Identification of microRNAs and their target genes associated with chasmogamous and cleistogamous flower development in Viola prionantha.

MAIN CONCLUSION: Differentially expressed microRNAs were found associated with the development of chasmogamous and cleistogamous flowers in Viola prionantha, revealing potential roles of microRNAs in the developmental evolution of dimorphic flowers. In Viola prionantha, chasmogamous (CH) flowers are induced by short daylight, while cleistogamous (CL) flowers are triggered by long daylight. How environmental factors and microRNAs (miRNAs) affect dimorphic flower formation remains unknown. In this study, small RNA sequencing was performed on CH and CL floral buds at different developmental stages in V. prionantha, differentially expressed miRNAs (DEmiRNAs) were identified, and their target genes were predicted. In CL flowers, Viola prionantha miR393 (vpr-miR393a/b) and vpr-miRN3366 were highly expressed, while in CH flowers, vpr-miRN2005, vpr-miR172e-2, vpr-miR166m-3, vpr-miR396f-2, and vpr-miR482d-2 were highly expressed. In the auxin-activated signaling pathway, vpr-miR393a/b and vpr-miRN2005 could target Vpr-TIR1/AFB and Vpr-ARF2, respectively, and other DEmiRNAs could target genes involved in the regulation of transcription, e.g., Vpr-AP2-7. Moreover, Vpr-UFO and Vpr-YAB5, the main regulators in petal and stamen development, were co-expressed with Vpr-TIR1/AFB and Vpr-ARF2 and showed lower expression in CL flowers than in CH flowers. Some V. prionantha genes relating to the stress/defense responses were co-expressed with Vpr-TIR1/AFB, Vpr-ARF2, and Vpr-AP2-7 and highly expressed in CL flowers. Therefore, in V. prionantha, CH-CL flower development may be regulated by the identified DEmiRNAs and their target genes, thus providing the first insight into the formation of dimorphic flowers in Viola.

Effects of Viola odorata syrup on sleep quality in menopausal women: a randomized, triple-blind, controlled trial.

PURPOSE: The present study aimed to assess the effects of Viola odorata syrup on the sleep quality of postmenopausal women. METHODS: This triple-blinded randomized clinical trial was conducted on menopausal women presenting to the healthcare centers in Mashhad, Iran, in 2019. The participants were selected using simple random sampling. Participants received 5 ml syrup V. odorata or placebo twice a day for 1 month. Data were collected using the Pittsburgh Sleep Quality Index. The data were analyzed using SPSS version 25. RESULTS: The 118 eligible women enrolled in the study were divided into two groups of V. odorata syrup and placebo (n = 59 each). The analysis was conducted on only 84 menopausal women (42 in each group) due to exclusions. Exclusions consisted of 12 participants who withdrew from the study due to unwillingness to cooperate, 8 who had irregular consumption of the therapeutic syrup, 6 with inaccurate completion of the questionnaire, and 8 due to lack of accessibility. The two study groups were homogenous in terms of demographic characteristics. Before the intervention, no significant difference was observed in the mean PSQI score between the two groups (9.2 ± 2.9 vs. 8.4 ± 2.5) (P = 0.18). However, a significant difference was seen in the mean PSQI score between the two groups (4.9 ± 1.9 vs. 8.1 ± 2.1, P < 0.001) after the intervention. CONCLUSIONS: The findings of this study suggest that V. odorata syrup may be a useful therapeutic agent to improve the sleep quality of menopausal women. REGISTRATION CODE: IRCT20180514039660N1.

Preliminary Feasibility and Acceptability Examination of Using a Novel Ergonomic Chinrest with a Low Shoulder Rest by a Viola Player: A Case Report.

