Fine mapping and candidate gene analysis of a major QTL for oil content in the seed of Brassica napus.

KEY MESSAGE: By integrating QTL fine mapping and transcriptomics, a candidate gene responsible for oil content in rapeseed was identified. The gene is anticipated to primarily function in photosynthesis and photosystem metabolism pathways. Brassica napus is one of the most important oil crops in the world, and enhancing seed oil content is an important goal in its genetic improvement. However, the underlying genetic basis for the important trait remains poorly understood in this crop. We previously identified a major locus, OILA5 responsible for seed oil content on chromosome A5 through genome-wide association study. To better understand the genetics of the QTL, we performed fine mapping of OILA5 with a double haploid population and a BC3F2 segregation population consisting of 6227 individuals. We narrowed down the QTL to an approximate 43 kb region with twelve annotated genes, flanked by markers ZDM389 and ZDM337. To unveil the potential candidate gene responsible for OILA5, we integrated fine mapping data with transcriptome profiling using high and low oil content near-isogenic lines. Among the candidate genes, BnaA05G0439400ZS was identified with high expression levels in both seed and silique tissues. This gene exhibited homology with AT3G09840 in Arabidopsis that was annotated as cell division cycle 48. We designed a site-specific marker based on resequencing data and confirmed its effectiveness in both natural and segregating populations. Our comprehensive results provide valuable genetic information not only enhancing our understanding of the genetic control of seed oil content but also novel germplasm for advancing high seed oil content breeding in B. napus and other oil crops.

Advanced network pharmacology study reveals multi-pathway and multi-gene regulatory molecular mechanism of Bacopa monnieri in liver cancer based on data mining, molecular modeling, and microarray data analysis.

Liver cancer is a malignant tumor that grows on the surface or inside the liver. The leading cause is a viral infection with hepatitis B or C virus. Natural products and their structural analogues have historically made a major contribution to pharmacotherapy, especially for cancer. A list of studies evidences the therapeutic efficacy of Bacopa monnieri against liver cancer, but the precise molecular mechanism is yet to be discovered. This study combines data mining, network pharmacology, and molecular docking analysis to potentially revolutionize liver cancer treatment by identifying effective phytochemicals. Initially, the information on active constituents of B. monnieri and target genes of both liver cancer and B. monnieri were retrieved from literature as well as from publicly available databases. Based on the matching results between B. monnieri potential targets and liver cancer targets, the protein-protein interaction (PPI) network was constructed using the STRING database and imported into Cytoscape for screening of hub genes based on their degree of connectivity. Later, the interactions network between compounds and overlapping genes was constructed using Cytoscape software to analyze the network pharmacological prospective effects of B. monnieri on liver cancer. Gene Ontology (GO) and KEGG pathway analysis of hub genes revealed that these genes are involved in the cancer-related pathway. Lastly, the expression level of core targets was analyzed using microarray data (GSE39791, GSE76427, GSE22058, GSE87630, and GSE112790). Further, the GEPIA server and PyRx software were used for survival and molecular docking analysis, respectively. In summary, we proposed that quercetin, luteolin, apigenin, catechin, epicatechin, stigmasterol, beta-sitosterol, celastrol, and betulic acid inhibit tumor growth by affecting tumor protein 53 (TP53), interleukin 6 (IL6), RAC-alpha serine/threonine protein kinases 1 (AKT1), caspase-3 (CASP3), tumor necrosis factor (TNF), jun proto-oncogene (JUN), heat shot protein 90 AA1 (HSP90AA1), vascular endothelial growth factor A (VEGFA), epidermal growth factor receptor (EGFR), and SRC proto-oncogene (SRC). Through, microarray data analysis, the expression level of JUN and IL6 were found to be upregulated while the expression level of HSP90AA1 was found to be downregulated. Kaplan-Meier survival analysis indicated that HSP90AA1 and JUN are promising candidate genes that can serve as diagnostic and prognostic biomarkers for liver cancer. Moreover, the molecular docking and molecular dynamic simulation of 60ns well complemented the binding affinity of the compound and revealed strong stability of predicted compounds at the docked site. Calculation of binding free energies using MMPBSA and MMGBSA validated the strong binding affinity between the compound and binding pockets of HSP90AA1 and JUN. Despite that, in vivo and in vitro studies are mandatory to unveil pharmacokinetics and biosafety profiles to completely track the candidature status of B. monnieri in liver cancer.

