Carotenoid Extraction from Plant Tissues.

Carotenoids are the natural pigments available in nature and exhibit different colors such as yellow, red, and orange. These are a class of phytonutrients that have anti-cancer, anti-inflammatory, anti-oxidant, immune-modulatory, and anti-aging properties. These were used in food, pharmaceutical, nutraceutical, and cosmetic industries. They are divided into two classes: carotenes and xanthophylls. The carotenes are non-oxygenated derivatives and xanthophylls are oxygenated derivatives. The major source of carotenoids are vegetables, fruits, and tissues. Carotenoids also perform the roles of photoprotection and photosynthesis. In addition to the roles mentioned above, they are also involved and act as precursor molecules for the biosynthesis of phytohormones such as strigolactone and abscisic acid. This chapter briefly introduces carotenoids and their extraction method from plant tissue. Proposed protocol describes the extraction of carotenoid using solvents chloroform and dichloromethane. Reverse-phase HPLC can be performed with C30 columns using gradient elution. The column C30 is preferred to the C18 column because the C30 column has salient features, which include selective nature in the separation of structural isomers and hydrophobic, long-chain compounds, and shows the best compatibility with highly aqueous mobile phases. A complete pipeline for the extraction of carotenoids from plant tissue is given in the present protocol.

Sample Preparation from Plant Tissue for Gas Chromatography-Mass Spectrometry (GC-MS)we.

Metabolites are intermediate products formed during metabolism. Metabolites play different roles, including providing energy, supporting structure, transmitting signals, catalyzing reactions, enhancing defense, and interacting with other species. Plant metabolomics research aims to detect precisely all metabolites found within tissues of plants through GC-MS. This chapter primarily focuses on extracting metabolites using chemicals such as methanol, chloroform, ribitol, MSTFA, and TMCS. The metabolic analysis method is frequently used according to the specific kind of sample or matrix being investigated and the analysis objective. Chromatography (LC, GC, and CE) with mass spectrometry and NMR spectroscopy is used in modern metabolomics to analyze metabolites from plant samples. The most frequently used method for metabolites analysis is the GC-MS. It is a powerful technique that combines gas chromatography's separation capabilities with mass spectrometry, offering detailed information, including structural identification of each metabolite. This chapter contains an easy-to-follow guide to extract plant-based metabolites. The current protocol provides all the information needed for extracting metabolites from a plant, precautions, and troubleshooting.

Identification of novel AKT1 inhibitors from Sapria himalayana bioactive compounds using structure-based virtual screening and molecular dynamics simulations.

Through the experimental and computational analyses, the present study sought to elucidate the chemical composition and anticancer potential of Sapria himalayana plant extract (SHPE). An in vitro analysis of the plant extract was carried out to determine the anticancer potential. Further, network pharmacology, molecular docking, and molecular dynamic simulation were employed to evaluate the potential phytochemical compounds for cervical cancer (CC) drug formulations. The SHPE exhibited anti-cancerous potential through inhibition properties against cancer cell lines. The LC-MS profiling showed the presence of 14 compounds in SHPE. Using network pharmacology analysis, AKT1 (AKT serine/threonine kinase 1) is identified as the possible potential target, and EGFR (Epidermal Growth Factor Receptor) is identified as the possible key signal pathway. The major targets were determined to be AKT1, EGFR by topological analysis and molecular docking. An in silico interaction of phytoconstituents employing molecular docking demonstrated a high binding inclination of ergoloid mesylate and Ergosta-5,7,9(11),22-tetraen-3-ol, (3.beta.,22E)- with binding affinities of -15.5 kcal/mol, and -11.3 kcal/mol respectively. Further, MD simulation and PCA analyses showed that the phytochemicals possessed significant binding efficacy with CC protein. These results point the way for more investigation into SHPE compound's potential as CC treatment.

Hepatoprotective and cardioprotective effect of Artemisia nilagirica leaf extract on E. coli challenged broiler chicken.