BACKGROUND: High-string players, such as violinists and violists, are prone to neck problems. One factor can be their body posture, with often a rotated and flexed neck position with the jaw placed on a flat chinrest. The Kréddle chinrest (EC) was specifically designed to promote a more neutral neck position and prevent musculoskeletal problems among high-string players. This case report aims to evaluate the preliminary feasibility and acceptability of using the EC, with a low shoulder rest, in a pain-free professional high-string player. A secondary aim was to register newly developed pain and fatigue. METHODS: A 32-year-old professional viola player was instructed to use the EC for most of her playing time during a 6-week trial period. Outcome measures such as compliance (% of use out of total playing time over the last 7 days), performance, comfort and acceptability were assessed at baseline and at the end of each trial week. The Rapid Upper Limb Assessment (RULA) tool was used to evaluate the body posture at baseline when using the EC versus her own chinrest and shoulder rest. RESULTS: The initial trial period was interrupted after 2 days due to pain. A second 6-week period was completed. While compliance was high and there was positive feedback on how the EC affected her performance and comfort while holding the instrument, the case report identified a challenge with the complex instruction material, which made it difficult to install and adjust the product. The viola player was positive towards using EC in the end despite pain and fatigue during the trial period. CONCLUSION: This case report examined a professional viola player's 6-week experience with an EC. While positive outcomes were observed in posture, performance, and product acceptance, issues with complex instructions were noted. This study marks the first exploration of individual challenges with EC use.

Cyclotides derived from Viola dalatensis Gagnep: A novel approach for enrichment and evaluation of antimicrobial activity.

Cyclotides, plant-derived cysteine-rich peptides, exhibit a wide range of beneficial biological activities and possess exceptional structural stability. Cyclotides are commonly distributed throughout the Violaceae family. Viola dalatensis Gagnep, a Vietnamese species, has not been well studied, especially for cyclotides. This pioneering research explores cyclotides from V. dalatensis as antimicrobials. This study used a novel approach to enhance cyclotides after extraction. The approach combined 30% ammonium sulfate salt precipitation and RP-HPLC. A comprehensive analysis was performed to ascertain the overall protein content, flavonoids content, polyphenol content, and free radical scavenging capacity of compounds derived from V. dalatensis. Six known cyclotides were sequenced utilizing MS tandem. Semi-purified cyclotide mixtures (M1, M2, and M3) exhibited antibacterial efficacy against Bacillus subtilis (inhibitory diameters: 19.67-23.50 mm), Pseudomonas aeruginosa (22.17-23.50 mm), and Aspergillus flavus (14.67-21.33 mm). The enriched cyclotide precipitate from the stem extract demonstrated a minimum inhibitory concentration (MIC) of 0.08 mg/mL against P. aeruginosa, showcasing significant antibacterial effectiveness compared to the stem extract (MIC: 12.50 mg/mL). Considerable advancements have been achieved in the realm of cyclotides, specifically in their application as antimicrobial agents.

The involvement of cyclotides in the heavy metal tolerance of Viola spp.

The Violaceae family is rich in metal-tolerant species and species producing cyclic peptides (cyclotides) that are linked to the resistance to biotic factors. Plants that inhabit areas polluted with heavy metals have developed various mechanisms of tolerance. To test the role of cyclotides in protection against abiotic factors, including heavy metals, cell suspension cultures of Viola species/genotypes (V. lutea ssp. westfalica, V. tricolor, V. arvensis, and V. uliginosa), representing different levels of tolerance to heavy metals (from the most tolerant-MET to the least tolerant populations/species-NMET), were used. The relative abundances of the cyclotides in the control, untreated cell suspensions of all the selected species/genotypes, and cells treated with Zn or Pb (200 µM or 2000 µM) for 24 h or 72 h were determined via MALDI-MS. Transmission electron microscopy with X-ray microanalysis was used to detect putative co-localization of the cyclotides with Zn or Pb in the cells of V. tricolor treated with the highest concentration of heavy metals for 72 h. Cyclotide biosynthesis was dependent on the type of heavy metal and its concentration, time of treatment, plant species, and population type (MET vs. NMET). It was positively correlated with the level of tolerance of particular Viola species. The increased production of cyclotides was observed in the cells of metallophyte species, mostly in Zn-treated cells. The nonmetallophyte-V. uliginosa presented a decrease in the production of cyclotides independent of the dose and duration of the metal treatment. Cyclotides co-localized with Pb more evidently than with Zn, suggesting that cyclotides have heavy metal affinity. V. lutea ssp. westfalica transcriptome mining yielded 100 cyclotide sequences, 16 known and 84 novel named viwe 1-84. These findings support the hypothesis that cyclotides are involved in certain mechanisms of plant tolerance to heavy metals.

