Phytohormones and related genes function as physiological and molecular switches regulating water stress response in the sunflower.

Water deficit stress reduces crop yield in field crops, including sunflowers, at any growth stage. In response, most plants activate hormonal and gene expression patterns to mitigate damage. In this study, we evaluated changes in the physiological and gene transcription levels of two sunflower (Helianthus annuus L.) inbred lines -one sensitive (B59 line) and one water stress-tolerant (B71)-in response to water stress, by using mannitol to simulate water deficit conditions, which provides moderate stress in both sunflower lines. The analyses of the accumulation of various phytohormones under this stress revealed that Jasmonic acid (JA) significantly increased in the shoots of both lines. Similarly, Salicylic acid (SA) increased in the shoots of both lines, although it also accumulated in B71 roots. In addition, Abscisic acid (ABA) and Indole-3-acetic acid (IAA) showed a considerable increase in the B59 shoots. Regarding the JA and SA pathways, the WRKY70 transcription levels were higher in the shoots of both lines and the roots of B71. The B59 line showed overtranscription of a gene related to the ABA pathway (XERICO) and genes associated with IAA (ARF9 and ARF16 genes). The B71 line, on the other hand, simultaneously triggered the JA, SA and ABA hormonal pathways in response to this stress condition. The ABA and JA hormonal pathways activated different TFs, such as RD20, RD22, RD26, ANAC19 and ANAC29, through MYC2. Both the JA and SA hormonal pathways activated the WRKY70 transcription factor. Altogether, each line triggered the hormonal and transcriptional pathways in response to water stress, although at varying intensities. The results suggest that the hormonal pathways of JA, SA, IAA and ABA, along with their primary associated genes, are activated in response to water deficit at the early growth stage in sunflower seedlings, which mitigates damage. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-024-01497-8.

Testicular damage without clinical manifestations in BALB/c mice experimentally infected with Zika virus.

In 2015-2016, the Zika virus (ZIKV) caused a major epidemic in the Americas, increasing cases of microcephaly and Guillain-Barré syndrome. During this period, the discovery of ZIKV sexual transmission intensified studies on the impact of this virus on the reproductive organs. For this study, 2-month-old male BALB/c mice were infected with 1.26 x 106 PFU/mL of ZIKV in solution via the intravenous route. After three, seven, and fourteen days post-infection (DPI), blood and testicle samples were obtained to detect ZIKV RNA. The authors observed that the infected animals had slower weight gain than the control group. Viremia occurred only at 3DPI, and the ZIKV RNA was detected in one testis sample at 7DPI. The histopathological analysis of this organ revealed intense disorganization of the seminiferous tubules' structure, inflammatory infiltrate, necrosis, hemorrhage, fluid accumulation, congestion of blood vessels, and reduced sperm count. Ultrastructural analysis showed nuclear changes in tubule cells, activation of interstitial cells, and morphological changes in spermatozoa, in addition to fragmentation and decreased electron density of the genetic material of these cells. Thus, despite causing predominantly asymptomatic infections, ZIKV can cause significant subclinical and transient damage, including to male reproductive organs.

Zebrafish (Danio rerio) Gynogenetic Production by Heat Shock: Comparison Between Mitotic and Meiotic Treatment.

Inbred species are useful resources for a variety of biomedical research applications. To create isogenic zebrafish, it is feasible to stop meiosis II (repeatedly) or mitosis (two times) in a haploid embryo by applying pressure or by delivering a heat shock, respectively. In this study, to improve the repeatability, we suggest a less complicated approach based on sperm ultraviolet-C (UV-C) exposure for a shorter period followed by heat shock at various temperatures, eliminating the use of pressure in meiotic therapy since heat shock is more accessible to laboratories. In this study, the survivability rates of meiotic (Mei) and mitotic (Mit) gynogenesis offspring produced by various combinations of irradiation (28.5, 105, and 210 mJ/cm2) and temperature (Mei: 40.40°C, 40.60°C, or 40.90°C; Mt: 41.40°C, 41.90°C, or 42.45°C) were compared with diploid (C) and haploid (H) controls. Our findings demonstrated that 40.60°C and 41.90°C were the most suitable temperatures to produce meiotic and mitotic gynogenesis, respectively, whereas 28.5 mJ/cm2 was more successful in ensuring haploid embryos. As a result, we deduced that meiotic gynogenesis produces more viable offspring than the mitotic approach and requires a lower temperature to maintain the second polar body.

