Bioassay Guided Fractionation of Senna singueana and Its Potential for Development of Poultry Phytogenic Feed Additives

There has been burgeoning interest in plant-based feed additives following restrictions placed on the use of antibiotic feed additives in many countries. Phytogenic feed additives are recommended to have a range of useful properties to support the growth and development of poultry to a similar level as that obtained by supplementing feed with antibiotics. The aim of this study was to evaluate the antibacterial, anti-lipoxygenase and antioxidant activity, and in vitro safety of fractions and isolated compounds from leaves of Senna singueana. Antibacterial activities of the fractions and isolated compounds were determined against a panel of bacteria using a two-fold serial microdilution assay and qualitative bioautography assays. Anti-lipoxygenase activity was evaluated using the ferrous oxidation-xylenol orange (FOX) method. Antioxidant activity was assessed qualitatively and quantitatively using radical scavenging assays. Dichloromethane and ethyl acetate fractions from solvent-solvent partitioning had the best antibacterial activity with MIC values ranging from 156 to 313 μg/ml. Fractions obtained from column chromatography had significant to weak antibacterial activity with MIC values ranging from 50 to 1,250 μg/ml. Bioautography showed clear bands of bacterial inhibition, indicating the presence of a number of active compounds in several fractions. The ethyl acetate fraction and all the tested column fractions had potent anti-lipoxygenase activity with IC50 values of ≤2.5 μg/ml which were lower than that of quercetin (positive control), indicating anti-inflammatory potential. The ethyl acetate fraction and several column fractions had powerful antioxidant activity with IC50 values of ≤5 μg/ml in the ABTS assay. Cytotoxicity values against Vero kidney cells ranged from LC50 = 40.0–989.3 μg/ml. Bioassay-guided fractionation led to the isolation and identification of a known bioactive compound, luteolin. S. singueana is a promising candidate for the development of poultry phytogenic feed additives.

Molecular docking study on columbin isolated from Tinospora cordifolia as a cholinesterase inhibitor

Purpose: To investigate the acetylcholinesterase (AChE) inhibitory potential of columbin and also to assess its binding affinity against AChE protein. Methods: Crystals of columbin were isolated from the ethyl acetate fraction of Tinospora cordifolia using column chromatography and its structure was determined using x-ray crystallography. Ellman colorimetric assay was used to determine the AChE inhibitory effect in vitro while molecular docking was performed using the MOE 2015.010 software. The selected protein data bank (PDB) was modeled using PDB ID: 10CE (pacific electric ray). Results: The crystal and structure refinement data of columbin were: C20H22O6, Orthorhombic, P212121, a = 7.4951(2) Å (α = 90°), b = 11.6451(3) Å (β = 90°), c = 19.5882(5) Å (γ = 90°), V=1709.68(8) Å3, Z = 4, Density (calculated) = 1.392 Mg/m3, absorption coefficient = 0.851 mm-1, goodness-of-fit on F2 =1.091, T = 100(2) K. Columbin demonstrated good AChE inhibitory effect with half-maximal inhibitory concentration (IC50) of 1.2993 ± 0.17 mg/mL. Molecular docking data revealed that it exhibited hydrophobic and hydrogen bonding interactions with the surrounding residues, and this accelerated complexation between the ligands and the active site of the enzyme. Conclusion: Columbin may be useful in the management of neurodegenerative conditions such as Alzheimer’s disease.

Cirsium japonicum var. Maackii Improves Cognitive Impairment under Amyloid Beta25-35-Induced Alzheimer’s Disease Model

Abnormal production and degradation of amyloid beta (Aβ) in the brain lead to oxidative stress and cognitive impairment in Alzheimer’s disease (AD). Cirsium japonicum var. maackii (CJM) is widely used as an herbal medicine and has antibacterial and anti-inflammatory properties. This study focused on the protective effect of the ethyl acetate fraction from CJM (ECJM) on Aβ25-35-induced control mice. In the T-maze and novel object recognition test, ECJM provided higher spatial memory and object recognition compared to Aβ25-35 treatment alone. In the Morris water maze test, ECJM-administered mice showed greater learning and memory abilities than Aβ25-35-induced control mice. Additionally, ECJM-administered mice experienced inhibited lipid peroxidation and nitric oxide production in a dose-dependent manner. The present study indicates that ECJM improves cognitive impairment by inhibiting oxidative stress in Aβ25-35-induced mice. Therefore, CJM may be useful for the treatment of AD and may be a potential material for functional foods.

A comparative study on the antioxidant activity of Gladiolus dalenii Van Geel and nine commonly used substances to compare the antioxidant activity of foods and medicinal plants

Reactive species, such as the free radicals produced during cell metabolism, are described as the main cause of oxidative stress, a process responsible for the development of degenerative diseases. Thus, the investigation of natural products containing chemical components with antioxidant capacity becomes necessary, since the frequent ingestion of these products may prevent the occurrence of this adverse event. In this perspective, total phenols (<b>TPC</b>) and total flavonoids (<b>TFC</b>) of the crude methanolic extract (<b>MCE</b>) and ethyl acetate fraction (<b>EAF</b>) obtained from <i>Gladiolus dalenii</i> bulbs were quantified and their antioxidant capacity evaluated and compared with that of gallic acid (<b>GA</b>), tannic acid (<b>TA</b>), pyrogallol (<b>PyG</b>), n-propyl gallate (<b>nPG</b>), quercetin (<b>Qtn</b>), rutin (<b>Rut</b>), butylated hydroxytoluene (<b>BHT</b>), 6-hydroxy-2,5,7-tetramethyl-chroman-2-carboxylic acid (<b>Trolox</b>) and ascorbic acid (<b>Asc)</b> using 1,1-diphenyl-2-picryl hydroxyl (DPPH) free radical scavenging method. The study revealed that the antioxidant activity of <b>MCE</b> (EC₅₀=6.550 ± 0.31 µg/mL) and <b>EAF</b> (EC₅₀=5.960 ± 0.61 µg/mL) was higher effect than <b>Rut</b> (EC₅₀=9.110 ± 0.04 µg/mL) and <b>BHT</b> (EC₅₀=11.18 ± 0.03 µg/mL), and in turn lower than that of the other substances analyzed. The antioxidant activity found for <i>G. dalenii</i> extracts may be due to the high level of TPC found in this species.