Sarcococca saligna extract attenuates formaldehyde-induced arthritis in Wistar rats via modulation of pro-inflammatory and inflammatory biomarkers.

Sarcococca saligna plant is commonly used as traditional therapy for arthritis especially in Asian countries. The current study is designed to explore the anti-arthritic potential of S. saligna aqueous methanolic extract (SSME). Preliminary proximate study and HPLC analysis were performed to investigate the phytochemical characterization and quality control. The safety of the SSME was evaluated by performing an acute oral toxicity study (OECD guidelines 425). The anti-arthritic potential of SSME was explored by in vivo formaldehyde-induced arthritis model. The antiarthritic effect of the SSME was determined through paw diameter, arthritic index, body weight, biochemical and haematological parameters. Radiographic and histopathological studies were also carried out to evaluate the results. qRT-PCR was performed to determine the upregulation and downregulation of anti- and pro-inflammatory cytokines in rats while ELISA was done to determine the concentration of HSP-70, IL-6 and TNF-α in the serum. Results of acute oral toxicity showed no abnormality and mortality. There was no noticeable change in haematological and biochemical parameters. Histopathological examination exhibited the normal structure of vital organs. So, SSME might be safe at a 2000 mg/kg dose, proposing that LD50 was higher than 2000 mg/kg body weight. Gallic acid, catechin, hydroxyl benzoic acid, sinapic acid, caffeic acid, ferulic acid and p-cumaric acid were identified by HPLC. The outcomes of in vivo formaldehyde-induced arthritic model showed that SSME significantly reduced paw inflammation and arthritic index and improved haematological and biochemical parameters. Moreover, the SSME influentially down-regulated the gene expression of IL-1ß, IL-6, COX-2, PGE2, TNF-α and NF-κB, and up-regulated the expression of IL-4, and IL-10. The results of the undertaken study suggest that S. saligna have strong anti-arthritic activity supporting its conventional application as the remedy of rheumatoid arthritis.

Anti-Parkinson's evaluation of Brassica juncea leaf extract and underlying mechanism of its phytochemicals.

Background: Parkinson's disease (PD) is associated with progressive neuronal damage and dysfunction. Oxidative stress helps to regulate neurodegenerative and neuronal dysfunction. Natural compounds could attenuate oxidative stress in a variety of neurological disorders. B. juncea is a rich source of antioxidants. The present study aimed to evaluate the therapeutic potential of B. juncea leaves for the treatment of PD by applying behavioral, in vivo and in silico studies. For in vivo studies rats were divided into six groups (n = 6). Group-I served as normal control (vehicle control). Group-II was disease control (haloperidol 1 mg/kg). Group-III was kept as a standard group (L-Dopa 100 mg/kg + carbidopa 25 mg/kg). Groups (IV-VI) were the treatment groups, receiving extract at 200-, 400- and 600 mg/kg doses respectively, for 21 days orally. Results: In vivo study results showed that the extract was found to improve muscles strength, motor coordination, and balance in PD. These behavioral outcomes were consistent with the recovery of endogenous antioxidant defence in biochemical analysis which was further corroborated with histopathological ameliorations. Dopamine levels increased and monoamine oxidase B (MAO-B) levels decreased dose-dependently in the brain during the study. Herein, we performed molecular docking analysis of the proposed extracted phytochemicals has explained that four putative phytochemicals (sinapic acid, rutin, ferulic acid, and caffeic acid) have presented very good results in terms of protein-ligand binding interactions as well as absorption, distribution, metabolism, excretion & toxicity (ADMET) profile estimations. Conclusion: The undertaken study concluded the anti-Parkinson activity of B. juncea and further suggests developments on its isolated compounds in PD therapeutics.

Integrated Core Proteomics, Subtractive Proteomics, and Immunoinformatics Investigation to Unveil a Potential Multi-Epitope Vaccine against Schistosomiasis.

