Phyllanthus muellerianus and Ficus exasperata exhibit anti-proliferative and pro-apoptotic activities in human prostate cancer PC-3 cells by modulating calcium influx and activating caspases

Purpose Phyllanthus muellerianus (PM) and Ficus exasperata (FE) are plants used against cancers. We evaluated the phytochemical profiles and in vitro antioxidant potentials of PM and FE, and investigate their effects on cell proliferation, intracellular calcium ([Ca2+]i), caspases 3/9, apoptosis, oxidative stress markers, and Bax/cytochrome C expression in PC-3 cells. Methods The phytochemical profiles were evaluated by liquid chromatography-mass spectrometry (LC-MS), and the antioxidant by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals scavenging method. The cells were incubated for 24 hours with 3% tween 80, paclitaxel (5 nM), PM (800 and 1200 µg/ml), and FE (800 and 1200 µg/ml). After treatments, [Ca2+]i, caspases 3/9, apoptosis and oxidative stress parameters were measured using colorimetric kits, while the mRNA levels of Bax and cytochrome C were quantified by RT‐qPCR. Results Nitidine, phloridzin and linoleic acid were identified in PM, while docosane, cardanol and chlorogenic acid were revealed in FE. The in vitro antioxidant potential of PM was greater than that of FE. Both plants inhibited the growth of PC-3 cells in a dose-dependent manner, but significantly (p<0.5-0.001) increased [Ca2+]i, apoptosis level, caspase 3/9 activities, reactive oxygen species production and lipid peroxidation, compared with control. Moreover, the activities of superoxide dismutase, catalase and glutathione peroxidase were significantly decreased in the cells incubated with the plant extracts, PM being the most effective. Paclitaxel, PM and FE upregulated Bax and cytochrome C genes in PC-3 cells. Conclusion PM and FE inhibited the growth of PC-3 cells by modulating the [Ca2+]i and inducing apoptosis through Bax/Cytochrome C/Caspase 3-9 signaling pathway.

Effects of Different Extracting Solvents on Non-Phenolic Phytochemical Profiles of Selected Nigerian Spices and Spice-treated Foods

The aim of this research was to investigate the impact of extraction solvents on the non-phenolic phytochemical profiles of selected spices (Ocimum viride, Monodora myristica, Monodora tenuifolia and Tetrapleura tetraptera) and spice-treated foods in southern part of Nigeria. The spice samples were processed into powder for antioxidant screening. The spice extracts were obtained from the samples using 5 extracting solvents [distilled water, 95 % methanol, acetone / hexane (1:1 v/v), n-hexane / methanol / acetone (2:1:1, v/v/v) and acetone / water / acetic acid (70:29.5:0.5, v/v/v)]. Water extracts were obtained from beef, pork and fluted pumpkin leaves. The alkaloid, saponin, oxalate and phytate components of the spice extracts and the spice-treated foods were evaluated using standard methods. The laboratory analyses were performed at analytical laboratory, National Centre for Energy Research and Development, University of Nigeria, Nsukka during the 3rd quarter of 2020. The percentage yield of the extracts were low (0.32 - 0.96 %) and varied widely among extracting solvents, spices and spice-treated foods. M. myristica and T. tetrpleura had the highest yield, 0.96, in methanol extracts. Phytochemical contents differed significantly (p < 0.05) among spices, extracts of the same spice and among spice-treated foods. Spices had high contents of oxalate (2.0 – 7.0 mg/100 g), alkaloid (0.8 – 5.76 mg/100 g) and phytate (2.14 – 3.88 mg/100 g) but relatively low content of saponin (0.03 – 0.736 mg/100g). Methanol alone or in combination with other solvents extracted higher amounts of phytochemicals (0.96 %) than other solvent mixtures from the spices. Phytochemical contents of spice-treated foods were in the order: vegetable > rice > pork > beef.

A Review of Ethnoveterinary Knowledge, Biological Activities and Secondary Metabolites of Medicinal Woody Plants Used for Managing Animal Health in South Africa

Globally, the use of ethnoveterinary medicine as remedies for animal health among different ethnic groups justify the need for a systematic exploration to enhance their potential. In addition, the increasing popularity and utilisation of woody plants remain common in traditional medicine, which may be attributed to their inherent benefits. The current review was aimed at analysing ethnoveterinary surveys, biological activities, and secondary metabolites/phytochemical profiles of the woody plants of South Africa. Eligible literature (period: 2000 to 2020) were retrieved from different databases such as Google Scholar, PubMed, Sabinet, and Science Direct. Based on the inclusion and exclusion criteria, 20 ethnoveterinary surveys were eligible and were subjected to further analysis. We identified 104 woody plant species from 44 plant families that are used in the treatment of different diseases in animals, particularly cattle (70%) and goats (20%). The most mentioned (with six citations) woody plants were Terminalia sericea Burch. ex DC and Ziziphus mucronata Willd., which were followed by plants with five (Cussonia spicata Thunb., Pterocarpus angolensis DC and Vachellia karroo (Hayne) Banfi & Galasso) or four (Acokanthera oppositifolia (Lam.) Codd, Cassia abbreviata Oliv., and Strychnos henningsii Gilg) individual mentions. The most dominant families were Fabaceae (19%), Apocynaceae (5.8%), Rubiaceae (5.8%), Anacardiaceae (4.8%), Combretaceae (4.8%), Euphorbiaceae (4.8%), Malvaceae (4.8%), Rhamnaceae (4.8%), and Celastraceae (3.8%). Bark (33%), leaves (29%), and roots (19%) were the plant parts dominantly used to prepare remedies for ethnoveterinary medicine. An estimated 20% of woody plants have been screened for antimicrobial, anthelmintic, antioxidant, and cytotoxicity effects. Phytochemical profiles established a rich pool of valuable secondary metabolites (phenolic, flavonoids and condensed tannins) that may be responsible for the exerted biological activities. Overall, the significant portion of woody plants lacking empirical evidence on their biological effects indicates a major knowledge gap that requires more research efforts.

