Neuroinflammation, blood-brain barrier dysfunction, hippocampal atrophy and delayed neurodevelopment: Contributions for a rat model of congenital Zika syndrome.

The congenital Zika syndrome (CZS) has been characterized as a set of several brain changes, such as reduced brain volume and subcortical calcifications, in addition to cognitive deficits. Microcephaly is one of the possible complications found in newborns exposed to Zika virus (ZIKV) during pregnancy, although it is an impacting clinical sign. This study aimed to investigate the consequences of a model of congenital ZIKV infection by evaluating the histopathology, blood-brain barrier, and neuroinflammation in pup rats 24 h after birth, and neurodevelopment of the offspring. Pregnant rats were inoculated subcutaneously with ZIKV-BR at the dose 1 × 107 plaque-forming unit (PFU mL-1) of ZIKV isolated in Brazil (ZIKV-BR) on gestational day 18 (G18). A set of pups, 24 h after birth, was euthanized. The brain was collected and later evaluated for the histopathology of brain structures through histological analysis. Additionally, analyses of the blood-brain barrier were conducted using western blotting, and neuroinflammation was assessed using ELISA. Another set of animals was evaluated on postnatal days 3, 6, 9, and 12 for neurodevelopment by observing the developmental milestones. Our results revealed hippocampal atrophy in ZIKV animals, in addition to changes in the blood-brain barrier structure and pro-inflammatory cytokines expression increase. Regarding neurodevelopment, a delay in important reflexes during the neonatal period in ZIKV animals was observed. These findings advance the understanding of the pathophysiology of CZS and contribute to enhancing the rat model of CZS.

Zika Virus affects neurobehavioral development, and causes oxidative stress associated to blood-brain barrier disruption in a rat model of congenital infection.

Zika virus (ZIKV) is a mosquito-borne flavivirus associated with several neurodevelopmental outcomes after in utero infection. Here, we studied a congenital ZIKV infection model with immunocompetent Wistar rats, able to predict disabilities and that could pave the way for proposing new effective therapies. We identified neurodevelopmental milestones disabilities in congenital ZIKV animals. Also, on 22nd postnatal day (PND), blood-brain barrier (BBB) proteins disturbances were detected in the hippocampus with immunocontent reduction of ß_Catenin, Occludin and Conexin-43. Besides, oxidative stress imbalance on hippocampus and cortex were identified, without neuronal reduction in these structures. In conclusion, even without pups' microcephaly-like phenotype, congenital ZIKV infection resulted in neurobehavioral dysfunction associated with BBB and oxidative stress disturbances in young rats. Therefore, our findings highlighted the multiple impact of the congenital ZIKV infection on the neurodevelopment, which reinforces the continuity of studies to understand the spectrum of this impairment and to provide support to future treatment development for patients affected by congenital ZIKV.

Resistance exercise was safe for the pregnancy and offspring's development and partially protected rats against early life stress-induced effects.

Promising evidence points to gestational physical exercise as the key to preventing various disorders that affect the offspring neurodevelopment, but there are no studies showing the impact of resistance exercise on offspring health. Thus, the aim of this study was to investigate whether resistance exercise during pregnancy is able to prevent or to alleviate the possible deleterious effects on offspring, caused by early life-stress (ELS). Pregnant rats performed resistance exercise throughout the gestational period:they climbed a sloping ladder with a weight attached to their tail, 3 times a week. Male and female pups, on the day of birth (P0), were divided into 4 experimental groups: 1) rats of sedentary mothers (SED group); 2) rats of exercised mothers (EXE group); 3) rats of sedentary mothers and submitted to maternal separation (ELS group) and 4) rats of exercised mothers and submitted to MS (EXE + ELS group). From P1 to P10, pups from groups 3 and 4 were separated from their mothers for 3 h/day. Maternal behavior was assessed. From P30, behavioral tests were performed and on P38 the animals were euthanized and prefrontal cortex samples were collected. Oxidative stress and tissue damage analysis by Nissl staining were performed. Our results demonstrate that male rats are more susceptible to ELS than females, showing impulsive and hyperactive behavior similar to that seen in children with ADHD. This behavior was attenuated by the gestational resistance exercise. Our results demonstrate, for the first time, that resistance exercise performed during pregnancy seems to be safe for the pregnancy and offspring's neurodevelopment and are effective in preventing ELS-induced damage only in male rats. Interestingly, resistance exercise during pregnancy improved maternal care and it is reasonable to propose that this finding may be related to the protective role on the animals neurodevelopment, observed in our study.

