Colebrookea Research Articles

Phytochemical Characterization and Synergistic Antibacterial Effects of Colebrookea Oppositifolia Essential Oil as Adjuvants to Modern Antibiotics in Combating Drug Resistance.

The global threat of multi-drug-resistant bacteria has severely limited the options available for effective antibiotics. This study focuses on the antimicrobial activity and phytochemical characterization of C. oppositifolia extracts, aiming to identify novel plant-based therapeutic agents. C. oppositifolia specimens-leaves and inflorescence. Specimens were cleaned, sterilized, dried, and ground into a fine powder. Extracts were obtained using methanol and petroleum ether via a Soxhlet apparatus, followed by fractionation with chloroform, n-butanol, and ethyl acetate. Volatile oil was extracted through hydro distillation using a Clevenger apparatus. Phytochemical analysis was conducted to identify bioactive compounds. Biophysical techniques, including UV-visible spectrophotometry, TLC, HPLC, GC-MS, FTIR, and NMR, were employed for characterization. Antimicrobial activity was tested against S. aureus ATCC25922 and E. coli ATCC25922 using agar well and disc diffusion methods, and synergistic effects were assessed with erythromycin and amoxicillin. Methanol extract exhibited bacteriostatic activity with inhibition zones of 13.0 ± 0.2 mm for both S. aureus and E. coli. Petroleum ether, chloroform, n-butanol, and ethyl acetate fractions showed varying inhibition zones. Erythromycin demonstrated bactericidal activity, which was enhanced synergistically when combined with methanol extract and volatile oil, increasing inhibition zones against S. aureus. Phytochemical analysis identified phenols, flavonoids, tannins, coumarins, alkaloids, terpenoids, saponins, and glycosides. FTIR analysis revealed functional groups such as amines, aldehydes, nitriles, alkenes, and sulfones. GC-MS identified 24 compounds, with α-pinene, caryophyllene, and carene as major components. NMR spectra indicated no complex formation between oils and antibiotics, suggesting the compounds act as synergists. The C. oppositifolia extracts possess significant antimicrobial activity and synergistic potential, particularly against S. aureus. The presence of various bioactive compounds suggests a promising role in developing new plant-based therapeutics.

Exploring the therapeutic potential of Colebrookea oppositifolia in protecting against H2O2 induced oxidative stress in drosophila

Oxidative stress plays a pivotal role in the pathogenesis of various diseases. In the quest for effective therapeutic agents, natural plants with antioxidant properties have gained significant attention. Colebrookea oppositifolia has historically been used as a folk remedy for problems caused by oxidative stress

Diversity and Distribution of 100 Plant Species of a Community Managed Forest from the Mid-Hills of Nepal: A Phytosociological Approach

Phytosociology is the study of interrelationship among plant species which classifies the vegetation in a meaningful manner. The aim of this study was to study diversity and distribution of plant species in Gobankholi Community Forest in the Mid-Hills of Nepal which is dominated by Pinus roxburghii. Trees were enumerated in a 400 sq. m. plot, shrubs and climber in a 100 sq. m plot and herbs in a 4 sq. m. plot. Density, frequency, abundance, and important value index of individual species were calculated. Similarly, Simpson's diversity index, Shannon’s diversity index, and Pielou's evenness index were also calculated, based on life form viz. herbs, shrubs and trees. 66 herbs, 8 Pteridophytes, 8 shrubs, 3 climbers and 15 trees were found, among a total of 100 species. The rare orchid, Satyrium nepalense, was found at an altitude of 1275 m a.s.l. Herbs were more diverse and even than trees, shrubs and climbers. Aleuritopteris bicolor, Colebrookea oppositifolia and Pinus roxburghii were the most dominant herb, shrub and tree respectively.

Phytochemistry and Polypharmacological Potential of Colebrookea oppositifolia Smith.

