Effect of arginine, glycine + serine concentrations, and guanidinoacetic acid supplementation in vegetable-based diets for chickens.

The study investigated guanidinoacetic acid (GAA) supplementation with varying dietary digestible arginine (Arg) and glycine+serine (Gly+Ser) concentrations in the starter phase, exploring respective carry-over effects on growth performance, blood chemistry, incidence of pectoral myopathies and proximate composition in broilers. A total of 2,800 one-day-old male broiler chicks were distributed in a central composite design with 2 factors and double experimental mesh, represented by supplementation or omission of 0.6 g per kg of GAA, with a central point represented by 107% of Arg and 147% of Gly+Ser, 4 factorial points (combinations of Arg/Gly+Ser concentrations: 96.4/132.5%; 117.6/132.5%; 96.4/161.5%, and 117.6/132.5%), and 4 axial points (combinations of axial points estimated for Arg and Gly+Ser, with the central points of 92/147%; 122/147%; 107/126.5, and 107/167.5%), totaling 18 treatments, 4 repetitions to factorial and axial points, 24 replicates to the central point, and 25 birds per pen. Feed conversion ratio (FCR) from d 1 to 10 had a linear response (P = 0.009) for the decreasing Arg content and a quadratic response (P = 0.047) for Gly+Ser concentrations. Broilers supplemented GAA had lower FCR compared with nonsupplemented groups from d 1 to 10 (P = 0.048) and d 1 to 42 (P = 0.026). Aspartate aminotransferase (AST) exhibited increasing and decreasing linear effects as a function of Arg (P = 0.008) and Gly+Ser (P = 0.020) concentrations, respectively. Guanidinoacetic acid decreased serum AST (P = 0.028). Guanidinoacetic acid reduced moderate + severe (P = 0.039) and mild (P = 0.015) Wooden Breast scores. The occurrence of normal White Striping increased (P = 0.002), while severe score was reduced (P = 0.029) with GAA supplementation. In conclusion, increased digestible Arg:Lys and 14% and 6% above the recommendations (107% and 147%), respectively, provided improved FCR during the starter phase. Dietary GAA supplementation (0.6 g per kg) improved FCR, reduced severity of breast myopathies and appears to have reduced muscle damage in broilers fed plant-based diets.

Glyphosate at low doses changes the physiology and increases the productivity of common bean as affected by sowing seasons.

Glyphosate applied at low doses can stimulate photosynthesis and yield. The objective of this study was to evaluate the application of low doses of glyphosate and sowing seasons in physiological characteristics and grain yield of common bean of early cycle. Two experiments were conducted in the field, the first in winter season and the second in wet season. The experimental design was a randomized complete block design, consisting of five and seven low doses of glyphosate and one period of application, with four replications. Glyphosate low dose of 108.0 g a.e. ha-1 impaired net CO2 assimilation rate, stomatal conductance, transpiration rate, instantaneous carboxylation efficiency, number of pods per plant, number of grains per plant and number of grains per pod. Glyphosate dose of 7.2 g a.e. ha-1 provided a 23% increase in grain yield in winter season, and the dose of 36.0 g a.e. ha-1 provided a 109% increase in grain yield in wet season. To our knowledge, this is the first report on effect of glyphosate at low doses and sowing season to obtain yield increases in common bean of early cycle.

Antioxidant status and their enhancements strategies for water stress tolerance in chickpea / Status antioxidante e suas estratégias de aprimoramento para tolerância ao estresse hídrico no grão-de-bico

Water stress executes severe influences on the plant growth and development through modifying physio-chemical properties. Therefore, a field experiment was designed to evaluate the antioxidant status and their enhancements strategies for water stress tolerance in chickpea on loam and clay loam soils under agro-ecological conditions of Arid Zone Research Institute, Bahawalpur (29.3871 °N, 71.653 °E) and Cholistan farm near Derawer (28.19°N, 71.80°E) of Southern Punjab, Pakistan during winter 2014-15. Experimental treatments comprised of two chickpea cultivars i.e. Bhakhar 2011 (drought tolerant) and DUSHT (drought sensitive), two water stress levels i.e. water stress at flowering stage and water stress at flowering + pod formation + grain filling stage including well watered (control) and two exogenous application of osmoprotectants i.e. glycine betaine (GB) 20 ppm and proline 10 uM including distilled water (control). Results indicated that water stress at various growth stages adversely affects the growth, yield and quality attributes of both chickpea cultivars. Exogenous application of GB and proline improved the growth, yield and quality parameters of both chickpea cultivars even under water stress conditions. However, superior results were obtained with exogenously applied GB on Bhakhar 2011 under well-watered conditions. Similarly, foliar spray of GB on chickpea cultivar Bhakhar 2011 under stress at flowering + pod formation + grain filling stage produced maximum superoxide dismutase, peroxidase and catalase contents. These results suggested that application of GB mitigates the adverse effects of water stress and enhanced tolerance in chickpea mainly due to higher antioxidant enzymes activity, demonstrating the protective measures of plant cells in stress condition. Hence, antioxidants status might be a suitable method for illustrating water stress tolerance in chickpea.(AU)

