Phytochemical and proximate content of Carapa procera bark and its antimicrobial potential against selected pathogens.

Medicinal plants represent a great source of antimicrobial and phytochemical constituents which are increasingly used to treat microbial infections and other ailments such as tuberculosis, anemia, and trachoma. Despite the use of antibiotics, antimicrobial resistance continues to be a world issue, in as much as nutrition. This study investigated the presence of phytochemicals, proximate compositions, and antimicrobial activity of methanolic extract of Carapa procera bark. The bark of Carapa procera was collected, cleaned and air dried for 72 h. The powder obtained was treated with diethyl ether and soaked in methanol (99%) for 72 h to obtain crude extract. The extract was used to test for the presence of phytochemicals and antimicrobial activities. The raw bark was used for proximate analysis. The result showed presence of steroids, tannins and saponins, but no alkaloids present. The 100 mg/mL extract had the highest inhibition zone on all tested organisms from 24.00 ± 0.94 to 26.67 ± 1.18, and 50 mg/mL showed the least (16.67 ± 1.24) on Candida albicans. Staphylococcus aureus showed the lowest minimum inhibition concentration (MIC) of 3.12 mg/mL, whereas the Gram-negative bacteria exhibited variations in their sensitivity with E. coli having the highest MIC of 25 mg/mL. The extract had high MIC (6.25 mg/mL) on Candida albicans than clotrimazole (50 mg/mL). The proximate compositions of Carapa procera were moisture (6.07 ± 0.07%), ash (12.46 ± 0.46%), crude protein (9.54 ± 0.12%), crude fat (1.42 ± 0.06%) and carbohydrate (70.50 ± 0.35%). The energy value was 1413.17 kj. Thus, Carapa procera possesses both antimicrobial and nutritional potentials worth exploring and domesticating for sustainable management and conservation.

The Effects of Tormentic Acid and Extracts from Callistemon citrinus on Candida albicans and Candida tropicalis Growth and Inhibition of Ergosterol Biosynthesis in Candida albicans.

Candida albicans and Candida tropicalis are the leading causes of human fungal infections worldwide. There is an increase in resistance of Candida pathogens to existing antifungal drugs leading to a need to find new sources of antifungal agents. Tormentic acid has been isolated from different plants including Callistemon citrinus and has been found to possess antimicrobial properties, including antifungal activity. The study aimed to determine the effects of tormentic and extracts from C. citrinus on C. albicans and C. tropicalis and a possible mode of action. The extracts and tormentic acid were screened for antifungal activity using the broth microdilution method. The growth of both species was inhibited by the extracts, and C. albicans was more susceptible to the extract compared to C. tropicalis. The growth of C. albicans was inhibited by 80% at 100 µg/ml of both the DCM: methanol extract and the ethanol: water extract. Tormentic acid reduced the growth of C. albicans by 72% at 100 µg/ml. The effects of the extracts and tormentic acid on ergosterol content in C. albicans were determined using a UV/Vis scanning spectrophotometer. At concentrations of tormentic acid of 25 µg/ml, 50 µg/ml, 100 µg/ml, and 200 µg/ml, the content of ergosterol was decreased by 22%, 36%, 48%, and 78%, respectively. Similarly, the DCM: methanol extract at 100 µg/ml and 200 µg/ml decreased the content by 78% and 88%, respectively. A dose-dependent decrease in ergosterol content was observed in cells exposed to miconazole with a 25 µg/ml concentration causing a 100% decrease in ergosterol content. Therefore, tormentic acid inhibits the synthesis of ergosterol in C. albicans. Modifications of the structure of tormentic acid to increase its antifungal potency may be explored in further studies.

Bruguiera gymnorhiza (L.) Lam. at the Forefront of Pharma to Confront Zika Virus and Microbial Infections-An In Vitro and In Silico Perspective.