Colebrookea oppositifolia Smith. is a valuable traditional therapeutic plant belonging to the family Lamiaceae. It is a dense and wool-like shrub that is mostly found in subtropical regions of some countries of Asia, such as China and India. It has been widely used for the mitigation of nervous system disorders like epilepsy. The active constituents of the plant have exhibited antioxidant, anti-microbial, and antifungal properties, which are considered due to the presence of polyphenols and flavonoids as chief chemical constituents. Flavonoids like quercetin, landenein, chrysin, and 5, 6, 7-trimethoxy flavones cause protein denaturation of the microbial cell wall. To comprehend and assemble the fragmented pieces of evidence presented on the traditional uses, botany, phytochemistry, and pharmacology of the plant to reconnoiter its therapeutic perspective and forthcoming research opportunities. The available information on Colebrookea oppositifolia has been established by electronically searching peer-reviewed literature from PubMed, Google Scholar, Springer, Scopus, Web of Science, and Science Direct over the earlier few years. The plant has been greatly used for the preparation of many herbal medicines which are used for treating traumatic injuries, fever, rheumatoid arthritis, headache, and gastric problems. From the aerial parts of the plant, a phenylethanoid glycoside named acteoside has been isolated and evaluated for its therapeutic potential viz. immunomodulatory, neuroprotective, hepatoprotective, analgesic, anti-tumour, antispasmodic, antioxidant, antibacterial, free radical scavenger, and improving sexual function. Acteoside showed neuroprotective activities against Aβ-peptide, which is neurotoxic and causes apoptosis. The petroleum ether extract of the plant leaves offers many active compounds like sitosterol, n-triacontane, hydroxydotriacontyl ferulate, acetyl alcohol, and 3,7,4,2-tetramethoxyflavones which have shown hepatoprotective potential. The plant should be evaluated further for the estimation of some other health benefits. The consequences of restricted pharmacological screening and reported phytomolecules of Colebrookea oppositifolia Smith. advocate that there is still an exigent requisite for in-depth pharmacological studies of the plant.

Colebrookea Oppositifolia Anti Arthritic Potential Vs Methotrexate in Pristane Induced Rat Arthritis

Introduction: Rheumatoid arthritis (RA) is a chronic inflammatory disease of joints with 0.24% global prevalence. Numerous pharmacological agents are available for the management of RA, but they are associated with many adverse effects. Alternative therapies for disease management are urgently needed. Colebrookea oppositifolia (CO) is an important herb with various traditional uses and pharmacological actions. In the present study, anti-arthritic effect of ethanolic extract of Colebrookea oppositifolia (EECO) leaves was evaluated in pristane induced arthritic rats (PIA) and compared to methotrexate by assessing body weight, clinical score of inflammation and histopathology.
 Aims & Objectives: To study anti-arthritic effect of C. oppositifolia on a rat model of pristane induced arthritis.
 Place and duration of study: This experimental study was conducted in Animal House of Post Graduate Medical Institute, Lahore from March to September 2019.
 Material & Methods: Total forty female Sprague Dawley rats were categorized into five equal groups (n=8). Group A (normal control), group B (disease control). Group C and D belonged to low dose (250mg/kg) and high dose (500mg/kg) EECO treated groups respectively, while group E was methotrexate treated group. Arthritis was induced within two weeks by single intradermal injection of pristane on day 0 in groups B, C, D and E. At Day 15, treatment was initiated and at day 28 paw joint sections were taken for histopathology. Data input and analysis was done by using IBM SPSS version
 
 p value ?0.05 was considered significant.
 
 Results: At week 4, significant increase (approximately 16%) in body weight was observed in all treated groups as compared to disease control. A significant reduction (more than 50%) in clinical score of arthritis was observed in all treated groups compared to diseased control group in which clinical score was 14.50±0.2. All extract and MTX treated groups showed significant improvement (p<0.001) in total histological score of arthritis (no rat was having severe disease) as compared to disease control group (75% of the rats were having severe disease).
 Conclusion: This study supported anti-arthritic effect of EECO as illustrated by reduction in inflammatory and histopathological score.