O estresse hídrico exerce fortes influências no crescimento e no desenvolvimento das plantas, modificando as propriedades físico-químicas. Portanto, a presente atividade de pesquisa foi projetada para avaliar o status antioxidante e suas estratégias de aprimoramento para tolerância ao estresse hídrico no grão-de-bico em condiçõesa groecológicas, no Instituto de Pesquisa da Zona Árida, Bahawalpur (29.3871 ° N, 71.653 ° E) e fazenda do Cholistan, perto de Derawer (28.19 ° N, 71,80 ° E), no sul de Punjab, Paquistão, durante Rabi 2014-15. Tratamentos experimentais compostos de dois genótipos de grão-de-bico, como Bhakhar 2011 (tolerante à seca) e DUSHT (sensível à seca), dois níveis de estresse hídrico, ou seja, estresse hídrico no estágio de floração, estresse hídrico na fase de floração e estresse hídrico na fase de floração + formação de vagem + estágio de enchimento de grãos, incluindo água bem controlada (controle) e duas aplicações exógenas de osmoprotetores, isto é, glicina betaína 20 ppm e prolina 10 uM, incluindo água destilada (controle). Os resultados indicaram que o estresse hídrico em vários estágios de crescimento afeta negativamente os atributos de crescimento, rendimento e qualidade de ambas as cultivares de grão-de-bico. A aplicação exógena de glicina betaína e prolina melhorou os parâmetros de crescimento, rendimento e qualidade de ambos os genótipos de grão- de-bico, mesmo sob condições de estresse hídrico. No entanto, resultados superiores foram obtidos com glicina betaína aplicada exogenamente em Bhakhar 2011, em condições bem regadas. Além disso, o spray foliar de glicina betaína na cultivar de grão-de-bico Bhakhar 2011, sob estresse na fase de floração + formação de vagem + enchimento de grãos, produziu o máximo de superóxido dismutase, peroxidase e catalase. Esses resultados sugeriram que a aplicação de glicina betaína atenua os efeitos adversos do estresse hídrico e aumenta a [...].(AU)

Antioxidant status and their enhancements strategies for water stress tolerance in chickpea / Status antioxidante e suas estratégias de aprimoramento para tolerância ao estresse hídrico no grão-de-bico

Water stress executes severe influences on the plant growth and development through modifying physio-chemical properties. Therefore, a field experiment was designed to evaluate the antioxidant status and their enhancements strategies for water stress tolerance in chickpea on loam and clay loam soils under agro-ecological conditions of Arid Zone Research Institute, Bahawalpur (29.3871 °N, 71.653 °E) and Cholistan farm near Derawer (28.19°N, 71.80°E) of Southern Punjab, Pakistan during winter 2014-15. Experimental treatments comprised of two chickpea cultivars i.e. Bhakhar 2011 (drought tolerant) and DUSHT (drought sensitive), two water stress levels i.e. water stress at flowering stage and water stress at flowering + pod formation + grain filling stage including well watered (control) and two exogenous application of osmoprotectants i.e. glycine betaine (GB) 20 ppm and proline 10 uM including distilled water (control). Results indicated that water stress at various growth stages adversely affects the growth, yield and quality attributes of both chickpea cultivars. Exogenous application of GB and proline improved the growth, yield and quality parameters of both chickpea cultivars even under water stress conditions. However, superior results were obtained with exogenously applied GB on Bhakhar 2011 under well-watered conditions. Similarly, foliar spray of GB on chickpea cultivar Bhakhar 2011 under stress at flowering + pod formation + grain filling stage produced maximum superoxide dismutase, peroxidase and catalase contents. These results suggested that application of GB mitigates the adverse effects of water stress and enhanced tolerance in chickpea mainly due to higher antioxidant enzymes activity, demonstrating the protective measures of plant cells in stress condition. Hence, antioxidants status might be a suitable method for illustrating water stress tolerance in chickpea.