Vitamin D supplementation combined with aerobic physical exercise restores the cell density in hypothalamic nuclei of rats exposed to monosodium glutamate.

BACKGROUND & AIMS: In view of the increase in the prevalence of obesity and metabolic syndrome in childhood and adolescence, this study proposed the early and combined use of treatments to restore brain areas related to satiety. The vitamin D supplementation, aerobic exercise and the combination of these interventions on the structure of arcuate (ARC) and ventromedial (VMH) nuclei of hypothalamus were investigated in monosodium glutamate (MSG)-treated rats. METHODS: Wistar rats were separated into five groups: Control group (CT); Obese group injected with MSG (OB); Obese group supplemented with vitamin D (OBvd); Obese group submitted to forced swimming training (OBexe) and Obese group treated with vitamin D supplementation and forced swimming training (OBvd + exe). RESULTS: In the OB group, the visceral fat weight was significantly higher, there was a reduction in the number of glial cells in the ARC nucleus and also in the number of neurons in the ARC and VMH nuclei. Aerobic exercise was able to reduce the visceral fat weight in the OBexe group. The combination of treatments used in the OBvd + exe group reversed the loss of neurons and glial cells produced by MSG in the ARC nucleus. All treated groups exhibited a higher number of neurons in VMH nucleus, but an increase in the glial cells were observed only in the OBexe and OBvd + exe groups. CONCLUSIONS: The effectiveness of obesity treatment can be favored through the early and combined use of vitamin D supplementation and aerobic exercise, since these therapies are able to restore brain nuclei involved in the control of food intake.

Acrobatic exercise recovers object recognition memory impairment in hypoxic-ischemic rats.

Neonatal hypoxia-ischemia (HI) can lead to cognitive impairments and motor dysfunction. Acrobatic exercises (AE) were proposing as therapeutic option to manage HI motor deficits, however, the cognitive effects after this treatment are still poorly understood. Therefore, we evaluated the effects of AE protocol on memory impairments and brain plasticity markers after Rice-Vannucci HI rodent model. Wistar rats on the 7th postnatal day (PND) were submitted to HI model and after weaning (PND22) were trained for 5 weeks with AE protocol, then subsequently submitted to cognitive tests. Our results showed recovery in novel object recognition (NOR) memory, but not, spatial Morris Water Maze (WM) memory after AE treatment in HI rats. BDNF and synaptophysin neuroplasticity markers indicate plastic alterations in the hippocampus and striatum, with maintenance of synaptophysin despite the reduction of total volume tissue, besides, hippocampal HI-induced ipsilateral BDNF increased, and striatum contralateral BDNF decreased were noted. Nevertheless, the exercise promoted functional recovery and seems to be a promising strategy for HI treatment, however, future studies identifying neuroplastic pathway for this improvement are needed.

The interplay between irrigation and fruiting on branch growth and mortality, gas exchange and water relations of coffee trees.

The overall coordination between gas exchanges and plant hydraulics may be affected by soil water availability and source-to-sink relationships. Here we evaluated how branch growth and mortality, leaf gas exchange and metabolism are affected in coffee (Coffea arabica L.) trees by drought and fruiting. Field-grown plants were irrigated or not, and maintained with full or no fruit load. Under mild water deficit, irrigation per se did not significantly impact growth but markedly reduced branch mortality in fruiting trees, despite similar leaf assimilate pools and water status. Fruiting increased net photosynthetic rate in parallel with an enhanced stomatal conductance, particularly in irrigated plants. Mesophyll conductance and maximum RuBisCO carboxylation rate remained unchanged across treatments. The increased stomatal conductance in fruiting trees over nonfruiting ones was unrelated to internal CO2 concentration, foliar abscisic acid (ABA) levels or differential ABA sensitivity. However, stomatal conductance was associated with higher stomatal density, lower stomatal sensitivity to vapor pressure deficit, and higher leaf hydraulic conductance and capacitance. Increased leaf transpiration rate in fruiting trees was supported by coordinated alterations in plant hydraulics, which explained the maintenance of plant water status. Finally, by preventing branch mortality, irrigation can mitigate biennial production fluctuations and improve the sustainability of coffee plantations.

Methylphenidate treatment increases hippocampal BDNF levels but does not improve memory deficits in hypoxic-ischemic rats.