Ripening Regulation of Banana Cv. Malbhog Using Different Ripening Inducers

This experiment was carried out under the study entitled “Ripening regulation of the banana Cv. Malbhog using different ripening inducers” for controlling the ripening of the banana, for improvement of quality attributes and post-harvest life of banana. The experiment was conducted at laboratory of horticulture, Agriculture and Forestry University, Rampur, Chitwan, Nepal from 14th March to 16th April 2019. These experiments were laid out in completely randomized design in which the first experiment comprised of seven treatments consisting of distilled water spray, ethephon @ 250 ppm, ethephon @ 500 ppm, ethephon @750 ppm, ethephon@1000 ppm, Dhurseli (Colebrookea oppositifolia) leaves and Ripe banana replicated thrice. Different post-harvest parameters were recorded at the interval of two days for the experiment till any one of the treatment attained score 6 in the color chart. In this experiment, the CI-6 stage was reached earlier on the 9th day with the use of ethephon @ 1000 ppm but ethephon @ 500 ppm was found more effective regarding quality parameters with TSS (21˚B), and TSS/TA (34.66). The maximum physiological loss in weight (12.927%) and pulp peel ratio (3.65) was observed with ethephon@1000ppm and the minimum was achieved in banana sprayed with distilled water. The shelf life of banana was seen minimum (13.33 days) in ethephon @ 1000 ppm and maximum (20.33 days) in banana sprayed with distilled water.

Dereplication Based Strategy for Rapid Identification and Isolation of a Novel Anti-inflammatory Flavonoid by LCMS/MS from Colebrookea oppositifolia.

Colebrookea oppositifolia is a folkloric medicinal plant, well known for its tremendous medicinal properties such as curing epilepsy, ulcers, and urinary problems. The aim of the present study was to apply the dereplication strategy on the ethanol extract of C. oppositifolia with potent anti-inflammatory activity for the rapid identification and isolation of novel bioactive molecules to aid the drug discovery process. An integrated approach using liquid chromatography–mass spectrometry (LCMS) followed by preparative high-performance liquid chromatography (HPLC) was used for the isolation of potent molecules from the anti-inflammatory extract of C. oppositifolia. Purity of the compounds (>98.5%) was established by HPLC, and identification was carried out by NMR and ESI-MS. 5,6,7-Trihydroxyflavone-3-O-glucuronide methyl ester (compound III) isolated from C. oppositifolia was extensively studied for anti-inflammatory potential in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells and the mice model. Compound III significantly repressed various proinflammatory mediators and upregulated the release of anti-inflammatory cytokine IL-10. Compound III reduced inflammation when studied for parameters such as the phagocytic index, carrageenan-induced paw edema in mice, and effect on organ weight. It reduced inflammation in a dose-dependent manner both in vitro and in vivo. Further molecular insights into the study revealed that compound III blocks the phosphorylation of I kappa b kinase α/β (IKKα/β), IκBα, and nuclear factor kB p65 (NF-κBp65) which is a key controller of inflammation, thereby showing anti-inflammatory potential. Hence, this study permits further investigation to develop compound III as an anti-inflammatory drug.

Acteoside isolated from Colebrookea oppositifolia attenuates I/R brain injury in Wistar rats via modulation of HIF-1α, NF-κB, and VEGF pathways.

The objective of this study was to assess the anti-stroke activity of acteoside isolated from methanolic root extract of C. oppositifolia METHODS: Ischemia-reperfusion(I/R) brain injury was induced in Wistar rats to assess the anti-stroke activity of acteoside. Rats were pretreated with acteoside (10, 25 & 50mg/kg, p.o.) before the induction of I/R injury. Parameters such as neurological, motor-cognitive functionswere evaluated along with morphological (brain volume, infarct size), biochemical (SOD, Catalase, GSH, lipid peroxidation, TNF-α, IL-6, IL-10, ICAM-1, HIF-1α, VEGF, and NF-κB), histopathological, and gene expression studies (HIF-1α, VEGF) were performed to study the protective effect of acteoside against I/R induced brain injury. I/R injury caused significant deterioration of neurological (p < 0.01), motor (p < 0.01) and cognitive (p < 0.01) functions, associated with increase in the brain volume (p < 0.01), and infarct size (p < 0.01); increase in the levels of MDA, TNF-α, IL-6, ICAM-1, HIF-1α, VEGF, and NF-κBalong with significant decrease in SOD, catalase, GSH, and IL-10 (p < 0.01 for all parameters) compared to Sham control group. Histology of brain tissue of disease control group exhibited significant vascular changes, neutrophil infiltration, cerebral oedema, and necrosis of the neuronal cells. Further, the gene-expression studies showed significant increase in the HIF-1α (p < 0.01) and VEGF (p < 0.01) mRNA levels in the I/R control compared to Sham control. Interestingly, the acteoside (10, 25 & 50mg/kg) has prevented the neurological, motor and cognitive dysfunctions, along with inhibiting the morphological, biochemical, histological and gene expression changes induced by I/R-injury (p < 0.05 for 10mg; p < 0.01 for 25 & 50mg/kg of acteoside for all the parameters). These findings suggest that acteoside possess potent anti-stroke activity through modulation of HIF-1α, NF-κB, and VEGF pathway along with its potent antioxidant activity.