O estresse hídrico exerce fortes influências no crescimento e no desenvolvimento das plantas, modificando as propriedades físico-químicas. Portanto, a presente atividade de pesquisa foi projetada para avaliar o status antioxidante e suas estratégias de aprimoramento para tolerância ao estresse hídrico no grão-de-bico em condiçõesa groecológicas, no Instituto de Pesquisa da Zona Árida, Bahawalpur (29.3871 ° N, 71.653 ° E) e fazenda do Cholistan, perto de Derawer (28.19 ° N, 71,80 ° E), no sul de Punjab, Paquistão, durante Rabi 2014-15. Tratamentos experimentais compostos de dois genótipos de grão-de-bico, como Bhakhar 2011 (tolerante à seca) e DUSHT (sensível à seca), dois níveis de estresse hídrico, ou seja, estresse hídrico no estágio de floração, estresse hídrico na fase de floração e estresse hídrico na fase de floração + formação de vagem + estágio de enchimento de grãos, incluindo água bem controlada (controle) e duas aplicações exógenas de osmoprotetores, isto é, glicina betaína 20 ppm e prolina 10 uM, incluindo água destilada (controle). Os resultados indicaram que o estresse hídrico em vários estágios de crescimento afeta negativamente os atributos de crescimento, rendimento e qualidade de ambas as cultivares de grão-de-bico. A aplicação exógena de glicina betaína e prolina melhorou os parâmetros de crescimento, rendimento e qualidade de ambos os genótipos de grão- de-bico, mesmo sob condições de estresse hídrico. No entanto, resultados superiores foram obtidos com glicina betaína aplicada exogenamente em Bhakhar 2011, em condições bem regadas. Além disso, o spray foliar de glicina betaína na cultivar de grão-de-bico Bhakhar 2011, sob estresse na fase de floração + formação de vagem + enchimento de grãos, produziu o máximo de superóxido dismutase, peroxidase e catalase. Esses resultados sugeriram que a aplicação de glicina betaína atenua os efeitos adversos do estresse hídrico e aumenta a [...].

Glyphosate exposure induces synaptic impairment in hippocampal neurons and cognitive deficits in developing rats.

Glyphosate is the active ingredient of several widely used herbicide formulations. Studies based on Glyphosate exposure in different experimental models have suggested that the nervous system represented a key target for its toxicity. Previously, we demonstrated that exposure to glyphosate during gestation induces deficits on behavioral and cognitive function in rats. The aim of the present work was to examine whether cognitive dysfunction induced by Glyphosate was connected to changes on synapse formation and maturation. To understand how glyphosate affects synaptic assembly, we performed in vitro assays on cultured hippocampal neurons that were exposed to the herbicide (0.5 or 1 mg/mL) for 5 or 10 days. Biochemical and immunocytochemical approaches revealed that Glyphosate treated neurons showed a decrease on dendritic complexity and synaptic spine formation and maturation. Moreover, results indicated that Glyphosate decreased synapse formation in hippocampal neurons. To evaluate these effects in vivo, pup rats were treated with 35 or 70 mg/kg of Glyphosate from PND 7 to PND 27, every 48 h. Results indicated that Glyphosate postnatal exposure induced cognitive impairments, since recognition and spatial memory were altered. To go further, we evaluated synaptic protein expression and synaptic organization in hippocampus. Images revealed that Glyphosate treatment downregulates synapsin-1, PSD-95, and CaMKII expression, and also decreased PSD-95 clustering in hippocampus. Taken together, these findings demonstrate for the first time that Glyphosate exposure affects synaptic assembly and reduced synaptic protein expression in hippocampus and that likely triggers the impairment of cognitive function and neuronal connectivity.

Low Doses of Glyphosate/Roundup Alter Blood-Testis Barrier Integrity in Juvenile Rats.