Expansion of the basic knowledge on the inheritance of characters that allow the development of new maize breeding techniques / Expansión de los conocimientos básicos de la herencia de caracteres que posibiliten desarrollar nuevas técnicas de mejoramiento en maíz

ABSTRACT In order to increase the efficiency of maize hybrid seed production it is necessary to achieve a high grain yield to reduce production costs. This goal requires an expansion of the basic knowledge of the inheritance of characters in order to develop new breeding techniques to improve experimental materials with hard endosperm (flint). The balanced lethal system allows to study the relative contribution of different chromosome segments to hybrid vigour due to the heterozygosity of certain chromosome segments while the rest of the genome becomes homocygotic through continuous selfing. In this way, these segments can be transferred to inbred lines in order to increase grain yield or tassel size (to increase pollen production). The goal of this study was to transfer a heterotic segment by using a balanced lethal system regulated line (BLS14), through crosses and backcrosses, to S5 flint lines derived from two commercial hybrids, ACA 2000 and Cóndor with closed pedigree, with the objective of increase grain yield or tassel size for pollen production. The analysis of variance (ANOVA) and principal components analysis (PCA) showed a significant improvement in grain yield and tassel size in the S5 flint lines of both commercial hybrids, carrying the heterotic segment of the BLS14 line.

RESUMEN Para incrementar la eficiencia en la producción de semillas híbridas en maíz se necesitan altos rendimientos con un correlativo bajo costo de producción. Esto requiere una expansión de los conocimientos básicos de la herencia de caracteres que posibiliten desarrollar nuevas técnicas de mejoramiento sobre materiales experimentales de textura dura (flint). Los sistemas de letales balanceados permiten estudiar la contribución relativa de distintos segmentos heteróticos en el vigor híbrido dado que permiten lograr la heterocigosis cuasipermanente de una porción del genomio, mientras el resto del genoma se vuelve homocigota por autofecundaciones sucesivas. Estos sistemas pueden ser transferidos a líneas endocriadas para incrementar el rendimiento del grano y el tamaño de la panoja para aumentar la producción de polen. El objetivo de este trabajo fue incorporar un segmento heterótico de una línea regulada por un sistema de letales balanceados (BLS14), mediante cruzamiento y retrocruza, a líneas S5 derivadas de dos híbridos comerciales, ACA 2000 y Cóndor, de pedigree cerrado, con textura dura (flint), con la finalidad de aumentar el rendimiento en grano o el tamaño de la panoja para la producción de polen. Los análisis de variancia (ANOVA) y de componentes principales (ACP) mostraron un incremento significativo en el rendimiento del grano y el tamaño de la panoja de las líneas S5 de ambos híbridos comerciales, portando cada una de ellas el segmento heterótico proveniente de la línea BLS14.

Molecular characteristics and transcriptional regulatory of spermatogenesis-related gene RFX2 in adult Banna mini-pig inbred line (BMI).

RFX2 plays critical roles in mammalian spermatogenesis and cilium maturation. Here, the testes of 12-month-old adult boars of Banna mini-pig inbred line (BMI) were subjected to whole-transcriptome sequencing. The results indicated that the average expression (raw count) of RFX2 gene in BMI testes was 16138.25, and the average expression value of the corresponding transcript ENSSSCT00000043271.2 was 123.1898. The CDS of RFX2 obtained from BMI testes was 2,817 bp (GenBank accession number: OL362242). Gene structure analysis showed that RFX2 was located on chromosome 2 of the pig genome with 19 exons. Protein structure analysis indicated that RFX2 contains 728 amino acids with two conserved domains. Phylogenetic analysis revealed that RFX2 was highly conserved with evolutionary homologies among mammalian species. Other analyses, including PPI networks, KEGG, and GO, indicated that BMI RFX2 had interactions with 43 proteins involving various functions, such as in cell cycle, spermatid development, spermatid differentiation, cilium assembly, and cilium organization, etc. Correlation analysis between these proteins and the transcriptome data implied that RFX2 was significantly associated with FOXJ1, DNAH9, TMEM138, E2F7, and ATR, and particularly showed the highest correlation with ATR, demonstrating the importance of RFX2 and ART in spermatogenesis. Functional annotation implied that RFX2 was involved in 17 GO terms, including three cellular components (CC), six molecular functions (MF), and eight biological processes (BP). The analysis of miRNA-gene targeting indicated that BMI RFX2 was mainly regulated by two miRNAs, among which four lncRNAs and five lncRNAs competitively bound ssc-miR-365-5p and ssc-miR-744 with RFX2, respectively. Further, the dual-luciferase report assay indicated that the ssc-miR-365-5p and ssc-miR-744 significantly reduced luciferase activity of RFX2 3'UTR in the 293T cells, suggesting that these two miRNAs regulated the expression of RFX2. Our results revealed the important role of RFX2 in BMI spermatogenesis, making it an intriguing candidate for follow-up studies.

Limited bedding nesting paradigm alters maternal behavior and pup's early developmental milestones but did not induce anxiety- or depressive-like behavior in two different inbred mice.