Artemisia nilagirica is an important medicinal plant found to exhibit several medicinal properties but the use of its leaves for combating E. coli infection has not been scientifically validated in poultry. The present study was conducted to evaluate the protective effects of methanol leaf extract of A. nilagirica (ANE) on E. coli challenged broiler chickens. Three hundred and thirty, day-old broiler chickens, were divided into 6 groups of 55 each, with group EX infected intraperitoneally (I/P) with LD50 dose of 1 × 107 cfu/ml of E. coli; group(s) EA1, EA2 and EA3 infected I/P with 1 × 107 cfu/ml of E. coli and supplemented with ANE @ 0.5, 1.0 and 2.0 g/L of drinking water, respectively; group AX were only given ANE @ 2.0 g/L in the drinking water. ANE treatment was started from day 4 and was continuously given in the drinking water up to day 21. E. coli infection was given to the birds on day 7 of their age. The effect of the plant extract was evaluated on the basis of gross, microscopic and ultrastructural alterations in E. coli challenged broiler chickens. The extract of A. nilagirica was found to show antibacterial, cardioprotective and hepatoprotective properties in a dose-dependent manner on the basis of gross and microscopic examination. The methanol extract of A. nilagirica leaves revealed no toxic effect on the hepatocytes on ultrastructural evaluation. This study demonstrates the antimicrobial, hepatoprotective and cardioprotective activities of ANE in broiler chickens infected with E. coli organism.

Cancer cell plasticity: from cellular, molecular, and genetic mechanisms to tumor heterogeneity and drug resistance.

Cancer is a complex disease displaying a variety of cell states and phenotypes. This diversity, known as cancer cell plasticity, confers cancer cells the ability to change in response to their environment, leading to increased tumor diversity and drug resistance. This review explores the intricate landscape of cancer cell plasticity, offering a deep dive into the cellular, molecular, and genetic mechanisms that underlie this phenomenon. Cancer cell plasticity is intertwined with processes such as epithelial-mesenchymal transition and the acquisition of stem cell-like features. These processes are pivotal in the development and progression of tumors, contributing to the multifaceted nature of cancer and the challenges associated with its treatment. Despite significant advancements in targeted therapies, cancer cell adaptability and subsequent therapy-induced resistance remain persistent obstacles in achieving consistent, successful cancer treatment outcomes. Our review delves into the array of mechanisms cancer cells exploit to maintain plasticity, including epigenetic modifications, alterations in signaling pathways, and environmental interactions. We discuss strategies to counteract cancer cell plasticity, such as targeting specific cellular pathways and employing combination therapies. These strategies promise to enhance the efficacy of cancer treatments and mitigate therapy resistance. In conclusion, this review offers a holistic, detailed exploration of cancer cell plasticity, aiming to bolster the understanding and approach toward tackling the challenges posed by tumor heterogeneity and drug resistance. As articulated in this review, the delineation of cellular, molecular, and genetic mechanisms underlying tumor heterogeneity and drug resistance seeks to contribute substantially to the progress in cancer therapeutics and the advancement of precision medicine, ultimately enhancing the prospects for effective cancer treatment and patient outcomes.

Modulation of mammary tumour progression using murine model by ethanol root extract of Saussurea costus (falc.) lipsch.

ETHNOPHARMACOLOGICAL RELEVANCE: Breast cancer is a major cause of death among human females across the globe. The anti-neoplastic agents or therapies used for the treatment of cancers can enhance longevity but are subsequently observed to deteriorate the quality of life due to the extensive side effects produced. Saussurea costus is a potential medicinal plant of the Himalayas with noticeable ethnopharmacological properties. The phytochemicals present in Saussurea costus are responsible for anti-carcinogenic potential and warranted nil or minimal side effects of Saussurea costus and directed to use this plant as a preventive or therapeutic drug candidate against cancers. AIM OF THE STUDY: The present study was planned to evaluate the anti-neoplastic activity of Saussurea costus root extract (SL) in rat mammary tumour model. MATERIALS AND METHODS: The anti-neoplastic activity of SL root extract at 3 different doses (100, 250 and 500 mg/kg BW) for 18 weeks against 12-dimethylbenz (a) anthracene (DMBA)-induced mammary tumours in Sprague Dawley (SD) female rats was analyzed through serum biochemistry (ALT, AST, ALP, Total protein, Creatinine and BUN), oxidative stress parameters (Lipid peroxidation, Catalase and Reduced glutathione), pro-inflammatory cytokines (TNF-α and NF-κB), immunohistochemical markers (Ki-67, MMP-9 and VEGF), real-time PCR (PCNA, p53, bax, bcl-2 and caspase-3, genes) and molecular docking. RESULTS: Inhibition of tumour parameters, minimal alteration in the liver (ALT, AST and ALP) and kidney enzymes (Creatinine and BUN), decreased activity of MDA, elevated levels of GSH and catalase, reduction in the levels of pro-inflammatory cytokines i.e. TNF-α and NF-κB, reduced gross and histomorphological changes, declined expression of Ki-67, MMP-9 and VEGF in vivo rat model, mRNA expression of cancer-related genes and docking of dehydrocostus lactone and costunolide with NF-κB and TNF-α demonstrated the chemopreventive action of SL root extract. CONCLUSIONS: The in-vivo trial elucidates anti-neoplastic activity of Saussurea costus root extract as demonstrated through the reduction of biochemical indices, oxidative stress parameters, histological changes, pro-inflammatory cytokines (NF-κB and TNF-α), cellular proliferation (Ki-67), metastases (MMP-9) and neovascularization (VEGF) markers with highest anti-neoplastic effect of SL extract at the dose of 500 mg/kg body weight. Therefore, the present study signifies the need to use the active principles present in the root extract of Saussurea costus against breast cancer as a therapeutic regimen.