It has been postulated that glyphosate (G) or its commercial formulation Roundup (R) might lead to male fertility impairment. In this study, we investigated the possible effects of G or R treatment of juvenile male rats on blood-testis barrier function and on adult male sperm production. Pups were randomly assigned to the following groups: control group (C), receiving water; G2 and G50 groups, receiving 2 and 50 mg/kg/day G respectively; and R2 and R50 groups receiving 2 and 50 mg/kg/day R respectively. Treatments were performed orally from postnatal day (PND) 14 to 30, period of life that is essential to complete a functional blood-testis barrier. Evaluation was done on PND 31. No differences in body and testis weight were observed between groups. Testis histological analysis showed disorganized seminiferous epithelium, with apparent low cellular adhesion in treated animals. Blood-testis barrier permeability to a biotin tracer was examined. A significant increase in permeable tubules was observed in treated groups. To evaluate possible mechanisms that could explain the effects on blood-testis barrier permeability, intratesticular testosterone levels, androgen receptor expression, thiobarbituric acid reactive substances (TBARS) and the expression of intercellular junction proteins (claudin11, occludin, ZO-1, connexin43, 46, and 50 which are components of the blood-testis barrier) were examined. No modifications in the above-mentioned parameters were detected. To evaluate whether juvenile exposure to G and R could have consequences during adulthood, a set of animals of the R50 group was allowed to grow up until PND 90. Histological analysis showed that control and R50 groups had normal cellular associations and complete spermatogenesis. Also, blood-testis barrier function was recovered and testicular weight, daily sperm production, and epididymal sperm motility and morphology did not seem to be modified by juvenile treatment. In conclusion, the results presented herein show that continuous exposure to low doses of G or R alters blood-testis barrier permeability in juvenile rats. However, considering that adult animals treated during the juvenile stage showed no differences in daily sperm production compared with control animals, it is feasible to think that blood-testis barrier impairment is a reversible phenomenon. More studies are needed to determine possible damage in the reproductive function of human juvenile populations exposed to low doses of G or R.

Can low doses of glyphosate stimulate common bean growth?

Plant growth can be stimulated by low doses of glyphosate. The objective of this work was to evaluate the effect of low doses of glyphosate and sowing season on the growth of the early cycle common bean. Two experiments were conducted in the field, the first in the winter and the second in the wet season, with the early cycle common bean cultivar IAC Imperador. The experimental design was a randomized complete block design, consisting of low doses of glyphosate applied on phenological stage V4, with four replications. Environmental conditions, such as air temperature, interfered in the early cycle common bean response to low doses of glyphosate. In the winter season, a dose of 36 g a.e. ha-1 promoted growth in the common bean, and a dose of 7.2 g a.e. ha-1 improved the harvest index. In the wet season, there was no growth stimulus, and the harvest index increased with a dose of 36 g a.e. ha-1. The harvest index was the only characteristic improved in both seasons, but with different doses. Our study indicates that growth characteristics of early cycle common bean are stimulated by low doses of glyphosate, but this response is dependent on the growing environment.

Sleep in honey bees is affected by the herbicide glyphosate.

Sleep plays an essential role in both neural and energetic homeostasis of animals. Honey bees (Apis mellifera) manifest the sleep state as a reduction in muscle tone and antennal movements, which is susceptible to physical or chemical disturbances. This social insect is one of the most important pollinators in agricultural ecosystems, being exposed to a great variety of agrochemicals, which might affect its sleep behaviour. The intake of glyphosate (GLY), the herbicide most widely used worldwide, impairs learning, gustatory responsiveness and navigation in honey bees. In general, these cognitive abilities are linked with the amount and quality of sleep. Furthermore, it has been reported that animals exposed to sleep disturbances show impairments in both metabolism and memory consolidation. Consequently, we assessed the sleep pattern of bees fed with a sugar solution containing GLY (0, 25, 50 and 100 ng) by quantifying their antennal activity during the scotophase. We found that the ingestion of 50 ng of GLY decreased both antennal activity and sleep bout frequency. This sleep deepening after GLY intake could be explained as a consequence of the regenerative function of sleep and the metabolic stress induced by the herbicide.

Intranasal glyphosate-based herbicide administration alters the redox balance and the cholinergic system in the mouse brain.