Animal models are crucial to understanding the mechanisms underlying the deleterious consequences of early-life stress. Here, we aimed to examine the effect of the limited bedding nesting (LBN) paradigm on early life development milestones and anxiety- and/or depression-like behavior in adolescent and adult mice from two inbred mice of both sexes. C57BL/6NCrl and BALB/c litters were exposed to the LBN paradigm postnatal day (PND) 2-9. Maternal behavior recording occurred on PND 3-9, and pups were weighed daily and examined to verify the eye-opening on PND 10-22. The male and female offspring underwent evaluation in the open field test, elevated plus-maze, and the forced swimming test during adolescence (PND 45-49) and adulthood (PND 75-79). We found that LBN impaired the maternal behavior patterns of both strain dams, mainly on C57BL/6NCrl strain. Also, LBN delayed the pup's eye-opening time and reduced body weight gain, impacting C57BL/6NCrl pups more. We also found that LBN decreased anxiety-related indices in adolescent and adult male but not female mice of both strains. Furthermore, LBN decreased depression-related indices only adolescent female and male BALB/c and female but not male C57BL/6NCrl mice. These findings reinforce the evidence that the LBN paradigm impairs the maternal behavior pattern and pup's early developmental milestones but does not induce anxiety- or depressive-like behavior outcomes during later life.

Molecular characteristics and transcriptional regulatory of spermatogenesis-related gene RFX2 in adult Banna mini-pig inbred line (BMI)

RFX2 plays critical roles in mammalian spermatogenesis and cilium maturation. Here, the testes of 12-month-old adult boars of Banna mini-pig inbred line (BMI) were subjected to whole-transcriptome sequencing. The results indicated that the average expression (raw count) of RFX2 gene in BMI testes was 16138.25, and the average expression value of the corresponding transcript ENSSSCT00000043271.2 was 123.1898. The CDS of RFX2 obtained from BMI testes was 2,817 bp (GenBank accession number: OL362242). Gene structure analysis showed that RFX2 was located on chromosome 2 of the pig genome with 19 exons. Protein structure analysis indicated that RFX2 contains 728 amino acids with two conserved domains. Phylogenetic analysis revealed that RFX2 was highly conserved with evolutionary homologies among mammalian species. Other analyses, including PPI networks, KEGG, and GO, indicated that BMI RFX2 had interactions with 43 proteins involving various functions, such as in cell cycle, spermatid development, spermatid differentiation, cilium assembly, and cilium organization, etc. Correlation analysis between these proteins and the transcriptome data implied that RFX2 was significantly associated with FOXJ1, DNAH9, TMEM138, E2F7, and ATR, and particularly showed the highest correlation with ATR, demonstrating the importance of RFX2 and ART in spermatogenesis. Functional annotation implied that RFX2 was involved in 17 GO terms, including three cellular components (CC), six molecular functions (MF), and eight biological processes (BP). The analysis of miRNA-gene targeting indicated that BMI RFX2 was mainly regulated by two miRNAs, among which four lncRNAs and five lncRNAs competitively bound ssc-miR-365-5p and ssc-miR-744 with RFX2, respectively. Further, the dual-luciferase report assay indicated that the ssc-miR-365-5p and ssc-miR-744 significantly reduced luciferase activity of RFX2 3'UTR in the 293T cells, suggesting that these two miRNAs regulated the expression of RFX2. Our results revealed the important role of RFX2 in BMI spermatogenesis, making it an intriguing candidate for follow-up studies.(AU)

Combining datasets for maize root seedling traits increases the power of GWAS and genomic prediction accuracies.

The identification of genomic regions associated with root traits and the genomic prediction of untested genotypes can increase the rate of genetic gain in maize breeding programs targeting roots traits. Here, we combined two maize association panels with different genetic backgrounds to identify single nucleotide polymorphisms (SNPs) associated with root traits, and used a genome-wide association study (GWAS) and to assess the potential of genomic prediction for these traits in maize. For this, we evaluated 377 lines from the Ames panel and 302 from the Backcrossed Germplasm Enhancement of Maize (BGEM) panel in a combined panel of 679 lines. The lines were genotyped with 232 460 SNPs, and four root traits were collected from 14-day-old seedlings. We identified 30 SNPs significantly associated with root traits in the combined panel, whereas only two and six SNPs were detected in the Ames and BGEM panels, respectively. Those 38 SNPs were in linkage disequilibrium with 35 candidate genes. In addition, we found higher prediction accuracy in the combined panel than in the Ames or BGEM panel. We conclude that combining association panels appears to be a useful strategy to identify candidate genes associated with root traits in maize and improve the efficiency of genomic prediction.

Behavioral genetics of alcohol's effects in three zebrafish (Danio rerio) populations.

