Phytochemical Studies on Two Unexplored Endemic Medicinal Plants of India, Barleria terminalis and Calacanthus grandiflorus

Barleria terminalis Nees and Calacanthus grandiflorus (Dalzell) Radlk. are endemic medicinal plants of the Western Ghats of India. The aim of the present research work was to investigate phytochemical profile, potent bioactives using RP-HPLC, LC-MS and GC-MS and to evaluate their bioactivities. Acetone was found to be the best extraction medium for separating phytochemicals. Similarly, acetone and methanol extracts exhibited potential antioxidant properties. Ethanol extract of B. terminalis stem showed potent acetylcholinesterase (AChE) (89.10 ± 0.26%) inhibitory activity. Inhibition of α-amylase (36.96 ± 2.96%) activity was observed the best in ethanol extract of B. terminalis leaves and α-glucosidase inhibitory activity (94.33 ± 0.73%) in ethanol extract of C. grandiflorus stem. RP-HPLC analysis confirmed the presence of several phenolic compounds (gallic acid, hydroxybenzoic acid, vanillic acid, chlorogenic acid and coumaric acid) and phenylethanoid glycoside (verbascoside). The highest phenolics content were observed in B. terminalis (GA (4.17 ± 0.002), HBA (3.88 ± 0.001), VA (4.54 ± 0.001), CHLA (0.55 ± 0.004) mg/g DW, respectively). Similarly, LC-MS and GC-MS revealed the presence of phenolics, glycosides, terpenes, steroids, fatty acids, etc. Moreover, positive correlation between studied phytochemicals and antioxidants was observed in principal component analysis. Based on the present investigation, we conclude that B. terminalis and C. grandiflorus can be further explored for their active principles particularly, phenylethanoid glycosides and iridoids and their use in drug industry for pharmaceutical purposes.

UJI AKTIVITAS ANTIBAKTERI EKSTRAK DAUN TEMBELEKAN (Lantana camara Linn) TERHADAP PERTUMBUHAN Staphylococcus aureus dan Escherichia coli

This study aimed to determine the effect of concentration ethanol extract from tembelakan leaf (Lantana camara Linn) on bacteria growth of Staphylococcus aureus and Escherichia coli. Extraction was done by maceration using ethanol 96% and then separated using rotary evaporator. Antibacterial activity test of the ethanol extract by Well agar diffusion method. Variation in crude extract saponin used in this study was 5%, 10%, 15%, 20%, 25% and positive controls were used for comparison with Amoxicilin and Chloramphenicole concentration of 25 µg/mL and DMSO as a negative control. The results of antibacterial activity test is indicated by the formation of growth inhibitory region S. aureus and E. coli. The result of growth inhibitory regions was analyzed by One way ANOVA. One way ANOVA test results indicate that there are effects of ethanol extract concentration of tembelekan leaf (L. camara Linn) against S. aureus and E. coli. Effective concentration of ethanol extract tembelekan leaf (L. camara Linn) when compared with positive control to inhibit the growth of S. aureus and E. coli is at 25% with a relatively strong antibacterial activity. Test with phytochemicals screening method which is showed that tembelekan leaf contains the flavanoid, saponins, and tannins compounds as antibacterial

EFEKTIVITAS ANALGETIK KRIM TIPE M/A EKSTRAK ETANOL DAUN RANDU (Ceibapentandra (L) Gaertn) PADA HEWAN UJI MENCIT JANTAN

Randu plant leaves (Ceiba pentandra (L) Gaertn) are commonly used in traditional medicine, one of which is an analgesic with secondary metabolites which are thought to be analgesics, namely flavonoids. The purpose of this study was to determine the analgesic effectiveness of the ethanol extract cream of randu leaves (Ceiba pentandra (L) Gaertn) in white male mice (Mus musculus). The research on the analgesic effectiveness of the ethanol extract cream of randum leaves used the hot plate method and used male white mice as the experimental animal media. The soles of the hind legs of the mice were heated over hot plated with a temperature of 550C then the test animals were given the treatment which was divided into 5 treatment groups, namely positive control (Conterpain cream), negative control (F0), F1 (10%), F2 (15%), F3 (20%) and 1 normal group. The response of mice was calculated every 30 minutes in 120 minutes. The data obtained were analyzed using the one-way ANOVA statistical test followed by the Duncan test with a confidence level of 99%. The results of the one-way ANOVA test showed that there were significant differences between treatment groups (0.00 <0.05). The results of the Duncan test showed that the group that provided the best analgesic effectiveness was F2 (15%) but not better than Positive Control.