151 Intake, Growth, and Insectifugal Outcomes Associated with Free-choice Supplementation of Garlic Products to Feedlot Steers

Conflicting anecdotal accounts from producers about their cattle performance and fly-repellence outcomes while receiving garlic-infused supplements have limited feeding decisions. This study evaluated differences in animal performance and insectifugal outcomes among four groups (26 steers per group) over two years (total = 208 steers). The steers either received non-garlic mineral supplement (MS), MS+0.3% garlic oil-based premix (GOil), MS+2.5% garlic powder (LGP), or MS+5% garlic powder (HGP). A completely randomized design was implemented where each steer was assigned to each treatment group while balancing for bodyweight and breed composition. Steers received a feedlot ration and free-choice mineral supplement for 86 and 108 d in the first and second year, respectively. Individual feed and supplement consumption were measured with automatic feeding units. Fly abundance on each animal was determined using digital images collected during weekly fly surveys. The ADG, feed efficiency, ultrasound body composition, and intake-related activities were also evaluated during the study. Data were analyzed using linear- or generalized linear mixed models in SAS. A significant year and Group×year interactions were observed for daily supplement intake (P &lt; 0.02). Similar interactions (P &lt; 0.04) were observed for feeding behaviors associated with supplement intake. Fly abundance during the peak months was below the economic threshold and was not different among the groups (P = 0.25), but the year and Group×year interactions were significant (P &lt; 0.02). There were no differences among the groups for DMI (P = 0.81), ADG (P = 0.54) and gain to feed ratio (P = 0.34). The Group×year interactions were present for the change in ultrasound rib-fat-thickness. The reasons behind the differences in supplement intake are not clear but may be related to the garlic products’ phytochemical profiles. These results suggest that further studies are required to identify the specific bioactive compounds in garlic that affect supplement intake.

Nutritional and phytochemical profiles of common pepper (Capsicum spp.) foliage consumed as leafy vegetables in Southeast Nigeria

Pepper foliage is consumed as a secondary plant part and as a traditional leafy vegetable in some climes. The six commonly consumed pepper species namely Cayenne, Anaheim, Bell, Bird’s eye, Scotch Bonnet and Habanero were analysed for their nutrient and phytochemical contents on a fresh weight basis. Proximate composition, vitamins were evaluated while minerals were analysed by the dry ash extraction method. Phytochemicals namely alkaloids, saponins and flavonoids were analysed by gravimetry; tannin and phenols by spectrometry; oxalate by titrimetry and hydrogen cyanide were analysed by alkaline picrate assay. Results indicated a significant difference (p<0.05) in the nutrient and phytochemical contents of the pepper foliage. Moisture ranged between 86.90 to 88.6% while their energy densities ranged between 34.33 to 45.66 kcal/100 g. Anahiem had the highest crude protein while Scotch Bonnet had the highest carbohydrate contents of 3.50% and 2.70% respectively. Bird's eye, Bell and Scotch Bonnet had the highest ash, fat and crude fibre contents respectively. The order of concentration of vitamins in the pepper foliage was in this order: Vitamin C > Vitamin A > Vitamin E > Vitamin B2 > Vitamin B1 > Vitamin B3. Scotch Bonnet had the highest calcium and magnesium contents, Cayenne had the highest phosphorus, potassium and iron contents while Anahiem and Bell had the highest zinc and sodium contents respectively. The phytochemical profile indicated that oxalate content of the foliage ranged between 0.14 to 0.28 mg/100 g and this classifies them as low oxalate vegetables. Bird's eye had the highest hydrogen cyanide content (9.45 mg/100 g) while Cayenne had the highest phytate content (12.72%). Results indicated the presence of tannins, alkaloids, phytosterols, phenols, flavonoids and saponins. These phytochemicals in the pepper foliage can be beneficial for human wellbeing based on their health-promoting effects.

Phenological and Geographical Effects on Phenolic and Triterpenoid Content in Vaccinium vitis-idaea L. Leaves

Lingonberry leaves have been proposed as a potential raw material for nutraceutical products and functional food due to the richness of phenolic and triterpenic compounds. However, contents of these bioactive compounds tend to vary greatly with physiological, climatic, and edaphic conditions, resulting in lingonberry leaves’ nutritional-pharmaceutical quality changes. In this context, we examined the effects of seasonal and geographical factors on phenolic and triterpenoid contents in lingonberry leaves. Quantitative and qualitative differences between samples were determined using validated HPLC-PDA methods. A total of 43 bioactive compounds were found at a detectable level throughout the year in young and old lingonberry leaves, with the highest contents of most compounds observed in samples collected in autumn–first half of spring. This suggests the potential to exploit the continuous biosynthesis for a longer harvesting season. Considerable variations in phytochemical profiles of lingonberry leaves, obtained from 28 locations in Lithuania, were found. Correlation analyses revealed significant negative correlations between contents of particular constituents and sunshine duration, temperature, and precipitation, and positive correlation with air humidity, longitudes, and altitudes of collecting locations and macronutrients in soil. These results suggest that harsh weather is favorable for most identified compounds and it may be possible to achieve appropriate accumulation of secondary metabolites by adjusting edaphic conditions. Taken together, the accumulation of phenolics and triterpenoids in lingonberry leaves highly depends on phenological and geographical factors and the influence of both variables differ for the particular compounds due to different metabolic processes in response to stresses.