Viola yedoensis Makino alleviates heat stress-induced inflammation, oxidative stress, and cell apoptosis in the spleen and thymus of broilers.

ETHNOPHARMACOLOGICAL RELEVANCE: Viola yedoensis Makino (VYM) is a traditional Chinese herbal medicine widely distributed in China. It has many pharmacological effects such as anti-inflammatory, immune regulation and anti-oxidation. However, the protective effect of VYM on the spleen and thymus of broilers induced by heat stress has rarely been reported. AIM OF THE STUDY: We established a heat stress model of broilers to explore the protective effect of VYM on spleen and thymus of broilers. MATERIALS AND METHODS: In this experiment, a heat stress model was made by adjusting the feeding temperature of broilers. The protective effect of VYM on the spleen and thymus of heat-stressed broilers were evaluated by detecting immune organ coefficient, histological observation, Enzyme-Linked Immunosorbent Assay, production of antioxidant enzymes and peroxides, TUNEL Staining, Quantitative Real-time PCR. RESULTS: In this study, 60 healthy male AA broilers were divided into 6 groups: Control, 4.5% VYM, HS, HS + 0.5% VYM, HS + 1.5% VYM, HS + 4.5% VYM. After 42 days of feeding, serum, spleen and thymus were collected for detection and analysis. The study revealed that heat stress can lead to pathological damage in the spleen and thymus of broilers, reduce the content of immunoglobulin and newcastle disease (ND), infectious bursal disease (IBD) antibody levels, increase the expression of inflammatory factors IL-1ß, INF-γ, heat shock 70 kDa protein (HSP70), heat shock 90 kDa protein (HSP90). Heat stress inhibits the activity of antioxidant enzymes CAT and SOD, promotes the production of MDA, and then lead to oxidative damage of the spleen and thymus. In addition, apoptotic cells and the ratio of Bax/Bcl-2 was increased. However, the addition of VYM to the feed can alleviate the adverse effects of heat stress on the spleen and thymus of broilers. CONCLUSIONS: This study showed that the addition of VYM to the diet could inhibit oxidative stress and apoptosis, and reduce the inflammatory damage of heat stress on the spleen and thymus of broilers. This study provides a basis for further exploring the regulatory role of VYM in heat stress-induced immune imbalance in broilers. In addition, this study also provides a theoretical basis for the development of VYM as a feed additive with immunomodulatory effects.

Preparation and adjuvanticity against PCV2 of Viola philippica polysaccharide loaded in Chitosan-Gold nanoparticle.

The present Porcine circovirus type 2 virus (PCV2) vaccine adjuvants suffer from numerous limitations, such as adverse effects, deficient cell-mediated immune responses, and inadequate antibody production. In this study, we explored the potential of a novel nanoparticle (CS-Au NPs) based on gold nanoparticles (Au NPs) and chitosan (CS) that modified Viola philippica polysaccharide (VPP) as efficient adjuvants for PCV2 vaccine. The characterization demonstrated that CS-Au-VPP NPs had a mean particle size of 507.42 nm and a zeta potential value of -21.93 mV. CS-Au-VPP NPs also exhibited good dispersion and a stable structure, which did not alter the polysaccharide properties. Additionally, the CS-Au-VPP NPs showed easy absorption and utilization by the organism. To investigate their immune-enhancing potential, mice were immunized with a mixture of CS-Au-VPP NPs and PCV2 vaccine. The evaluation of relevant immunological indicators, including specific IgG antibodies and their subclasses, cytokines, and T cell subpopulations, confirmed their immune-boosting effects. The in vivo experiments revealed that the medium-dose CS-Au-VPP NPs significantly elevated the levels of specific IgG antibodies and their subclasses, cytokines, and T cell subpopulations in PCV2-immunized mice. These findings suggest that CS-Au-VPP NPs can serve as a promising vaccine adjuvant due to their stable structure and immunoenhancement capabilities.