Alcohol abuse is one of the most dangerous and serious problems for patients and society. Interpopulation studies are important in understanding how genetic background contributes to the effects of alcohol. In this study, we applied a chronic alcohol exposure protocol in three zebrafish populations (Danio rerio; both sexes; AB, TU, and outbred fish - OB). We analyzed the behavioral responses and mRNA expression involved in neurotransmitter metabolism - th1, tph1, ache, ada1, gaba1, gad1b, and bdnf. Locomotion patterns were similar between populations (increased speed after acute alcohol and unaltered locomotion after chronic and withdrawal treatments). All populations exhibited increased expression of genes associated with locomotion (th1, gad1b, and gaba1) after acute alcohol exposure. Anxiety-like responses increased in AB and TU fish during withdrawal and decreased in AB fish after acute alcohol exposure. Genes related to anxiety-like behavior (tph1 and ada1) were overexpressed in AB and TU fish after acute and withdrawal treatments, while OB fish exhibited unaltered responses. Bdnf levels decreased during withdrawal in AB and OB fish, while TU showed upregulated levels in both chronic and withdrawal treatments. Our results suggest that zebrafish populations respond differently to alcohol exposure, which may contribute to understanding the mechanisms underlying alcohol use and dependence. Moreover, we found that a more diverse genetic background (OB) was related to higher variability in behavioral and mRNA expression, demonstrating that inbred populations (AB and TU) may be useful tools in identifying alcohol use and abuse mechanisms.

Maize topcrosses for yield performance by additive main effects and multiplicative interaction analysis / Seleção de topcrosses de milho para rendimento de grãos por meio da análise de efeitos principais aditivos e interação multiplicativa

The analysis of main additive effects and multiplicative interaction is commonly used in the evaluation of the genotype x environment interaction, however, its application can be used for other purposes, as it is performed in the presentresearch, which uses this technique in the selection of inbred lines, testers and hybrids in maize topcrosses. Thisresearch determined the effect of the inbred lines x testers interaction through the analysis of main additive effects and multiplicative interaction, verifying their efficiency in the selection of inbred lines, testers and hybrid combinations in topcrosses. The trials were carried out in the 2015/16 and 2016/17 crop seasons, with a complete block design, with three replications. Thirty S3 maize inbred lines were evaluated in crosses with the AG8025, P30B39, MLP102, 60.H23.1 and 70.H26.1 testers forming 150 hybrids topcrosses. The trait evaluated was grain yield. The adaptability and stability of testers and inbred lines were evaluated by the methodology of analysis of main additive effects and multiplicative interaction directed to the interaction of testers x inbred lines. The 96.3 inbred line has the most homogeneous performance and the highest grain yield, considering the crossing with all testers in both environments. The 70.H26.1 tester is considered the most stable and the most recommended for topcrosses. The best specific combinations were 96.3 x 70.H26.1 and 96.3 x 60.H23.1.

A análise dos principais efeitos aditivos e interação multiplicativa é comumente utilizada na avaliação da interação genótipo x ambiente, porém, sua aplicação pode ser estendida para outros propósitos assim como realizado no presente trabalho em que utiliza esta técnica na seleção de linhagens, testadores e híbridos em topcrosses de milho. O objetivo deste trabalho foi determinar o efeito da interação linhagens x testadores por meio da análise dos principais efeitos aditivos e da interação multiplicativa, verificando sua eficiência na seleção de linhagens, testadoras e combinações híbridas em topcrosses. Os ensaios foram realizados nas safras 2015/16 e 2016/17, com delineamento em blocos completos, com três repetições. Trinta linhagens de milho S3 foram avaliadas em cruzamentos com os testadores AG8025, P30B39, MLP102, 60.H23.1 e 70.H26.1 formando 150 topcrosses híbridos. A característica avaliada foi a produtividade de grãos. A adaptabilidade e estabilidade de testadores e linhagens foram avaliadas pela metodologia de análise de efeitos aditivos principais e interação multiplicativa direcionada à interação testadores x linhagens. A linhagem 96.3 apresentou o desempenho mais homogêneo e o maior rendimento de grãos, considerando o cruzamento com todos os testadores em ambos os ambientes. O testador 70.H26.1 é considerado o mais estável e o mais recomendado para topcrosses. As melhores combinações específicas foram 96.3 x 70.H26.1 e 96.3 x 60.H23.1.