Schistosomiasis is a parasitic infection that causes considerable morbidity and mortality in the world. Infections of parasitic blood flukes, known as schistosomes, cause the disease. No vaccine is available yet and thus there is a need to design an effective vaccine against schistosomiasis. Schistosoma japonicum, Schistosoma mansoni, and Schistosoma haematobium are the main pathogenic species that infect humans. In this research, core proteomics was combined with a subtractive proteomics pipeline to identify suitable antigenic proteins for the construction of a multi-epitope vaccine (MEV) against human-infecting Schistosoma species. The pipeline revealed two antigenic proteins-calcium binding and mycosubtilin synthase subunit C-as promising vaccine targets. T and B cell epitopes from the targeted proteins were predicted using multiple bioinformatics and immunoinformatics databases. Seven cytotoxic T cell lymphocytes (CTL), three helper T cell lymphocytes (HTL), and four linear B cell lymphocytes (LBL) epitopes were fused with a suitable adjuvant and linkers to design a 217 amino-acid-long MEV. The vaccine was coupled with a TLR-4 agonist (RS-09; Sequence: APPHALS) adjuvant to enhance the immune responses. The designed MEV was stable, highly antigenic, and non-allergenic to human use. Molecular docking, molecular dynamics (MD) simulations, and molecular mechanics/generalized Born surface area (MMGBSA) analysis were performed to study the binding affinity and molecular interactions of the MEV with human immune receptors (TLR2 and TLR4) and MHC molecules (MHC I and MHC II). The MEV expression capability was tested in an Escherichia coli (strain-K12) plasmid vector pET-28a(+). Findings of these computer assays proved the MEV as highly promising in establishing protective immunity against the pathogens; nevertheless, additional validation by in vivo and in vitro experiments is required to discuss its real immune-protective efficacy.

Green Synthesis of Copper Oxide Nanoparticles Using Aerva javanica Leaf Extract and Their Characterization and Investigation of In Vitro Antimicrobial Potential and Cytotoxic Activities.

The development of green technology is creating great interest for researchers towards low-cost and environmentally friendly methods for the synthesis of nanoparticles. Copper oxide nanoparticles (CuO-NPs) attracted many researchers due to their electric, catalytic, optical, textile, photonic, monofluid, and pharmacological activities that depend on the shape and size of the nanoparticles. This investigation aims copper oxide nanoparticles synthesis using Aerva javanica plant leaf extract. Characterization of copper oxide nanoparticles synthesized by green route was performed by three different techniques: X-Ray Diffraction (XRD), Fourier Transform Infrared (FTIR) Spectroscopy, and Scanning Electron Microscopy (SEM). X-ray diffraction (XRD) reveals the crystalline morphology of CuO-NPs and the average crystal size obtained is 15 nm. SEM images showed the spherical nature of the particles and size is lying in the 15-23 nm range. FTIR analysis confirms the functional groups of active components present in the extract which are responsible for reducing and capping agents for the synthesis of CuO-NPs. The synthesized CuO-NPs were studied for their antimicrobial potential against different bacterial as well as fungal pathogens. The results indicated that CuO-NPs show maximum antimicrobial activities against all the selected bacterial and fungal pathogens. Antimicrobial activities of copper oxide nanoparticles were compared with standard drugs Norfloxacin and amphotericin B antibiotics. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of copper oxide nanoparticles were 128 µg/mL against all selected bacterial pathogens. MIC of fungus and minimum fungicidal concentration (MFC) of CuO-NPs were 160 µg/mL. Thus, CuO-NPs can be utilized as a broad-spectrum antimicrobial agent. The cytotoxic activity of the synthesized CuO-NPs suggested that toxicity was negligible at concentrations below 60 µg/mL.

Saponin toxicity as key player in plant defense against pathogens.

Microbial pathogens attack every plant tissue, including leaves, roots, shoots, and flowers during all growth stages. Thus, they cause several diseases resulting in a plant's failure or loss of the whole crop in severe cases. To combat the pathogens attack, plants produce some biologically active toxic compounds known as saponins. The saponins are secondary metabolic compounds produced in healthy plants with potential anti-pathogenic activity and serve as potential chemical barriers against pathogens. Saponins are classified into two major groups the steroidal and terpenoid saponins. Here, we reported the significance of saponin toxins in the war against insect pests, fungal, and bacterial pathogens. Saponins are present in both cultivated (chilies, spinach, soybean, quinoa, onion, oat, tea, etc.) and wild plant species. As they are natural toxic constituents of plant defense, breeders and plant researchers aiming to boost plant imm unity should focus on transferring these compounds in cash crops.