The recent emergence of Zika virus (ZIKV) in Brazil and the increasing resistance developed by pathogenic bacteria to nearly all existing antibiotics should be taken as a wakeup call for the international authority as this represents a risk for global public health. The lack of antiviral drugs and effective antibiotics on the market triggers the need to search for safe therapeutics from medicinal plants to fight viral and microbial infections. In the present study, we investigated whether a mangrove plant, Bruguiera gymnorhiza (L.) Lam. (B. gymnorhiza) collected in Mauritius, possesses antimicrobial and antibiotic potentiating abilities and exerts anti-ZIKV activity at non-cytotoxic doses. Microorganisms Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Klebsiella pneumoniae ATCC 70603, methicillin-resistant Staphylococcus aureus ATCC 43300 (MRSA), Salmonella enteritidis ATCC 13076, Sarcina lutea ATCC 9341, Proteus mirabilis ATCC 25933, Bacillus cereus ATCC 11778 and Candida albicans ATCC 26555 were used to evaluate the antimicrobial properties. Ciprofloxacin, chloramphenicol and streptomycin antibiotics were used for assessing antibiotic potentiating activity. ZIKVMC-MR766NIID (ZIKVGFP) was used for assessing anti-ZIKV activity. In silico docking (Autodock 4) and ADME (SwissADME) analyses were performed on collected data. Antimicrobial results revealed that Bruguiera twig ethyl acetate (BTE) was the most potent extract inhibiting the growth of all nine microbes tested, with minimum inhibitory concentrations ranging from 0.19-0.39 mg/mL. BTE showed partial synergy effects against MRSA and Pseudomonas aeruginosa when applied in combination with streptomycin and ciprofloxacin, respectively. By using a recombinant ZIKV-expressing reporter GFP protein, we identified both Bruguiera root aqueous and Bruguiera fruit aqueous extracts as potent inhibitors of ZIKV infection in human epithelial A549 cells. The mechanisms by which such extracts prevented ZIKV infection are linked to the inability of the virus to bind to the host cell surface. In silico docking showed that ZIKV E protein, which is involved in cell receptor binding, could be a target for cryptochlorogenic acid, a chemical compound identified in B. gymnorhiza. From ADME results, cryptochlorogenic acid is predicted to be not orally bioavailable because it is too polar. Scientific data collected in this present work can open a new avenue for the development of potential inhibitors from B. gymnorhiza to fight ZIKV and microbial infections in the future.

Polyphenolics profile effects upon the antioxidant and antimicrobial activity of propolis extracts.

Propolis, a complex bee product, is a source of numerous bioactive principles, beneficial for human health, therefore it is intensively studied. In the present work, extracts of propolis from Bihor Romanian County were studied to identify the relationship between the polyphenolic derivatives profile and their antioxidant and antimicrobial activity. Extracts were obtained using water and 25%, 50%, and 70% ethanolic solutions (w/w), at 2:1, 4:1, and 6:1 liquid: solid ratios (w/w). 21 polyphenolic derivatives were quantified by UHPLC-MS, proving that the extracts composition strongly depends on the solvent. The sum of quantified polyphenolics extracted varied between 1.5 and 91.2 mg/g propolis. The antioxidant capacity was evaluated using the free radicals 2,2'-azino-bis (3-ethylbenzothiazoline-6 sulfonic acid) diammonium salt (ABTS) and 1-diphenyl-2-picryl-hydrazyl (DPPH) scavenging methods. Antimicrobial efficiency was tested against Gram-positive (B. subtilis), Gram-negative bacteria (E. coli), and fungi (C. albicans) by disc-diffusion method. All extracts, even the aqueous ones, demonstrated antibacterial and antifungal activity. Chemometric methods (partial least squares) and a saturation-type model were used to evaluate the contribution of various bioactive principles in building the antioxidant capacity of extracts. Both experimental and modelling results show that 50% ethanolic extracts provide a rich polyphenolics profile and ensure a good antioxidant capacity.

Cationic Clitoria ternatea Seed Peptide as a Potential Novel Bioactive Molecule.

