Ganoderma lucidum methyl ganoderate E extends lifespan and modulates aging-related indicators in Caenorhabditis elegans.

Methyl Ganoderate E (MGE) is a triterpenoid derived from Ganoderma lucidum (Reishi), an edible mushroom, commonly processed into food forms such as soups, drinks, culinary dishes, and supplements. MGE has been shown to inhibit 3T3-L1 murine adipocyte differentiation when combined with other G. lucidum triterpenes. However, the specific effect of MGE on biological processes remains unknown. In this study, we present the first evidence of MGE's anti-aging effect in Caenorhabditis elegans. Through our screening process using the UPRER regulation ability, we evaluated a library of 74 pure compounds isolated from G. lucidum, and MGE exhibited the most promising results. Subsequent experiments demonstrated that MGE extended the lifespan by 26% at 10 µg ml-1 through daf-16, hsf-1, and skn-1-dependent pathways. MGE also enhanced resistance to various molecular stressors, improved healthspan, increased fertility, and reduced the aggregation of alpha-synuclein and amyloid-beta. Transcriptome data revealed that MGE promoted processes associated with proteolysis and neural activity, while not promoting cell death processes. Collectively, our findings suggest that G. lucidum MGE could be considered as a potential anti-aging intervention, adding to the growing list of such interventions.

Quantitative determination of selected heavy metals and micronutrients in branded dairy products sold in Nigeria using atomic absorption spectroscopy (AAS)

Quantitative analysis of heavy metals and nutrients in food helps indicate the safety and quality of food for final consumers. The present study was conducted to assess the presence of heavy metals (arsenic, copper, mercury, chromium, and lead) and the nutritional value of calcium in branded milk and yogurt to evaluate health risks for consumers. Ten (10) samples of branded milk and dairy products manufactured in Nigeria were purchased. The metal contents of the samples were determined using atomic absorption spectroscopy. The concentrations of calcium in the milk samples were between 9.33 ± 0.0023 and 18 ± 0.0071 ppm and were detected in all samples. Arsenic concentrations ranged from 0.45 ± 0.00042 to 2.48 ± 0.00064 ppm in eight branded samples but were undetected in two samples. Chromium levels were undetected in most samples, except for two with concentrations of 0.12±0.00049 ppm and 0.23±0.00021 ppm, respectively. Copper ranged from 0.032±0.00021 ppm to 0.129±0.00021 ppm in six samples. Mercury levels were detected in six samples at a concentration of 1.0±1.0 ppm. Lead concentrations ranged from 0.15±0.00064 to 0.29±0.00028 ppm in three samples. The study found heavy metals above the ideal concentration in branded milk and dairy products in Nigeria, highlighting the need for quality control measures during production to prevent contamination.

Bioactive Phytochemicals with Anti-Aging and Lifespan Extending Potentials in Caenorhabditis elegans.

In the forms of either herbs or functional foods, plants and their products have attracted medicinal, culinary, and nutraceutical applications due to their abundance in bioactive phytochemicals. Human beings and other animals have employed those bioactive phytochemicals to improve health quality based on their broad potentials as antioxidant, anti-microbial, anti-carcinogenic, anti-inflammatory, neuroprotective, and anti-aging effects, amongst others. For the past decade and half, efforts to discover bioactive phytochemicals both in pure and crude forms have been intensified using the Caenorhabditis elegans aging model, in which various metabolic pathways in humans are highly conserved. In this review, we summarized the aging and longevity pathways that are common to C. elegans and humans and collated some of the bioactive phytochemicals with health benefits and lifespan extending effects that have been studied in C. elegans. This simple animal model is not only a perfect system for discovering bioactive compounds but is also a research shortcut for elucidating the amelioration mechanisms of aging risk factors and associated diseases.

Antiviral activity of Salidroside from the leaves of Nigerian mistletoe (Loranthus micranthus Linn) parasitic on Hevea brasiliensis against respiratory syncytial virus.

Isolated Salidroside from the leaves of Nigerian mistletoe (Loranthus micranthus Linn) parasitic on Hevea brasiliensis was evaluated for its antiviral activity against respiratory syncytial virus. Semi- preparative HPLC separation of the ethyl acetate fraction of the leave extract of Loranthus micranthus Linn parasitic on Hevea brasiliensis led to the isolation of a polyphenol. Using spectroscopic methods (1D and 2D NMR and mass spectroscopic data) as well as by comparison with literature data the structure of the compound was determined as 6-O-galloyl salidroside. The antiviral activity of the isolated compound was evaluated against the respiratory syncytial virus. The isolated Salidroside showed potent inhibition towards a recombinant straining respiratory syncytial virus with Inhibitory Concentration (IC 50) value of 10.3±1.50 µg/mL. The result indicates that Salidroside is an efficient antiviral agent against RSV infection and might be useful for the management of RSV pathogenesis.