RP-HPLC-based phytochemical analysis and diverse pharmacological evaluation of Quercus floribunda Lindl. ex A. Camus nuts extracts.

Quercus floribunda Lindl. ex A. Camus nuts have important folklore uses, assessed for underexplored biological potential. Nuts galls or cores and coats were utilized for the preparation of extracts using 14 solvent systems. Antioxidant, antimicrobial, cytotoxic and enzyme inhibition assays along with phytochemical profiling was carried out. Distilled water cores extract demonstrated maximum percent yield, phenolics content and total antioxidant capacity. Methanol: ethyl acetate cores extract showed maximum flavonoids content, total reducing power and protein kinase inhibition. Highest percentage radical scavenging and brine shrimp lethality was revealed by acetone: distilled water cores extract. Ethyl acetate cores extract indicated maximum α-amylase inhibition. Methanol: water coats extract showed substantial leishmanial growth inhibition. n-Hexane and chloroform coats extracts showed maximum cytotoxicity against HepG2 and THP-1 cell lines, respectively. Polyphenols quantified through RP-HPLC analysis were quercetin, pyrocatechol, gallic acid, catechin and chlorogenic acid ranging from 0.003 ± 0.001 to 1.785 ± 0.5 µg/mg extract.

Antiarthritic Potential of Comprehensively Standardized Extract of Alternanthera bettzickiana: In Vitro and In Vivo Studies.

Alternanthera bettzickiana is being used as a folk remedy for treating arthritis by conventional healers in Thailand. The current research was undertaken to explore the antiarthritic potential of A. bettzickiana ethanolic extract (ABEE). Plant characterization, molecular docking, and in vitro and in vivo (ABEE at 250, 500, and 1000 mg/kg was administered orally to rats once daily for 28 days) studies to explore the antiarthritic effect and enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (RT-PCR) analyses were performed. Oxidative stress biomarkers (superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA)) in the serum and histopathological and radiographic assessment of joints were also carried out. Gallic acid, catechin, chlorogenic acid, sinapic acid, quercetin, and γ- and α-tocopherol were identified in high-performance liquid chromatography (HPLC). Molecular docking revealed a strong interaction between these compounds and cyclooxygenase (COX) enzymes. The extract significantly subdued paw swelling and arthritic scoring, inhibited cachexia, and considerably improved biochemical and hematological modifications. SOD and CAT levels increased and the MDA level decreased in ABEE-treated rats dose-dependently. Radiographic and histopathological analyses also supported the antiarthritic effect of ABEE, which was linked with the downregulation of nuclear factor (NF)-kB, COX-2, interleukin (IL)-6, tumour necrosis factor (TNF)-α, and IL-1ß and upregulation of IL-10, I-kB, and IL-4 as compared to disease control rats. Results suggested that A. bettzickiana possessed antiarthritic potential, supporting its folkloric use for treating rheumatoid arthritis.

Calotropis procera (root) escalates functions rehabilitation and attenuates oxidative stress in a mouse model of peripheral nerve injury.

Peripheral nerve injuries result in sensorimotor functional loss, leading to permanent disability and physical dependency with immense cost and reduced quality of life. These injuries are among those complicated medical situations which still are waiting for their first-line treatment. This study was designed to investigate the role of Calotropis procera (crude roots) in accelerating functional retrieval following mechanically induced sciatic nerve injury in healthy albino male mice. Following acclimatization, mice were grouped equally as "Control" fed on normal chow and "Root" fed on C. procera root (100mg/kg/day) mixed chow. A mechanical crush was induced in right sciatic nerve of animals. Behavioral analyses (grip strength, SFI, pinprick and hot plate tests) were conducted for assessing sensorimotor function reclamation and blood was collected for oxidative stress assessment. Significantly earlier retrieval of sensorimotor activities (p<0.05), reduced total oxidant status, increased total antioxidant capacity with prominently enhanced arylesterase and paraoxonase activities (p<0.001) in treatment group suggested positive impact of C. procera roots on quickening functional recovery and combating oxidative stress following nerve injury. Thus C. procera root can be considered as potential candidate drug for further investigation to seek bioactive compound/s that may actually responsible for ameliorative functional recovery following nerve injury.

