Abstract Background The distinct arterial and venous cell fates are dictated by a combination of various genetic factors which form diverse types of blood vessels such as arteries, veins, and capillaries. We report here that YULINK protein is involved in vasculogenesis, especially venous formation. Methods In this manuscript, we employed gene knockdown, yeast two-hybrid, FLIM-FRET, immunoprecipitation, and various imaging technologies to investigate the role of YULINK gene in zebrafish and human umbilical vein endothelial cells (HUVECs). Results Knockdown of YULINK during the arterial-venous developmental stage of zebrafish embryos led to the defective venous formation and abnormal vascular plexus formation. Knockdown of YULINK in HUVECs impaired their ability to undergo cell migration and differentiation into a capillary-like tube formation. In addition, the phosphorylated EPHB4 was decreased in YULINK knockdown HUVECs. Yeast two-hybrid, FLIM-FRET, immunoprecipitation, as well as imaging technologies showed that YULINK colocalized with endosome related proteins (EPS15, RAB33B or TICAM2) and markers (Clathrin and RHOB). VEGF-induced VEGFR2 internalization was also compromised in YULINK knockdown HUVECs, demonstrating to the involvement of YULINK. Conclusion This study suggests that YULINK regulates vasculogenesis, possibly through endocytosis in zebrafish and HUVECs. Key points Knockdown of YULINK with morpholino in embryos of double transgenic zebrafish exhibited abnormal venous formation. Tube formation and phosphorylated EPHB4 were decreased in YULINK knockdown HUVECs. FLIM-FRET, immunoprecipitation, as well as other imaging technologies showed that YULINK colocalized with endosome related proteins (EPS15, RAB33B and TICAM2) and endosome markers (Clathrin and RHOB). Knockdown of YULINK decreased the internalization of VEGF and VEGFR2 in HUVECs.

Abstract Lychnophora pinaster, known as arnica-mineira, is endemic to campos rupestres, at risk of extinction. The present study aimed to characterize the ecogeography and phenolic constituents of 11 L. pinaster populations collected in the mesoregions North, Jequitinhonha, Metropolitan of Belo Horizonte and Campos das Vertentes in the state of Minas Gerais, Brazil. Phenolic constituents were identified and quantified by Ultra-high performance liquid chromatography- mass spectrometry. Lychnophora pinaster occurs in sites at high altitude (700 and 1498 m), annual rainfall of up to 1455 m, soils with low fertility and predominantly loamy texture. Therefore, it can be considered tolerant to acidic soils, with low availability of nutrient. The most abundant substances in all populations were vitexin (18 – 1345 ng/g) and chlorogenic acid (60 – 767 ng/g). The 11 populations formed four groups in relation to the phenolic constituents, with group 1 consisting of the populations of the North Mesoregion (GM, OD) and Jetiquinhonha (DIMa), group 2, the Metropolitan of Belo Horizonte (SRM, NLSC, SM, RPS, CTRA), group 3, the North Mesoregion (ODMa and DI), and group 4, the Campos das Vertentes (CC). Among the populations, only those from the Metropolitan of Belo Horizonte showed correlation of soil properties with phenolic constituents.