BACKGROUND: While several biologics have been reported from different parts of Clitoria ternatea, a herbaceous climber of the family Fabaceae, specific production of cationic peptides other than cyclotides (<3.7 kDa) has barely been investigated, or their bioactive potential been looked into. OBJECTIVE: The study aims to uncover potential bioactivities and characteristics of novel cationic peptides from C. ternatea seeds. METHODS: C. ternatea seed cationic peptide purified by simple and cost-effective procedures was analyzed by electrophoresis and mass spectrometry. Antimicrobial efficacy was evaluated against bacterial and fungal pathogens. Antioxidant potential was quantified by in vitro antioxidant assays. Physicochemical characterization and Tandem mass spectrometry were performed. RESULTS: An 8.5 kDa cationic peptide purified from C. ternatea seeds was active against Candida albicans, Staphylococcus aureus, Aeromonas hydrophila and Escherichia coli at a minimum inhibitory concentration in the range of 8-32 µg/ml. This activity was totally uncompromised at pH 5-8 or after 1 h of heat treatment at 70-80ºC, but was sensitive to protease treatment. Concentration-dependent free-radical scavenging activity and ferric-reducing capacity demonstrated the antioxidant potential of the peptide. Tandem MS analysis of trypsin-digested peptide based on shotgun proteomics detected matching peptide sequences with one or two cysteine residues but had low sequence coverage (≤17%) to known sequences in the C. ternatea protein database. Taken together, the distinct characteristics of this novel 8.5 kDa peptide clearly distinguish it from known cyclotides of C. ternatea. CONCLUSIONS: Insights have been obtained into the functional characteristics of what appears to be a novel cationic peptide from C. ternatea seeds, exhibiting significant antimicrobial and antioxidant activities.

Chemical Constituents of the Essential Oil from Ecuadorian Endemic Species Croton ferrugineus and Its Antimicrobial, Antioxidant and α-Glucosidase Inhibitory Activity.

Croton ferrugineus Kunth is an endemic species of Ecuador used in traditional medicine both for wound healing and as an antiseptic. In this study, fresh Croton ferrugineus leaves were collected and subjected to hydrodistillation for extraction of the essential oil. The chemical composition of the essential oil was determined by gas chromatography equipped with a flame ionization detector and gas chromatography coupled to a mass spectrometer using a non-polar and a polar chromatographic column. The antibacterial activity was assayed against three Gram-positive bacteria, one Gram-negative bacterium and one dermatophyte fungus. The radical scavenging properties of the essential oil was evaluated by means of DPPH and ABTS assays. The chemical analysis allowed us to identify thirty-five compounds representing more than 99.95% of the total composition. Aliphatic sesquiterpene hydrocarbon trans-caryophyllene was the main constituent with 20.47 ± 1.25%. Other main compounds were myrcene (11.47 ± 1.56%), ß-phellandrene (10.55 ± 0.02%), germacrene D (7.60 ± 0.60%), and α-humulene (5.49 ± 0.38%). The essential oil from Croton ferrugineus presented moderate activity against Candida albicans (ATCC 10231) with an MIC of 1000 µg/mL, a scavenging capacity SC50 of 901 ± 20 µg/mL with the ABTS method, and very strong antiglucosidase activity with an IC50 of 146 ± 20 µg/mL.

Antimicrobial and antioxidant activity of Evernia prunastri extracts and their isolates.

Lichens are symbiotic organisms formed by a fungus and one or more photosynthetic partners which are usually alga or cyanobacterium. Their diverse and scarcely studied metabolites facilitate adaptability to extreme living conditions. We investigated Evernia prunastri (L.) Ach., a widely distributed lichen, for its antimicrobial and antioxidant potential. E. prunastri was sequentially extracted by hexane (Hex), dichloromethane (DCM) and acetonitrile (ACN) that were screened for their antioxidant and antimicrobial (against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Candida albicans) activities. The Hex extract possessed the highest antioxidant capacity (87 mg ascorbic acid/g extract) corresponding to the highest content of phenols (73 mg gallic acid/g extract). The DCM and Hex extracts were both active against S. aureus (MICs of 4 and 21 µg/ml, respectively) but were less active against Gram-negative bacteria and yeast. The ACN extract exhibited activity on both S. aureus (MIC 14 µg/ml) and C. albicans (MIC 38 µg/ml) and was therefore further fractionated by silica gel column chromatography. The active compound of the most potent fraction was subsequently characterized by 1H and 13C-NMR spectroscopy and identified as evernic acid. Structural similarity analyses were performed between compounds from E. prunastri and known antibiotics from different classes. The structural similarity was not present. Antioxidant and antimicrobial activities of E. prunastri extracts originate from multiple chemical compounds; besides usnic acid, most notably evernic acid and derivatives thereof. Evernic acid and its derivatives represent possible candidates for a new class of antibiotics.