Network pharmacology on mechanistic role of Thymus linearis Benth. against gastrointestinal and neurological diseases.

BACKGROUND: Thymus linearis Benth. (TL) is native to the Himalayas and has been used traditionally to cure various diseases. Thymus is a well-known aromatic perennial herb commonly known as Van ajwain or Himalayan Thyme. The main components of the TL essential oil are thymol, terpinene, and p-cymene. There are many biological properties that TL has been reported to exhibit, including antioxidant, analgesic, anti-inflammatory, antipyretic, antibacterial, anticancerogenic, and neuroprotective effects. PURPOSE: In this study, the network pharmacology and molecular docking were used to explore the potential compounds of TL and their interaction mechanism toward gastrointestinal complications and neurological diseases. METHODS: Network pharmacology was used to analyze the active compounds and protein targets of TL on gastrointestinal and neurological related diseases. Protein-protein interaction and Kyoto Encyclopaedia of Genes and Genomes analysis were used to enrich and evaluate key pathways of target proteins. To ensure the reliability of the network pharmacology prediction estimates, molecular docking was used to confirm the relationships between the core components and targets of TL. RESULT: 77 physiologically active compounds and their 717 predicted protein targets with high association to the neuroactive ligand-receptor interaction pathway were derived from TL. Beta-citronellol, piperitol, p-cymen-8-ol, and alpha-humulene were found to have a role in gastrointestinal diseases associated with neurological diseases. These compounds showed significant levels of multitargeting cluster regulatory activity. The molecular docking results showed regulatory activity of highlighted multi-targeting compounds and the highest docking energy was reported in piperitol. CONCLUSION: The study shows that Thymus linearis Benth., a medicinal plant with traditional use, possesses valuable biologically active compounds. It emphasizes the potential of Thymus in treating gastrointestinal and neurological diseases by regulating oxidative stress pathways. This research opens up possibilities for discovering antioxidant molecules for future drug development. It is an interesting study with promising implications for further research.

Fatty Acids Reverse the Supramolecular Chirality of Insulin Fibrils.

Long-chain unsaturated and polyunsaturated fatty acids (LCUFAs and LCPUFAs, respectively) are the essential components of phospholipids and sphingolipids, major building blocks of plasma and organelle membranes. These molecules are also involved in cell signaling and energy metabolism. Hence, both LCUFAs and LCPUFAs are broadly used as food supplements. However, the role of these fatty acids (FAs) in the self-assembly of misfolded proteins remains unclear. In this study, we investigated the effect of LCUFAs and LCPUFAs, as well as their saturated analogue, on insulin aggregation. Using vibrational circular dichroism, we found that all analyzed FAs reversed the supramolecular chirality of insulin fibrils. Molecular dynamics simulations showed that strong hydrophobic interactions were formed between the long aliphatic tails of FAs and hydrophobic amino acid residues of insulin. We infer that such insulin:FA complexes had different self-assembly mechanisms compared to that of insulin alone, which resulted in the observed reversal of the supramolecular chirality of the amyloid fibrils.

