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.

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.

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.

Magnesium and manganese induced changes on chemical, nutritional, antioxidant and antimicrobial properties of the pansy and Viola edible flowers.

Pansy and viola edible flowers were grown hydroponically with different levels of Mg and Mn. The nutritional composition was determined using standard methods. Free sugars, fatty acids, organic acids, tocopherols, and phenolic compounds were analyzed using various HPLC and GC devises. The extract's antimicrobial, antioxidant, cytotoxicity, and anti-inflammatory activity were assessed. The results indicated that Mg enrichment negatively affected plant growth and mineral accumulation but improved photosynthetic performance. The edible flowers contained significant amounts of protein, low levels of fat, and varying sugar contents, such as glucose and fructose. Various fatty acids and phenolic compounds were identified, with different concentrations depending on the treatment. The flowers exhibited antioxidant potential, antimicrobial activity, cytotoxic effects, and anti-inflammatory properties. The correlations between the investigated parameters not only expand knowledge on Mg and Mn interaction but also catalyze significant advancements in sustainable agriculture and food health, fostering a healthier and more conscious future.

UPLC-Q-TOF-MS profiling of Viola stocksii Boiss. and evaluation of aphrodisiac potential and risk factors associated with erectile dysfunction.

ETHNOPHARMACOLOGICAL RELEVANCE: Viola stocksii Boiss. locally known as makhni or makhanr booti, is an important medicinal food plant with multiple therapeutic applications, including erectile dysfunction (ED). It is mixed with butter and used for boosting energy and sexual health in the subcontinent. AIMS OF THE STUDY: This study was designed to evaluate the chemical composition, aphrodisiac potential and effect of V. stocksii on the risk factors associated with ED. METHODOLOGY: The hydroethanolic extract of V. stocksii (HEEVS) was prepared through the microwave-assisted extraction (MAE) technique. The chemical composition was evaluated using preliminary phytochemical screening and UPLC-Q-TOF-MS analysis. Metals and minerals analysis was performed by an atomic absorption spectrophotometer. The aphrodisiac activity of HEEVS was evaluated using an in vivo aphrodisiac model established in male albino rats and the effect on various sexual parameters such as mount, intromission, ejaculation frequencies and mount, intromission, ejaculation latencies, postejaculatory interval, penile reflexes and serum hormone concentration were analyzed. The effect of HEEVS on various risk factors associated with ED, including prostate cancer (PC), bacterial infections, diabetes and obesity, was evaluated using various in vitro assays. Moreover, four compounds were selected from the UPLC-Q-TOF-MS profile and evaluated for in silico computational analysis against phosphodiesterase-5 (PDE-5) for possible interaction. FINDINGS: The phytochemical screening revealed the presence of various secondary metabolites in HEEVS, while 58 compounds were tentatively identified in the UPLC-Q-TOF-MS analysis. Various important minerals and metals such as zinc, calcium, cadmium and magnesium were detected in the atomic absorption spectrometry analysis. The in vivo aphrodisiac evaluation showed a significant (p < 0.05) increase in the mount, intromission and ejaculation frequencies and a decrease in the mount, intromission latencies and post-ejaculatory intervals at a dose of 300 mg/kg. A marked (p < 0.05) increase was observed in the concentration of serum testosterone and luteinizing hormones in HEEVS treated animals with a significant increase in total penile reflexes. The extract displayed significant anti-prostate cancer activity and a potential antibacterial spectrum against E. coli and S. aureus, with MIC50 values of 215.72 µg/mL and 139.05 µg/mL, respectively. Similarly, HEEVS was found active towards pancreatic lipase (67.34 ± 1.03%), α-glucosidase (3.87 ± 0.54 mmol ACAE/g d.w.) and α-amylase (6.98 ± 1.63 mmol ACAE/g d.w.). The in silico docking study presented a potential interaction between the selected compounds and residues of the active site of PDE-5. CONCLUSION: This report highlights the aphrodisiac potential of V. stocksii and provides experimental support for its traditional use in ED with an attenuative effect on the risk factors associated with ED. Moreover, the chemical composition displayed the presence of functional phytoconstituents and minerals in HEEVS and paves the way for the isolation of compounds with potent aphrodisiac activity.

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.

High-throughput microfluidic chip with silica gel-C18 channels for cyclotide separation.