Epidemiology and antibiogram of common mastitis-causing bacteria in Beetal goats.

BACKGROUND AND AIM: Mastitis has been identified as the most prevalent and economically imperative disease among dairy animals. Thus, understanding its common bacterial pathogens and risk factors is necessary to improve udder health at herd, region, or country level. However, scientific research on caprine mastitis, especially on Beetal breed, has remained to be insufficient in Pakistan. Therefore, this study aimed to evaluate the epidemiology and antibiogram assay of common mastitis-causing bacterial agents, that is, Staphylococcus, Streptococcus, and Escherichia coli, in dairy goats. MATERIALS AND METHODS: In total, 500 Beetal goats, irrespective of age and those that were not treated with any kind of antimicrobial agents during the past 120 h, were screened using California Mastitis Test in Pattoki, Kasur District, whereas epidemiological factors were recorded. The milk samples of mastitic goats were then collected and processed using standard methods. Each sample was primarily cultured on nutrient agar. Using a specific medium, each bacterial colony was separated using several streak methods. Six antibiotic disks belonging to different antibiotic groups were used for antibiogram profiling of bacterial isolates. Chi-square test was used to assess the association of baseline characteristics and mastitis occurrence. Meanwhile, multivariable logistic regression (p<0.001) was utilized to determine the risk factors associated with positive and negative dichotomous outcome of mastitis. RESULTS: The results revealed that the overall prevalence of goat mastitis was 309 (61.8%), in which 260 (52%) and 49 (9.8%) cases were positive for subclinical mastitis (SCM) and clinical mastitis (CM), respectively. Streptococcus and E. coli were found to be the predominant isolates causing SCM and CM, respectively (p<0.001). It was observed that amoxicillin+clavulanic acid was highly sensitive to isolates of Staphylococcus and Streptococcus and ceftiofur sodium to isolates of Streptococcus and E. coli., while enrofloxacin was found to be sensitive to isolates of Streptococcus and E. coli. Risk factors such as herd structure, deworming, vaccination, presence of ticks, use of teat dip and mineral supplements, feeding type, age, parity, housing, blood in the milk, milk leakage, milk taste, and milk yield were found to have the strongest association with mastitis occurrence, while ease of milking has moderate association. CONCLUSION: In the area examined, cases of SCM were found to be higher compared with that of CM, and ceftiofur sodium has been identified as the preferred treatment in both clinical and subclinical forms of caprine mastitis in Beetal goats. Risk factors for mastitis that was identified in this study can form the basis for the creation of an udder health control program specific for dairy goats. We hope our findings could raise awareness of the risk factors and treatment approaches for common mastitis-causing bacterial agents.

Structural diversity and functional variability of gut microbial communities associated with honey bees.

Microbial consortia accompanied to all eukaryotes can be inherited from ancestors, environment, and/or from various food source. Gut microbiota study is an emerging discipline of biological sciences that expands our understanding of the ecological and functional dynamics of gut environments. Microorganisms associated with honey bees play an important role in food digestion, colony performance, immunity, pollination, antagonistic effect against different pathogens, amelioration of food and many more. Although, many repots about honey bee gut microbiota are well documented, microbiome with other key components of honey bees such as larvae, adults, their food (pollen, beebread, and honey), honey combs, and floral nectar are poorly understood. Mutual interactions and extent of the roles of microbial communities associated with honey bees are still unclear and demand for more research on the nutritional physiology and health benefits of this ecologically and economically important group. Here in this study, we highlighted all the honey bee microbiome that harbored from different life stages and other relevant components. The anatomical parts of honey bee (larvae, adults), food source (pollen, beebread, and honey), honey combs, and floral nectar were highly flourished by numerous microorganisms like bacteria (Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Actinobacteria, Actinomycetes, Bacilli, Bacteroidetes, Cocci, Clostridia, Coliforms, Firmicutes, Flavobacteriia, Mollicutes) and fungi (Dothideomycetes, Eurotiomycetes, Mucormycotina, Saccharomycetes, Zygomycetes, Yeasts, Molds). Some distinctive microbial communities of a taxonomically constrained species have coevolved with social bees. This contribution is to enhance the understanding of honey bee gut microbiota, to accelerate bee microbiota and microbiome research in general and to aid design of future experiments in this growing field.