Effects of supplemented multicomponent mycotoxin detoxifying agent in laying hens fed aflatoxin B1 and T2-toxin contaminated feeds.

The present study was conducted to determine the ability of multicomponent mycotoxin detoxifying agent (MMDA) in feed to prevent the gastrointestinal absorption of aflatoxin B1 (AFB1) and T2-toxin supplemented via spiked maize. For comparisons, hens were fed with uncontaminated basal diet without or with addition of MMDA at 2 g/kg feed. The trial consisted of 105 laying hens (Lohmann Brown) without obvious signs of disease allocated to 7 treatment groups in 35 pens. Responses were demonstrated on laying performance and health status throughout the 42 d experimental period. The results of laying performance indicated significantly decreased egg mass with increasing mycotoxin (AFB1 and T2-toxin) levels up to the maximum tolerated dosage, however simultaneous presence of MMDA laying performance was slightly modified linearly to increasing application. Dose-dependent pathological changes in liver and kidneys and their relative weights, changes in blood parameters and reduced eggshell weights were observed in the hens fed AFB1 and T2-toxin. The pathological changes in the hens fed with diets containing AFB1 and T2-toxin without MMDA were significantly higher as compared with the control group, but eggshell stability was not affected. The contents of AFB1, T2-toxin and their metabolites in liver and kidney tissues were significantly decreased in the hens supplemented with MMDA at 2 and 3 g/kg in feed. MMDA supplementation significantly reduced the deposition of AFB1, T2-toxin and their metabolites in liver and kidneys at the maximum tolerated dosage (2 and 3 g/kg) indicating specific binding to AFB1 and T2-toxin in the digestive tract as compared to the corresponding diets without MMDA. Exposure of AFB1 and T2-toxin indicated significantly decreased egg mass with increasing mycotoxin levels up to the maximum tolerated dosage because of the significantly reduced egg production. Therefore, in this study, MMDA could reduce negative effects of feeding AFB1 and T-2 to laying hens.

Comparative Genomics and Physiological Investigations Supported Multifaceted Plant Growth-Promoting Activities in Two Hypericum perforatum L.-Associated Plant Growth-Promoting Rhizobacteria for Microbe-Assisted Cultivation.

Plants are no longer considered standalone entities; instead, they harbor a diverse community of plant growth-promoting rhizobacteria (PGPR) that aid them in nutrient acquisition and can also deliver resilience. Host plants recognize PGPR in a strain-specific manner; therefore, introducing untargeted PGPR might produce unsatisfactory crop yields. Consequently, to develop a microbe-assisted Hypericum perforatum L. cultivation technique, 31 rhizobacteria were isolated from the plant's high-altitude Indian western Himalayan natural habitat and in vitro characterized for multiple plant growth-promoting attributes. Among 31 rhizobacterial isolates, 26 produced 0.59 to 85.29 µg mL-1 indole-3-acetic acid and solubilized 15.77 to 71.43 µg mL-1 inorganic phosphate; 21 produced 63.12 to 99.92% siderophore units, and 15 exhibited 103.60 to 1,296.42 nmol α-ketobutyrate mg-1 protein h-1 1-aminocyclopropane-1-carboxylate deaminase (ACCD) activity. Based on superior plant growth-promoting attributes, eight statistically significant multifarious PGPR were further evaluated for an in planta plant growth-promotion assay under poly greenhouse conditions. Plants treated with Kosakonia cowanii HypNH10 and Rahnella variigena HypNH18 showed, by significant amounts, the highest photosynthetic pigments and performance, eventually leading to the highest biomass accumulation. Comparative genome analysis and comprehensive genome mining unraveled their unique genetic features, such as adaptation to the host plant's immune system and specialized metabolites. Moreover, the strains harbor several functional genes regulating direct and indirect plant growth-promotion mechanisms through nutrient acquisition, phytohormone production, and stress alleviation. In essence, the current study endorsed strains HypNH10 and HypNH18 as cogent candidates for microbe-assisted H. perforatum cultivation by highlighting their exclusive genomic signatures, which suggest their unison, compatibility, and multifaceted beneficial interactions with their host and support the excellent plant growth-promotion performance observed in the greenhouse trial. IMPORTANCE Hypericum perforatum L. (St. John's wort) herbal preparations are among the top-selling products to treat depression worldwide. A significant portion of the overall Hypericum supply is sourced through wild collection, prompting a rapid decline in their natural stands. Crop cultivation seems lucrative, although cultivable land and its existing rhizomicrobiome are well suited for traditional crops, and its sudden introduction can create soil microbiome dysbiosis. Also, the conventional plant domestication procedures with increased reliance on agrochemicals can reduce the diversity of the associated rhizomicrobiome and plants' ability to interact with plant growth-promoting microorganisms, leading to unsatisfactory crop production alongside harmful environmental effects. Cultivating H. perforatum with crop-associated beneficial rhizobacteria can reconcile such concerns. Based on a combinatorial in vitro, in vivo plant growth-promotion assay and in silico prediction of plant growth-promoting traits, here we recommend two H. perforatum-associated PGPR, Kosakonia cowanii HypNH10 and Rahnella variigena HypNH18, to extrapolate as functional bioinoculants for H. perforatum sustainable cultivation.