Abstract LaoXiangHuang (LXH) is a medical pickled fruit made of fresh finger citron (Citrus medica L. var. sarcodactylis) in Chaozhou, China. Aging is a process to strengthen the efficacy of Laoxianghuang (LXH), which is related to the activity of microorganisms. In this study, the changes of the microbial community of LXH within the three-years aging process were analyzed by high-throughput sequencing (HTS). According to alpha diversity results, bacterial abundance and diversity were higher than those of fungi. A total of 3 phyla and 13 bacterial genera were identified whereas 2 phyla and 4 fungal genera were identified. Meta-taxonomic of the overall bacterial communities in samples revealed that Halomonas spp. was the most abundant genus in all samples, which showed the first decresed, and then increased and decreased finally. The followed by Nitriliuptor spp., Nesterenkonia spp., Weissella spp., Notinibacter spp., Streptococcus spp., Nocardiopsis spp., Acinetobacter spp., Ralstonia spp., Lactobacillus spp. and unassigned genus. The unassigned genera (through the whole three-years fermentation period), Ustilago spp., Malassezia spp., Aspergillus spp. and Schizophyllum spp. were the predominant fungal genera. The result of PCOA and T-test analysis exhibited that the samples with different aging times have significant differences and are mainly reflected in Halomonas, unassigned bacterial genera, Nitriliruptor, Nesterenkonia, Weissella, Intestinibacter, Streptococcus and Nocardiopsis, Lactobacillus, Acinetobacter, Ralstonia, unassigned fungal genera, Ustilago spp. Malassezia spp. Aspergillus spp. and Schizophyllum spp., which may have important potential in promoting the aging process of LXH. This study helps us to understand the dynamic change of microorganisms of LXH during the aging process and provides a research basis for shortening the aging time of LXH in the future.

Abstract Aflatoxin B1 (AFB1), a widespread mycotoxin found in food, poses a significant threat to the food industry and production. The primary target of AFB1 is the liver. Probiotic-mediated antitoxicity has been proposed to overcome aflatoxin toxicity. In the present study, to investigate the protective effects and molecular mechanisms of Lactobacillus plantarum SCS2 (L. plantarum SCS2) against HepG2 cells oxidative stress induced by AFB1. HepG2 cells were cultured and treated with different concentrations of AFB1 (0-10 μmol/L) to induce oxidative stress and L. plantarum SCS2 (total protein, 0.4 mg/mL) was pretreated HepG2 for 2 h and then supplemented with AFB1 (10 μmol/L) for 22 h to observe cellular oxidative stress. The cytotoxicity, reactive oxygen species, malondialdehyde, activities and mRNA expressions of antioxidant enzymes and activation of the Nrf2 signaling pathway were measured. The results showed that AFB1 exposure caused oxidative stress in HepG2. Supplementation of L. plantarum SCS2 prevented AFB1-induced HepG2 cells activity decreased and activated antioxidant enzymes activity by inhibiting Nrf2 over expression. The results of in vitro experiments revealed that L. plantarum SCS2 evidently protected against AFB1-induced oxidative stress.

Abstract Several technological applications of Lactococcus lactis are attributed to their high metabolite-secreting abilities. Among these, bacteriocins can be used as food preservatives or additives in cheese manufacture. The objective was to evaluate the technological characteristics and antibacterial activity of 10 Lactococcus lactis isolates from traditional fermented dairy products, including acidifying ability, proteolytic ability, diacetyl production and autolytic activity. L. lactis subsp. cremoris Y14 and Y15 showed significantly (P < 0.05) higher ΔpH than other strains. L. lactis subsp. cremoris Y17 exhibited the highest proteolytic, L. lactis subsp. cremoris Y15 and L. lactis subsp. cremoris Y18 were next in terms of proteolytic activity. All strains possessed the ability of diacetyl production, with the highest production of 10.39 mg/g observed in L. lactis subsp. cremoris Y15. L. lactis subsp. cremoris Y14 had the highest (P < 0.05) autolytic ability (8.96%), followed by L. lactis subsp. cremoris Y15 (8.05%). L. lactis Y15 exhibited discernible antibacterial activity with the largest zone of inhibition diameters (25.56 mm). Based on the above results, L. lactis subsp. cremoris Y15 was selected as the starter to manufacture cheese. There were no significant differences in texture and sensory acceptability of Cheddar cheese manufactured by L. lactis subsp. cremoris Y15 with L. lactis subsp. lactis KLDS4.0325 or commercial starter. L. lactis subsp. cremoris Y15 can be used as a starter to produce Cheddar cheese.