Exposure to glyphosate-based herbicide during early stages of development increases insulin sensitivity and causes liver inflammation in adult mice offspring

ABSTRACT Objective: To investigate the effect of pre and postnatal exposure to a glyphosate-based herbicide on glucose metabolism and liver histology in adult F1 mice offspring. Methods: Female mice (C57Bl/6) received 0.5% of glyphosate (Roundup Original DI®) in drinking water or purified water (Glyphosate Group and Control Group respectively) during pregnancy and lactation. Offspring (F1) were submitted to glucose and insulin tolerance tests and euthanized on postnatal day 150. Body and plasma parameters, and liver histology were analyzed. Results: Exposure to glyphosate reduced maternal body weight gain during pregnancy and lactation, with no impacts on litter size. Pre and postnatal exposure to glyphosate did not affect body parameters but increased glucose tolerance on postnatal day 60. In spite of glucose tolerance normalization by postnatal day 143, this effect was associated with higher insulin sensitivity relative to mice in the Control-F1 Group. Mice in the Glyphosate-F1 Group had mild and moderate lobular inflammation in the liver. Conclusion: Maternal exposure to glyphosate affected insulin sensitivity and caused hepatic inflammation in adult F1 mice offspring.

Comparative hormonal and metabolic profile analysis based on mass spectrometry provides information on the regulation of water-deficit stress response of sunflower (Helianthus annuus L.) inbred lines with different water-deficit stress sensitivity.

Water-deficit stress is the most important abiotic stress restricting plant growth, development and yield. The effects of this stress, however, depend on genotypes, among other factors. This study assembles morpho-physiological and metabolic approaches to assess hormonal and metabolic profile changes, upon water-deficit stress, in the shoot and roots of two contrasting sunflower inbred lines, B59 (water-deficit stress sensitive) and B71 (water-deficit stress tolerant). The analyses were carried out using mass spectrometry and performing a multivariate statistical analysis to identify relationships between the analyzed variables. Water-deficit stress reduced all morpho-physiological parameters, except for root length in the tolerant inbred line. The hormonal pathways were active in mediating the seedling performance to imposed water-deficit stress in both lines, although with some differences between lines at the organ level. B59 displayed a diverse metabolite battery, including organic acids, organic compounds as well as sugars, mainly in the shoot, whereas B71 showed primary amino acids, organic acids and organic compounds predominantly in its roots. The discrimination between control and water-deficit stress conditions was possible thanks to potential biomarkers of stress treatment, e.g., proline, maleic acid and malonic acid. This study indicated that the studied organs of sunflower seedlings have different mechanisms of regulation under water-deficit stress. These findings could help to better understand the physio-biochemical pathways underlying stress tolerance in sunflower at early-growth stage.

Identification of genetic modifiers of murine hepatic ß-glucocerebrosidase activity.

The acid ß-glucocerebrosidase (GCase) enzyme cleaves glucosylceramide into glucose and ceramide. Loss of function variants in the gene encoding for GCase can lead to Gaucher disease and Parkinson's disease. Therapeutic strategies aimed at increasing GCase activity by targeting a modulating factor are attractive and poorly explored. To identify genetic modifiers, we measured hepatic GCase activity in 27 inbred mouse strains. A genome-wide association study (GWAS) using GCase activity as a trait identified several candidate modifier genes, including Dmrtc2 and Arhgef1 (p=2.1x10-7), and Grik5 (p=2.1x10-7). Bayesian integration of the gene mapping with transcriptomics was used to build integrative networks. The analysis uncovered additional candidate GCase regulators, highlighting modules of the acute phase response (p=1.01x10-8), acute inflammatory response (p=1.01x10-8), fatty acid beta-oxidation (p=7.43x10-5), among others. Our study revealed previously unknown candidate modulators of GCase activity, which may facilitate the design of therapies for diseases with GCase dysfunction.

The morphine/heroin vaccine decreased the heroin-induced antinociceptive and reinforcing effects in three inbred strains mouse.

Clinical trials have indicated that a vaccine must be immunogenic in genetically diverse human populations and that immunogenicity and protective efficacy in animal models are two key indices required for the approval of a new vaccine. Additionally, the immune response (immunogenicity) and immunoprotection are dependent on the mouse strain. Therefore, the objective of the present study was to determine the immune response (immunogenicity) and the protective efficacy (behavioral response) in three inbred mouse strains immunized with the M6TT vaccine. Female BALB/c, C57Bl/6, and DBA/2 inbred mice were immunized with the M6-TT vaccine. A solid-phase antibody-capture ELISA was used to monitor antibody titer responses after each booster dose in vaccinated animals. The study used tail-flick testing to evaluate the antinociceptive effects induced by heroin. Additionally, heroin-induced locomotor activity and place preference were evaluated. The M6-TT vaccine was able to generate a specific antibody titer in the three inbred mouse strains evaluated. The antibodies reduced the antinociceptive effect of different doses of heroin. In addition, they decreased the heroin-induced locomotor activity and place preference. These findings suggest that the M6-TT vaccine generates a powerful immunogenic response capable of reducing the antinociceptive and reinforcing effects of heroin in different inbred mouse strains, which supports its possible future use in clinical trials in genetically diverse human populations.

