probably not related to the DNA damage observed, suggesting that the genotoxic effect observed may be due to a different mechanism instead of ROS production.

RANK/RANKL/OPG Expression in Rapid Maxillary Expansion

Abstract The aim of this study was to evaluate osteoclastogenesis signaling in midpalatal suture after rapid maxillary expansion (RME) in rats. Thirty male Wistar rats were randomly assigned to two groups with 15 animals each: control (C) and RME group. RME was performed by inserting a 1.5-mm-thick circular metal ring between the maxillary incisors. The animals were euthanized at 3, 7 and 10 days after RME. qRT-PCR was used to evaluate expression of Tnfsf11 (RANKL), Tnfrsf11a (RANK) and Tnfrsf11b (OPG). Data were submitted to statistical analysis using two-way ANOVA followed by Tukey test (a=0.05). There was an upregulation of RANK and RANKL genes at 7 and 10 days and an upregulation of the OPG gene at 3 and 7 days of healing. Interestingly, an increased in expression of all genes was observed over time in both RME and C groups. The RANKL/OPG ratio showed an increased signaling favoring bone resorption on RME compared to C at 3 and 7 days. Signaling against bone resorption was observed, as well as an upregulation of OPG gene expression in RME group, compared to C group at 10 days. The results of this study concluded that the RANK, RANK-L and OPG system participates in bone remodeling after RME.

Resumo O objetivo deste estudo foi avaliar a sinalização osteoclastogenese na sutura palatina após a expansão rápida da maxila (ERM) em ratos. Um total de 30 ratos Wistar machos foram divididos aleatoriamente em dois grupos com 15 animais cada: controle (C) e grupo ERM. ERM foi realizada através da inserção de um anel de metal circular de 1,5 mm de espessura entre os incisivos superiores. Os animais foram sacrificados aos 3, 7 e 10 dias após a RME. qRT-PCR foi utilizado para avaliar a expressão de Tnfsf11 (RANKL), Tnfrsf11a (RANK) e TNFRSF11b (OPG). Os dados foram submetidos à análise de variância de duas vias, seguido pelo teste de Tukey (a=0,05). Houve uma regulação positiva de genes RANK e RANKL aos 7 e 10 dias e uma regulação positiva do gene OPG aos 3 e 7 dias de tratamento. Curiosamente, foi observado um aumento na expressão de todos os genes ao longo do tempo nos grupos ERM e C. O RANKL/OPG mostrou um aumento na sinalização favorecendo a reabsorção óssea no ERM em comparação com o C nos períodos de 3 e 7 dias. Foi observada uma sinalização contra a reabsorção óssea, assim como, uma regulação favorável da expressão do gene OPG no grupo ERM, comparado ao grupo C aos 10 dias. Os resultados deste estudo permitem concluir que o sistema RANK, RANK-L e OPG participa de remodelação óssea após a ERM.

RANK/RANKL/OPG Expression in Rapid Maxillary Expansion.

The aim of this study was to evaluate osteoclastogenesis signaling in midpalatal suture after rapid maxillary expansion (RME) in rats. Thirty male Wistar rats were randomly assigned to two groups with 15 animals each: control (C) and RME group. RME was performed by inserting a 1.5-mm-thick circular metal ring between the maxillary incisors. The animals were euthanized at 3, 7 and 10 days after RME. qRT-PCR was used to evaluate expression of Tnfsf11 (RANKL), Tnfrsf11a (RANK) and Tnfrsf11b (OPG). Data were submitted to statistical analysis using two-way ANOVA followed by Tukey test (a=0.05). There was an upregulation of RANK and RANKL genes at 7 and 10 days and an upregulation of the OPG gene at 3 and 7 days of healing. Interestingly, an increased in expression of all genes was observed over time in both RME and C groups. The RANKL/OPG ratio showed an increased signaling favoring bone resorption on RME compared to C at 3 and 7 days. Signaling against bone resorption was observed, as well as an upregulation of OPG gene expression in RME group, compared to C group at 10 days. The results of this study concluded that the RANK, RANK-L and OPG system participates in bone remodeling after RME.

A apropriação de conceitos científicos no contexto escolar e as pedagogias do aprender a aprender / The appropriation of scientific concepts in school context and the pedagogies of learning to learn / La apropiación de conceptos científicos en el contexto escolar y las pedagogías del aprender a aprender

O artigo objetiva discutir teoricamente como a escola atua na apropriação de conceitos científicos e como essa atuação pode se alienar. Justifica-se este estudo por trazer acréscimos à Psicologia e à Educação, evidenciando o pensamento por conceitos como elemento cognitivo essencial do processo de elaboração do conhecimento. A partir da Psicologia Histórico-Cultural evidencia-se o papel da escola na mediação do desenvolvimento dos conceitos espontâneos e científicos no sistema interfuncional da consciência. Além disso, como este papel mediador e este desenvolvimento podem se alienar por conta das tendências pedagógicas atuais, como as Pedagogias do Aprender a Aprender, consideradas a expressão ideológica da materialidade da sociedade de classes contemporânea. Por último, defende que a escola deve se ater ao conhecimento sistematizado e não ao conhecimento espontâneo; ao conhecimento fruto da prática e da teorização histórico-social humana e não ao conhecimento pragmático e construído individualmente; ao saber totalizante e não ao saber fragmentado.