Over the past two decades, microfluidic-based separations have been used for the purification, isolation, and separation of biomolecules to overcome difficulties encountered by conventional chromatography-based methods including high cost, long processing times, sample volumes, and low separation efficiency. Cyclotides, or cyclic peptides used by some plant families as defense agents, have attracted the interest of scientists because of their biological activities varying from antimicrobial to anticancer properties. The separation process has a critical impact in terms of obtaining pure cyclotides for drug development strategies. Here, for the first time, a mimic of the high-performance liquid chromatography (HPLC) on microfluidic chip strategy was used to separate the cyclotides. In this regard, silica gel-C18 was synthesized and characterized by Fourier-transform infrared spectroscopy (FTIR) and proton nuclear magnetic resonance (1H-NMR) and then filled inside the microchannel to prepare an HPLC C18 column-like structure inside the microchannel. Cyclotide extract was obtained from Viola ignobilis by a low voltage electric field extraction method and characterized by HPLC and matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF). The extract that contained vigno 1, 2, 3, 4, 5, and varv A cyclotides was added to the microchannel where distilled water was used as a mobile phase with 1 µL/min flow rate and then samples were collected in 2-min intervals until 10 min. Results show that cyclotides can be successfully separated from each other and collected from the microchannel at different periods of time. These findings demonstrate that the use of microfluidic channels has a high impact on the separation of cyclotides as a rapid, cost-effective, and simple method and the device can find widespread applications in drug discovery research.

Exploring Immune Modulatory Effects of Cyclotide-Enriched Viola tricolor Preparations.

Viola tricolor is a medicinal plant with documented application as an anti-inflammatory herb. The standard of care for the treatment of inflammatory bowel disease is immunosuppressive therapeutics or biologics, which often have undesired effects. We explored V. tricolor herbal preparations that are rich in an emerging class of phytochemicals with drug-like properties, so-called cyclotides. As an alternative to existing inflammatory bowel disease medications, cyclotides have immunomodulatory properties, and their intrinsic stability allows for application in the gastrointestinal tract, for instance, via oral administration. We optimized the isolation procedure to improve the yield of cyclotides and compared the cellular effects of violet-derived organic solvent-extracts, aqueous preparations, and an isolated cyclotide from this plant on primary human T lymphocytes and macrophages, i.e., cells that are crucial for the initiation and progression of inflammatory bowel disease. The hot water herbal decoctions have a stronger immunosuppressive activity towards proliferation, interferon-γ, and interleukin-21 secretion of primary human T cells than a DCM/MeOH cyclotide-enriched extract, and the isolated cyclotide kalata S appears as one of the active components responsible for the observed effects. This effect was increased by a longer boiling duration. In contrast, the DCM/MeOH cyclotide-enriched extract was more effective in reducing the levels of cytokines interleukin-6, interleukin-12, interleukin-23, tumor necrosis factor-α, and C - X-C motif chemokine ligand 10, secreted by human monocyte-derived macrophages. Defined cyclotide preparations of V. tricolor have promising pharmacological effects in modulating immune cell responses at the cytokine levels. This is important towards understanding the role of cyclotide-containing herbal drug preparations for future applications in immune disorders, such as inflammatory bowel disease.

Intelligent double-layer polymers based on carboxymethyl cellulose-cellulose nanocrystals film and poly(lactic acid)-Viola odorata petal anthocyanins nanofibers to monitor food freshness.

The present study was conducted with the aim of fabricating smart bilayer polymers based on carboxymethyl cellulose-cellulose nanocrystals film and poly(lactic acid)-Viola odorata extract nanofibers (CMC-CNC and PLA-VOE) for freshness monitoring of Pacific white shrimps, minced lamb meat, chicken fillets, and rainbow trout fillets, during refrigerated storage conditions. The fabricated indicators based on CMC-PLA-VOE 5%, CMC-CNC 1%-PLA-VOE 5%, and CMC-CNC 3%-PLA-VOE 5% presented remarkable color changes in pH 1-12 buffer solutions, including red at pH 1-6, violet at pH 7-8, green at pH 9-10, and brown at pH 11-12. Significantly lower water vapor permeability and oxygen transmission rate of prepared polymers were found in comparison with the control groups (P < 0.05). Regarding the monitoring of food samples in real-time, the samples spoiled after 3 days, evidenced by total viable count, psychrotrophic bacterial count, total volatile basic nitrogen, and pH values of 7.17-7.54 log CFU/g, 5.68-6.23 log CFU/g, 25.14-28.12 mg N/100 g, and 7.10-7.66, respectively. Meanwhile, the noticeable color change of prepared indicators from white to violet (day 3) and finally dark violet (day 7) was observed, indicating a potential application in intelligent packaging for real-time control of the freshness of perishable food samples.