Pesticide exposure is associated with cognitive and psychomotor disorders. Glyphosate-based herbicides (GlyBH) are among the most used agrochemicals, and inhalation of GlyBH sprays may arise from frequent aerial pulverizations. Previously, we described that intranasal (IN) administration of GlyBH in mice decreases locomotor activity, increases anxiety, and impairs recognition memory. Then, the aim of the present study was to investigate the mechanisms involved in GlyBH neurotoxicity after IN administration. Adult male CF-1 mice were exposed to GlyBH IN administration (equivalent to 50 mg/kg/day of Gly acid, 3 days a week, during 4 weeks). Total thiol content and the activity of the enzymes catalase, acetylcholinesterase and transaminases were evaluated in different brain areas. In addition, markers of the cholinergic and the nigrostriatal pathways, as well as of astrocytes were evaluated by fluorescence microscopy in coronal brain sections. The brain areas chosen for analysis were those seen to be affected in our previous study. GlyBH IN administration impaired the redox balance of the brain and modified the activities of enzymes involved in cholinergic and glutamatergic pathways. Moreover, GlyBH treatment decreased the number of cholinergic neurons in the medial septum as well as the expression of the α7-acetylcholine receptor in the hippocampus. Also, the number of astrocytes increased in the anterior olfactory nucleus of the exposed mice. Taken together, these disturbances may contribute to the neurobehavioural impairments reported previously by us after IN GlyBH administration in mice.

Hormetic effect of glyphosate on Urochloa decumbens plants.

Urochloa decumbens plants may be reached by herbicide drift from applications of glyphosate from neighboring areas or by variations during applications. Considering the different phenological stages and size of plants in these areas, the amount of active ingredient that reaches the plants probably varies. The objective of this study was to evaluate the effects of the application of different doses of glyphosate on U. decumbens plants. Two greenhouse experiments were conducted with two replications at different times. The first experiment evaluated the biological response of U. decumbens plants to glyphosate doses (0, 2.81, 5.63, 11.25, 22.5, 45, 90, 180, 360, 720, and 1,440 g a.e. ha-1), with six replications. The second experiment evaluated the response of U. decumbens plants to the application of a selected low dose of 11.25 g a.e. ha-1. Evaluations of injury were performed at 0, 7, 14, and 21 days after application, and dry weight of plants was determined for each evaluation period. U. decumbens plants increased in dry weight when using the glyphosate dose of 11.25 g a.e. ha-1. However, plants had different responses to the application of this low dose. It can promote both stimulation and inhibition of plant growth.

Effect of day time climatic conditions associated with different 2,4-D formulations on spray deposition and weed control.

The aim of this study was to evaluate the effect of time of the day and their associated climatic conditions on spray deposition of two 2,4-D formulations, as well as the influence on weed control. The experiment was installed in the field in complete randomized design. Treatments were arranged in factorial design 8 × 2, with 20 repetitions. First factor corresponded to different application time (1:00, 4:00, 7:00, 10:00, 13:00, 16:00, 19:00, and 22:00) with their respective climatic conditions. The second factor consisted of two formulations of 2,4-D applied at 776 g a.e. ha-1 (2,4-D amine and 2,4-D choline salt with Colex-D™ Technology) + glyphosate (816 g a.e. ha-1). There was more spray deposition when 2,4-D choline formulation was used, and such differences were more evident for applications performed under adverse climatic conditions. More spray deposition was found in applications performed at times of day with more favorable temperature and humidity of the air conditions. Only the initial control of the evaluated species was affected by the time of application.

The effects of Roundup® in embryo development and energy metabolism of the zebrafish (Danio rerio).

Roundup® is currently the most widely used and sold agricultural pesticide in the world. The objective of this work was to investigate the effects of Roundup® on energy metabolism during zebrafish (Danio rerio) embryogenesis. The embryo toxicity test was performed for 96 h post-fertilisation and the sublethal concentration of Roundup® was defined as 58.3 mg/L, which resulted in failure to inflate the swim bladder. Biochemical assays were performed with viable embryos following glyphosate exposure, and no significant effects on protein, glucose, glycogen, triglyceride levels or the enzymatic activities of alanine aminotransferase and aspartate aminotransferase were observed. However, the activity of hexokinase was significantly altered following exposure to 11.7 mg/L Roundup®. Through molecular docking we have shown for the first time that the interactions of glucokinase and hexokinases 1 and 2 with glyphosate showed significant interactions in the active sites, corroborating the biochemical results of hexokinase activity in zebrafish exposed to the chemical. From the results of molecular docking interactions carried out on the Zfishglucok, ZfishHK1 and ZfishHK2 models with the glyphosate linker, it can be concluded that there are significant interactions between glyphosate and active sites of glucokinase and hexokinase 1 and 2 proteins. The present work suggests that Roundup® can induce problems in fish embryogenesis relating to the incapacity of swim bladder to inflate. This represents the first study demonstrating the interaction of glyphosate with hexokinase and its isoforms.

