Phytocompounds of Bistorta macrophylla (D. Don) Sojak. as bioavailability enhancers of fluconazole and amphotericin B to better manage Candida species infections

Bistorta macrophylla (D. Don) Sojak. is a medicinal plant of high altitude and so far, not been scientifically explored? Since prehistoric times, B. macrophylla has been used to cure stomach pain, pyretic fever, flu, lungs infections, diarrhea, vomiting. The present research was aimed to examine the phytochemicals, antifungal, and synergistic potential of methanolic extracts of B. macrophylla. Methanolic extract of B. macrophylla was found to have high phenolic (191.18 ± 29.18 mg g−1 GAE) and flavonoid (26.71 ± 3.21 mg g−1 RE) content. Methanolic extract also demonstrate strong antifungal action with diameter of zone of inhibition of 17.5±0.5 mm (fungicidal) against both the strains of C. albicans (MTCC277 and ATCC90028). The minimal inhibitory concentration (MIC) of methanolic extract was found to be 62.5 µg ml−1 against C. albicans (MTCC277 and ATCC90028). In addition, the combination of methanolic extract of B. macrophylla with antifungal antibiotics (fluconazole and amphotericin B) showed synergistic interaction with MIC reduction from 4-128 folds against both candida strains. GC-MS analysis of methanolic extract revealed the presence of 15 major phytocompounds with area more than 1%. Molecular docking showed that sucrose and 9,9-Dimethoxybicyclo [ 3.3.1] nona-2,4-dione has highest binding energy of -6.3 and -5.1 KJ/mol against Cytochrome P450 14 alpha-sterol Demethylase (PDB ID: 1EA1) protein respectively. Combination of methanolic extract of B. macrophylla with antifungal antibiotics (fluconazole, amphotericin B) can be used to treat drug-resistant candida.

Antifungal Activity of N-(4-Halobenzyl)amides against Candida spp. and Molecular Modeling Studies

Fungal infections remain a high-incidence worldwide health problem that is aggravated by limited therapeutic options and the emergence of drug-resistant strains. Cinnamic and benzoic acid amides have previously shown bioactivity against different species belonging to the Candida genus. Here, 20 cinnamic and benzoic acid amides were synthesized and tested for inhibition of C. krusei ATCC 14243 and C. parapsilosis ATCC 22019. Five compounds inhibited the Candida strains tested, with compound 16 (MIC = 7.8 µg/mL) producing stronger antifungal activity than fluconazole (MIC = 16 µg/mL) against C. krusei ATCC 14243. It was also tested against eight Candida strains, including five clinical strains resistant to fluconazole, and showed an inhibitory effect against all strains tested (MIC = 85.3–341.3 µg/mL). The MIC value against C. krusei ATCC 6258 was 85.3 mcg/mL, while against C. krusei ATCC 14243, it was 10.9 times smaller. This strain had greater sensitivity to the antifungal action of compound 16. The inhibition of C. krusei ATCC 14243 and C. parapsilosis ATCC 22019 was also achieved by compounds 2, 9, 12, 14 and 15. Computational experiments combining target fishing, molecular docking and molecular dynamics simulations were performed to study the potential mechanism of action of compound 16 against C. krusei. From these, a multi-target mechanism of action is proposed for this compound that involves proteins related to critical cellular processes such as the redox balance, kinases-mediated signaling, protein folding and cell wall synthesis. The modeling results might guide future experiments focusing on the wet-lab investigation of the mechanism of action of this series of compounds, as well as on the optimization of their inhibitory potency.