Essential oil composition and antimicrobial potential of aromatic plants grown in the mid-hill conditions of the Western Himalayas.

Essential oils are highly concentrated natural extracts obtained from plants, rich in bioactive constituents with antimicrobial properties, but the distinctive climate of the Western Himalayan region influences the same. Aromatic and medicinal plants, viz., Origanum majorana, Origanum vulgare, Cymbopogon winterianus, Pelargonium graveolens, and Nepeta cataria were grown in the foothills of the Western Himalayan condition and evaluated for essential oil content, composition, and their effect on some of the most common pathogenic microorganisms. The essential oil content (%) was 0.77, 0.45, 1.37, 0.15 and 0.17% in O. majorana, O. vulgare, C. winterianus, P. graveolens, and N. cataria, respectively. The major essential oil constituents of the isolated oils were terpinen-4-ol, thymol, citronellal, citronellol, and nepetalactone, contributing 41.24%, 31.81%, 43.13%, 43.35% and 91.43% in O. majorana, O. vulgare, C. winterianus, P. graveolens, and N. cataria, respectively. Well-diffusion assay revealed that the essential oil of O. majorana and O. vulgare was active against both the tested Gram-positive, viz., Bacillus subtilis MTCC 121, Micrococcus luteus MTCC 2470, and Staphylococcus aureus MTCC 96; and Gram-negative, viz., Escherichia coli MTCC 43, Klebsiella pneumoniae MTCC 109, and Pseudomonas aeruginosa MTCC 2453 bacteria, while the essential oil of C. winterianus, P. graveolens, and N. cataria showed activity against only some Gram-positive bacteria. Minimum inhibitory concentration (v/v) values indicated the highest efficacy of O. majorana essential oil against B. subtilis (0.5%), M. luteus (1%), and S. aureus (1%), while O. vulgare was most efficient to E. coli (2%) and K. pneumoniae (2%). C. winterianus essential oil did not inhibit any bacterial strains. M. luteus was susceptible to the essential oil of P. graveolens (1%) and N. cataria (0.5%) at low concentrations. Present findings showed the association between the chemical constituents' profile of isolated essential oils from the Himalayan region and their antimicrobial activity, indicating their perspective to be utilized as antibacterial means.

Development of Reverse Transcription Recombinase Polymerase Amplification (RT-RPA): A Methodology for Quick Diagnosis of Potato Leafroll Viral Disease in Potato.