Traditional uses, phytochemistry, and ethnopharmacology of Colebrookea oppositifolia Smith: a mini-review

This review was focused to collect and exemplify the data on traditional uses, phytochemistry, and pharmacological outline of C. oppositifolia, and thereby recognize the current obscurities and highlight the scope and opportunities for future research on the plant. A thorough literature search was performed on C. oppositifolia in the online databases such as Scopus, PubMed/Medline, Science direct, Scifinder, EMBASE, Web of Science, Google scholar, and Elsevier. C. oppositifolia belongs to family Lamiaceae, was widely used in the traditional system of medicine for treating various human ailments. Further, phytochemicals such as acteoside, chrysin, negletein, ladanein, betulonic acid, apigenin, caffeic acid, quercetin, anisofolin A, apigetrin, forsythoside A, β-sitosterol, ferulic acid, and its derivatives were isolated from various parts of C. oppositifolia. Additionally, the plant has been scientifically validated to possess antifertility, anti-ulcer, hepatoprotective, antimicrobial, anti-oxidant, cardioprotective, anti-inflammatory, wound healing, anti-convulsant, and antihypertensive activities in various experimental models. With the available literature we can conclude that despite of enormous traditional uses, the C. oppositifolia has not been explored for its beneficial effects on cataract, rheumatism, joint pain, bleeding disorders, and dysentery. Also, there is a need for clinical studies to explore the efficacy, safety, and toxicity in humans.

Acteoside Isolated from Colebrookea oppositifolia Smith Attenuates Epilepsy in Mice Via Modulation of Gamma-Aminobutyric Acid Pathways.

The present study was aimed to evaluate the anticonvulsant activity of acteoside and explore its mechanism of action. Initially, the acteoside was evaluated in maximal electroshock (MES) and pentylenetetrazole (PTZ)-induced convulsions, and later it was evaluated against N-methyl-D-aspartic acid (NMDA)-induced mortality in Swiss albino mice. Based on the response in these models, further evaluations were performed to explore the mechanism of action. In the results, the acteoside (10, 25, and 50mg/kg) has shown significant anticonvulsant activity in the PTZ model (p < 0.01 for all doses); however, there was no protection observed in MES and NMDA models. Therefore, further mechanism-based studies were performed on the PTZ model, and the outcomes have revealed that there was a significant reduction in GABA (p < 0.01 for both regions) and elevation of glutamate (p < 0.01 for both regions) in the cortex and hippocampus regions of PTZ-treated animals. Further, the antioxidant levels (SOD, catalase, GPx, GR, GSH, LPO) were altered significantly (p < 0.01 for all parameters), with reduced GABAA mRNA levels (p < 0.01) in the PTZ control compared with the normalcontrol. Interestingly, co-administration of acteoside (25mg/kg) (p < 0.01 for all parameters) has restored all the PTZ-induced alterations compared to PTZ-control. Moreover, the anti-PTZ action of acteoside was completely blocked in the presence of flumazenil, and thus confirmed the GABAergic mechanism behind the anticonvulsant activity of acteoside. Besides, actophotometer and rotarod tests have confirmed that the acteoside is free from central side effects like motor incoordination and locomotor deficits.

Comprehensive Characterization of Secondary Metabolites from Colebrookea oppositifolia (Smith) Leaves from Nepal and Assessment of Cytotoxic Effect and Anti-Nf-κB and AP-1 Activities In Vitro.

Here we report the comprehensive characterization of the secondary metabolites from the leaves of Colebrookea oppositifolia Smith, a species used as medicinal plant in the traditional medicine of Nepal. Phytochemical screening of bioactives was performed using an integrated LC-MSn and high resolution MS (Mass Spectrometry) approach. Forty-three compounds were tentatively identified, mainly aglyconic and glycosilated flavonoids and phenolic acids, as well as other bioactives such as coumarins and terpenes were detected. Furthermore, the NF-κB and AP-1 inhibitory activity of C. oppositifolia extract were evaluated, as well as its cytotoxicity against THP-1 cells, in order to assess the potential use of this herb as a source of anti-inflammatory and cytotoxic compounds. The results so far obtained indicate that C. oppositifolia leaves extract could significantly reduce the viability of THP-1 cells (IC50 = 6.2 ± 1.2 µg/mL), as well as the activation of both NF-κB and AP-1 at the concentration of 2 μg/mL. Our results indicate that Nepalese C. oppositifolia is a valuable source of anti-inflammatory and cytotoxic compounds. The phytochemical composition reported here can partially justify the traditional uses of C. oppositifolia in Nepal, especially in the treatment of inflammatory diseases, although further research will be needed to assess the full potential of this species.

