Amit Kumar Tyagi

Liquid and vapour-phase antifungal activities of selected essential oils against candida albicans: Microscopic observations and chemical characterization of cymbopogon citratus

BackgroundUse of essential oils for controlling Candida albicans growth has gained significance due to the resistance acquired by pathogens towards a number of widely-used drugs. The aim of this study was to test the antifungal activity of selected essential oils against Candida albicans in liquid and vapour phase and to determine the chemical composition and mechanism of action of most potent essential oil.MethodsMinimum Inhibitory concentration (MIC) of different essential oils in liquid phase, assayed through agar plate dilution, broth dilution & 96-well micro plate dilution method and vapour phase activity evaluated through disc volatilization method. Reduction of C. albicans cells with vapour exposure was estimated by kill time assay. Morphological alteration in treated/untreated C. albicans cells was observed by the Scanning electron microscopy (SEM)/Atomic force microscopy (AFM) and chemical analysis of the strongest antifungal agent/essential oil has been done by GC, GC-MS.ResultsLemon grass (Cymbopogon citratus) essential oil exhibited the strongest antifungal effect followed by mentha (Mentha piperita) and eucalyptus (Eucalyptus globulus) essential oil. The MIC of lemon grass essential oil in liquid phase (288 mg/l) was significantly higher than that in the vapour phase (32.7 mg/l) and a 4 h exposure was sufficient to cause 100% loss in viability of C. albicans cells. SEM/AFM of C. albicans cells treated with lemon grass essential oil at MIC level in liquid and vapour phase showed prominent shrinkage and partial degradation, respectively, confirming higher efficacy of vapour phase. GC-MS analysis revealed that lemon grass essential oil was dominated by oxygenated monoterpenes (78.2%); α-citral or geranial (36.2%) and β-citral or neral (26.5%), monoterpene hydrocarbons (7.9%) and sesquiterpene hydrocarbons (3.8%).ConclusionLemon grass essential oil is highly effective in vapour phase against C. albicans, leading to deleterious morphological changes in cellular structures and cell surface alterations.

In situ SEM, TEM and AFM studies of the antimicrobial activity of lemon grass oil in liquid and vapour phase against Candida albicans

Inhibition of Candida albicans growth was shown by lemon grass oil (LGO) and lemon grass oil vapour (LGO vapour) at 288microg/ml and 32.7microg/ml concentration, respectively. The assessment of cell damage by LGO and LGO vapour was done through scanning electron microscope (SEM), transmission electron microscope (TEM) and atomic force microscope (AFM) observations. SEM analysis showed complete rupture of C. albicans cells treated with LGO vapour while in those treated with LGO in broth, only shrinkage was observed. TEM study showed the alterations in morphology upon treatment with LGO while complete degradation of the Candida cells was observed in case of LGO vapour. Further three dimensional morphological changes and roughness of the cells have also been evaluated with AFM after the treatment with LGO & LGO vapour. Roughness (root mean square value) was significantly higher in control C. albicans cells (211.97nm) than LGO (143nm) and LGO vapour (5.981nm) treated cells. The results for the first time demonstrate relatively higher efficacy of LGO vapours for inhibition and cellular damage of C. albicans cells as compared to the LGO in liquid phase. This suggests the potential application of LGO vapour phase against infections caused by C. albicans.

Reactive oxygen species (ROS) and cancer: Role of antioxidative nutraceuticals

Extensive research over the past half a century indicates that reactive oxygen species (ROS) play an important role in cancer. Although low levels of ROS can be beneficial, excessive accumulation can promote cancer. One characteristic of cancer cells that distinguishes them from normal cells is their ability to produce increased numbers of ROS and their increased dependence on an antioxidant defense system. ROS are produced as a byproduct intracellularly by mitochondria and other cellular elements and exogenously by pollutants, tobacco, smoke, drugs, xenobiotics, and radiation. ROS modulate various cell signaling pathways, which are primarily mediated through the transcription factors NF-κB and STAT3, hypoxia-inducible factor-1α, kinases, growth factors, cytokines and other proteins, and enzymes; these pathways have been linked to cellular transformation, inflammation, tumor survival, proliferation, invasion, angiogenesis, and metastasis of cancer. ROS are also associated with epigenetic changes in genes, which is helpful in diagnosing diseases. This review considers the role of ROS in the various stages of cancer development. Finally, we provide evidence that nutraceuticals derived from Mother Nature are highly effective in eliminating cancer cells.