A novel galactoxylan derived from Viola diffusa alleviates LPS-induced acute lung injury via antagonizing P-selectin-mediated adhesion function.

Acute lung injury (ALI) greatly threatens human health worldwide. P-selectin is a potential target for the treatment of acute inflammatory diseases, and natural polysaccharides exhibit high-affinity for P-selectin. Viola diffusa, a traditional Chinese herbal, shows strong anti-inflammatory effects, but pharmacodynamic substances and underlying mechanisms are still unclear. In this study, a galactoxylan polysaccharide (VDPS) derived from Viola diffusa was isolated and characterized, evaluated the protective effect on LPS induced ALI and underlying mechanism. VDPS significantly alleviated LPS-induced pathological lung injury, and decreased the numbers of total cells and neutrophils as well as the total protein contents in the bronchoalveolar lavage fluid (BALF). Moreover, VDPS reduced proinflammatory cytokine production both in BALF and lung. Interestingly, VDPS significantly restrained the activation of NF-κB signaling in the lung of LPS-exposed mice, but it cannot inhibit LPS-induced inflammation in human pulmonary microvascular endothelial cells (HPMECs) in vitro. Additionally, VDPS disrupted neutrophil adhesion and rolling on the activated HPMECs. VDPS cannot impact the expression or cytomembrane translocation of endothelial P-selectin, but remarkably interrupt the binding of P-selectin and PSGL-1. Overall, this study demonstrated that VDPS can alleviate LPS-induced ALI via inhibiting P-selectin-dependent adhesion and recruitment of neutrophils on the activated endothelium, providing a potential treatment strategy for ALI.

Diversity analysis of endophytes with antimicrobial and antioxidant potential from Viola odorata: an endemic plant species of the Himalayas.

Viola odorata, also known as "Banafshah" in high altitudes of Himalayas, is well known for its pharmaceutical importance in Ayurvedic and Unani medicinal system. The plant is a source of various drugs for its anti-inflammatory, diaphoretic, diuretic, emollient, expectorant, antipyretic, and laxative properties. The endophytes of plants have been reported for their role in modulating various physiological and biological processes of the host plants. In the present study, a total of 244 endophytes were isolated in pure cultures from the roots of Viola odorata, and genetic diversity was evaluated using amplified ribosomal DNA restriction analysis (ARDRA) and enterobacterial repetitive intergenic consensus (ERIC). The molecular fingerprinting revealed variation among various rRNA types among morphologically different endophytes based on ARDRA and ERIC-PCR. The screening of endophytes showed antimicrobial activity of 11 bacterial isolates and one actinomycete SGA9 against various pathogens Bacillus cereus, Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Staphylococcus aureus, and Staphylococcus epidermidis. The antioxidant activity revealed the majority of the bacterial isolates able to scavenge the free radical in the range of 10-50% and 8 bacterial isolates in the range of 50-85%. Principal component analysis separated eight isolates away from the central eclipse and form a separate group based on antimicrobial and antioxidant potential. The identification of these eight isolates showed affiliation with different species of the genus Enterobacter, Microbacterium, Pseudomonas, Rhizobium, and Streptomyces. This is the first report on the characterization of endophytic bacteria and actinomycetes from endemic Viola odorata. Results suggested that these endophytes could be explored for the production of antimicrobial and antioxidant products.

Effect of environmental factors on seed germination and seedling emergence of Viola prionantha, a cleistogamous plant.