Potatoes are developed vegetatively from tubers, and therefore potato virus transmission is always a possibility. The potato leafroll virus (PLRV) is a highly devastating virus of the genus Polerovirus and family Luteoviridae and is regarded as the second-most destructive virus after Potato virus Y. Multiple species of aphids are responsible for the persistent and non-propagating transmission of PLRV. Due to intrinsic tuber damage (net necrosis), the yield and quality are drastically diminished. PLRV is mostly found in phloem cells and in extremely low amounts. Therefore, we have attempted to detect PLRV in both potato tuber and leaves using a highly sensitive, reliable and cheap method of one-step reverse transcription-recombinase polymerase amplification (RT-RPA). In this study, an isothermal amplification and detection approach was used for efficient results. Out of the three tested primer sets, one efficiently amplified a 153-bp product based on the coat protein gene. In the present study, there was no cross-reactivity with other potato viruses and the optimal amplification reaction time was thirty minutes. The products of RT-RPA were amplified at a temperature between 38 and 42 °C using a simple heating block/water bath. The present developed protocol of one-step RT-RPA was reported to be highly sensitive for both leaves and tuber tissues equally in comparison to the conventional reverse transcription-polymerase chain reaction (RT-PCR) method. By using template RNA extracted employing a cellular disc paper-based extraction procedure, the method was not only simplified but it detected the virus as effectively as purified total RNA. The simplified one-step RT-RPA test was proven to be successful by detecting PLRV in 129 samples of various potato cultivars (each consisting of leaves and tubers). According to our knowledge, this is the first report of a one-step RT-RPA performed using simple RNA extracted from cellular disc paper that is equally sensitive and specific for detecting PLRV in potatoes. In terms of versatility, durability and the freedom of a highly purified RNA template, the one-step RT-RPA assay exceeds the RT-PCR assay, making it an effective alternative for the certification of planting materials, breeding for virus resistance and disease monitoring.

An insight into the interaction between Argulus siamensis and Labeo rohita offers future therapeutic strategy to combat argulosis.

Aquaculture and fisheries are salient flourishing sectors in the world but their sustainability is often afflicted by several pathogenic diseases. Among all the pathogenic diseases of fish, parasitic diseases are found to be a major cause of concern. Argulosis is one of the dominant parasitic problems encountered in Indian aquaculture practices. Argulus siamensis is the most prevalent argulid species harming the Indian major carp species including Labeo rohita. The major carps respond to parasitic infestation by elevating various immune relevant genes. The therapeutic chemicals, synthetic drugs and other plant extracts have made a progress in the fight against argulosis. However, there is no effective vaccine and drugs are available for this disease. Thus, designing efficient, cost-effective and eco-friendly control and treatment strategies for argulosis is presently needed. Keeping the aforementioned facts in mind, the current review elaborated the immunological interaction between A. siamensis and L. rohita, available combat tactics, highlighted the already identified vaccine candidates to design effective control measures and illustrated the use of omics technology in future to combat argulosis.

Linseed oil in boar's diet improved in vivo fertility and antioxidant status.

The reactive oxygen species (ROS) which are produced during storage of boar semen are causing oxidative stress and leads to poor fertility. Also, tropical and sub-tropical weather condition adversely impacts the physicomorphological quality and fertility of boar sperm. The aim of this study was to examine the effects of feeding linseed oil to boar on its seminal attributes, sperm kinetics, biomarkers of antioxidant, fatty acid profile of seminal plasma (SP) and sperm and in vivo fertility. Six Hampshire crossbreed boars were fed with 90 ml linseed oil (LIN) whereas six Hampshire crossbreed boars were fed 90 ml canola oil (CON) for 16 weeks. Sperm quality was evaluated (60 ejaculates for each group; a total of 120 ejaculates) for motility, livability, abnormal morphology, acrosomal membrane integrity, hypo-osmotic swelling test (HOST) and sperm kinetic parameters by computer assisted semen analysis (CASA) at 0 h and at 72 h of storage at 17°C. Biomarkers of antioxidant (glutathione peroxidase; GPx, catalase; CAT, total antioxidant capacity; TAC) and malondialdehyde (MDA) were measured in SP and serum. Gas chromatography-mass spectrometry (GC-MS) was used for the estimation of fatty acid composition of SP and sperm. Boars fed with linseed oil had higher semen volume (p < .01) and more total sperm numbers (p < .01). Feeding linseed oil to boar enhanced seminal attributes (p < .05) at 0 h as well as at 72 h of storage. Linseed oil feeding (p < .01) improved biomarkers of antioxidants and significantly (p < .01) lowered the lipid peroxidation in serum and SP. Linseed oil feeding (p < .05) increased the proportion of alpha linolenic (ALA), arachidonic and docosahexaenoic (DHA) fatty acids in SP. The ratio of n-6 to n-3 fatty acids in sperm increased significantly (p < .01) in treatment group. Farrowing rate was significantly (p < .05) higher in treatment group. In conclusion, feeding linseed oil to boar improved the in vivo fertility, enhanced antioxidant capacity and increased the DHA content of SP and sperm.