Knockdown of vps54 aggravates tamoxifen-induced cytotoxicity in fission yeast

Tamoxifen (TAM) is an anticancer drug used to treat estrogen receptor (ER)‒positive breast cancer. However, its ER-independent cytotoxic and antifungal activities have prompted debates on its mechanism of action. To achieve a better understanding of the ER-independent antifungal action mechanisms of TAM, we systematically identified TAM-sensitive genes through microarray screening of the heterozygous gene deletion library in fission yeast (Schizosaccharomyces pombe). Secondary confirmation was followed by a spotting assay, finally yielding 13 TAM-sensitive genes under the drug-induced haploinsufficient condition. For these 13 TAM-sensitive genes, we conducted a comparative analysis of their Gene Ontology (GO) ‘biological process’ terms identified from other genome-wide screenings of the budding yeast deletion library and the MCF7breast cancer cell line. Several TAM-sensitive genes overlapped between the yeast strains and MCF7 in GO terms including ‘cell cycle’ (cdc2, rik1, pas1, and leo1), ‘signaling’ (sck2, oga1, and cki3), and ‘vesicle-mediated transport’ (SPCC126.08c, vps54, sec72, and tvp15), suggesting their roles in the ER-independent cytotoxic effects of TAM. We recently reported that the cki3 gene with the ‘signaling’ GO term was related to the ER-independent antifungal action mechanisms of TAM in yeast. In this study, we report that haploinsufficiency of the essential vps54 gene, which encodes the GARP complex subunit, significantly aggravated TAM sensitivity and led to an enlarged vesicle structure in comparison with the SP286 control strain. These results strongly suggest that the vesicle-mediated transport process might be another action mechanism of the ER-independent antifungal or cytotoxic effects of TAM.

Effect of Trachyspermum ammi essential oil, fractions and its derivatives on resistant fungal Strains

Persistentantimicrobial drugs treatmenthas resulted in antimicrobial resistance in fungi. There is always a gap for newer antifungal agent. As fungi are associated with multiple health risks in humans and many diseases in crops as well.Objective: To find alternate natural antimicrobial agent as compared to the synthetic one. Method:Essential oil of Trachyspermumammi was isolated, fractionated, and subjected to GC-MS analysis. Components from fractions were derivatized to check their antimicrobial potential against fungal resistant strains. Results:Analysis showed γ -terpinene (39%), α-phellandrene (1.3%), α-pinene (0.5%), Sabinene (0.15%), β-pinene (4.40%), β-myrcene (1.14%), O-cymene (15.78%), p-cymefne (38.78%), and other components were less than 1%. Fractional components were derivatised and their antifungal action was studied. Conclusion: Ajwain oil components found to be good against resistant fungal strains. While some derivatives showed more and some less antimicrobial action.

Study of wound healing in rat skin treated with extract of Hedera helix, L / Estudo da cura de feridas na pele de rato tratada com extrato de Hedera helix, L

The Hera (Hedera helix L) is part of the ARALIACEAE family, is included in the group of plants that produce saponins, has antifungal action, is hypocholesterolemic, has anti-inflammatory activity, is expectorant, antispasmodic and purifying. It was evaluated, by histological studies, the wound healing action of the extract of Hera leaves on skin wounds, as well as changes in the epithelial tissue and wound healing period. 75 Wistar rats were used and divided into five groups, according to the treatment: negative control (PBS), Hera 10 mg/ml, Hera 30 mg/ml, Hera 50 mg/ml and the positive control (Nebacetin). The animals were anesthetized and undergone through a cut of 4 cm in the dorsal region, exposing their muscle fascias. Right after, daily, the suitable substance to each group was applied in the lesion. After periods of 3, 7, 14, 21 days after the surgery, the animals were killed to collect fragments of the lesion. The material was prepared in stained slides with HE and toluidine blue for histologic analysis. The results showed that at day 14 of treatment, the animals under effect of 30 mg/ml and 50 mg/ml of Hera extract did not present edema. It was also observed a reduction in vascular congestion in the Hera of 30 mg/ml, Hera 50 mg/ml and Nebacetin groups compared to the other groups analyzed. All groups treated with Hera extract showed a reduction of inflammatory cells in day 14 post-lesion, besides the increase of fibroblast this period, showing acceleration in the chronicity of the lesion. Regarding the number of mast cells, a significant increase in the early lesion, in the 3 and 7 days periods was observed, in Hera 50 mg/ml and 30 mg/ml groups, respectively. According to the results, the Hera extract, especially at concentrations of 30 mg/ml and 50 mg/ml accelerated the healing process; based on the decreased permanence period of the edema and congestion of the vessels, as well as changes in the number of cells related to inflammation of lesions.