Gene-guided identifications of a structure-chimeric cyclotide viphi I from Viola philippica: Potential functions against cadmium and nematodes.

The cyclic peptides, cyclotides, are identified mostly with 29-31-aa (amino acid residues) but rarely with ≥ 34-aa in plants. Viola philippica is a well-known medicinal plant but a rare metallophyte with cyclotides. A hypothesis was hence raised that the potential novel 34-aa cyclotide of Viola philippica would clearly broaden the structural and functional diversities of plant cyclotides. After homology-cloning the cyclotide precursor gene of VpCP5, a 34-aa cyclotide (viphi I) was identified to be larger than 22 other known cyclotides in V. philippica. It had a chimeric primary structure, due to its unusual loop structures (8 residues in loop 2 and 6 residues in loop 5) and aa composition (3 E and 5 R), by using phylogenetic analyses and an in-house cyclotide analysis tool, CyExcel_V1. A plasmid pCYC-viphi_I and a lab-used recombinant process were specially constructed for preparing viphi I. Typically, 0.12 or 0.25 mg ml-1 co-exposed viphi I could significantly remain cell activities with elevating Cd2+-exposed doses from 10-8 to 10-6 mol l-1 in MCF7 cells. In the model nematode Caenorhabditis elegans, IC50 values of viphi I to inhibit adult ratios and to induce death ratios, were 184.7 and 585.9 µg ml-1, respectively; the median lifespan of adult worms decreased from 14 to 2 d at viphi I doses ranging from 0.05 to 2 mg ml-1. Taken together, the newly identified viphi I exhibits functional potentials against cadmium and nematodes, providing new insights into structural and functional diversity of chimeric cyclotides in plants.

Characterization and evaluation of cytotoxic and antimicrobial activities of cyclotides from Viola japonica.

Cyclotides are a type of defense peptide most commonly found in the Violaceae family of plants, exhibiting various biological activities. In this study, we focused on the Viola japonica as our research subject and conducted transcriptome sequencing and analysis using high-throughput transcriptomics techniques. During this process, we identified 61 cyclotides, among which 25 were previously documented, while the remaining 36 were designated as vija 1 to vija 36. Mass spectrometry detection showed that 21 putative cyclotides were found in the extract of V. japonica. Through isolation, purification and tandem mass spectrometry, we characterized and investigated the activities of five cyclotides. Our results demonstrated inhibitory effects of these cyclotides on the growth of Acinetobacter baumannii and Bacillus subtilis, with minimum inhibitory concentrations (MICs) of 4.2 µM and 2.1 µM, respectively. Furthermore, time killing kinetic assays revealed that cyclotides at concentration of 4 MICs achieved completely bactericidal effects within 2 h. Additionally, fluorescence staining experiments confirmed that cyclotides disrupt microbial membranes. Moreover, cytotoxicity studies showed that cyclotides possess cytotoxic effects, with IC50 values ranging from 0.1 to 3.5 µM. In summary, the discovery of new cyclotide sequences enhances our understanding of peptide diversity and the exploration of their activity lays the foundation for a deeper investigation into the mechanisms of action of cyclotides.