Microscopic Identification and Determination of Total Flavonoid Content of Moringa Leaves Extract and Ethyl Acetate Fraction (Moringa oleifera L.)

This research was conducted to identify simplicia microscopically, phytochemical screening and determination of total flavonoid content of extract and ethyl acetate fraction from Moringa (Moringa oleifera L.) leaves using UV-Visible Spectrophotometry method. The experimental design used in this study was to perform microscopic identification of Moringa leaf powder simplicia, make 96% and 70% ethanol extract and ethyl acetate fraction of Moringa leaves from 70% ethanol extract, then carry out phytochemical screening and determination of total flavonoid content with quercetin standards. Phytochemical screening on the ethyl acetate fraction of Moringa leaves included tests for the content of flavonoids, saponins, tannins and alkaloids. The results of microscopic identification of Moringa leaf simplicia showed the presence of calcium oxalate crystals in the form of rosettes, mesophyll and stomata. The result of determination of total flavonoid content in 96% ethanol extract was 16.69 ± 0.74% (w/w), 70% ethanol extract was 10.84 ± 0.49% (w/w), Moringa leaf ethyl acetate fraction 14 .45 ± 0.90% (w/w). The highest total flavonoid content was found in the 96% ethanol extract of Moringa leaves in accordance with the 2017 Indonesian Herbal Pharmacopoeia, that the thick extract of Moringa leaves containing no less than 6.30% (w/w) total flavonoids was calculated as quercetin.

Antinociceptive and anti-inflammatory activities of Hymenaea martiana Hayne (Fabaceae) in mice

Hymenaea martiana is a species popularly known in Northeastern Brazil as “jatobá” and used in folk medicine to treat pain and inflammation. The aim of this work was to evaluate the antinociceptive and anti-inflammatory activity of H. martiana. In the present study, we carried out an investigation about the effects of the crude ethanolic extract (Hm-EtOH) and the ethyl acetate fraction (Hm-AcOEt) in models of nociception and inflammation in mice. Chemical (acetic acid-induced writhing and formalin) and thermal stimuli (hot plate) were used for the evaluation of antinociceptive activity, while for the anti-inflammatory profile paw edema induced by carrageenan was used, along with leukocyte migration to the peritoneal cavity. The presence of the flavonoid astilbin in the samples was characterized through HPLC-DAD-MS analysis. Hm-EtOH and Hm-AcOEt (100, 200 and 400 mg.kg-1, i.p.) significantly reduced the number of abdominal contortions and decreased the paw licking time in the formalin test. In the hot plate, the extract increased the latency time of animals. Hm-EtOH and Hm-AcOEt inhibited significantly the increase in the edema after the administration of carrageenan. Hm-EtOH and Hm-AcOEt inhibited leukocyte migration in the peritonitis test. HPLC-DAD-MS analysis of Hm-EtOH and Hm-AcOEt revealed the presence of the flavonoid astilbin in the samples. According to the results of this study, both Hm-EtOH and Hm-AcOEt have antinociceptive and anti-inflammatory activities, which could be related with the presence of flavonoid in the extracts. The results reinforce the popular use of this plant.

Immunomodulatory effect of petroleum ether extract and ethyl acetate fraction of bengkoang (Pachyrhizus erosus (L.) Urban) in vitro

Bengkoang (Pachyrhizus erosus (L.) Urban) contains phytosterol and the isoflavone daidzein, which are thought to have immunomodulatory activity. There have been no studies reporting on the immunomodulatory effects of bengkoang extract containing polar and semi-polar compounds, such as phytosterols and isoflavone-like compounds. The objective of this study was to evaluate the immunomodulatory effects of bengkoang extracts, including petroleum ether extract (PEE), methanol extract (ME), and the ethyl acetate fraction (EAF) of bengkoang, in vitro. The immunomodulatory effects of PEE, ME, and EAF of bengkoang were determined according to the phagocytic activity of macrophages based on phagocytosis of latex beads, lymphocyte proliferation, and detection of cytokine production of tumor necrosis factor-α (TNF-α) interleukin-6 (IL-6), and interleukin-10 (IL-10) levels. Results: The phagocytic index and phagocytic capacity of ME, PEE, and EAF of bengkoang on macrophage cells were significantly increased (p < 0.05), whereas lymphocyte proliferation was unchanged compared with the control (p > 0.05), and ME of bengkoang enhanced the levels of the cytokines TNF-α and IL-6. In contrast, PEE and EAF of bengkoang decreased TNF-α and IL-6 levels compared with the control group. All of the bengkoang extracts decrease the production of the anti-inflammatory cytokine IL-10. In conclusion, this study showed that PEE, ME, and EAF of bengkoang could increase the non-specific immune response (phagocytic activity) but had a lesser effect on the specific immune response (lymphocyte proliferation). The ME of bengkoang acts as an immunostimulant by increasing the levels of the inflammatory cytokines TNF-α and IL-6 and decreasing those of the anti-inflammatory cytokine IL-10.