Management of house fly, Musca domestica L. (Muscidae: Diptera), through botanical baits

ABSTRACT The house fly, Musca domestica L. (Diptera: Muscidae), is a major pest of all aspects of life, like the domestic, medical and veterinary and causal agent of several pathogenic diseases. The present study was conducted to evaluate the potential of different insecticide-free baits against house fly by incorporating flower methanol extract of Helianthus annuus (sunflower) and Tegetes erecta (marigold) at 10%, 20% and 30% bait formulation of corn syrup, dried milk and water. However, imidacloprid and thiacloprid (each at 5% concentration) were also included in the study for comparison. Results showed that insecticide baits were superior in causing mortality of adult house fly but dependent upon syrup. Overall, 20% baits of both extracts caused more than half population death of house fly within 48h. On the other hand, the mortality rate by 30% baits (from sunflower and marigold) had a similar impact as observed in case of imidacloprid and thiacloprid baits. Therefore, biological baits could play a more active and safer role in the management of house fly as compared to synthetic insecticides.

Enhancement of nutraceutical and antioxidant potential of sunflower hybrid seed varieties through chemical priming.

Pathogenic agents cause an increased risk of various fatal diseases and there is a need to reduce this risk using medicinal plants and their seeds. The present research work was designed to study the efficacy of different sunflower seed hybrid varieties (i.e. FH622, FH620, FH615, FH613 and FH545) chemically primed with potassium nitrate as natural antioxidant and antimicrobial agent. Antioxidant potential was determined using DPPH test, reducing power, TPC and TFC. Antibacterial activity was determined against Gram positive and Gram negative bacterial species. After one week, the germination data including mean germination and percentage of final emergence was calculated. It was found that seed varieties FH620 and FH615 have higher values of mean germination as compared to FH545 while FH615 has higher percentage of final emergence as compared to FH620 and FH545. High phenolic and flavonoid contents were observed in FH620 and FH615 as compared to FH545 variety. It was also observed that seed variety FH615 when treated with KNO3 solution had significantly high germination as well as antioxidant parameters and antibacterial activity as compared to other varieties. Similarly FH615 showed high antibacterial activities against Gram positive Staphylococcus aureus and Gram-negative Escherichia coli bacteria. This study showed that all selected sunflower hybrids have good antioxidant and antibacterial potentials that would further be used for different trials to cure different pathogen related diseases, and these are natural sources of antioxidants for commercial and therapeutic applications.

TH1/TH2 chemokines/cytokines profile in rats treated with tetanus toxoid and Euphorbia tirucalli.

Natural products, including their purified materials, play a remarkable role in drug development. The Euphorbiaceae family, mainly Euphorbia tirucalli, is used in some traditional medicine, and has evidence that its latex comprises immunomodulatory properties and cytokine production. This study aimed to measure the in vivo production of chemokines (IL-1α, IL-1ß, IL-12, and RANTES), TH1 cytokines (IFN-γ, TNF-α, GM-CSF, and IL-2) and TH2 cytokines (IL-4, IL-6, IL-10, and IL-13) in rats after treatments with ethanol latex extract of E. tirucalli. Vaccine treated and untreated rats were divided into seven groups to assess antimicrobial activities of the extracted components. After completion of the treatment schedule, blood was withdrawn and sera were collected. The results showed that the main component of the extract was a euphol compound. The extract showed antimicrobial activity and had the ability to modulate innate and adaptive immunity. Animals treated with extract for only 7 days before vaccination showed higher levels of antibody production. The extract showed antibacterial and antifungal activities. The extract could stimulate both adaptive and innate immunity. Pre-treatment with the extract increased immune responses in vaccinated animals, indicating the usefulness of the extract before immunization.