Characterization and Anti-stroke Activity of Stem Extract of Colebrookea oppositifolia Smith

This study was aimed to investigate the cerebroprotective potential of methanolic stem extract of Colebrookea oppositifolia (MsCO). The MsCO was evaluated against global ischemia and reperfusion (I/R) induced brain injury in Wistar rats. In the outcomes, the I/R insult caused significant deterioration of neurological (p<0.01), cognitive (p<0.01), and motor functions (p<0.01), along with significant alterations in the antioxidant parameters (decreased SOD and catalase enzyme activities, along with derease in GSH and increase in LPO; p<0.01 for all parameters), morphology (cerebral edema and infarct area) (p<0.01) and histology of the brain tissue in the I/R control group compared to normal control. Interestingly, the MsCO (200 and 400 mg/kg) pretreatment for seven days has attenuated all the I/R-induced alterations significantly and dose-dependently compared to I/R control group. Moreover, the LC-MS/MS analysis of MsCO revealed the presence of Acteoside, Pentagalloyl glucose, and Quercetin as major phytoconstituents. Based on these outcomes we conclude that MsCO possesses potent cerebroprotective activity against ischemia-reperfusion-induced brain injury in Wistar rats.

Methanolic Stem Extract of Colebrookea oppositifolia Attenuates Epilepsy in Experimental Animal Models: Possible Role of GABA Pathways

The current study aims to evaluate the methanolic stem extract of Colebrookea oppositifolia (MsCO) for its anticonvulsant property and its role in GABA pathways. Initially, the PsCO (petroleum ether), MsCo, and AsCO (aqueous) extracts of C. oppositifolia were evaluated in Six -hertz seizure test. Later, the potent extract was evaluated on MES, PTZ and NMDA models to explore the possible mechanisms of action. In addition, the potent extract was evaluated for its locomotor deficit inducing potential in actophotometer test. In the Six-hertz (6-Hz) seizure test, MsCO showed the most potent and dose-dependent anticonvulsant activity, and as per the pre-defined criteria, it was further evaluated in other models to explore its mechanisms of action. Interestingly, the MsCO at doses 25 (p<0.05), 50 (p<0.01), 100 (p<0.01) and 200 mg/kg (p<0.01), doses showed significant protection in both MES and PTZ models; however, there was no protection in the NMDA-induced mortality test. Based on these outcomes, the MsCO (25, 50, 100 and 200 mg/kg) was further evaluated in the PTZ model to confirm the involvement of GABA pathways, the levels of GABA in the cortex and hippocampus were measured, and also to confirm the GABA receptor mediated action, the anti-PTZ action of MsCO (100 mg/kg) was evaluated in presence of flumazenil, which is a GABAA receptor antagonist. In the outcomes, the MsCO (100 and 200 mg/kg) administration showed a significant increase in GABA levels in both cortex (p<0.01) and hippocampus (p<0.01), compared to PTZ control. Also, the anticonvulsant action of MsCO was blocked in the presence of flumazenil, thus confirmed the possible role of GABA mechanism. Moreover, actophotometer test revealed that MsCO (25, 50, 100 and 200 mg/kg) is free from locomotor deficit-inducing potential. Thus, the results of the study indicate that MsCO possesses potent anticonvulsant activity through GABA mediated mechanism.

Antioxidant potential and chemical characterization of bioactive compounds from a medicinal plant Colebrokea oppositifolia Sm.