Study of muscle fibers of the extensor digitorium longus and soleus muscles of C57BL/6 females exposed to glyphosate during pregnancy and lactation / Estudo das fibras musculares dos músculos extensor longo dos dedos e sóleo de fêmeas C57BL/6 expostas ao glifosato durante a prenhez e a lactação

ABSTRACT Objective To evaluate the morphology and morphometry of the muscles extensor digitorium longus and soleus of C57BL/6 females, who were exposed to glyphosate during pregnancy and lactation. Methods Twelve female mice from the C57BL/6 lineage were used. After detection of pregnancy, they were divided into a Control Group, which received only water, and a Glyphosate Group, which received water with 0.5% glyphosate during pregnancy and lactation. Both groups received ad libitum standard diet. After weaning, the females were euthanized and weighed; naso-anal length was measured, and fats were collected and weighed. The muscles extensor digitorium longus and soleus were collected, and their length and weight were measured. Then, the muscles were fixed in Methacarn to perform the histological study of muscle fibers. Results Glyphosate Group presented lower weight gain during pregnancy and also lower final body weight and naso-anal length; however, the other body parameters evaluated did not present a significant difference in relation to the Control Group. Significant differences were also not observed in the analysis of muscle fibers and connective tissue. Conclusion Exposure to 0.5% glyphosate during pregnancy and lactation resulted in lower weight gain during pregnancy, final weight, and naso-anal length. Despite not directly altering the morphology of muscle tissue, these results may indicate enough exposure to interfere with animal metabolism.

RESUMO Objetivo Avaliar a morfologia e a morfometria dos músculos extensor longo dos dedos e sóleo de fêmeas C57BL/6 expostas ao glifosato durante a prenhez e lactação. Métodos Foram utilizados 12 camundongos fêmeas da linhagem C57BL/6. Após detecção da prenhez, foram separadas em Grupo Controle, que recebeu somente água, e Grupo Glifosato, que recebeu água com 0,5% de glifosato durante a prenhez e lactação. Ambos os grupos receberam dieta padrão ad libitum. Após o desmame, as fêmeas foram eutanasiadas e pesadas; o comprimento nasoanal foi mensurado, e as gorduras foram coletadas e pesadas. Os músculos extensor longo dos dedos e sóleo foram coletados, e seu comprimento e peso foram mensurados. Em seguida, os músculos foram fixados em Methacarn para a realização do estudo histológico das fibras musculares. Resultados O Grupo Glifosato apresentou menor ganho de peso durante a prenhez e também menor peso corporal final e comprimento nasoanal, entretanto os demais parâmetros corporais avaliados não apresentaram diferença significativa em relação ao Grupo Controle. Na análise das fibras musculares e do tecido conjuntivo, também não foram observadas diferenças significativas. Conclusão A exposição a 0,5% de glifosato durante a prenhez e lactação resultou em menor ganho de peso na gestação, peso final e comprimento nasoanal, o que pode indicar que, apesar de não alterar a morfologia do tecido muscular diretamente, a exposição foi suficiente para interferir no metabolismo dos animais.

Host Genetics Background Influence in the Intragastric Trypanosoma cruzi Infection.

Background: Considering the complexity of the factors involved in the immunopathology of Chagas disease, which influence the Chagas' disease pathogenesis, anti-T. cruzi immune response, and chemotherapy outcome, further studies are needed to improve our understanding about these relationships. On this way, in this article we analyzed the host genetic influence on hematological, histopathological and immunological aspects after T. cruzi infection. Methods: BALB/c and A mice were intragastrically infected with T. cruzi SC2005 strain, isolated from a patient of an outbreak of Chagas disease. Parameters such as parasite load, survival rates, cytokines production, macrophages, T and B cell frequencies, and histopathology analysis were carried out. Results: BALB/c mice presented higher parasitemia and mortality rates than A mice. Both mouse lineages exhibited hematological alterations suggestive of microcytic hypochromic anemia and histopathological alterations in stomach, heart and liver. The increase of CD8+ T cells, in heart, liver and blood, and the increase of CD19+ B cells, in liver, associated with a high level of proinflammatory cytokines (IL-6, TNF-α, IFN-γ), confer a resistance profile to the host. Although BALB/c animals exhibited the same findings observed in A mice, the response to infection occurred later, after a considerable parasitemia increase. By developing an early response to the infection, A mice were found to be less susceptible to T. cruzi SC2005 infection. Conclusions: Host genetics background shaping the response to infection. The early development of a cytotoxic cellular response profile with the production of proinflammatory cytokines is important to lead a less severe manifestation of Chagas disease.