Resumo Ensaios práticos de enzimas para detectar contaminação subletal de peixes neotropicais são da maior importância. Assim, ensaiamos a atividade da glutationa S-transferase (GST) em citosóis de fígado e de rim de Piaractus mesopotamicus injetados com metilparation ou benzo[a]pireno usando 1-cloro-2,4-dinitrobenzeno (CDNB), o substrato usual para ensaiar uma denominada atividade geral de GST. Desde que para indicar alterações na atividade de uma isoenzima, é necessário o substrato mais reativo, também usamos dois substratos alternativos, o 1,2-dicloro-4-nitrobenzeno (DCNB) e o ácido etacrínico (ETHA). As atividades com CDNB ou ETHA não mudaram. Entretanto, ensaios com DCNB mostraram que metilparation decresceu a atividade de GST no fígado em 24, 48 e 96 horas depois da injeção. O DCNB também revelou que sete dias depois da injeção de benzo[a]pireno a atividade da GST aumentou no fígado, normalizando depois de 14 dias. Benzo[a]pireno, mas não metilparation, aumentou as atividades com DCNB no citosol dos rins sete e 14 dias depois da injeção. Assim, uma atividade de GST ensaiada com DCNB diminuída no fígado correspondeu à contaminação de P. mesopotamicus com metilparation. O aumento dessa atividade de GST no fígado e nos rins está correlacionada com contaminação do pacu por hidrocarbonetos policíclicos aromáticos.

Abstract Functional enzyme assays to detect sublethal poisoning of Neotropical fish are paramount. Accordingly, we assayed a glutathione S-transferase (GST) activity in liver and kidney cytosols from Piaractus mesopotamicus injected with methyl parathion or benzo[a]pyrene using the substrate 1-chloro-2,4-dinitrobenzene (CDNB), which is the usual substrate for assaying a known general activity of GST. Since the most reactive substrate is required to reveal specific changes in enzyme activity, we also used two alternative substrates, 1,2-dichloro-4-nitrobenzene (DCNB) and ethacrynic acid (ETHA). Activities with CDNB or ETHA did not change. However, assays with DCNB showed that methyl parathion caused a decrease in GST activity in the liver on the 24th, 48th and 96th hour after the injection. DCNB also revealed that GST activity in the liver increased seven days after benzo[a]pyrene injection, coming down to normal after fourteen days. Benzo[a]pyrene, but not methyl parathion, increased the activities with DCNB in cytosol from the kidney seven and fourteen days after the injection. Thus, a decreased liver GST activity assayed with DCNB corresponded to contamination of P. mesopotamicus with methyl parathion. The increase of this GST activity in the liver and the kidney correlates to pacu contamination with polycyclic aromatic hydrocarbons.

Malt is the mature fruit of Hordeum vulgare L. after germination and drying and has been applied for treatment female abnormal galactorrhea. Previous studies have showed total alkaloids in malt have anti-HPRL effect. However, total alkaloids of malt change with the growth cycle, and the specified levels of total alkaloids in different bud length of malt have not been decided. To determine the definitive level of total alkaloids in different buds of malt and the most suitable bud length for clinical application by comparing effects on hyperprolactinemia rat. During the budding of malt, the content of total alkaloids first increased and then decreased, and it peaked at a bud length of 0.75 cm. Treated the HPRL model rats with different buds of malt, the PRL level was decreased, the number of PRLpositive cells and the mRNA expression level in the pituitary were significantly declined, and the number of dopamine D1 and D2 receptors in the hypothalamus was increased. The above changes were most significant in 0.75 cm bud. These results suggest that in terms of the content of effective substance and the effects on HPRL model rats, a malt bud length of 0.75 cm is optimal for clinical application.