Environmental conditions during seed development and maturation can affect seed traits and germination behavior, yet systematic research on the effects of seed maturation time on seed traits, germination behavior and seedling emergence of cleistogamy plants is lacking. Here, we determined the difference in phenotypic characteristics of CH and CL (namely CL1, CL2 and CL3 based on maturation time, respectively) fruits/seeds that were collected from Viola prionantha Bunge, a cleistogamous perennial plant, and evaluated the effects of various environmental factors on seed germination and seedling emergence. The fruit mass, width, seed number per fruit and mean seed mass of CL1 and CL3 were greater than that of CH and CL2, while seed setting of CH was lower than that of CL1, CL2 and CL3. Germination of CH, CL1, CL2, and CL3 seeds was < 10% in the dark at 15/5 and 20/10 â„ƒ, whereas germination (0%-99.2%) of CH, CL1, CL2, and CL3 seeds changed significantly under light conditions. In contrast, more than 71% (from 71.7 to 94.2%) germination of both CH, CL1, CL2 and CL3 seeds occurred under both light/dark conditions and continuous darkness at 30/20 â„ƒ. Germination of CH, CL1, CL2 and CL3 seeds was sensitive to osmotic potential, but CL1 seeds were more resistant to osmotic stress, compared with CH, CL2 and CL3. Seedling emergence of CH seeds was more than 67% (from 67.8 to 73.3%) at a burial depth of 0 cm-2 cm, while all types of CL seeds were below 15% at a burial depth of 2 cm. Information gathered from this study indicates that CH and CL seeds of V. prionantha were different in fruit size, seed mass, thermoperiod and photoperiod sensitivity, osmotic potential tolerance and seedling emergence, especially, maturation time significantly affect phenotypic characteristics and germination behavior of CL seeds matured at different periods. These results indicate that V. prionantha adapts to unpredictable environmental conditions by developing a variety of adaptation strategies, and ensures the survival and reproduction of the populations.

[Research progress on chemical constituents and pharmacological activities of Viola plants].

There are 500 species of Viola(Violaceae) worldwide, among which 111 species are widely distributed in China and have a long medicinal history and wide varieties. According to the authors' statistics, a total of 410 compounds have been isolated and identified from plants of this genus, including flavonoids, terpenoids, phenylpropanoids, organic acids, nitrogenous compounds, sterols, saccharides and their derivatives, volatile oils and cyclotides. The medicinal materials from these plants boast anti-microbial, anti-viral, anti-oxidant and anti-tumor activities. This study systematically reviewed the chemical constituents and pharmacological activities of Viola plants to provide a basis for further research and clinical application.

Cow dung putrefaction via vermicomposting using Eisenia fetida and its influence on seed sprouting and vegetative growth of Viola wittrockiana (pansy).

The current research was conducted at Vermi-tech Unit, Muzaffarabad in 2018 to evaluate the efficacy of cow dung and vermicompost on seed sprouting, seedlings, and vegetative developmental parameters of Viola x wittrokiana (pansy). In the current study, vermicompost was produced using Eisenia fetida. Physicochemical parameters of vermicompost and organic manure were recorded before each experimentation. The potting experiment was designed and comprised of eight germination mediums containing different combinations of soil, sand, cow dung, and various concentrations of vermicompost such as 10% VC, 15% VC, 20% VC, 25% VC, 30% VC, and 35% VC. Seed sprouting and seedling developmental parameters were observed for 28 days while vegetative plant growth parameters were recorded after 10 weeks of transplantation in various vermicompost amended germination media. Pre and post-physicochemical analysis of germination media were also recorded to check their quality and permanency. The current findings showed that 30% VC germination media was an effective dose for early seed germination initiation and all seed germination parameters. However, the significant vegetative plant growth and flowering parameters of pansy occurred at 35% VC. Findings revealed that vermicompost not only enhanced the seed germination and growth of pansy but also improved soil health. These results indicate that vermicompost can be exploited as a potent bio-fertilizer for ornamental plant production.

Commercially Available Viola odorata Oil, Chemical Variability and Antimicrobial Activity.