Colebrookea oppositifolia is a highly used medicinal plant and an enriched source of essential oils. Therefore, the present study was designed with the aim to extract the chemical constituents and to evaluate its antioxidant potential. Fresh plant parts were subjected to the extraction of volatile chemical constituents by maceration using n-hexane as the menstruum. The resulting n-hexane fractions were purified and then subjected to GC-MS and FTIR analysis. In-vitro antioxidant abilities were evaluated by, DPPH, total phenolic content (TPC), total flavonoid content (TFC) method against the standard solutions of (Gallic acid, Quercetin) as a positive control. The GC-MS analysis of leaves, stem and inflorescence showed a total of 100, 98 and 48 components out of which 47, 16 and 17 peaks were identified representing the 67.64 %, 73.16 % and 61.93 % of the total oily fractions, respectively. The FTIR spectrum indicated the presence of various functional groups. In-vitro antioxidant results exhibited that leaves showed the highest antioxidant potential by DPPH (3.365 ± 0.002), and the highest total phenolic content by FC method (203.00 ± 0.091). Foliar micromorphological features were found significant in the authentication of C. oppositifolia. Further pharmacognostic studies of this plant are recommended to evaluate its therapeutic potential.

Pharmacognostical, Phytochemical and Pharmacological profile of Colebrookea oppositifolia Smith

Colebrookea oppositifolia commonly known as ‘Bhaman’ is distributed throughout India from the Himalayas down to Deccan. The plant is used traditionally as such as dermatitis, dysentery, fever, headache, peptic ulcer, haemostatic, wounds, as anti-fertility agent, fungicide, and the roots of the plant has been most widely used for the treatment of epilepsy.. Medicinally, it has been proven to possess various pharmacological activities like treating corneal opacity or conjunctivitis, sore eyes due to its anti-inflammatory properties, cardioprotective, hepatoprotective, anti-inflammatory, antihelmintic, antifungal, antioxidant, antimicrobial, antinociceptive, cytotoxic activity, anticonvulsant, antiulcer, antimicrobial, anti-fertility, antipyretic and insecticide. Further, studies reveal the presence of various phytochemical constituents mainly flavone glycosides viz. chrysin, negletein, landenein; leaves contain 5,6,7- tri-methoxyflavone, 5,6,7,4'-tetramethoxyflavone, acteoside, and quercetin in the bark; root contains stearic, palmitic, oleic acids, triacontanol, flavone glycoside echioidin, 5,6,7-trimethoxyflavone and 4',5,6,7- tetra methoxy flavone; sugars and vitamins have also been isolated from this plant. These studies reveal that Colebrookea oppositifolia is a source of medicinally active compounds and have various pharmacological effects; hence, this drug encourage finding its new therapeutic uses.&#x0D; Keywords: Colebrookea oppositifolia, wound healing, anticonvulsant, Lamiaceae

Anticonvulsant Activity Guided Isolation of Acteoside from Methanolic Root Extracts of Colebrookea oppositifolia Smith

Present study was aimed to isolate the anticonvulsant principle/s from methanolic root extract of Colebrookea oppositifolia (MCO). The MCO was subjected for fractionation, and column chromatography using various solvents, and the fractions obtained were evaluated using Six-hertz seizure test. The potent fraction obtained was subjected for detailed spectral analysis to identify the possible chemical structure. Initially, the MCO was made into 3 three major fractions, namely n-butanol soluble fraction, ethyl acetate soluble fraction and residue fraction. The n-butanol fraction has shown better anticonvulsant activity; therefore, it was subjected for column chromatography. A total of 72 fractions (Fr-1 to Fr-72) of 100 ml each were collected and the fractions with similar TLC pattern were pooled together to form 11 pooled fractions (Fr-I (1-5), Fr-II (6-10), Fr-III (11-21), Fr-IV (22-29), Fr-V (30-36), Fr-VI (37-44), Fr-VII (45-52), Fr-VIII (53), Fr-IX (54-60), Fr-X (61-66), Fr-XI (67-72)). In six-hertz seizure test, the Fr-X (61 to 66) was identified as a most potent fraction, and upon crystallization from methanol/chloroform, yielded a light brown colored amorphous powder (COP-I - 1.6 g) soluble in water. Furthermore, the COP-I was subjected for detailed spectral analysis (UV, FT-IR, 1H-NMR, 13C-NMR and Mass), and based on the spectral analysis, it was identified as Acteoside (Verbascoside), belongs to phenylethanoid class of glycosides. Based on bioactivity guided isolation process, COP-I was isolated from MCO, and based on detailed spectral analysis the COP-I was identified as Acteoside.

Foliar epidermal anatomy of Lamiaceae with special emphasis on their trichomes diversity using scanning electron microscopy.