Electrocardiographic Abnormalities in Hypertension Models

Abstract Background: Hypertensive condition can lead to abnormalities in heart structure and electrical activity. The electrocardiogram (ECG) is a recording of the electrical activity of the heart and widely used to diagnose and detect heart problem. Objective: We conducted a comparative ECG analysis between two hypertension models (L-NAME and SHR) and their controls (Wistar and Wistar-Kyoto) at six and 15 th week of age. Methods: Blood pressure was measured at the end of the 15 th week, and electrocardiography was performed at six and 15 weeks of age in anaesthetized rats. Data normality was confirmed by Kolmogorov-Smirnov test followed by unpaired Student's t-test and the Mann-Whitney for parametric and non-parametric data, respectively. Results are expressed as mean ± SD. The accepted level of significance was set at p < 0.05. Results: L-NAME exhibited prolongation of JT and QT intervals and SHR showed a decrease in heart rate when compared to Wistar-Kyoto and L-NAME. Wistar-Kyoto exhibited short PR interval with increased QRS complex, and only QT prolongation at 15 weeks compared to Wistar. Conclusions: All the hypertension models used in this study featured an increase in blood pressure. However, while SHR showed cardiac dysfunction, L-NAME exhibited changes in ventricular performance. These results may guide future studies on different types and models of hypertension.

Efecto de la endocría sobre habilidad combinatoria del rendimiento y calidad en zapallo ( Cucurbita moschata Duchesne) / Inbreeding effect in combinatorial ability of yield and quality on butternut squash ( Cucurbita moschata Duchesne)

RESUMEN El zapallo Cucurbita moschata Duchesne es una hortaliza para diversos procesos culinarios y agroindustriales, por su color, sabor y textura; sin embargo, estas características cambian dependiendo del cultivar. Con base en 6 progenitores, 15 híbridos F1, sin incluir los recíprocos, se estudió el efecto de la endocría en la habilidad combinatoria (HC), de caracteres asociados con el rendimiento y la calidad del fruto del zapallo, en dos localidades, Candelaria y Buga, Valle del Cauca, Colombia. Se usó cruzamientos dialélicos y progenitores con diferente grado de endocría (S0xS0, S1xS1 y S2xS2). Se utilizó el método 2 y modelo I, propuesto por Griffing y el análisis de Hallauer & Miranda, para evaluar la habilidad combinatoria general (HCG) y específica (HCE) del rendimiento y la calidad del fruto, con fines agroindustriales. Todos los caracteres evaluados variaron entre generaciones de endocría. Se detectó interacción entre generación de endocría por localidad para todos los caracteres (P<0,01), con excepción de almidón (AL) y caroteno total (CT) en fruto. La HCG y HCE no se ajustaron a un patrón de comportamiento, debido a la variación de los progenitores en la susceptibilidad a la endocría. Los progenitores con HCG altamente significativa (P<0,01) para producción, fueron: 79S0, 79S1, 80S0 y 34S1, en Candelaria y 79S0, 79S1, 79S2, 34S0, 6S0, 6S1, 2S1 y 2S2, en Buga. Los mejores híbridos con alta HCE y alto rendimiento y calidad, para la localidad Candelaria, fueron: 2x28-S0, 2x80-S0, 28x80-S0 y 80x34 y para la localidad Buga: 2x6-S1, 80x34-S1, 2x80-S2 y 79x34-S2.

ABSTRACT Butternut squash, Cucubita moschata Duchesne, is a vegetable used for various culinary and agro-industrial processes due to its colour, flavour and texture; however, these characteristics change depending on the cultivar. Based on 6 parents, 15 F1 hybrids, not including reciprocals, the effect of inbreeding on the combinatorial ability (HC) of characters associated with the performance and quality of Butternut squash fruit was studied; in two locations, Candelaria and Buga, Valle del Cauca-Colombia. Diallel and progenitor crosses with different degrees of inbreeding (S0xS0, S1xS1 and S2xS2) were used. Method 2 and model I proposed by Griffing and the Hallauer & Miranda analysis were applied to assess the general combinatorial (HCG) and specific combinatorial ability (HCE) of the yield and quality of the fruit, for agroindustrial purposes. All evaluated characters varied between inbreeding generations. Interaction between inbreeding generation by locality was detected for all characters (P <0.01), with the exception of starch (AL) and total carotene (CT) in fruit. The HCG and HCE did not adjust to expected behaviour pattern, due to the variation of parents to inbreeding susceptibility. The parents with highly significant HCG (P <0.01) for production were 79S0, 79S1, 80S0 y 34S1 in Candelaria and 79S0, 79S1, 79S2, 34S0, 6S0, 6S1, 2S1 y 2S2 to Buga. The best hybrids with high HCE and high performance and quality, for Candelaria locality were: 2x28-S0, 2x80-S0, 28x80-S0 and 80x34 and for locality Buga: 2x6-S1, 80x34-S1, 2x80-S2 and 79x34-S2.

Gastrointestinal motility and morphology in mice: Strain-dependent differences.