Viola odorata L. oil is frequently recommended in the aromatherapeutic literature for treating respiratory, urinary, and skin infections; however, antimicrobial evidence is lacking. In addition, in aromatherapy, combinations of essential oils are predominantly utilized with the goal of achieving therapeutic synergy, yet no studies investigating the interaction of essential oil combinations with V. odorata oil exists. This study thus aimed to address these gaps by investigating the antimicrobial activity of three Viola odorata oil samples, sourced from different suppliers, independently and in combination with 20 different commercial essential oils, against micro-organisms involved in respiratory, skin, and urinary tract infections associated with global resistance trends. These pathogens include several of the ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter spp.) The chemical profile of the oils was determined using gas chromatography coupled with mass spectrometry. The minimum inhibitory concentrations (MIC) were determined using the broth micro-dilution method. The interactive profiles for the combinations were assessed by calculating the fractional inhibitory concentration index (ΣFIC). The main compounds varied across the three samples, and included phenethyl alcohol, isopropyl myristate, 2-nonynoic acid, methyl ester, α-terpineol, α-cetone, and benzyl acetate. The V. odorata oil samples displayed overall poor antimicrobial activity when tested alone; however, the antimicrobial activity of the combinations resulted in 55 synergistic interactions where the combination with Santalum austrocaledonicum resulted in the lowest MIC values as low as 0.13 mg/mL. The frequency of the synergistic interactions predominantly occurred against Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, and Enterococcus faecium with noteworthy MIC values ranging from 0.25-1.00 mg/mL. This study also reports on the variability of V. odorata oils sold commercially. While this warrants caution, the antimicrobial benefit in combination provides an impetus for further studies to investigate the therapeutic potential.

First detection of endopolyploidy in tapetal cells and chromosomal anomalies in meiocytes of Viola pilosa cytotypes (2n=20) from Pir Panjal (Himalayas).

The nine Viola pilosa Blume populations studied from Pir Panjal contained 20 chromosomes. This count is not reported so far in Indian populations. Currently, comparison of tapetal and meiotic cells revealed the existence of synchrony in different developmental phases. Young tapetal cells at prometaphase co-occurred with the pollen mother cells (PMCs) at diakinesis to metaphase, mature tapetal cells with disintegrated chromatin material co-occurred with tetrads and no tapetal cells were found at mature pollen stage. Cytological studies in young tapetal cells revealed most of these to be endopolyploid, with each having 40 chromosomes. While outnumbering somatic cells contained clear 40 chromosomes which seemed to be the outcome of endomitosis, a sizeable number of cells possessed 40 sticky chromosomes at metaphase. Later chromosomes are likely to form restitution nucleus. Mature tapetal cells, occurring singly/cytomictically connected (3.2-26.31%) or showing coalescence (10.5-22.8%), did not contain recognizable chromosomes. Instead, they were characterized by disintegrated nuclear content. Further, meiotic studies revealed that the present population contained all/outnumbering euploid cells (2n=20); many of which exhibited nearly regular behaviour. However, 6.5-26.9% meiocytes of eight populations and 47% cells of P-Khe population depicted aneuploidy/contained quadri-octavalents, with per cent pollen viabilities of these ranging from 38.6 to 49.9. Going by the normal tapetal development in V. pilosa, existence of various chromosomal anomalies seems to have accounted for the reduction in gametic fertility of this taxon.

Volumetric imaging of optically cleared and fluorescently labeled animal tissue (VIOLA) for quantifying the 3D biodistribution of nanoparticles at cellular resolution in tumor tissue.

Nanoparticle (NP) technology holds significant promise to mediate targeted drug delivery to specific organs in the body. Understanding the 3D biodistribution of NPs in heterogeneous environments such as the tumor tissue can provide crucial information on efficacy, safety and potential clinical outcomes. Here we present a novel end-to-end workflow, VIOLA, which makes use of tissue clearing methodology in conjunction with high resolution imaging and advanced 3D image processing to quantify the spatiotemporal 3D biodistribution of fluorescently labeled ACCURIN® NPs. Specifically, we investigate the spatiotemporal biodistribution of NPs in three different murine tumor models (CT26, EMT6, and KPC-GEM) of increasing complexity and translational relevance. We have developed new endpoints to characterize NP biodistribution at multiple length scales. Our observations reveal that the macroscale NP biodistribution is spatially heterogeneous and exhibits a gradient with relatively high accumulation at the tumor periphery that progressively decreases towards the tumor core in all the tumor models. Microscale analysis revealed that NP extravasation from blood vessels increases in a time dependent manner and plateaus at 72 h post injection. Volumetric analysis and pharmacokinetic modeling of NP biodistribution in the vicinity of the blood vessels revealed that the local NP density exhibits a distance dependent spatiotemporal biodistribution which provide insights into the dynamics of NP extravasation in the tumor tissue. Our data represents a comprehensive analysis of NP biodistribution at multiple length scales in different tumor models providing unique insights into their spatiotemporal dynamics. Specifically, our results show that NPs exhibit a dynamic equilibrium with macroscale heterogeneity combined with microscale homogeneity.