Modulation of intracellular expression of IFNγ and IL-2 in culture of splenic T lymphocytes by some flavonoid glycosides of Alchornea floribunda.

Context Alchornea floribunda Müll. Arg. (Euphorbiaceae) leaves are widely used in ethnomedicine for the management of rheumatism, arthritis and toothache. Objective In this study, flavonoid glycosides isolated from Alchornea floribunda were screened for their effect on the intracellular expression of interferon-gamma (IFNγ) and interleukin-2 (IL-2) type-1 cytokines. Materials and methods Chromatographic purification of the ethyl acetate fraction of the methanol leaf extract led to the isolation of seven flavonoid glycosides (1-7). Their structures were elucidated by 1D and 2D nuclear magnetic resonance and mass spectrometry. Splenocytes were treated with graded concentrations of the compounds (6.25-25 µg/mL) and incubated for 24 h. Thereafter, their effect on the expression of IFNγ and IL-2 by CD4(+ )and CD8(+ )T-lymphocytes was evaluated using intracellular cytokine staining and FACS analysis. Results Compounds 1-7 (6.25-25 µg/mL) caused the up-regulation of activated CD8(+ )(57.85-72.45% versus 57.85% for untreated control) and, to a lesser extent, activated CD4(+ )(3.21-7.21% versus 2.75% for the untreated control) T-lymphocytes that were both largely interferon-gamma-releasing in treated mouse T lymphocytes relative to untreated control. FACS data analysis showed that stimulation with all the compounds increased the proportion of CD8(+)/IFNγ(+ )and CD4(+)/IFNγ(+ )T lymphocytes up to two-fold when compared with the cells in untreated control wells. Intracellular IL-2 secretion by treated T cells was not detected. Conclusion This recorded T-lymphocyte-specific immune-modulatory property may contribute to explain in part the dynamics associated with the ethnomedicine of Alchornea floribunda, and may find relevance as a necessary cellular immune response precursor to infection-associated disease management.

New neolignan glycoside and an unusual benzoyl malic acid derivative from Maytenus senegalensis leaves.

Further investigation of the methanol leaf extract of Maytenus senegalensis led to the isolation of six compounds, including mayselignoside (1) and an unusual benzoyl malic acid derivative, benzoyl R-(+)-malic acid (2). Two known lignan derivatives (+)-lyoniresinol (3) and (-)-isolariciresinol (4), a known neolignan derivative dihydrodehydrodiconiferyl alcohol (5) and the triterpenoid, ß-amyrin (6) were also isolated. The structures of these compounds were elucidated by a combination of 1D and 2D NMR and mass spectroscopy. All compounds were tested for cytotoxicity against mouse lymphoma cell line (L5178Y) and for antimicrobial activity against strains of bacteria and fungi. None of the compounds showed promising cytotoxic and/or antimicrobial activities.

Isolation and structure elucidaton of polyphenols from Loranthus micranthus Linn. parasitic on Hevea brasiliensis with antiinflammatory property.

The present study was carried out to evaluate the anti-inflammatory activities of polyphenols isolated from the leaves of mistletoe (Loranthus micranthus Linn.) parasitic on Hevea brasiliensis. The anti-inflammatory properties of the isolated compounds were evaluated on the basis of their ability to inhibit the production of nitric oxide (NO) and tumuor necrosis factor-α (TNF-α) in lipopolysaccharide (LPS) activated RAW 264.7 mouse macrophages. Semi-preparative HPLC separation of the ethyl acetate (EtOAc) and butanol (n-BuOH) fractions of the leaves of mistletoe (Loranthus micranthus Linn) parasitic on Hevea brasiliensis led to the isolation of four polyphenols: 3-O-(3,4,5-trimethoxybenzoyl)-(-)-epicatechin (TMECG) (1); (-)-epicatechin-3-O-(3″-O-methyl)-gallate (ECG3″Me) (2); rutin (3) and peltatoside (4). Compounds 1-4 were isolated for the first time from this plant while 1 was isolated for the first time in nature. These compounds (1-4) were readily identified by comparison of their spectroscopic data with those reported in the literature. The polyphenols proved to have anti-inflammatory activity as evidenced by the suppression of inducible nitric oxide (iNO) and cytokine (TNF-α) levels in the culture supernatant of lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophages. However, the study showed that the quercetin diglycosides showed stronger inhibition of proinflammatory mediators than the epicatechin derivates. These data provide evidence that polyphenolic compounds isolated from the mistletoe parasitic on Hevea brasiliensis may contribute to its anti-inflammatory properties by inhibiting the expression of inducible nitric oxide and proinflammatory cytokines such as tumour necrosis factor-α.

Analysis of constituents of the eastern Nigeria mistletoe, Loranthus micranthus linn revealed presence of new classes of osteogenic compounds.