Characterization of Citrusnobilis Peel Methanolic Extract for Antioxidant, Antimicrobial, and Anti-Inflammatory Activity.

Currently, the potential utilization of fruits and vegetable waste as a source of micronutrients and antioxidants has increased. The present study, therefore, aimed to determine the antimicrobial and anti-inflammatory activities of Citrus nobilis peel extract. A modified solvent evaporation technique was employed for peel extract preparation. For effective utilization of the natural product, quantitative analysis of phenolic compounds was carried out using liquid chromatography and mass spectroscopy technique. Phenolic and flavonoids were present in high amounts, while ß-carotene and lycopene were present in vestigial amounts. The antimicrobial efficiency of peel extract was evaluated against four bacterial strains including Staphylococcus aureus (MTCC 3160), Klebsiella pneumoniae (MTCC 3384), Pseudomonas aeruginosa (MTCC 2295), and Salmonella typhimurium (MTCC 1254), and one fungal strain Candida albicans (MTCC 183), and zone of inhibition was comparable to the positive control streptomycin and amphotericin B, respectively. The extract of Citrus nobilis peels showed effective anti-inflammatory activity during human red blood cell membrane stabilization (HRBC) and albumin denaturation assay. The extracts also exhibited 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity ranging from 53.46 to 81.13%. Therefore, the obtained results suggest that Citrus nobilis peel could be used as an excellent source of polyphenols and transformed into value-added products.

Associating chitosan and microemulsion as a topical vehicle for the administration of herbal medicines.

A viscous solution of low molecular weight chitosan (CH) at 5% w/v (10.2 kDa, 75 % deacetylated, 1451 cP at 25 °C) was associated with a microemulsion (ME) that undergoes a phase transition after water absorption in situ (≈28 % w/w), forming a more viscous liquid crystal, which was potentially evaluated as a topical vehicle. The ME was selected from a phase diagram, selecting a composition based on Tween® 80 (52 %), myristate isopropyl (28 %), and the aqueous phase (water and polyethylene glycol 400, 60:40 w/w) (20 %), which was after replaced by CH and herbal medicines (HM). HM are alternatives to treat candidiasis, and Stryphnodendron adstringens shell extract, characterized by molecular networking, and Melaleuca alternifolia Chell essential oil (46 % of terpinen-4-ol), showed in vitro activity against Candida albicans. Associating CH in ME improved the mechanical properties of the topical formulation, as adhesiveness, which is an advantageous feature for the topical treatment of vulvovaginal candidiasis.

Therapeutic potential of labdane diterpene isolated from Alpinia nigra: detailed hemato-compatibility and antimicrobial studies.

(E)-labda-8(17), 12-diene-15,16-dial has been isolated from the seeds of Alpinia nigra that is unsuitable for oral administration and evident from in silico studies. The present investigation therefore deals with understanding the effect of this compound on RBCs for intravenous administration. No prominent hemolytic effect of compound at a concentration of ≤0.4 mg/ml was found whereas higher concentrations perforated RBC membrane. The molecule showed remarkable inhibitory potential against Gram negative bacteria (concentration ≥0.025 mg/ml) causing cell lysis. In case of pathogenic yeast Candida albicans although growth was inhibited (concentration ≥ 0.0025 mg/ml), growth kinetic study revealed that the diterpene significantly delayed fungal growth (concentration 0.005-0.020 mg/ml) by preventing substrate uptake and was able to extend its lag phase in a dose-dependent manner. This study tries to unveil the mechanism of action of this diterpene on microorganisms with differential cell wall compositions.