Bezafibrate Prevents Glycine-Induced Increase of Antioxidant Enzyme Activities in Rat Striatum.

Non-ketotic hyperglycinemia (NKH) is a severe neurological disorder caused by defects in glycine (GLY) catabolism and characterized by a high cerebrospinal fluid/plasma GLY ratio. Treatment is often ineffective and limited to the control of symptoms and detoxification of GLY. In the present work, we investigated the in vivo effects of GLY intracerebroventricular administration on oxidative stress parameters in rat striatum, cerebral cortex, and hippocampus. In vitro effects of GLY were also evaluated in striatum. The effects of bezafibrate (BEZ), a potential neuroprotective agent, on the possible alterations caused by GLY administration were further evaluated. Our in vivo results showed that GLY increased the activities of the antioxidant enzymes superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), and glucose-6-phosphate dehydrogenase (G6PDH) in striatum. Furthermore, GLY decreased the concentrations of total glutathione and reduced glutathione (GSH), as well as GSH/oxidized glutathione ratio in vivo in hippocampus. In vitro data also showed that GLY induced lipid peroxidation and decreased GSH in striatum. Regarding the effects of BEZ, we found that GLY-induced increase of GPx, SOD, and GR activities was attenuated or prevented by this compound. However, BEZ did not alter GLY-induced decrease of GSH in hippocampus. We hypothesize that GLY-induced increase of the activities of antioxidant enzymes in striatum occurs as a mechanism to avoid accumulation of reactive oxygen species and consequent oxidative damage. Furthermore, since BEZ prevented GLY-induced alterations, it might be considered as an adjuvant therapy for NKH.

Supplementation of L-glycine and L-glutamate to Japanese quails from 01 to 36 days of age using the ideal protein concept.

Experiment I: T1-1 =  basal diet with 25% crude protein (CP) + limiting amino acids (LA); T1-2  = 20% CP + LA; T1-3  = 20% CP + LA + L-glycine; T1-4  = 20% CP + LA + L-glutamate; T1-5  = 20% CP + LA + L-glycine + L-glutamate. Experiment II: T2-1  = basal diet with 22% CP + LA; T2-2  = 20% CP + LA; T2-3  = 17.6% CP + LA + L-glycine; T2-4  = 17.6% CP + LA + L-glutamate; T2-5  = 17.6% CP + LA + L-glycine + L-glutamate. The reduction of dietary protein based on the concept of ideal protein decreases nitrogen excretion in quails when L-glycine is added to the diets. Quails fed diets supplemented with L-glutamate as the non-specific nitrogen source equivalent to the nitrogen level of the control diet had increased nitrogen excretion. However, quails had reduced nitrogen excretion in both experiments when L-glycine was added to diets with L-glutamate. Carcass fat was increased by reducing dietary protein, but fat deposition was reduced by adding L-glutamate and L-glycine, or both. The dietary addition of L-glutamate and L-glycine in quails based on the ideal protein concept is not necessary (Exp. I). Although the total nitrogen, electrolytic balance and glycine level were adjusted in diets, quails had decreased performance. Therefore, other hypotheses besides protein reduction need to be studied (Exp. II). Protein reduction with supplementation of only limiting essential amino acids does not affect quail performance. Dietary addition of L-glycine reduces nitrogen excretion.

Male mammary gland development and methylation status of estrogen receptor alpha in Wistar rats are modified by the developmental exposure to a glyphosate-based herbicide.

Postnatal treatment with glyphosate-based herbicides (GBHs) induces endocrine-disrupting effects on the male rat mammary gland. In this study, the effects of developmental exposure to GBH on mammary gland growth and development, and the possible molecular mechanisms involved, were evaluated in pre- and post-pubertal male rats. To this end, pregnant rats were orally exposed through the food to 0, 3.5 or 350 mg GBH/kg bw/day from gestational day 9 until weaning. Mammary gland development and estradiol (E2) and testosterone (T) serum levels of male offspring were evaluated on postnatal day (PND)21 and PND60. Besides, prolactin (PRL) serum levels, proliferation index, androgen (AR) and estrogen receptor alpha (ESR1) expression, ESR1 alternative transcript mRNA levels, and DNA methylation status of ESR1 promoters were assessed on PND60. No differences between groups were observed in mammary gland development at PND21 or in E2 and T levels on both PNDs studied. On PND60, GBH3.5-exposed animals presented similar mammary gland histology but higher AR protein expression than controls, whereas GBH350-exposed males presented a less developed mammary gland, accompanied by a lower proliferation index, similar AR levels, and slightly increased PRL serum levels than controls. In both exposed groups, ESR1 expression was lower than in control rats, being lower in GBH350-exposed rats. GBH also altered the abundance of ESR1 transcript variants by hypermethylation of ESR1 promoters. GHB3.5 decreased only ESR1-OS expression, whereas GBH350 affected ESR1-O, OT and E1 expression. Our results show that developmental exposure to GBH induces epigenetic changes in ESR1, which could be responsible for the altered male mammary gland development observed in GBH350-exposed animals.