BACKGROUND: Methylphenidate (MPH) is a stimulant drug mainly prescribed to treat cognitive impairments in attention-deficit/hyperactivity disorder (ADHD). We demonstrated that neonatal hypoxia-ischemia (HI) induced attentional deficits in rats and MPH administration reversed these deficits. However, MPH effects on memory deficits after the HI procedure have not been evaluated yet. AIMS: We aimed to analyze learning and memory performance of young hypoxic-ischemic rats after MPH administration and associate their performance with brain-derived neurotrophic factor (BDNF) levels in the prefrontal cortex and hippocampus. METHODS: Male Wistar rats were divided into four groups (n=11-13/group): control saline (CTS), control MPH (CTMPH), HI saline (HIS) and HIMPH. The HI procedure was conducted at post-natal day (PND) 7 and memory tasks between PND 30 and 45. MPH administration (2.5 mg/kg, i.p.) occurred 30 min prior to each behavioral session and daily, for 15 days, for the BDNF assay (n=5-7/group). RESULTS: As expected, hypoxic-ischemic animals demonstrated learning and memory deficits in the novel-object recognition (NOR) and Morris water maze (MWM) tasks. However, MPH treatment did not improve learning and memory deficits of these animals in the MWM-and even disrupted the animals' performance in the NOR task. Increased BDNF levels were found in the hippocampus of HIMPH animals, which seem to have been insufficient to improve memory deficits observed in this group. CONCLUSIONS: The MPH treatment was not able to improve memory deficits resulting from the HI procedure considering a dose of 2.5 mg/kg. Further studies investigating different MPH doses would be necessary to determine a dose-response relationship in this model.

Neonatal hypoxia-ischemia caused mild motor dysfunction, recovered by acrobatic training, without affecting morphological structures involved in motor control in rats.

The aim of this study was to evaluated motor function and morphological aspects of the components involved in motor control (sensorimotor cortex, spinal cord, sciatic nerve, neuromuscular junctions and skeletal muscle) in male Wistar rats exposed to a model of neonatal hypoxic-ischemic encephalopathy (HIE) and the possible influence of different physical exercise protocols - treadmill and acrobatic. Male Wistar rats at the 7th post-natal day (PND) were submitted to the HIE model and from the 22nd until 60th PND the exercise protocols (treadmill or acrobatic training) were running. After the training, the animals were evaluated in Open Field, Ladder Rung Walking and Rotarod tasks and after samples of the motor control components were collected. Our results evidenced that the acrobatic training reversed the hyperactivity and anxiety, caused locomotion improvement and decreased brain atrophy in HIE animals. We did not find morphological differences on sensorimotor cortex, spinal cord, sciatic nerve, neuromuscular junctions and skeletal muscle in the animals submitted to HIE model. These intriguing data support the statement of the Rice-Vannucci model does not seem to reproduce, in structures involved in control function, the damage found in humans that suffer HIE. Regarding the protocols of exercise, we proposed that the acrobatic exercise could be a good therapeutic option especially in children affected by neonatal HIE and can be responsible for good results in cognitive and motor aspects.

Characterization of a cerebral palsy-like model in rats: Analysis of gait pattern and of brain and spinal cord motor areas.

In an attempt to propose an animal model that reproduces in rats the phenotype of cerebral palsy, this study evaluated the effects of maternal exposure to bacterial endotoxin associated with perinatal asphyxia and sensorimotor restriction on gait pattern, brain and spinal cord morphology. Two experimental groups were used: Control Group (CTG) - offspring of rats injected with saline during pregnancy and Cerebral Palsy Group (CPG) - offspring of rats injected with lipopolysaccharide during pregnancy, submitted to perinatal asphyxia and sensorimotor restriction for 30days. At 29days of age, the CPG exhibited coordination between limbs, weight-supported dorsal steps or weight-supported plantar steps with paw rotation. At 45days of age, CPG exhibited plantar stepping with the paw rotated in the balance phase. An increase in the number of glial cells in the primary somatosensory cortex and dorsal striatum were observed in the CPG, but the corpus callosum thickness and cross-sectional area of lateral ventricle were similar between studied groups. No changes were found in the number of motoneurons, glial cells and soma area of the motoneurons in the ventral horn of spinal cord. The combination of insults in the pre, peri and postnatal periods produced changes in hindlimbs gait pattern of animals similar to those observed in diplegic patients, but motor impairments were attenuated over time. Besides, the greater number of glial cells observed seems to be related to the formation of a glial scar in important sensorimotor brain areas.