The Antimicrobial Activity of Pechuel-Loeschea leubnitziae Leaf Extract and its Effect on The Expression Level of Methicillin Resistant Staphylococcus Aureus and Candida albicans Virulence-Associated Genes

The complete halt in the synthesis of new effective antimicrobial compounds is a global concern. Pathogenic microorganisms' virulence mechanisms seem to have a significant impact on their pathogenesis. The purpose of this study was to examine the antimicrobial activity of the ethanol and methanol fractions of Pechuel-Loeschea leubnitziae leaf extract, as well as its effect on the expression level of virulence-associated genes.The extract's fractions were evaluated for antimicrobial activity against Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Klebsiella pneumoniae (clinical), Methicillin-resistant Staphylococcus aureus, and Candida albicans ATCC 90029. The test organism's antibiogram pattern was determined. The extracts' attenuation effect on the target genes of the susceptible organisms was investigated employing relative quantification using RT-qPCR. The test organism's antibiogram pattern revealed that it was drug-resistant, intermediate, and sensitive. The extracts tested positive for antimicrobial activity against Methicillin-resistant Staphylococcus aureus and Candida albicans ATCC 90029, with zones of inhibition varying from 20.33 to 29 mm. The lowest recorded MIC value was 4.688 mg/ml, while the highest was 37.5 mg/ml. In contrast to the methanol extract, the ethanol extract had a cidal action at a lower dose. The ethanol extract's Sub-MIC (18.25 mg/ml) merely reduced the expression of the hly gene in MRSA. The MRSA virulence genes were not suppressed by the sub-MIC of methanol extract (18.25 mg/ml). Notably, the expression of als1, pbl1, and sap1 in Candida albicans ATCC 90029 was significantly attenuated when exposed to sub-MICs of ethanol extract (2,344 mg/ml) and methanol extract (9.375 mg/ml). Per the findings of this research, the leaves of P. leubnitziae could be a source of an effective antimicrobial agent in the therapy of MRSA/Candida-related disorders.

In Vitro Antibacterial Activity, Preliminary Phytochemical Screening Profile, and In Vivo Toxicity of Seven Traditional Medicinal Plants in Ethiopia

The threat of antibiotic-resistance calls for novel antibacterial agents. This study was aimed at screening medicinal plants for their antibacterial properties, phytochemical content and safety. Leaves of Allophylus abyssinicus (Hochst.) Radlk., Dicliptera laxata C.B.Clarke, Ligustrum vulgare L., Solanecio gigas (Vatke) c. Jeffrey and Gymnanthemum myrianthum (Hook.f.) H.Rob.; leaf and stem-bark of Olinia rochetiana A. Juss. and the seed of Cucurbita pepo L. were used. Chloroform and ethanol were used to extract G. myrianthum, D. laxata and O. rochetiana; ethyl acetate and methanol for the rest, and water for all. The extracts were tested against clinical/standard strains of Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi and Staphylococcus aureus by the agar-diffusion method. The minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) were determined. Acute toxicity to mice was checked and preliminary phytochemical screening was done. Thirteen extracts, out of 24, were active (inhibition zone >7 mm) at differing levels (9.67±0.33-25.66±0.57 mm) against at least one bacterial strain. The MICs and MBCs were 1.95-15.6 mg/mL and 7.8-125 mg/mL respectively. The aqueous extract of S. gigas, methanol extracts of L. vulgare and A. abyssinicus, and ethanol extract of O. rochetiana leaf were the most active (MIC 1.95mg/ml) against S. aureus. Ethyl acetate extracts of A. abyssinicus, L. vulgare and S. gigas; aqueous of C. pepo, O. rochetiana and G. myrianthum; and all D. laxata had no antibacterial activity. P. aeruginosa was the least susceptible to any extract, although the methanol and aqueous extracts of S. gigas performed better against it. Preliminary phytochemical screening of selected extracts for phenols, flavonoids, tannins, steroids, terpenoids, steroidal glycosides, alkaloids, saponins, resins and glycosides showed positivity at least for four of these phytochemicals with glycoside and terpenoids in nearly all extracts and resin in none. The plants were not toxic to mice at 2000 mg/kg. Further consideration of S. gigas, L. vulgare, A. abyssinicus and O. rochetiana is recommended in light of their promising potential and safety.