Toxicity and Antimicrobial Activities of Amaranthus caudatus L. (Amaranthaceae) Harvested From Formulated Soils at Different Growth Stages.

This study examined the toxicity and antimicrobial effects of ethanol and aqueous extracts from Amaranthus caudatus grown on soils formulated from parent particles of silt, sand and clay in a glasshouse. Four different soils namely; sandy clay loam, loam, clayey loam and silty clay loam from were formulated were used for cultivation with the unfractionated soil which was the control. Crude extracts obtained from the plant shoots harvested at different growth stages were tested on some certain gram-negative and gram-positive bacteria and some fungi via agar dilution assay. The toxicity of the water and ethanol extracts was also examined via Artemia salina assay and the level of lethality was measured against Clarkson's lethality scale. All aqueous samples, as well as ethanol extracts of flowering and pre-flowering harvests of control soil tested, were non-toxic (LC50 > 1 mg/mL). At post flowering, the ethanolic extracts were highly toxic mostly in clayey loam, control, sandy-clayey loam soils (LC50 < 0.5 mg/mL). Also, antifungal effects of the plant revealed that extracts inhibited the growth of Candida albicans significantly with mild effect on Candida glabrata, Penicillium chrysogenum and Penicillium aurantiogriseum suggesting that the plant is a promising pharmacological candidate in the treatment of candidiasis. For an optimal yield of non-toxic supplement for household consumption which may also serve as pharmacological precursors, clayey loam soil is recommended for cultivation and harvesting may occur at pre-flowering or flowering stage using ethanol and water as solvents of extraction.

Human Mesenchymal Stromal Cell Secretome Promotes the Immunoregulatory Phenotype and Phagocytosis Activity in Human Macrophages.

Human mesenchymal stromal/stem cells (hMSCs) show great promise in cell therapy due to their immunomodulatory properties. The overall immunomodulatory response of hMSCs resembles the resolution of inflammation, in which lipid mediators and regulatory macrophages (Mregs) play key roles. We investigated the effect of hMSC cell-cell contact and secretome on macrophages polarized and activated toward Mreg phenotype. Moreover, we studied the effect of supplemented polyunsaturated fatty acids (PUFAs): docosahexaenoic acid (DHA) and arachidonic acid, the precursors of lipid mediators, on hMSC immunomodulation. Our results show that unlike hMSC cell-cell contact, the hMSC secretome markedly increased the CD206 expression in both Mreg-polarized and Mreg-activated macrophages. Moreover, the secretome enhanced the expression of programmed death-ligand 1 on Mreg-polarized macrophages and Mer receptor tyrosine kinase on Mreg-activated macrophages. Remarkably, these changes were translated into improved Candida albicans phagocytosis activity of macrophages. Taken together, these results demonstrate that the hMSC secretome promotes the immunoregulatory and proresolving phenotype of Mregs. Intriguingly, DHA supplementation to hMSCs resulted in a more potentiated immunomodulation with increased CD163 expression and decreased gene expression of matrix metalloproteinase 2 in Mreg-polarized macrophages. These findings highlight the potential of PUFA supplementations as an easy and safe method to improve the hMSC therapeutic potential.

Bioguided isolation, identification and activity evaluation of antifungal compounds from Acorus tatarinowii Schott.