ETHNOPHARMACOLOGICAL RELEVANCE: Mistletoe extracts (decoctions) are used traditionally in eastern Nigeria for the management of bone pain, post menopausal syndrome and diabetes amongst several other ailments. While scientific evidence supporting its folkloric use as an antidiabetic agent has been documented, the age-long practice of its use in treatment of post menopausal syndrome has not been scientifically validated. Postmenopausal osteoporosis accounts for one of the prevalent disease conditions in aging population globally. This situation is exacerbated by the lack of osteogenic therapy. In search for plants of Nigerian origin with osteogenic potential, we evaluated eastern Nigerian mistletoe, having ethnotraditional claims of anti-diabetic, anti-hypertensive and anti-cancer activities as well as preventive effect in various post-menopausal syndromes. MATERIALS AND METHODS: Methanolic extracts of mistletoe leaves harvested from three host tress - Kola acuminata (KM), Citrus spp (CM) and Garcinia kola (GKM) - were evaluated for osteoblast viability and osteogenic activities using primary rat calvaria culture. Lupeol (1) was isolated from the stem bark of Bombax ciba and its congener, dihydoxylupeol palmitate (2) in addition to three other compounds; 3-methoxy quercetin (3), 3,4,5-trimethoxy gallate (4), and friedelin (5) were isolated from the leaves of mistletoes species. Following their chemical characterization, the compounds were evaluated for osteogenic potential using validated models including alkaline phosphatase (ALP) assay, mineralization assay and expression of osteogenic genes - bone morphogenetic protein-2 (BMP2) and osteoblast transcription factor (RUNX2) - in primary calvarial cultures harvested from neonatal rats. Uterine estrogenicity of the extracts was tested in adult female Sprague Dawley rats. RESULTS: Methanol extracts of mistletoe from three hosts exhibited increase in ALP activity (a marker of osteoblast differentiation) at lower concentrations (0.2-0.8 µg/ml) and either no or inhibitory effect at higher concentrations (1.6 and 3.2 µg/ml). None of the extract had cytotoxicity to osteoblasts at the concentrations tested. Five compounds viz. 1 from Bombax ciba, and 2-5 were isolated from the mistletoe leaves. Out of these, 5 exhibited significant loss of osteoblast viability and hence it was not considered further. All four compounds exhibited stimulatory effects on osteoblast differentiation as assessed by ALP assay and determination of osteogenic gene expression. Compound 2 was relatively more potent than its precursor, compound 1 in stimulating BMP2 upregulation. KM did not show uterine estrogenicity. CONCLUSION: Methanolic extracts from the three mistletoes species possess in vitro osteogenic activity, and from these extracts three new classes of compounds have been found to promote osteoblast differentiation in vitro. In light of these findings, we propose that mistletoe species may be developed as safer alternative(s) in the management of diseases where lack of bone formation is the pathology.

Three hydroxylated lupeol-based triterpenoid esters isolated from the Eastern Nigeria mistletoe parasitic on Kola acuminata.

Column fractionation and purification of an n-hexane fraction led to the isolation of three lupeol-based triterpenoid esters from the leaves of the Eastern Nigeria mistletoe, Loranthus micranthus Linn parasitic on a local kola nut tree, Kola acuminata. These three compounds were adequately characterised using a combination of UV/visible, IR, NMR (¹³C-NMR and ¹H-NMR), DEPT, MS and two-dimensional correlation (H-H COSY, Hetero-nuclear Single Quantum Correlation (HSQC), HMBC, NOE and NOESY) studies as 7ß,15α-dihydroxyl-lup-20(29)-ene-3ß-esters of palmitic (I), stearic (II) and eicosanoic acids (III). The characterisation of other isolated compounds is ongoing. Remarkably, this is the first report of the existence of fatty acid esters of an unusual 7ß,15α-dihydroxylated lupeol in the Eastern Nigeria mistletoe. These isolated compounds might contribute in part to the numerous established bio-activities of the Eastern Nigeria mistletoes.

A novel sesquiterpene acid and an alkaloid from leaves of the Eastern Nigeria mistletoe, Loranthus micranthus with potent immunostimulatory activity on C57BL6 mice splenocytes and CD69 molecule.