Discovery and identification of potential anti-melanogenic active constituents of Bletilla striata by zebrafish model and molecular docking

Background Bletilla striata is the main medicine of many skin whitening classic formulas in traditional Chinese medicine (TCM) and is widely used in cosmetic industry recently. However, its active ingredients are still unclear and its fibrous roots are not used effectively. The aim of the present study is to discover and identify its potential anti-melanogenic active constituents by zebrafish model and molecular docking. Methods The antioxidant activities were evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, 2,2′-azino-bis-(3-ethylbenthiazoline-6-sulphonic acid) (ABTS) radical scavenging activity and ferric reducing antioxidant power (FRAP) assay. The anti-melanogenic activity was assessed by tyrosinase inhibitory activity in vitro and melanin inhibitory in zebrafish. The chemical profiles were performed by ultra-high-performance liquid chromatography combined with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS). Meanwhile, the potential anti-melanogenic active constituents were temporary identified by molecular docking. Results The 95% ethanol extract of B. striata fibrous roots (EFB) possessed the strongest DPPH, ABTS, FRAP and tyrosinase inhibitory activities, with IC50 5.94 mg/L, 11.69 mg/L, 6.92 mmol FeSO4/g, and 58.92 mg/L, respectively. In addition, EFB and 95% ethanol extract of B. striata tuber (ETB) significantly reduced the melanin synthesis of zebrafish embryos in a dose-dependent manner. 39 chemical compositions, including 24 stilbenoids were tentatively identified from EFB and ETB. Molecular docking indicated that there were 83 (including 60 stilbenoids) and 85 (including 70 stilbenoids) compounds exhibited stronger binding affinities toward tyrosinase and adenylate cyclase. Conclusion The present findings supported the rationale for the use of EFB and ETB as natural skin-whitening agents in pharmaceutical and cosmetic industries.

Persimmon Leaves (Diospyros kaki) Extract Enhances the Viability of Human Corneal Endothelial Cells by Improving Na+-K+-ATPase Activity

The Na+/K+-ATPase, present in the basolateral membrane of human corneal endothelial cells (HCECs), is known to play an important role for corneal transparency. Na+/K+-ATPase dysfunction is one of the major causes of corneal decompensation. The ethanol extract of Diospyros kaki (EEDK) has been reported to increase corneal cell viability. Thus, we treated HCECs with EEDK and studied its effects on HCECs survival and Na+/K+-ATPase against cytotoxic drugs like staurosporine (ST) and ouabain (OU). Firstly, survival assays, (MTT assay and live dead-imaging) showed that decreased HCECs viability by ST and OU was significantly recovered by EEDK co-treatment. Secondly, Na+/K+-ATPase activity assays revealed that EEDK enhanced Na+/K+-ATPase enzymatic activity (* p < 0.01) with/without ST and OU. Finally, Na+/K+-ATPase expression analysis (Western Blot and confocal microscopy) demonstrated that EEDK treatment with/without ST and OU facilitates Na+/K+-ATPase expression in HCECs. Taken together, our findings led us to the conclusion that EEDK might aid HCECs survival in vitro by increasing the activity and expression of Na+/K+-ATPase enzyme. Since Na+/K+-ATPase activity is important to maintain cellular function of HCECs, we suggest that EEDK can be a potential effective agent against corneal edema and related corneal disorders.

Rhynchosia volubilis Promotes Cell Survival via cAMP-PKA/ERK-CREB Pathway

Rhynchosia volubilis, a small black bean, has been used as a traditional remedy to treat diseases and maintain health in East Asia, but its cellular effects and molecular mechanisms are not fully understood. The purpose of this study was to investigate the effect of ethanol extract from Rhynchosia volubilis (EERV) on cell survival and to elucidate the biochemical signaling pathways. Our results showed that EERV stimulated the cyclic AMP (cAMP) signal revealed by a fluorescent protein (FP)-based intensiometric sensor. Using a Förster resonance energy transfer (FRET)-based sensor, we further revealed that EERV could activate PKA and ERK signals, which are downstream effectors of cAMP. In addition, we reported that EERV could induce the phosphorylation of CREB, a key signal for cell survival. Thus, our results suggested that EERV protects against apoptosis by activating the cell survival pathway through the cAMP-PKA/ERK-CREB pathway.