Perinatal exposure to a glyphosate-based herbicide impairs female reproductive outcomes and induces second-generation adverse effects in Wistar rats.

Glyphosate-based herbicides (GBHs) are the most globally used herbicides raising the risk of environmental exposition. Here, we investigated whether perinatal exposure to low doses of a GBH alters the female reproductive performance, and/or induced second-generation effects related to congenital anomalies or growth alterations. Pregnant rats (F0) received a GBH through food, in a dose of 2 mg (GBH-LD: GBH-low dose group) or 200 mg (GBH-HD: GBH-high dose group) of glyphosate/kg bw/day from gestational day (GD) 9 until weaning. Body weight gain and vaginal canal-opening of F1 females were recorded. Sexually mature F1 females were mated to evaluate their reproductive performance by assessing the pregnancy rate, and on GD19, the number of corpora lutea, the implantation sites (IS) and resorption sites. To analyze second-generation effects on F2 offspring, we analyzed the fetal morphology on GD19, and assessed the fetal length and weight, and the placental weight. GBH exposure neither altered the body weight gain of F1 females, nor vaginal opening onset. Although all GBH-exposed F1 rats became pregnant, a lower number of IS was detected. F2 offspring from both GBH groups showed delayed growth, evidenced by lower fetal weight and length, associated with a higher incidence of small for gestational age fetuses. In addition, higher placental weight and placental index were found in F2 offspring from GBH-HD dams. Surprisingly, structural congenital anomalies (conjoined fetuses and abnormally developed limbs) were detected in the F2 offspring from GBH-HD group. In conclusion, perinatal exposure to low doses of a GBH impaired female reproductive performance and induced fetal growth retardation and structural congenital anomalies in F2 offspring.

Glycine Administration Alters MAPK Signaling Pathways and Causes Neuronal Damage in Rat Brain: Putative Mechanisms Involved in the Neurological Dysfunction in Nonketotic Hyperglycinemia.

High glycine (GLY) levels have been suggested to induce neurotoxic effects in the central nervous system of patients with nonketotic hyperglycinemia (NKH). Since the mechanisms involved in the neuropathophysiology of NKH are not totally established, we evaluated the effect of a single intracerebroventricular administration of GLY on the content of proteins involved in neuronal damage and inflammatory response, as well as on the phosphorylation of the MAPK p38, ERK1/2, and JNK in rat striatum and cerebral cortex. We also examined glial fibrillary acidic protein (GFAP) staining, a marker of glial reactivity. The parameters were analyzed 30 min or 24 h after GLY administration. GLY decreased Tau phosphorylation in striatum and cerebral cortex 30 min and 24 h after its administration. On the other hand, synaptophysin levels were decreased in striatum at 30 min and in cerebral cortex at 24 h after GLY injection. GLY also decreased the phosphorylation of p38, ERK1/2, and JNK 30 min after its administration in both brain structures. Moreover, GLY-induced decrease of p38 phosphorylation in striatum was attenuated by N-methyl-D-aspartate receptor antagonist MK-801. In contrast, synuclein, NF-κB, iκB, inducible nitric oxide synthase and nitrotyrosine content, and GFAP immunostaining were not altered by GLY infusion. It may be presumed that the decreased phosphorylation of MAPK associated with alterations of markers of neuronal injury induced by GLY may contribute to the neurological dysfunction observed in NKH.

Thidiazuron (TDZ) increases fruit set and yield of 'Hosui' and 'Packham's Triumph' pear trees.