ETHNOPHARMACOLOGY RELEVANCE: As a traditional folk medicine, Acorus tatarinowii Schott was used to treat digestive diseases, such as diarrhea, which may be related to Candida albicans infection; however according to literature surveys, there have been few studies of A. tatarinowii focusing on its antimicrobial activity, and almost all describe investigations using crude extracts or fractions. AIM OF THE STUDY: The aims of the current study were to isolate and identify antifungal fractions of A. tatarinowii based on their antifungal activity, explore the preliminary mechanism of 60% ethanol elution (AT60) by metabonomics, and evaluate the antifungal activity of AT60 in vivo and in vitro, to provide natural resources against fungal infections. MATERIALS AND METHODS: As a pilot evaluation of activity, A. tatarinowii fractions and compounds with antifungal bioactivity were isolated by bioactive-guided column chromatography, and identified by LC-QTOF-MS/MS and NMR spectroscopy. The antifungal effects of the active ingredients against resistant C. albicans were evaluated by in vivo and in vitro colony forming unit assays. The mechanism underlying the activity of AT60 against C. albicans was explored using an LC-QTOF-based metabonomics approach and fluorescence microscopy imaging. RESULTS: AT60 showed better activity against C. albicans than the same dose of the first line antifungal drugs, fluconazole and itraconazole (positive control drugs). Subsequent phytochemical investigation of AT60 identified twenty-five known compounds, six of which were isolated: asaraldehyde (7), 1-(2,4,5-trimethoxyphenyl)-1,2-propanediol (12), α-asarone (14), ß-asarone (15), γ-asarone (18), acotatarone C (19). Further, the compounds α-asarone (14) and acotatarone C (19) may be responsible for the antifungal activity, and exhibit synergistic effects. Metabonomics analysis indicated that AT60 can inhibit biofilm formation by regulating the C. albicans protein kinase C pathway. CONCLUSIONS: Our results show that A. tatarinowii has potent bioactivity against C. albicans in vitro and in vivo, and can be considered an antifungal botanic agent.

Boesenbergia rotunda extract inhibits Candida albicans biofilm formation by pinostrobin and pinocembrin.

ETHNOPHARMACOLOGICAL RELEVANCE: Boesenbergia rotunda (L.) Mansf. (Zingiberaceae) is an indigenous plant of Southeast Asia. Based on ethnopharmacological use, the rhizome is recommended in the treatment of stomachache, leukoplakia, abscesses, and leukorrhea in Thailand primary health care system. Candida albicans often causes leukorrhea, and infection of many mucosal sites. Its infection leads to serious illness. AIM OF THE STUDY: This study aimed to investigate the effects of the ethanolic extract of the B. rotunda rhizome on C. albicans ATCC10231 in the stages of planktonic and biofilm formation and to explore the underlying mechanisms. MATERIALS AND METHODS: The chemical composition of the extract was determined using ultra-performance liquid chromatography (UPLC). The planktonic growth of C. albicans was evaluated by the microdilution method, following EUCAST guidelines. For each stage of biofilm formation, the biofilm was assessed by the MTT assay. The biofilm structure was examined under a light microscope. The degree of cell surface hydrophobicity was measured. The mRNA levels of ALS1, ALS3, and ACT1 were determined by RT-qPCR. RESULTS: The extract of B. rotunda consisted of 25% (w/w) pinostrobin and 12% (w/w) pinocembrin. All stages of C. albicans biofilm formation were significantly inhibited by the extract, whereas the planktonic growth did not change. Biofilm development greatly decreased due to the extract in a concentration-dependent manner, with an IC50 value of 17.7 µg/mL. Pinostrobin and pinocembrin demonstrated inhibitory effects during this stage. These results were in accordance with the microscopic evaluation. The filamentous form decreased with pinocembrin rather than pinostrobin. Moreover, the cell surface hydrophobicity was significantly decreased by 6.25 and 12.5 µg/mL of the extract and 100 µM of pinocembrin. The ALS3 mRNA level was noticeably decreased by 12.5 µg/mL of the extract, 100 µM of pinostrobin, and 100 µM of pinocembrin. The ACT1 mRNA level decreased significantly with pinocembrin. However, the ALS1 mRNA level was not altered following all treatments. CONCLUSION: The ethanolic extract of B. rotunda could inhibit biofilm formation of C. albicans, especially during the biofilm development stage, by means of reducing the cell surface hydrophobicity and suppressing the ALS3 mRNA expression. Pinocembrin had a stronger effect on ALS3 mRNA expression than pinostrobin. Only pinocembrin significantly decreased the ACT1 mRNA level.

Potential of Methylglyoxal-Conjugated Chitosan Nanoparticles in Treatment of Fluconazole-Resistant Candida albicans Infection in a Murine Model.