Cleaner production technologies for the amelioration of soil health, biomass and secondary metabolites in Ocimum basilicum L. under Indian Western Himalaya.

Ocimum basilicum L. and its derived products are primarily consumed by humans; hence, agrochemical use seems inappropriate for its cultivation. However, farmers are accustomed to using rampant inorganic fertilizers to augment crop productivity, which has unintendedly engendered severe environmental perturbations. Concomitantly, farmers will soon have to confront the challenges of growing crops under suboptimal conditions driven by global climate change. Consequently, to develop a clean, sustainable, and resilient production technology, field experiments spanning over two years (2020 and 2021) were conducted, comprising three biostimulants, viz., vermicompost (0, 4, and 8 Mg ha-1), biofertilizer (uninoculated and inoculated), and liquid seaweed extract (without and at 7 ml L-1) in the Indian western Himalaya for the first time. Soil health indicators, leaf photosynthetic pigments, gaseous exchange, mineral contents, essential oil (EO) quantity, and composition were evaluated. Soil microbial respiration (SMR), microbial biomass carbon (MBC), organic carbon (OC), dehydrogenase (DHA), alkaline phosphatase (ALP), and ß-glucosidase activities were increased by 36.23, 83.98, 30.61, 42.69, 34.00, and 40.57%, respectively, when compared with the initial soil status. The net photosynthetic rate (Pn) was significantly increased with the highest (8 Mg ha-1) and moderate (4 Mg ha-1) vermicompost dosages by 13.96% and 4.56%, respectively, as compared with the unfertilized control (0 Mg ha-1). Likewise, the biofertilizer and seaweed extract also enhanced Pn by 15.09% and 10.09%, respectively. The crop's key EO constituents, viz., methyl chavicol and linalool, were significantly improved with the highest and moderate vermicompost rates of 2.71, 9.85%, and 1.18, 5.03%, respectively. Similarly, biofertilization and seaweed application also boosted methyl chavicol and linalool by 3.29, 8.67%, and 1.93, 3.66%, respectively. In both years, significantly higher herbage (8.86 and 11.25 Mg ha-1) and EO yield (113.78 and 154.87 kg ha-1) were recorded with a congregate treatment of the highest vermicompost dose, biofertilizer, and liquid seaweed extract. In conclusion, the integrated use of biostimulants having complementary properties can sustainably maximize the quantity and quality of O. basilicum and concomitantly ameliorate soil health. This study can inspire scientific communities and industries to develop second-generation biostimulant products, delivering better sustainability and resilience for a renaissance in agriculture.

Essential Oil Content and Compositional Variability of Lavandula Species Cultivated in the Mid Hill Conditions of the Western Himalaya.

The increase in the utilization of Lavandula essential oil in industries led to an impressive rise in the demand for quality essential oils. However, a post-harvest drying of Lavandula species can be a decisive factor to determine the quantity and quality of essential oil. The study was conducted in western Himalayan conditions to assess the essential oil content and composition of two Lavandula species viz., lavender (Lavandula angustifolia Mill.), and lavandin (Lavandula × intermedia Emeric ex Loisel), at four different drying duration (0 h, 24 h, 48 h and 72 h after the harvest). The higher growth attributes viz., plant height (71.7 cm), ear length (8.8 cm), number of spikes (18.1), and number of flowers per ear (47.5) were higher in lavandin, while the number of branches (17.1) was higher in lavender. Essential oil content (%) and moisture reduction (%) were significantly higher at 72 h than at 0 h. The major components of lavender and lavandin essential oil were linalool (33.6-40.5%), linalyl acetate (10.8-13.6%), lavandulyl acetate (2.8-14.5%), and linalyl propionate (5.3-14.1%) in both the Lavandula species. There was a decreasing trend in linalool and an increasing trend in linalyl acetate content in lavandin, with an increase in drying duration up to 72 h; while in lavender, no regular trend was observed in linalool and linalyl acetate content. It was observed that linalool and linalyl acetate levels were the highest at 24 and 0 h of drying in lavender and lavandin, respectively, and essential oil extraction can be done according to the desire of the constituent at varied drying duration.