Caracterização de um modelo de paralisia cerebral em ratos: cognição e estrutura do hipocampo e amígdala / Characterization of a model of cerebral palsy in rats: cognition and structureof the hippocampus and amygdala

FUNDAMENTO: A paralisia cerebral (PC) é caracterizada por distúrbios do movimento e da postura, que podem estar associados a déficits cognitivos. Tais comprometimentos são atribuídos a lesões não progressivas ao encéfalo em desenvolvimento. No âmbito experimental, modelos animais dessa condição clínica capazes de reproduzir o fenótipo e as alterações estruturais vistas em humanos são escassos. OBJETICO: Investigar as repercussões da indução de um modelo de PC sobre a função cognitiva e estrutura do hipocampo e amígdala em ratos Wistar. MÉTODOS: Dois grupos experimentais foram utilizados: 1) Controle - filhotes de ratas injetadas com solução salina durante a gestação (n=8) e 2) Paralisia cerebral - filhotes de ratas injetadas com Lipopolissacarídeo (LPS) durante a gestação (n=8), submetidos à anóxia perinatal e restrição sensório-motora durante 30 dias. A memória espacial dos animais foi avaliada pela tarefa de reconhecimento da localização de objetos, enquanto o comportamento do tipo ansioso foi verificado pelo teste de labirinto em cruz elevado. Após a avaliação comportamental, os animais foram eutanasiados e os encéfalos dissecados para posterior processamento histológico. RESULTADOS: O grupo PC apresentou déficits de memória espacial e uma redução do número de neurônios granulares no giro denteado. Entretanto o comportamento do tipo ansioso e a histologia do núcleo central e complexo basolateral da amígdala foram semelhantes entre os grupos. CONCLUSÃO: Como observado em parte dos pacientes com PC, este modelo experimental prejudica a memória dependente do hipocampo. Entretanto, a combinação de intervenções não alterou a ansiedade e estrutura da amígdala.

BASIS: Cerebral palsy (CP) is a disorder of movement and posture, which may be associated with cognitive impairments. Such clinical condition is caused by non progressive injuries ocurred during the brain development. In the experimental context, animal models of this condition that can reproduce the phenotype and the structural changes seen in humans are scarce. OBJECTIVE: The present study investigated cognitive function and hippocampus and amygdala structure in rats submitted to a CP model. METHODS: Two experimental groups were used: 1) Control - offspring of rats injected with saline during pregnancy (n = 8) and 2) Cerebral Palsy - offspring of rats injected with lipopolysaccharide (LPS) during pregnancy (n = 8), submitted to perinatal anoxia and sensorimotor restriction for 30 days. The spatial memory was evaluated by the object-placement recog- nition task and anxiety like-behavior by elevated plus maze test. After the behavioral assessment, animals were euthanized and brains dissected for histological processing. RESULTS: The PC group showed spatial memory deficits and a reduction of granule neurons in the dentate gyrus. However, the anxiety like-behavior and the number of neurons in central nucleus and basolateral complex of the amygdala were similar between studied groups. CONCLUSION: This animal model affects the hippocampus dependent memory, a deficit seen in part of CP patients. However, the interventions used did not alter the anxiety like-behavior and amygdala structure.

Desempenho fotossintético e micorrização em plantas de soja Roundup Ready tratadas com diferentes formulações de glyphosate / Photosynthetic performance and mycorrhization in Roundup Ready soybean plants treated with different formulations of glyphosate

Com a expansão do cultivo de soja resistente ao glyphosate, observa-se aumento considerável tanto do uso desse herbicida como do número de formulações comerciais à base deste princípio ativo. Objetivou-se, portanto, avaliar o efeito de seis formulações de glyphosate (Roundup Original®, Trop®, Roundup Ultra®, Roundup WG®, Roundup Transorb R® e Zapp Qi®) em parâmetros fotossintéticos, colonização micorrízica e produtividade de soja (TMG 125 Roundup Ready), na região do Alto Paranaíba. Utilizou-se do delineamento em blocos casualizados, com quatro repetições. Uma única aplicação dos herbicidas (720 g e.a. ha-1) foi realizada entre os estádios V2-V3. Aos 3 e 7 dias após a aplicação dos tratamentos (DAT), foram feitas avaliações instantâneas de trocas gasosas entre 08:00 e 9:00 h, em folíolos completamente expandidos. Em seguida, o desempenho fotossintético foi avaliado por meio de curvas de luz (taxa de assimilação líquida de CO2 vs irradiância). A colonização micorrízica, altura de plantas e massa seca da parte aérea foram avaliadas quando as plantas de soja atingiram o estádio R2. Além disso, avaliaram-se o peso de sementes e produtividade de soja. As diferentes formulações de glyphosate não afetaram as trocas gasosas das plantas de soja, nem a eficiência dessas plantas na utilização da irradiância, razão pela qual também não se observou diferenças significativas na massa seca da parte aérea, colonização micorrízica e produtividade de soja.