CONTEXT: The Eastern Nigeria mistletoe, Loranthus micranthus Linn. (Loranthaceae), is used in the treatment of several diseases including immune-modifying diseases and thus there is a need to identify the immunoactive constituents. OBJECTIVE: This research isolated and characterized the immunoactive constituents in the Eastern Nigeria mistletoe. MATERIALS AND METHODS: Bioassay-guided fractionation was employed in the isolation and purification of the constituents. The characterized compounds were screened for immunostimulatory activities on isolated C57BL/6 mice splenocytes and early activation marker, CD69 at concentrations of 10, 25, and 100 µg/mL using flow cytometry techniques and compared to lipopolysaccharide (LPS; 10 µg/mL) and concanavalin A (ConA; 2 µg/mL) as standards. RESULTS: Two compounds, a novel sesquiterpene, 2, 3-dimethoxy-benzo [a, b] cyclopentenyl-3',3',5'-trimethyl pyran-4-carboxylic acid, and a known alkaloid, lupinine were isolated and characterized. The compounds (25 µg/mL) showed statistically significantly (p < 0.05) stimulatory activity on the splenocytes with values of 56.34 ± 0.26% and 69.84 ± 0.19%, respectively, compared to 7.58 ± 0.42% recorded for the unstimulated control. Similarly, the CD69 expression assay showed immunostimulation with statistically significant values (p < 0.05) of 2.31 ± 0.07% and 2.71 ± 0.03%, respectively, compared to 1.69 ± 0.05% recorded for the nonstimulated control. DISCUSSION: These data suggest that the isolated compounds possess immunomodifying abilities. In addition, the activation of the CD69 molecule is possibly one of its mechanisms of action. CONCLUSION: These compounds may be responsible in part, for the immunostimulatory activities already established for the Eastern Nigeria mistletoes.

Immunomodulatory activity of a lupane triterpenoid ester isolated from the eastern Nigeria mistletoe, Loranthus micranthus (Linn)

OBJECTIVE@#To provide further evidence for the ethnomedicinal use of the Eastern Nigeria mistletoe, Loranthus micranthus (L. micranthus), as an immunostimulant.@*METHODS@#Solvent fractions from the crude extract of the mistletoe plant was obtained and screened by the cell mediated delayed type hypersensitivity reaction (DTHR) model in mice. Then the immunomodulatory potentials of a major lupane triterpenoid ester isolated from an active hexane fraction of the Eastern Nigeria mistletoe was investigated. Three lupeol-based triterpenoid esters: 7β 15α-dihydroxyl-lup-20(29)-ene-3β-palmitate (I), 7β, 15α-dihydroxyl-lup-20(29)-ene-3β-stearate (II) and 7β, 15α-dihydroxyl-lup-20(29)-ene-3β-decadecanoate (III) were isolated from the plant leaves epiphyting on a local kola nut tree and were characterized. Compound 1 was subjected to cell proliferation studies using C57Bl/6 splenocytes at three dose levels (5, 25 and 100 μg/mL) in presence of controls. Furthermore, the effect of this compound on IL-8 receptor expression was evaluated at three doses (1, 5 and 10 μg/mL) using the real time polymerase chain reaction assay.@*RESULTS@#This triterpenoid ester produced some enhancement of the splenocytes at the tested doses but at doses higher than 5 μg/mL caused inhibition of the IL-8 receptor expression.@*CONCLUSIONS@#The present findings support the ethnomedicinal use of the Eastern Nigeria Mistletoe in the management of diseases affecting the immune system. The triterpenoid(s) have some immunomodulatory abilities on splenocytes and IL-8 receptors and may partly account for the overall immunomodulatory activity of this plant.

Comparative acute toxicities and immunomodulatory potentials of five Eastern Nigeria mistletoes.

ETHNOPHARMACOLOGICAL RELEVANCE: Traditionally, mistletoes of Eastern Nigeria origin, Loranthus micranthus Linn. have been used as immunostimulant for the management of certain diseases with high profile immune depleting potentials. This practice has remained till date without scientific validation. AIM OF STUDY: To obtain and validate evidence for or against its continued use as immunostimulant and afford data for further studies on this specie of mistletoe. The present work is an in vivo proof of ethnopharmacological concept of the age long immunomodulatory use of our local mistletoe. MATERIALS AND METHODS: Aqueous-methanol extracts of the plant leaves from five different host trees were evaluated for immunomodulatory activity using four in vivo models in mice or rats, namely; total and differential leukocyte count (TLC and DLC), the cellular mediated delayed-type hypersensitivity reaction (DTHR) test, the humoral mediated antibody titration (AT) test and the cyclophosphamide-induced myelosuppression (CIM) test at different dose levels (100, 200 and 400 or 50, 100 and 250 mg kg(-1); depending on model) against standard controls. Phytochemical and acute toxicity tests were equally carried out on all the extracts. RESULTS: Results obtained indicate that all the mistletoes contained the same phytochemical constituents, although in varying amounts. The mistletoes exhibited statistically significantly different (p < 0.05 or p < 0.001, ANOVA) immunomodulatory (up-regulatory) activities in the overall order of that from Kola acuminata > Citrus spp > Persia americana > Parkia biglobosa > Pentaclatra macrophylla. LD(50) values were generally greater than 5000 mg/kg. CONCLUSION: The present study confirms the Eastern Nigeria mistletoe as a potent and safe alternative or complementary medicine for the management of immunodeficiency diseases.