Foliar epidermal features were based on the micromorphology of trichomes types, epidermal cells and stomatal complex. Even though each feature has its own limited taxonomic value but collectively these characteristics may be systematically important especially for the discrimination and identification of complex and problematic taxa. The systematics significance of nonglandular (NGTs) and glandular trichomes (GTs), stomatal complex and epidermal cells of Lamiaceous flora were analyzed by using the light microscopy (LM) and scanning electron microscopy (SEM). Variations on the observed epidermal appendages were divided into two basic types: glandular and nonglandular. GTs can be divided into subtypes: sessile capitate, subsessile capitate, and barrel and sunken. NGTs were also divided into subtypes: dendritic, stellate, conical, falcate, simple and 1-6 cells long having granulate and smooth surface ornamentation. NGTs were the most dominant features of both adaxial and abaxial surfaces of all observed taxa. Vitex negundo, Isodon rugosus, Colebrookea oppositifolia, and Marrubium vulgare could be demarked because of their twisted like appearance of NGTs at the abaxial surface. The Lamiaceae had both hypostomatic and amphistomatic leaf. Stomata were observed as diacytic, anisocytic, and anomocytic. Epidermal cells were found to be irregular, isodiametric, and rectangular. Based on these characters a taxonomic key was developed to delimit the closely related taxa. Distribution and morphology of the foliar epidermal trichomes through SEM highlight an important taxonomic tool used by the taxonomists as an aid to the correct identification of problematic Lamiaceae taxa.

Biopesticidal potential of green chemicals against Callosobruchus analis (f.) (Coleoptera: Bruchidae):

Pulses have 20-27% proteins which is 2- 3 times higher than traditional cereals. These constitute the main source of proteins for developing countries, like India where per capita consumption of the animal protein is low, thus they are rightly considered the poor man’s meat. India is largest pulse consumer, importer and producer country of the world occupying an area of 228.47 lakh hectares with the production of 17380 million tones every year. With the United Nations declaration of 2016 as International Year of Pulses to replace the social evil of malnutrition by legume, the research pertaining to the biology and bio intensive management of bruchids pests has become increasingly important. Therefore, laboratory bioassay of essential oils which are regarded as “Green Chemicals” extracted from Zanthoxylum armatum DC., Rabdosia rugosa Wall. ex Benth, Artemisia maritima Linn. and Colebrookea oppositifolia Sm. by hydro distillation was carried out against Callosobruchus analis (F.) to evaluate biopesticidal potential in terms of oviposition and progeny deterrence and ovicidal activities. There was a significant difference in the number of eggs laid on treated and control sets and among the different treatments of essential oils. Z. armatum at 100 µl/ml allowed the bruchid to lay only 19.15±3.6 eggs as compared to 82.35±4.5 in control and proved to be most effective treatment with 76.74% oviposition deterrence. R. rugosa and A. maritima oil were found most effective in reducing the egg hatchability to 48.00±3.2 and 49.52±2.2% respectively at a lowest dose of 10 µl/ml. Egg hatching inhibition percentage increased with an increase in concentration of all the treatments. R. rugosa oil at 100 µl/ml proved to be most effective in reducing the adult emergence with 85.48% progeny deterrence followed by A. maritima showing 81.67% deterrence. All the tested essential oils revealed a wide range of bioactivities against the bruchid pest.

Larvicidal effects of eight essential oils against Plodia interpunctella and T. castaneum, serious pests of stored products worldwide

Fumigant toxicity of eight essential oils obtained by hydrodistillation from eight plant species viz. Artemisia maritima L., Cinnamomum zeylanicum Blume, Citrus hystrix DC., Colebrookea oppositifolia Sm., Pelargonium hortorum Bailey, Rabdosia rugosa Wall. ex Benth, Thuja occidentalis L. and Zanthoxylum armatum DC. were tested against different immature stages of Tribolium castaneum (Herbst) and Plodia interpunctella (Hubner) both are serious pests of stored products worldwide. 8-10 day and 18-20 day old larvae of P. interpunctella and T. castaneum were used for the bioassay. R. rugosa oil gave 80.03±5.6% mortality at 100 µl/ml followed by 72.28±4.9% at 50 µl/ml against 8-10 day old larvae. Whereas 65.47±2.9% mortality at 100 µl/ml followed by 62.09±1.8% mortality at a dose of 50 µl/ml was obtained against 18-20 day old larvae of T. castaneum after 120 hrs. At a lowest concentration of 10 µl/ml R. rugosa oil produced 53.8.33±2.2 % mortality for 8-10 day old larvae and a mortality of 40.55±3.9% respectively was obtained for 18-20 day old larvae of P. interpunctella after 120 hrs. Among eight essential oils tested R. rugosa had the highest toxicity followed by A. maritima, C. zeylanicum, Z. armatum, T. occidentalis, P. hortorum, C. oppositifolia and C. hystrix against both the insect pests. The results obtained also showed that tolerance increases as the immature stages grow older and 18-20 day old larvae were least susceptible to all the treatments as compared to 8-10 days old larvae. Larvae of T. castaneum were found to be most susceptible for all treatments than P. interpunctella.