Jogabilidade, aprendizados e emoções no jogo Violetas: cinema & ação no enfrentamento da violência contra a mulher / Gameplay, learning and emotions in the board game Violets: cinema & action in combating violence against women

Resumo O jogo de tabuleiro Violetas: cinema & ação no enfrentamento da violência contra a mulher, forjado a partir da priorização das características libertárias do lúdico, ambienta lutas para a conquista da cidadania. O objetivo do artigo foi analisar a jogabilidade do Violetas quanto ao entendimento das regras, ao envolvimento das(os) jogadoras(es), à mecânica e ao design do jogo; e avaliar comparativamente as dimensões da jogabilidade, das emoções e da aprendizagem como expressões da ludicidade. Pesquisa de métodos mistos, em etapas: a) aperfeiçoamento da jogabilidade: oficina com 12 especialistas; testes de usabilidade (33 participantes); análise de conteúdo; b) avaliação da ludicidade: questionários a 78 participantes, teste não paramétrico U de Mann-Whitney para comparação dos grupos de variáveis. Os aspectos da jogabilidade obtiveram graus de concordância elevados entre as/os participantes. As variáveis da jogabilidade assumiram valor igual em relação ao grupo da aprendizagem, ambos significativamente diferentes de um grupo 2, das emoções sentidas na partida. No Violetas, o entrelaçamento da jogabilidade com os componentes formativos da aprendizagem viabilizam a criação de um campo simbólico, desafiador e afetivo em que a imaginação, a interação, a tensão e o interesse das(os) jogadoras(es) se manifestam durante as partidas.

Abstract The board game Violets: cinema and action in combating violence against women was developed prioritising the liberating features of play to offer a setting for struggles to secure citizenship. The objective of the article was to examine the gameplay of Violets as regards players' understanding of the rules and engagement, and the game's mechanics and design; and to evaluate gameplay, emotions and learning comparatively as dimensions of play. This mixed method study proceeded in stages: a) perfecting gameplay: a workshop with 12 experts, usability tests with 33 participants and content analysis; and b) evaluating play: questionnaires for 78 participants and non-parametric Mann-Whitney U-test comparing groups of variables. Agreement among participants on aspects of gameplay was high. The group of gameplay variables returned values equal to those of the learning group; both differed significantly from the group for emotions felt while playing. In Violets, the interweave of gameplay with the formative, learning components set up a challenging, affective, symbolic field where players' imagination, interaction, tension and interest were expressed during play.

Um paradoxo entre o existir e o resistir: a moda de viola através dos tempos / A paradox between existing and resisting: viola fashion through the ages

RESUMO A moda de viola é considerada um dos gêneros mais tradicionais da música caipira, se aproximando enquanto gênero narrativo das antigas lendas medievais e dos romances de cavalaria ibéricos renascentistas, sendo ainda afluente da cultura moura. Suas temáticas são reveladoras de uma funcionalidade social que, ao transmitirem valores pedagógicos, morais e expressarem poética e musicalmente determinados contextos sociais, servem como ferramenta de denúncia e crítica a processos de exclusão e opressão social. Este artigo traz uma breve descrição de seus primeiros registros coletados por folcloristas no início do século XX, dos dilemas decorrentes de sua inserção no registro fonográfico a partir de 1929 e das contradições acerca da perpetuação do gênero através dos tempos, focalizando principalmente sua potencialidade no campo dos estudos sobre identidade.

ABSTRACT The moda-de-viola [folk guitar songs] is considered one of the most traditional genres of country music, a narrative genre approaching the old medieval legends and romances of the Iberian Renaissance when it was still an affluent of Moorish culture. Its themes reveal a social functionality that, by conveying pedagogical and moral values, and by expressing poetically and musically certain specific social contexts, serves as a tool to denounce and criticize processes of exclusion and social oppression. This article presents a brief description of the first records collected by folklorists in the early twentieth century, of the dilemmas arising from its insertion in the phonographic record from 1929 onward, and of the contradictions in the perpetuation of this genre over time, focusing mainly on its potentiality in the field of identity studies.