The article aims to discuss theoretically how the school acts in the appropriation of scientific concepts and how this action can be alienated. This study is justified because it brings contribution to Psychology and Education, evidencing thinking by concepts as an essential cognitive element of the process of knowledge elaboration. From Historical-Cultural Psychology the role of the school in the mediation of the development of spontaneous and scientific concepts in the inter-functional system of consciousness is evidenced. Moreover, how this mediating role and this development can be alienated by present pedagogical tendencies, such as the Learning to Learn Pedagogies, considered the ideological expression of the materiality of the contemporary class society. Finally, he argues that the school should stick to systematized knowledge and not to spontaneous knowledge; to the knowledge derived from human historical and social theorization and practice, and not from individual pragmatic knowledge; to totalizing knowledge and not to fragmented knowledge.

El presente artículo tiene por objetivo discutir teóricamente cómo la escuela actúa en la apropiación de conceptos científicos y cómo esa actuación pude alienarse. Se justifica este estudio por traer adicción a la Psicología y a la Educación, evidenciando el pensamiento por conceptos como elemento cognitivo esencial del proceso de elaboración del conocimiento. A partir de la Psicología Histórico-Cultural se evidencia el papel de la escuela en la mediación del desarrollo de los conceptos espontáneos y científicos en el sistema inter-funcional de la consciencia. Además de eso, como este papel mediador y este desarrollo pueden alienarse por cuenta de las tendencias pedagógicas actuales, como las Pedagogías del Aprender a Aprender, consideradas la expresión ideológica de la materialidad de la sociedad de clases contemporánea. Por último, defiende que la escuela debe atenerse al conocimiento sistematizado y no al conocimiento espontáneo; al conocimiento fruto de la práctica y de la teorización histórico-social humana y no al conocimiento pragmático y construido individualmente; al saber totalizante y no al saber fragmentado.

Molecular design, synthesis and biological evaluation of 1,4-dihydro-4-oxoquinoline ribonucleosides as TcGAPDH inhibitors with trypanocidal activity.

The 1,4-dihydro-4-oxoquinoline ribonucleoside, Neq135, is the first low micromolar trypanosomatidae inhibitor to show good ligand efficiency (0.28 kcal mol(-1)atom(-1)) and good ligand lipophilicity efficiency (0.37 kcal mol(-1)atom(-1)) when acting against Trypanosoma cruzi glyceraldehyde 3-phosphate dehydrogenase (TcGAPDH). This and other six ribonucleosides were synthesized using our in-house technology, and assayed against the GAPDH NAD(+) site using isothermal titration calorimetry (ITC). Compound Neq135 had acceptable in vitro cytotoxicity, inhibited TcGAPDH with a Ki(app) value of 16 µM and killed the trypomastigote form of Trypanosoma cruzi Tulahuen strain with a concentration similar to that displayed by the control drug benznidazole. Neq135 is tenfold lower kinetic affinity against hGAPDH and does not kill Balb-c fibroblast nor spleen mouse cells. These results emphasize the possibility of integrating ligand- and target-based designs to uncover potent and selective TcGAPDH inhibitors that expands the opportunity for further medicinal chemistry endeavor towards NAD(+) TcGAPDH site.

Adhesion of four endodontic sealers to root dentin conditioned with different chemical solutions / Adesão de quatro cimentos endodônticos em dentina condicionada com diferentes soluções químicas

Objective - To evaluate in vitro adhesion of endodontic sealers Rickert, AH Plus, Sealapex and Epiphany to dentin previously conditioned with one of two different chemical solutions. Methods - A total of 80 specimens were made up with bovine incisive teeth that were prepared by cutting predetermined sections and wearing them, producing standard experimental samples. Forty samples were treated with 2.5% sodium hypochlorite and divided into four groups with 10 specimens each, to be coated with the different sealers. Dentin of other forty samples was treated with 17% EDTA, and these samples were also divided into four groups. Prepared specimens were hydrated uniformly, dried, bonded and kept at 37°C and 100% relative humidity. After 72 hours, specimens were analyzed using an Instron universal testing machine in traction mode. Data were analyzed by Kruskal-Wallis test (?=0.05). Results - EDTA treatment led to the greatest adhesiveness in all experimentalgroups. Conclusion - It was concluded that conditioning of the surface with chelating agents increase adhesiveness of the tested sealers and similar behavior was observed in the adhesion of epoxy-based and methacrylate-based sealers.

Objetivo - Avaliar in vitro a adesão dos cimentos endodônticos Rickert, AH Plus, Sealapex e Epiphany à dentina previamente tratada por duas soluções irrigadoras. Métodos - Foram empregados 80 corpos de prova obtidos a partir de dentes incisivos bovinos que foram preparados por cortes e desgastes pré-estabelecidos, resultando em modelos experimentais padronizados. Quarenta dessas amostras foram tratadas com hipoclorito de sódio 2,5% e subdivididas em quatro grupos de 10 espécimes para cada cimento. As demais 40 amostras tiveram a dentina tratada com EDTA a 17% e foram divididos igualmente em 4 grupos. Os espécimes preparados foram uniformemente hidratados, secos, receberam os cimentos, e foram mantidos sob umidade relativa a 37ºC. Após 72 horas os espécimes foram submetidos a testes de tração com auxilio da máquina de ensaio universal Instron, sendo os dados analisados pelo teste de Kruskal-Wallis (?=0.05). Resultados - O tratamento com EDTA proporcionou maiores índices de adesão para todos os grupos experimentais (p<0.05). Conclusão - Pode-se concluir que o tratamento da superfície com agente quelante, aumenta a capacidade de adesão dos cimentos testados. Não houve diferença na adesão dos cimentos a base de resina epóxica e a base de metacrilatos.