Studies on antioxidant, hepatoprotective, and vasculoprotective potential of Viola odorata and Wrightia tinctoria.

We have reported the antidyslipidemic, antihypertensive, and Ca++ channel blocking activities of Viola odorata (VO) and Wrightia tinctoria (WT). This study extends our understanding of their therapeutic potential by exploring the effects on biomarkers of hepatic and vascular dysfunction together with phytochemical standardization and antioxidant potential. Total phenolic compounds, total flavonoids content, and proanthocyanins of the methanolic extracts were identified using HPLC. Antioxidant capacity was measured using the in vitro assays. Two studies of 6-week duration were conducted on a high-fat diet rat model to test the leaves and seed extracts of VO and WT (300 and 600 mg/kg) for their effect on biomarkers for hepatic and vascular dysfunction. The HPLC analysis showed high contents of total phenolic compounds, total flavonoids content, and proanthocyanins along with distinctive phenolic composition. Both extracts exhibited significant antioxidant potential in 1,1-diphenyl-2-picrylhydrazyl, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), fluorescence recovery after photobleaching, and total antioxidant capacity (TAC) assays, comparable with synthetic standard antioxidants. The in vivo studies indicated a significant reduction in the high-fat-diet-induced rise in serum uric acid, phosphorus, aspartate aminotransferase, alanine aminotransferase, and gamma-glutamyl transferase. This study indicates the potential of VO and WT to protect from vascular and hepatic damage and an antioxidant effect, thus making these herbs strong candidates for managing cardiometabolic disorders.

Role of Saponins in Plant Defense Against Specialist Herbivores.

The diamondback moth (DBM), Plutella xylostella (Lepidoptera: Plutellidae) is a very destructive crucifer-specialized pest that has resulted in significant crop losses worldwide. DBM is well attracted to glucosinolates (which act as fingerprints and essential for herbivores in host plant recognition) containing crucifers such as wintercress, Barbarea vulgaris (Brassicaceae) despite poor larval survival on it due to high-to-low concentration of saponins and generally to other plants in the genus Barbarea. B. vulgaris build up resistance against DBM and other herbivorous insects using glucosinulates which are used in plant defense. Aside glucosinolates, Barbarea genus also contains triterpenoid saponins, which are toxic to insects and act as feeding deterrents for plant specialist herbivores (such as DBM). Previous studies have found interesting relationship between the host plant and secondary metabolite contents, which indicate that attraction or resistance to specialist herbivore DBM, is due to higher concentrations of glucosinolates and saponins in younger leaves in contrast to the older leaves of Barbarea genus. As a response to this phenomenon, herbivores as DBM has developed a strategy of defense against these plant biochemicals. Because there is a lack of full knowledge in understanding bioactive molecules (such as saponins) role in plant defense against plant herbivores. Thus, in this review, we discuss the role of secondary plant metabolites in plant defense mechanisms against the specialist herbivores. In the future, trials by plant breeders could aim at transferring these bioactive molecules against herbivore to cash crops.

Comparative Transcriptome Analysis of Developing Seeds and Silique Wall Reveals Dynamic Transcription Networks for Effective Oil Production in Brassica napus L.

Vegetable oil is an essential constituent of the human diet and renewable raw material for industrial applications. Enhancing oil production by increasing seed oil content in oil crops is the most viable, environmentally friendly, and sustainable approach to meet the continuous demand for the supply of vegetable oil globally. An in-depth understanding of the gene networks involved in oil biosynthesis during seed development is a prerequisite for breeding high-oil-content varieties. Rapeseed (Brassica napus) is one of the most important oil crops cultivated on multiple continents, contributing more than 15% of the world's edible oil supply. To understand the phasic nature of oil biosynthesis and the dynamic regulation of key pathways for effective oil accumulation in B. napus, comparative transcriptomic profiling was performed with developing seeds and silique wall (SW) tissues of two contrasting inbred lines with ~13% difference in seed oil content. Differentially expressed genes (DEGs) between high- and low-oil content lines were identified across six key developmental stages, and gene enrichment analysis revealed that genes related to photosynthesis, metabolism, carbohydrates, lipids, phytohormones, transporters, and triacylglycerol and fatty acid synthesis tended to be upregulated in the high-oil-content line. Differentially regulated DEG patterns were revealed for the control of metabolite and photosynthate production in SW and oil biosynthesis and accumulation in seeds. Quantitative assays of carbohydrates and hormones during seed development together with gene expression profiling of relevant pathways revealed their fundamental effects on effective oil accumulation. Our results thus provide insights into the molecular basis of high seed oil content (SOC) and a new direction for developing high-SOC rapeseed and other oil crops.