Resumo O objetivo deste estudo foi avaliar a capacidade de sobrevivência de Lactiplantibacillus plantarum 299v encapsulado em esferas de alginato de cálcio revestidos com quitosana, com inulina como prebiótico, em condições gastrointestinais simuladas e suco de rosélia. A concentração de cloreto de cálcio e inulina para a microencapsulação de L. plantarum 299v foi otimizada e a sobrevivência de L. plantarum livre e microencapsulado foi avaliada em condições gastrointestinais simuladas. A estabilidade do encapsulado otimizado L. plantarum 299v-inulina durante armazenamento foi determinada ao longo de quatro semanas em suco de rosélia a 4 °C e 25 °C. A fórmula otimizada para L. plantarum 299v foi de 2,0% (w/v) de cloreto de cálcio e 3,0% (w/ v) de inulina. Microesferas otimizadas de alginato de cálcio-quitosana contendo L. plantarum 299v com inulina apresentaram diâmetro médio de 685,27 μm sem diferença significativa (p>0,05) em relação às partículas sem inulina, e eficiência de microencapsulação de 95%. L. plantarum 299v encapsulado com inulina mostrou maior capacidade de sobrevivência (> 107 CFU/mL) do que células livres e L. plantarum 299v encapsulado sem inulina em condições gastrointestinais simuladas e após quatro semanas de armazenamento em suco de rosélia a 4 °C. Os resultados indicam que a a encapsulação por coextrusão e a adição de inulina melhoraram a viabilidade de L. plantarum 299v em suco de rosélia, protegendo o probiótico contra condições gastrointestinais desfavoráveis e prolongando o armazenamento.

Abstract This study aimed to evaluate the survivability of Lactiplantibacillus plantarum 299v encapsulated in chitosan-coated calcium alginate beads with inulin as prebiotic in simulated gastrointestinal conditions and roselle juice. The concentration of calcium chloride and inulin for L. plantarum 299v microencapsulation was optimised and the survivability of free and microencapsulated L. plantarum was assessed under simulated gastrointestinal conditions. Storage stability of the optimised encapsulated L. plantarum 299v-inulin was determined throughout four (4) weeks of storage in roselle juice at 4 °C and 25 °C. The optimized formula for L. plantarum 299v was 2.0% (w/v) of calcium chloride and 3.0% (w/v) of inulin. Optimized calcium alginate-chitosan L. plantarum 299v microbeads with inulin did not affect (p > 0.05) the bead diameter, with a mean diameter of 685.27 μm, and microencapsulation efficiency of 95%. Encapsulated L. plantarum 299v with inulin showed higher survivability (>107 CFU/mL) than free cells and encapsulated L. plantarum 299v without inulin under simulated gastrointestinal conditions and after four (4) weeks of storage in roselle juice at 4 °C. The results indicate that co-extrusion encapsulation and addition of inulin had improved the viability of L. plantarum 299v in roselle juice by protecting probiotic against unfavourable gastrointestinal conditions and prolonged storage.

Abstract Background: Liriodendron chinense ranges widely in subtropical China and northern Vietnam; however, it inhabits several small, isolated populations and is now an endangered species due to its limited seed production. The objective of this study was to develop a set of nuclear SSR (simple sequence repeats) and multiple chloroplast genome markers for genetic studies in L. chinense and their characterization in diverse germplasm. Results: We performed low-coverage whole genome sequencing of the L. chinense from four genotypes, assembled the chloroplast genome and identified nuclear SSR loci by searching in contigs for SSR motifs. Comparative analysis of the four chloroplast genomes of L. chinense revealed 45 SNPs, 17 indels, 49 polymorphic SSR loci, and five small inversions. Most chloroplast intraspecific polymorphisms were located in the interspaces of single-copy regions. In total, 6147 SSR markers were isolated from low-coverage whole genome sequences. The most common SSR motifs were dinucleotide (70.09%), followed by trinucleotide motifs (23.10%). The motif AG/TC (33.51%) was the most abundant, followed by TC/AG (25.53%). A set of 13 SSR primer combinations were tested for amplification and their ability to detect polymorphisms in a set of 109 L. chinense individuals, representing distinct varieties or germplasm. The number of alleles per locus ranged from 8 to 28 with an average of 21 alleles. The expected heterozygosity (He) varied from 0.19 to 0.93 and the observed heterozygosity (Ho) ranged from 0.11 to 0.79. Conclusions: The genetic resources characterized and tested in this study provide a valuable tool to detect polymorphisms in L. chinense for future genetic studies and breeding programs.