The low fruit set is one of the main factors leading to poor yield of pear orchards in Brazil. The exogenous application of thidiazuron (TDZ) and aminoethoxyvinilglycine (AVG) has shown promising results in some pear cultivars and other temperate fruit trees. The objective of this study was to evaluate the effect of TDZ and AVG on fruit set, yield, and fruit quality of 'Hosui' and 'Packham's Triumph' pears. The study was performed in a commercial orchard located in São Joaquim, SC. Plant material consisted of 'Hosui' and 'Packham's Triumph' pear trees grafted on Pyrus calleryana. Treatments consisted on different rates of TDZ (0 mg L-1, 20 mg L-1, 40 mg L-1 and 60 mg L-1) sprayed at full bloom for both cultivars. An additional treatment of AVG 60 mg L-1 was sprayed one week after full bloom in 'Hosui'. The fruit set, number of fruit per tree, yield, fruit weight, seed number, and fruit quality attributes were assessed. Fruit set and yield of both cultivars are consistently increased by TDZ, within the rates of 20 to 60 mg L-1. Besides, its application increased fruit size of 'Hosui' and did not negatively affect fruit quality attributes of both cultivars.

Effect of oral glycine on the clinical, spirometric and inflammatory status in subjects with cystic fibrosis: a pilot randomized trial.

BACKGROUND: Patients with cystic fibrosis (CF) have airway inflammation that contributes to symptoms and to pulmonary function derangement. Current drugs used to diminish airway inflammation improve the clinical and spirometric status of patients with CF, but their use is limited due to their undesired side effects, for example, glucose intolerance, growth retardation, and cataracts with corticosteroids, gastrointestinal toxicity with ibuprofen, and macrolide resistance with azythromycin. Glycine is known to decrease activation of inflammatory cells, including alveolar macrophages and neutrophils, and is relatively inexpensive, palatable, and virtually devoid of untoward effects. These features make glycine a good candidate for antiinflammatory treatment of CF. Thus, we aimed to explore whether glycine can exert a beneficial effect in a population of patients with CF. METHODS: This was a randomized, double blinded, cross-over pilot clinical trial. Subjects with CF received, in random order, oral glycine (0.5 g/kg/day, dissolved in any liquid) and placebo (glass sugar), each during 8 weeks with an intermediate 2-week wash-out period. RESULTS: Thirteen subjects aged 6-23 years, 8 females, completed the two arms of the study. As compared with placebo, after glycine intake patients had better symptom questionnaire scores (p = 0.02), mainly regarding sputum features and dyspnea. While spirometric variables tended to decline during placebo intake, they remained stable or even increased during glycine treatment (p = 0.04 to p = 0.003). In this context, FEV1 declined 8.6% after placebo and increased 9.7% at the end of the glycine period. Pulse oximetry improved after glycine intake (p = 0.04 vs. placebo). TNF-α in serum and IL-6 and G-CSF in sputum tended to decline at the end of the glycine period (p = 0.061, p = 0.068 and p = 0.04, respectively, vs placebo). Glycine was remarkably well tolerated. CONCLUSIONS: The clinical, spirometric and inflammatory status of subjects with CF improved after just 8 weeks of glycine intake, suggesting that this amino acid might constitute a novel therapeutic tool for these patients. Thus, further studies are warranted. TRIAL REGISTRATION: www.clinicaltrials.gov , registration number: NCT01417481 , date of registration: March 12, 2012.

Thidiazuron (TDZ) increases fruit set and yield of 'Hosui' and 'Packham's Triumph' pear trees

ABSTRACT The low fruit set is one of the main factors leading to poor yield of pear orchards in Brazil. The exogenous application of thidiazuron (TDZ) and aminoethoxyvinilglycine (AVG) has shown promising results in some pear cultivars and other temperate fruit trees. The objective of this study was to evaluate the effect of TDZ and AVG on fruit set, yield, and fruit quality of 'Hosui' and 'Packham's Triumph' pears. The study was performed in a commercial orchard located in São Joaquim, SC. Plant material consisted of 'Hosui' and 'Packham's Triumph' pear trees grafted on Pyrus calleryana. Treatments consisted on different rates of TDZ (0 mg L-1, 20 mg L-1, 40 mg L-1 and 60 mg L-1) sprayed at full bloom for both cultivars. An additional treatment of AVG 60 mg L-1 was sprayed one week after full bloom in 'Hosui'. The fruit set, number of fruit per tree, yield, fruit weight, seed number, and fruit quality attributes were assessed. Fruit set and yield of both cultivars are consistently increased by TDZ, within the rates of 20 to 60 mg L-1. Besides, its application increased fruit size of 'Hosui' and did not negatively affect fruit quality attributes of both cultivars.