Effects of manganese on maturation of porcine oocytes in vitro and their subsequent embryo development after parthenogenetic activation and somatic cell nuclear transfer.

This study was carried out to examine the effects of manganese (Mn) on the developmental competence of porcine oocytes during in vitro maturation (IVM) after parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT). Upon treatment of porcine oocytes with different concentrations (0, 3, 6, and 12 ng/ml) of Mn during IVM, PA was performed to determine the optimum concentration. Following PA, the rate of blastocyst formation was higher significantly in treated porcine oocytes at 6 ng/ml of Mn than in other groups (P < 0.05). However, there was no substantial difference in the cleavage rate and total blastocyst cell numbers among all groups. SCNT was performed using the optimal concentration of Mn from PA, which showed an improved blastocyst formation rate in treated oocytes compared to that in control group (P < 0.05). However, the cleavage rate and total cell numbers per blastocyst were not different between the control and the Mn treated groups after SCNT. Additionally, oocyte nuclear maturation, intracellular glutathione (GSH), and reactive oxygen species (ROS) levels were assessed. There was no significant difference observed in nuclear maturation among all the groups. However, enhanced intracellular GSH levels while lower levels of ROS were seen in the Mn treated group compared to the control group (P < 0.05). Thus, these results indicate that Mn supplementation can improve the developmental competence of porcine PA and SCNT embryos by increasing GSH and decreasing ROS levels.

Bio-guided profiling and HPLC-DAD finger printing of Atriplex lasiantha Boiss.

BACKGROUND: Plants represent an intricate and innovative source for the discovery of novel therapeutic remedies for the management of various ailments. The current study has been aimed to validate the therapeutic potential of ethnomedicinally significant plant Atriplex lasiantha Boiss. METHODS: The polarity based extraction process was carried out using fourteen solvents to figure out best extraction solvent and bioactive fractions. Total phenolic-flavonoids contents were quantified colorimetrically and polyphenolics were measured using HPLC-DAD analysis. Moreover, the test samples were tested against several diseases targets following various assays including free radicals scavenging, antibacterial, antifungal, cytotoxic and antileishmanial assay. RESULTS: Among the solvent fractions, maximum yield was obtained with methanol-water extract i.e., 11 ± 0.49%. Maximum quantity of gallic acid equivalent phenolic content and quercetin equivalent flavonoid content were quantified in methanol-ethyl acetate extract of A. lasiantha. Significant quantity of rutin i.e., 0.3 µg/mg was quantified by HPLC analysis. The methanol-ethyl acetate extract of A. lasiantha exhibited maximum total antioxidant and total reducing power with 64.8 ± 1.16 AAE/mg extract respectively, while showing 59.8 ± 1.07% free radical scavenging potential. A significant antibacterial potential was exhibited by acetone-distilled water extract of A. lasiantha with 11 ± 0.65 mm zone of inhibition against B. subtilis. Considerable antifungal activity was exhibited by ethyl acetate-n-hexane extract of aerial part of A. lasiantha with 14 ± 1.94 mm zone of inhibition against A. fumigatus. Highest percentage of α-amylase inhibition (41.8 ± 1.09%) was observed in ethyl acetate-n-hexane extract. Methanol-acetone extract of A. lasiantha demonstrated significant inhibition of hyphae formation with 11 ± 0.49 mm bald zone of inhibition. Significant in-vitro cytotoxicity against Hep G2 cell line has been exhibited by methanol-chloroforms extract of A. lasiantha. CONCLUSION: The current study reveals the prospective potential of Atriplex lasiantha Boiss. for the discovery of biologically active compounds through bioassay guided isolation against various diseases.