The expansion of soybean resistant to glyphosate has caused considerable increase in the use of this herbicide as well as the number of formulations based on this active ingredient. The aim is evaluate the effect of six formulations of glyphosate (Roundup Original®, Trop®, Roundup Ultra®, Roundup WG®, Roundup Transorb R® e Zapp Qi®) in photosynthetic parameters, mycorrhizal colonization and soybean yield (TMG 125 Roundup Ready), in the Alto Paranaíba. It was used a randomized block design with four replications. A single application of herbicides (780 g a.e ha-1) was performed between stages V2-V3. At 3 and 7 days after application of the treatments were evaluated instantaneous gas exchange between 08:00 and 9:00 am, in fully expanded leaflets. Then, the photosynthetic performance was evaluated by means of light curves (CO2 assimilation rate vs. irradiance). The mycorrhizal colonization, plant height and shoot dry matter were assessed when the soybean plants reached the stage R2. In addition, we evaluated the weight of seeds and soybean yield. The different formulations of glyphosate did not affect gas exchange of soybean plants and the efficiency of these plants in the use of irradiance, therefore also no observed significant differences in shoot dry weight, mycorrhizal colonization and soybean yield.

Complete genome sequence of the sugarcane nitrogen-fixing endophyte Gluconacetobacter diazotrophicus Pal5.

BACKGROUND: Gluconacetobacter diazotrophicus Pal5 is an endophytic diazotrophic bacterium that lives in association with sugarcane plants. It has important biotechnological features such as nitrogen fixation, plant growth promotion, sugar metabolism pathways, secretion of organic acids, synthesis of auxin and the occurrence of bacteriocins. RESULTS: Gluconacetobacter diazotrophicus Pal5 is the third diazotrophic endophytic bacterium to be completely sequenced. Its genome is composed of a 3.9 Mb chromosome and 2 plasmids of 16.6 and 38.8 kb, respectively. We annotated 3,938 coding sequences which reveal several characteristics related to the endophytic lifestyle such as nitrogen fixation, plant growth promotion, sugar metabolism, transport systems, synthesis of auxin and the occurrence of bacteriocins. Genomic analysis identified a core component of 894 genes shared with phylogenetically related bacteria. Gene clusters for gum-like polysaccharide biosynthesis, tad pilus, quorum sensing, for modulation of plant growth by indole acetic acid and mechanisms involved in tolerance to acidic conditions were identified and may be related to the sugarcane endophytic and plant-growth promoting traits of G. diazotrophicus. An accessory component of at least 851 genes distributed in genome islands was identified, and was most likely acquired by horizontal gene transfer. This portion of the genome has likely contributed to adaptation to the plant habitat. CONCLUSION: The genome data offer an important resource of information that can be used to manipulate plant/bacterium interactions with the aim of improving sugarcane crop production and other biotechnological applications.

Cidadania ativa: a experiência dos pequenos produtores rurais / Active citizenship: the experience of small rural producers

Relata que, em meio ao quadro de extrema pobreza dos habitantes da cidade de Valente e da seca que devasta o território ao longo dos anos, a Associaçäo dos Pequenos Agricultures do Município de Valente (Apaeb), através de uma ampla açäo de desenvolvimento local, vem modificando sensivelmente o panorama da regiäo sisaleira da Bahia. Observa que, a partir de três projetos básicos interligados - Batedeira Comunitária, Cooperativa de Crédito e Fábrica de Sisal - a Apaeb demonstra toda a vitalidade e o poder transformador das soluçöes simples e concretas na melhoria da qualidade de vida das populaçöes empobrecidas e de suas geraçöes futuras. Descreve a entidade. Avalia os projetos desenvolvidos e seus resultados junto à comunidade.