Synthesis, Characterization of Some New 1,3,5-Trisubstituted Pyrazole Derivatives for Their Antifungal Potential

The aim of the study was to develop, synthesis, and characterise a novel 1,3,5-trisubstituted-2-pyrazolines derivative, as well as to evaluate its antifungal activity. The reaction of chalcone derivatives with succinic hydrazide in the presence of pyridine yielded the 1,3,5-tri-substituted-2-pyrazolines derivatives. Total 20 compounds has been synthesized and characterized by the IR, 1HNMR and mass spectral analysis. Antifungal activity of the compounds carried out onfour fungal strains i.e. Saccharomyces cerevisiae, A. Niger, C. Albicans and R. Oryzae in two different concentration i.e. 50 and 100 µg/ml by Agar-diffusion method using Cup-plate method. The usual antifungal medicine was ketoconazole. All of the synthesized 1,3,5-trisubstituted pyrazole compounds (ME1-ME8, CL1-CL8, BR1-BR4) showed medium to best action against examined organisms, according to antifungal activity data. The antifungal activity of compounds against fungal strains (Saccharomyces cerevisiae, A. Niger, Candida albicans, and R. Oryzae) revealed the following order of action: CL-4 > BR-4 > CL-3 > CL-2 > ME-3> ME-2> CL-5 > CL-6 > ME-4 > ME-5 > ME-6 > ME-7 > CL-7 > CL-8>ME-8 >CL-1 >ME-1 > CL-5 > CL-6 > ME-4 > ME-5 > ME-6 > ME-7 > CL-7 > CL-8>ME-8 >CL-1 >ME-1. Electronegative groups (Br, Cl, F, and NO2) must be present at the third and fifth positions of the 1,3,5-pyrazoline ring for significant antifungal action. The presence of an electronegative group at the third and fifth positions may be required for the best action against bacterial and fungal strains, however the addition of F, NO2 has demonstrated moderate activity, while the substitution of methyl and methoxy groups may reduce the activity. The synthesized compounds in the BR-1 through BR-4 class are the most active.

Encapsulation of agrichemicals by renewable cellulosic derivatives

<p>Cellulose derivatives, charged with fungicides, have been prepared as particles for use as a wood preservative. The particles were designed to encapsulate the current industry-standard chemical agents used to minimise wood degrading fungal action and to deter termites. A detailed study on the most effective methodology that would be suitable for scaled-up production was undertaken. The methods explored included: double emulsions, solvent diffusion by dialysis membrane and phase inversion emulsification. Particles formed by these methods were characterised by scanning electron microscopy, dynamic light scattering, nuclear magnetic resonance spectroscopy and infrared spectroscopy. The fungicide incorporation was confirmed by nuclear magnetic resonance studies and gas-chromatography analytical analysis. The phase inversion emulsion process was found to be highly effective and readily manipulated to modify particle formation. Particles were successfully prepared containing fungicides in a yield of 35-75% (method dependant), containing the biocide at approximately 50% mass of biocide to total particle mass. Thus, this process was optimised through modifying the addition time of the aqueous phase, as well as variation of the surfactant and salt concentrations. With an optimised particle forming method, three fungicides were incorporated into the formulation and subsequently analysed to demonstrate successful biocide incorporation. The biocide charged mesoparticles underwent testing for antifungal action by our industrial partners Lonza.</p>

Encapsulation of agrichemicals by renewable cellulosic derivatives

<p>Cellulose derivatives, charged with fungicides, have been prepared as particles for use as a wood preservative. The particles were designed to encapsulate the current industry-standard chemical agents used to minimise wood degrading fungal action and to deter termites. A detailed study on the most effective methodology that would be suitable for scaled-up production was undertaken. The methods explored included: double emulsions, solvent diffusion by dialysis membrane and phase inversion emulsification. Particles formed by these methods were characterised by scanning electron microscopy, dynamic light scattering, nuclear magnetic resonance spectroscopy and infrared spectroscopy. The fungicide incorporation was confirmed by nuclear magnetic resonance studies and gas-chromatography analytical analysis. The phase inversion emulsion process was found to be highly effective and readily manipulated to modify particle formation. Particles were successfully prepared containing fungicides in a yield of 35-75% (method dependant), containing the biocide at approximately 50% mass of biocide to total particle mass. Thus, this process was optimised through modifying the addition time of the aqueous phase, as well as variation of the surfactant and salt concentrations. With an optimised particle forming method, three fungicides were incorporated into the formulation and subsequently analysed to demonstrate successful biocide incorporation. The biocide charged mesoparticles underwent testing for antifungal action by our industrial partners Lonza.</p>