Linseed oil in boar's diet during high temperature humidity index (THI) period improves sperm quality characteristics, antioxidant status and fatty acid composition of sperm under hot humid sub-tropical climate.

Environmental heat stress in sub-tropical climates negatively impacts boar semen production and its quality. The present study aimed to examine the heat stress alleviating effects of dietary linseed oil on semen quality and antioxidant status of boar, in the summer and winter seasons in sub-tropical climate. Six Hampshire crossbreed boars were fed with 90 mL linseed oil (treatment) whereas six boars of the same breed were fed 90 mL vegetable oil (control) for sixteen weeks during both season. Sperm quality was assessed for motility, viability, abnormality, acrosomal integrity, and Hypo-osmotic swelling test (HOST). Sperm velocity attributes were assessed by computer-assisted semen analysis (CASA). Antioxidants (glutathione peroxidase; GPx, catalase; CAT, total antioxidant capacity; TAC and nitric oxide; NO) and lipid peroxidation (malondialdehyde; MDA) were measured in seminal plasma and serum. Gas chromatography-mass spectrometry (GC-MS) was used for the estimation of fatty acid composition of seminal plasma and spermatozoa. Feeding linseed oil to the boars significantly (p < 0.05) improved sperm quality at the fresh stage and after 72 h of liquid storage in both season. There was a significant (p < 0.01) effect of treatment and season on semen quality parameters. Significant boar (p < 0.05) effect was recorded on reaction time, semen volume, sperm abnormality, acrosomal integrity and HOST reactive sperm. There was a significant (p < 0.01) effect of treatment and season on the velocity attributes viz. VAP, VSL, VCL, ALH, BCF and STR%. Linseed oil supplementation significantly (p < 0.01) enhanced antioxidant and lowered MDA levels in serum as well as seminal plasma. The concentration of alpha-linolenic (ALA), arachidonic and docosahexaenoic (DHA) fatty acids were significantly (p < 0.01) increased in seminal plasma and sperm after linseed oil supplementation. In conclusion, linseed oil supplementation to boar during high THI months improved the semen quality parameters viz. semen volume, sperm concentration, and progressive motile sperm, along with enhanced antioxidant capacity.

Pharmacological evaluation of bromelain in mouse model of Alzheimer's disease.

The current study elucidates pharmacological evaluation of bromelain as a bioactive compound obtain from pineapple stem belongs to family Bromeliaceae in AlCl3 and D - galactose induced mice. In mice, co-administration of AlCl3 at dose 5 mg/kg b.w., via the oral route, and D - galactose at dose 60 mg/kg b.w., via intraperitoneal route for 90 days resulted in cognitive impairment, spatial learning, and memory deficits, as well as neurotoxicity. However, 30 consecutive days, treatments via an intraperitoneal route with bromelain low dose (Brm L) at dose 10 mg/kg b.w., bromelain high dose (Brm H) at dose 20 mg/kg b.w., donepezil (Dnpz) at dose 2 mg/kg b.w., and Brm L + Dnpz at doses 10, 2 mg/kg b.w. were considerably reversed the effect of AlCl3 and D - galactose induced AD mice. Consequences of behavioral parameters (Morris water maze, elevated plus maze and locomotor), biochemical estimation (MDA, GSH, SOD, CAT, Nitrite and AChE), and ELISA tests (mouse BACE, Aß1 - 42, TNF-α, IL-6, and BDNF) confirmed significant (p < 0.05) neuroprotective effect of treatments in AlCl3 and D - galactose induced mice. Additionally, hematoxylin and eosin staining of the cerebral cortex and the hippocampus exposed eosinophilic lesions and hyperchromatic nuclei in AD mice, but these neurodegenerative effects were eliminated by Brm L, Brm H, Dnpz, and Brm L + Dnpz treatments. Thus, bromelain alone and in combination with donepezil prevent AlCl3 and D - galactose induced spatial learning and memory deficits, as well as cognitive impairment, by increasing cholinergic activity and synaptic plasticity, as well as reducing oxidative damage, neuroinflammation, Aß 1-42 aggregations, and histopathological damage, according to our findings. The present study consequences indicate that bromelain alone and in combination with donepezil appears to have neuroprotective properties. Henceforward, this may be a promising treatment option for Alzheimer's disease.