Chemical characterization and cerebroprotective effect of methanolic root extract of Colebrookea oppositifolia in rats

Ethnopharmacological relevanceColebrookea oppositifolia Smith is one of the extensively used plants to treat neurological conditions such as epilepsy by the various ethnic communities in sub-Himalayan regions of India such as Bhoxa, Tharu and nomadic Gujjars. Aim of the studyThis study was conducted to evaluate the cerebroprotective effect of C. oppositifolia methanolic root (MeCO) extract in Wistar rats. Materials and methodsThe MeCO was characterized for total phenolic content and later subjected for detailed liquid chromatography–mass spectrometry analysis. Further, it was evaluated for in vitro antioxidant activity using 2, 2-diphenyl-1-picrylhydrazyl, ferric reducing antioxidant power and oxygen radical absorbance capacity assays. In addition, the MeCO was investigated on generation of ROS, nitrite, and TNF-α in LPS-stimulated RAW 264.7 cell lines. Finally, the cerebroprotective effect of MeCO was examined against global ischemia and reperfusion (I/R)–induced brain injury in Wistar rats. Male Wistar rats were allocated in to five groups (G-I to G-V, n = 10). G-I and G-II served as sham control and I/R control, respectively, and received only vehicle (0.5% w/v carboxy methyl cellulose, 10 ml/kg, p.o.). G-III served as reference standard and received quercetin (20 mg/kg, p.o.). G-IV and G-V animals received 200 and 400 mg/kg oral doses of MeCO, respectively. All the treatments were given for a period of seven days and the parameters such as neurobehavioral (neurological, and cognitive), and motor functions, biochemical (enzymatic and non-enzymatic antioxidants, TNF-α, IL-6, IL-10, ICAM-I), morphological (cerebral edema and infarct area) and histopathological evaluations were performed. ResultsThe MeCO showed a total phenolic content of 137.28 mg gallic acid equivalents/g, and LC-MS/MS analysis of MeCO showed presence of acteoside, gossypin, quercetin and ferulic acid as major ingredients (6680.3, 1.55, 3.52 and 431.1 ng/mg). In in vitro antioxidant assays, the MeCO exhibited potent activity with IC50 of 49.10 µg/ml in DPPH assay; FRAP and ORAC values of 1180.5 and 2983.5 respectively. Furthermore, the MeCO significantly inhibited generation of ROS, nitrite and TNF-α in LPS-stimulated RAW 264.7 cell lines. Sixty min of global ischemia with 24 h reperfusion produced substantial alterations in neurobehavioral functions in the I/R control group compared to sham control. In addition, a significant reduction in catalase and superoxide dismutase activities was observed. Moreover, lipid peroxidation increased and reduced glutathione levels decreased significantly. Furthermore, the levels of pro-inflammatory cytokines (TNF-α, IL-6, and ICAM-I) increased significantly and those of anti-inflammatory (IL-10) decreased. I/R insult increased the brain volume and aggravated cerebral infarct formation. Histopathological examination of the rat brain revealed vascular congestion, cerebral edema, leukocyte infiltration, and brain tissue necrosis. Interestingly, seven days pretreatment with MeCO (200 and 400 mg/kg) alleviated all the I/R-induced perturbances (neurobehavioral, and motor functions, biochemical, morphological and histopathological) compared with the I/R control. ConclusionsThe MeCO exhibit potent cerebroprotective activity through its potent antioxidant and anti-inflammatory mechanisms, and hence may be useful in the management of ischemic stroke and associated complications.