Abstract Background: Recent studies indicate that circular RNAs (circRNAs) may play important roles in the regulation of plant growth and development. Plant roots are the main organs of nutrient and water uptake. However, whether circRNAs involved in the regulation of plant root growth remains to be elucidated. Methods: LH9, XN979 and YN29 are three Chinese wheat varieties with contrasting root lengths. Here, the root circRNA expression profiles of LH9, XN979 and YN29 were examined by using high-throughput sequencing technology. Results: Thirty-three and twenty-two differentially expressed circRNAs (DECs) were identified in the YN29-LH9 comparison and YN29-XN979 comparison, respectively. Among them, ten DECs coexisted in both comparisons. As the roots of both LH9 and XN979 were significantly larger and deeper than YN29, the ten DECs coexisting in the two comparisons were highly likely to be involved in the regulation of wheat root length. Moreover, three of the ten DECs have potential miRNA binding sites. Real-time PCR analysis showed that the expression levels of the potential binding miRNAs exhibited significant differences between the long root plants and the short root plants. Conclusions: The expression levels of some circRNAs exhibited significant differences in wheat varieties with contrasting root phenotypes. Ten DECs involved in the regulation of wheat root length were successfully identified in which three of them have potential miRNAs binding sites. The expression levels of putative circRNA-binding miRNAs were correlated with their corresponding circRNAs. Our results provide new clues for studying the potential roles of circRNAs in the regulation of wheat root length.

Abstract Background: Drought is a major abiotic stress affecting global wheat (Triticum aestivum L.) production. Exploration of drought-tolerant genes is essential for the genetic improvement of drought tolerance in wheat. Previous studies have shown that some histone encoding genes are involved in plant drought tolerance. However, whether the H2B family genes are involved in drought stress response remains unclear. Methods: Here, we identified a wheat histone H2B family gene, TaH2B-7D, which was significantly up-regulated under drought stress conditions. Virus-induced gene silencing (VIGS) technology was used to further verify the function of TaH2B-7D in wheat drought tolerance. The phenotypic and physiological changes were examined in the TaH2B-7D knock-down plants. Results: In the TaH2B-7D knock-down plants, relative electrolyte leakage rate and malonaldehyde (MDA) content significantly increased, while relative water content (RWC) and proline content significantly decreased compared with those in the non-knocked-down plants under drought stress conditions. TaH2B-7D knock-down plants exhibited severe sagging, wilting and dwarf phenotypes under drought stress conditions, but not in the non-knocked-down plants, suggesting that the former were more sensitive to drought stress. Conclusion: These results indicate that TaH2B-7D potentially plays a vital role in conferring drought tolerance in wheat.

RESUMO: A salinidade do solo limita a produção agrícola. O trigo mole é uma das culturas mais importantes com característica allohexaploid e genoma altamente complexo. O mapeamento QTL é uma maneira muito útil de identificar genes para traços quantitativos, como a tolerância à salinidade em trigo hexaplóide. No presente estudo realizou-se um ensaio hidropónico para identificar locos de traços quantitativos (QTLs) associados à tolerância à salinidade do trigo sob concentração de NaCl 150 mM, usando uma população de linhagem consanguíneo recombinante (Xiaoyan 54 × Jing 411). Os valores dos traços de mudas de trigo, incluindo comprimento máximo da raiz (MRL), peso seco da raiz (RDW), ponha o peso seco (SDW), peso seco total (TDW) e a proporção das plantas de trigo TDW entre o estresse salgado e o controle (TDWR), foram avaliados ou calculados. Um total de 19QTLs para cinco traços foram detectados através do método de mapeamento de intervalo composto usando a versão 2.5 do cartógrafo QTL sob condições normais e de estresse salino. Estes QTLs distribuídos em 12 cromossomos explicaram a porcentagem de variação fenotípica por QTL individual variando de 7,9% a 19,0%. Entre eles, foram detectados 11 e 6 QTLs em condições de estresse normal e sal, respectivamente, e dois QTLs foram detectados para TDWR. Cromossoma 1A, 3A e 7A podem conter genes que são candidatos cruciais associados à tolerância ao sal de trigo. Nossos resultados seriam úteis para a seleção assistida por marcadores para produzir variedades de trigo com tolerância salina melhorada.