BACKGROUND: Fungal infections are becoming more prevalent and threatening because of the continuous emergence of azole-resistant fungal infections. The present study was aimed to assess the activity of free Methylglyoxal (MG) or MG-conjugated chitosan nanoparticles (MGCN) against fluconazole-resistant Candida albicans. MATERIALS AND METHODS: A novel formulation of MGCN was prepared and characterized to determine their size, shape and polydispersity index. Moreover, the efficacy of fluconazole or MG or MGCN was determined against intracellular C. albicans in macrophages and the systematic candidiasis in a murine model. The safety of MG or MGCN was tested in mice by analyzing the levels of hepatic and renal toxicity parameters. RESULTS: Candida albicans did not respond to fluconazole, even at the highest dose of 20 mg/kg, whereas MG and MGCN effectively eliminated C. albicans from the macrophages and infected mice. Mice in the group treated with MGCN at a dose of 10 mg/kg exhibited a 90% survival rate and showed the lowest fungal load in the kidney, whereas the mice treated with free MG at the same dose exhibited 50% survival rate. Moreover, the administration of MG or MGCN did not induce any liver and kidney toxicity in the treated mice. CONCLUSION: The findings of the present work suggest that MGCN may be proved a promising therapeutic formulation to treat azole-resistant C. albicans infections.

Antibiofilm efficacy of tea tree oil and of its main component terpinen-4-ol against Candida albicans.

Candida infection is an important cause of morbidity and mortality in immunocompromised patients. The increase in its incidence has been associated with resistance to antimicrobial therapy and biofilm formation. The aim of this study was to evaluate the efficacy of tea tree oil (TTO) and its main component - terpinen-4-ol - against resistant Candida albicans strains (genotypes A and B) identified by molecular typing and against C. albicans ATCC 90028 and SC 5314 reference strains in planktonic and biofilm cultures. The minimum inhibitory concentration, minimum fungicidal concentration, and rate of biofilm development were used to evaluate antifungal activity. Results were obtained from analysis of the biofilm using the cell proliferation assay 2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) and confocal laser scanning microscopy (CLSM). Terpinen-4-ol and TTO inhibited C. albicans growth. CLSM confirmed that 17.92 mg/mL of TTO and 8.86 mg/mL of terpinen-4-ol applied for 60 s (rinse simulation) interfered with biofilm formation. Hence, this in vitro study revealed that natural substances such as TTO and terpinen-4-ol present promising results for the treatment of oral candidiasis.

Effect of heat treatment on thermal resistance, transparency and antimicrobial activity of sonicated ginger cellulose film.

Transparent film with high thermal resistance and antimicrobial properties has many applications in the food packaging industry particularly packaging for reheatable food. This work investigates the effects of heat treatment on the thermal resistance, stability of transparency and antimicrobial activity of transparent cellulose film. The film from ginger nanocellulose fibers was prepared with chemicals and ultrasonication. The dried film was heated at 150 °C for 30, 60, 90, or 120 min. The unheated and sonicated film had the lowest crystallinity index and the lowest thermal properties. After heating, the film became brownish-yellow resulting from thermal oxidation. The reheated film had higher thermal resistance than unheated film. Heating led to further relaxation of cellulose network evidenced by shifting of the XRD peak positions toward lower values. The antimicrobial activity decreased due to heating. Average opacity value increases after short heating durations. It was relatively stable for further heating.

Fluconazole analogues with metal-binding motifs impact metal-dependent processes and demonstrate antifungal activity in Candida albicans.

Azole antifungals are an important class of antifungal drugs due to their low cost, ability to be administered orally, and broad-spectrum activity. However, their widespread and long-term use have given rise to adaptation mechanisms that render these compounds less effective against common fungal pathogens, including Candida albicans. New antifungals are desperately needed as drug-resistant strains become more prevalent. We recently showed that copper supplementation potentiates the activity of the azole antifungal fluconazole against the opportunistic fungal pathogen C. albicans. Here, we report eight new azole analogues derived from fluconazole in which one triazole group has been replaced with a metal-binding group, a strategy designed to enhance potentiation of azole antifungal activity by copper. The bioactivity of all eight compounds was tested and compared to that of fluconazole. Three of the analogues showed activity against C. albicans and two had lower levels of trailing growth. One compound, Flu-TSCZ, was found to impact the levels, speciation, and bioavailability of cellular metals.