BACKGROUND: BALB/c and C57BL/6 mice are widely used in biomedical research; however, the differences between strains are still underestimated. Our aims were to develop an experimental protocol to evaluate the duodenal contractility and gastrointestinal transit in mice using the Alternating Current Biosusceptometry (ACB) technique and to compare gastrointestinal motor function and morphology between BALB/c and C57BL/6 strains. METHODS: Male mice were used in experiments (a) duodenal contractility: animals which had a magnetic marker surgically fixed in the duodenum to determine the frequency and amplitude of contractions and (b) gastrointestinal transit: animals which ingested a magnetically marked chow to calculate the Oro-Anal Transit Time (OATT) and the Fecal Pellet Elimination Rate (FPER). The animals were killed after the experiments for organ collection and morphometric analysis. KEY RESULTS: BALB/c and C57BL/6 had two different duodenal frequencies (high and low) with similar amplitudes. After 10 hours of monitoring, BALB/c eliminated around 89% of the ingested marker and C57BL/6 eliminated 33%; OATT and FPER were slower for C57BL/6 compared with BALB/c. The OATT and amplitude of low frequency had a strong positive correlation in C57BL/6. For BALB/c, the gastric muscular layer was thicker compared to that measured for C57BL/6. CONCLUSIONS AND INFERENCES: The experimental protocol to evaluate duodenal contractility and fecal magnetic pellets output using the ACB technique in mice was successfully established. BALB/c strains had higher duodenal frequencies and a shorter time to eliminate the ingested marker. Our results showed differences in both motor function and gastrointestinal morphology between BALB/c and C57BL/6 strains.

Quantification of DNA Damage in Different Tissues in Rats with Heart Failure / Quantificação de Dano em DNA em Diferentes Tecidos em Ratos com Insuficiência Cardíaca

Abstract Background: Chronic heart failure (CHF) is a complex syndrome which comprises structural and functional alterations in the heart in maintaining the adequate blood demand to all tissues. Few investigations sought to evaluate oxidative DNA damage in CHF. Objective: To quantify the DNA damage using the comet assay in left ventricle (LV), lungs, diaphragm, gastrocnemius and soleus in rats with CHF. Methods: Twelve male Wistar rats (300 to 330 g) were selected for the study: Sham (n = 6) and CHF (n = 6). The animals underwent myocardial infarction by the ligation of the left coronary artery. After six weeks, the animals were euthanized. It was performed a cell suspension of the tissues. The comet assay was performed to evaluate single and double strand breaks in DNA. Significance level (p) considered < 0.05. Results: The CHF group showed higher values of left ventricle end-diastolic pressure (LVEDP), pulmonary congestion, cardiac hypertrophy and lower values of maximal positive and negative derivatives of LV pressure, LV systolic pressure (p < 0.05). CHF group showed higher DNA damage (% tail DNA, tail moment and Olive tail moment) compared to Sham (p < 0.001). The tissue with the highest damage was the soleus, compared to LV and gastrocnemius in CHF group (p < 0.05). Conclusion: Our results indicates that the CHF affects all tissues, both centrally and peripherically, being more affected in skeletal muscle (soleus) and is positively correlated with LV dysfunction.

Resumo Fundamento: A insuficiência cardíaca crônica (ICC) é uma síndrome complexa que compreende alterações estruturais e funcionais no coração, mantendo demanda sanguínea adequada a todos os tecidos. Poucas investigações procuraram avaliar o dano oxidativo ao DNA na ICC. Objetivo: Quantificar o dano ao DNA utilizando o ensaio cometa no ventrículo esquerdo (VE), pulmões, diafragma, gastrocnêmio e sóleo em ratos com ICC. Métodos: Doze ratos Wistar machos (300 a 330 g) foram selecionados para o estudo: placebo (n = 6) e ICC (n = 6). Os animais foram submetidos a infarto do miocárdio através de ligadura da artéria coronária esquerda. Após seis semanas, os animais foram sacrificados. Foi realizada uma suspensão celular dos tecidos. O ensaio cometa foi realizado para avaliar as quebras de fita simples e dupla no DNA. Nível de significância (p) < 0,05. Resultados: O grupo ICC apresentou maiores valores de pressão diastólica final do ventrículo esquerdo (PDFVE), congestão pulmonar, hipertrofia cardíaca e menores valores de derivados máximos positivos e negativos da pressão do VE, pressão sistólica do VE (p < 0,05). O grupo ICC apresentou maior dano ao DNA (% de DNA da cauda, momento da cauda e momento da cauda de Olive) em comparação ao placebo (p < 0,001). O tecido com maior dano foi o sóleo, comparado ao VE e ao gastrocnêmio no grupo ICC (p < 0,05). Conclusão: Nossos resultados indicam que a ICC afeta todos os tecidos, de maneira central e periférica, sendo mais afetada no músculo esquelético (sóleo) e está positivamente correlacionada com a disfunção do VE.