ABSTRACT: Soil salinity limits agricultural production and is a major obstacle for increasing crop yield. Common wheat is one of the most important crops with allohexaploid characteristic and a highly complex genome. QTL mapping is a useful way to identify genes for quantitative traits such as salinity tolerance in hexaploid wheat. In the present study, a hydroponic trial was carried out to identify quantitative trait loci (QTLs) associated with salinity tolerance of wheat under 150mM NaCl concentration using a recombinant inbred line population (Xiaoyan 54×Jing 411). Values of wheat seedling traits including maximum root length (MRL), root dry weight (RDW), shoot dry weight (SDW), total dry weight (TDW) and the ratio of TDW of wheat plants between salt stress and control (TDWR) were evaluated or calculated. A total of 19QTLs for five traits were detected through composite interval mapping method by using QTL Cartographer version 2.5 under normal and salt stress conditions. These QTLs distributed on 12 chromosomes explained the percentage of phenotypic variation by individual QTL varying from 7.9% to 19.0%. Among them, 11 and six QTLs were detected under normal and salt stress conditions, respectively and two QTLs were detected for TDWR. Some salt tolerance related loci may be pleiotropic. Chromosome 1A, 3A and 7A may harbor crucial candidate genes associated with wheat salt tolerance. Our results would be helpful for the marker assisted selection to breed wheat varieties with improved salt tolerance.

Abstract Demand for native fruits has been expanding, due to the increasing interest in foods with possible health benefits. The buriti (Mauritia flexuosa L. f.) is a native Brazilian fruit, appreciated by local populations, and its products are already on the market. This study investigated the physical characteristics and nutritional composition of buriti pulp, obtained from typical Cerrado and Amazon regions in Brazil. These regions influenced physical parameters and chemical composition. The fruits originating in the Cerrado were bigger and heavier than the ones from the Amazon. Nevertheless, protein, fatty acid and carbohydrate contents were higher in fruits from the Amazon region. The analyzed buriti fruits presented high monounsaturated fatty acid concentration (73.03 to 79.43 g/100 g), with the predominance of oleic acid (72.21 to 78.57 g/100 g). Buriti fruits may be a promising source of good nutritional quality vegetable oil and dietary fiber.

ABSTRACT Objective To screen for and characterize lactic acid bacteria strains with the ability to produce fermented milk and reduce cholesterol levels. Methods The strains were isolated from traditional fermented milk in China. In vitro and in vivo evaluation of cholesterol-reduction were used to identify and verify strains of interest. Characteristics were analyzed using spectrophotometry and plate counting assays. Results The isolate HLX37 consistently produced fermented milk with strong cholesterol-reducing properties was identified as Lactobacillus plantarum (accession number: KR105940) and was thus selected for further study. The cholesterol reduction by strain HLX37 was 45.84%. The isolates were acid-tolerant at pH 2.5 and bile-tolerant at 0.5% (w/v) in simulated gastric juice (pH 2.5) for 2 h and in simulated intestinal fluid (pH 8.0) for 3 h. The auto-aggregation rate increased to 87.74% after 24 h, while the co-aggregation with Escherichia coli DH5 was 27.76%. Strain HLX37 was intrinsically resistant to antibiotics such as penicillin, tobramycin, kanamycin, streptomycin, vancomycin and amikacin. Compared with rats in the model hyperlipidemia group, the total cholesterol content in the serum and the liver as well as the atherogenic index of rats in the viable fermented milk group significantly decreased by 23.33%, 32.37% and 40.23%, respectively. Fewer fat vacuoles and other lesions in liver tissue were present in both the inactivated and viable fermented milk groups compared to the model group. Conclusion These studies indicate that strain HLX37 of L. plantarum demonstrates probiotic potential, potential for use as a candidate for commercial use for promoting health.