The effects of clioquinol in morphogenesis, cell membrane and ion homeostasis in Candida albicans.

BACKGROUND: Candida albicans is the most prevalent opportunistic fungal pathogen. Development of antifungals with novel targets is necessary for limitations of current antifungal agents and the emergence of drug resistance. The antifungal activity of clioquinol was widely accepted while the precise mechanism was poorly understood. Hence, we aimed to seek for the possible mechanism of clioquinol against Candida albicans in the present study. RESULTS: Clioquinol could inhibit hyphae formation in a concentration-dependent manner in multiple liquid and solid media. The concentration and time-dependent anti-biofilm activities were observed in different incubation periods quantitatively and qualitatively. Further investigation found that clioquinol disrupted cell membrane directly in high concentration and induced depolarization of the membrane in low concentration. As for the influence on ion homeostasis, the antifungal effects of clioquinol could be reversed by exogenous addition of metal ions. Meanwhile, the minimum inhibitory concentration of clioquinol was increased in media supplemented with exogenous metal ions and decreased in media supplemented with exogenous metal chelators. We also found that the cellular labile ferrous iron level decreased when fungal cells were treated with clioquinol. CONCLUSION: These results indicated that clioquinol could inhibit yeast-hyphae transition and biofilm formation in Candida albicans. The effect on the cell membrane was different depending on different concentrations of clioquinol. Meanwhile, clioquinol could interfere with ion homeostasis as metal chelators for zinc, copper and iron, which was quite different with current common antifungal agents. All in all, clioquinol can be a new promising antifungal agent with novel target though more studies are needed to better understand the precise antifungal mechanism.

Dodonaea viscosa var angustifolia derived 5,6,8-trihydroxy-7,4' dimethoxy flavone inhibits ergosterol synthesis and the production of hyphae and biofilm in Candida albicans.

ETHNOPHARMACOLOGICAL RELEVANCE: Candida albicans is developing resistance to existing drugs increasing morbidity and mortality, which elevates an immediate need to explore new antifungal agents. Phytochemicals are an excellent source of therapeutic agents. We previously reported the antifungal activity of the crude extract of Dodonaea viscosa var. angustifolia Jacq. (DVA) from which a beneficial compound flavone: 5,6,8-trihydroxy-7,4' dimethoxy flavone (5,6,8-trihydroxy-7-methoxy-2-(4-methoxyphenyl)-4H-chromen-4-one) abbreviated as TMMC, was extracted. AIM OF THE STUDY: The present study evaluated the effect of a TMMC subfraction on biofilms, membrane stability, ergosterol biosynthesis and germ tube (GT) formation in Candida albicans. MATERIALS AND METHODS: Extracts were prepared and fractionated to obtain purified TMMC. Minimum inhibitory concentrations of TMMC were obtained and subinhibitory concentrations were selected for further studies. Confocal laser scanning microscopy (CLSM) was performed to assess the effect of TMMC on membrane permeability and sterol deposition using propidium iodide (PI) and filipin stains, respectively. RESULTS: Minimum inhibitory concentrations (MIC) and Minimum Fungicidal concentrations (MFC) of TMMC were 0.39 mg/mL and 1.56 mg/mL, respectively. TMMC inhibited biofilm formation and damaged mature biofilms at 0.39 mg/mL and 1.56 mg/mL, respectively. CLSM further confirmed the disruption and architectural changes in biofilms following treatment with TMMC. TMMC also inhibited GT formation and ergosterol biosynthesis in a concentration dependent manner, which was further confirmed by varying sterol distribution and membrane disruption in treated and untreated cells. CONCLUSIONS: With the multiple targets at different concentrations, TMMC warrants its potential use as antifungal drug against C. albicans. However further studies using animal models and more mechanistic approaches will be required.