Antimicrobial action of essential oil vapours and negative air ions against Pseudomonas fluorescens

The aim of this study was to investigate the antibacterial activity of essential oil (in liquid as well as in vapour phase) and negative air ions (NAI) against Pseudomonas fluorescens. The combined effect of NAI with essential oil vapour was also investigated to determine kill time and morphological changes in bacterial cells. The MIC of Cymbopogon citratus (0.567 mg/ml), Mentha arvensis (0.567 mg/ml), Mentha piperita (1.125 mg/ml) and Eucalyptus globulus (2.25 mg/ml) was studied via the agar dilution method. To estimate the antibacterial activity of essential oils in the vapour phase, agar plates inoculated with P. fluorescens were incubated with various concentrations of each essential oil vapour and zone of inhibition was recorded. Further, in order to assess the kill time, P. fluorescens inoculated agar plates were exposed to selected bactericidal essential oil vapour and NAI, separately, in an air-tight chamber. A continuous decrease in bacterial count was observed over time. A significant enhancement in the bactericidal action was observed by exposure to the combination of essential oil vapour and NAI as compared to their individual action. Scanning electron microscopy was used to study the alteration in morphology of P. fluorescens cells after exposure to C. citratus oil vapour, NAI, and combination of C. citratus oil vapour and NAI. Maximum morphological deformation was found due to the combined effect of C. citratus oil vapour and NAI. This study demonstrates that the use of essential oils in the vapour phase is more advantageous than the liquid phase. Further the antibacterial effect of the essential oil vapours can be significantly enhanced by the addition of NAI. The work described here offers a novel and efficient approach for control of bacterial contamination that could be applied for food stabilization practices.

Anti-yeast activity of mentha oil and vapours through in vitro and in vivo (real fruit juices) assays

The anti-yeast activity of mentha oil and vapours was evaluated against 8 food spoiling yeasts through disc diffusion, disc volatilisation and micro broth dilution method. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) varied from 0.28 to 2.25 and 1.13 to 4.5 mg/ml, respectively. Furthermore, the anti-yeast efficacy of mentha oil alone and in combination with thermal treatment was evaluated in a real food system i.e. mixed fruit juices. The samples treated with a combination of mentha oil at the MIC, ½ MIC and ¼ MIC levels and thermal treatment enhanced the reduction viability. Chemical characterisation of mentha oil by gas chromatography-mass spectrometry (GC-MS) revealed that the dominant compounds were cis-menthone (27.43%), menthol (24.3%), trans-menthone (9.23%), limonene (5.84%), menthofuran (4.44%) and isomenthol (3.21%). Present results established the superior performance of integrated treatment over individual exposure for fruit juice preservation.

Production of medium chain saturated fatty acids with enhanced antimicrobial activity from crude coconut fat by solid state cultivation of Yarrowia lipolytica.

Fatty acids profiles and antimicrobial activity of crude coconut fat hydrolysates obtained in solid-state cultivation system with a selected yeast strain Yarrowia lipolytica RO13 were performed. A preliminary step regarding extracellular lipase production and solid state enzymatic hydrolysis of crude fat at different water activity and time intervals up to 7 days was also applied. Gas chromatography-mass spectrometry analysis was used for quantification of medium chain saturated fatty acids (MCSFAs) and the results revealed a higher concentration of about 70% lauric acid from total fatty acids. Further, antimicrobial activity of fatty acids against some food-borne pathogens (Salmonella enteritidis, Escherichia coli, Listeria monocytogenes and Bacillus cereus) was evaluated. The minimum inhibitory concentration of the obtained hydrolysates varied from 12.5 to 1.56 ppm, significantly lower than values reported in literature. The results provide substantial evidence for obtaining biopreservative effects by coconut fat enzymatic hydrolysis.

Ursolic acid inhibits the growth of human pancreatic cancer and enhances the antitumor potential of gemcitabine in an orthotopic mouse model through suppression of the inflammatory microenvironment

The development of chemoresistance in human pancreatic cancer is one reason for the poor survival rate for patients with this cancer. Because multiple gene products are linked with chemoresistance, we investigated the ability of ursolic acid (UA) to sensitize pancreatic cancer cells to gemcitabine, a standard drug used for the treatment of pancreatic cancer. These investigations were done in AsPC-1, MIA PaCa-2, and Panc-28 cells and in nude mice orthotopically implanted with Panc-28 cells. In vitro, UA inhibited proliferation, induced apoptosis, suppressed NF-κB activation and its regulated proliferative, metastatic, and angiogenic proteins. UA (20 μM) also enhanced gemcitabine (200 nM)-induced apoptosis and suppressed the expression of NF-κB-regulated proteins. In the nude mouse model, oral administration of UA (250 mg/kg) suppressed tumor growth and enhanced the effect of gemcitabine (25 mg/kg). Furthermore, the combination of UA and gemcitabine suppressed the metastasis of cancer cells to distant organs such as liver and spleen. Immunohistochemical analysis showed that biomarkers of proliferation (Ki-67) and microvessel density (CD31) were suppressed by the combination of UA and gemcitabine. UA inhibited the activation of NF-κB and STAT3 and the expression of tumorigenic proteins regulated by these inflammatory transcription factors in tumor tissue. Furthermore, the combination of two agents decreased the expression of miR-29a, closely linked with tumorigenesis, in the tumor tissue. UA was found to be bioavailable in animal serum and tumor tissue. These results suggest that UA can inhibit the growth of human pancreatic tumors and sensitize them to gemcitabine by suppressing inflammatory biomarkers linked to proliferation, invasion, angiogenesis, and metastasis.

Synthesis, Characterization and In Vitro Anticancer Activity of C-5 Curcumin Analogues with Potential to Inhibit TNF-α-Induced NF-κB Activation

In a search of new compounds active against cancer, synthesis of a series of C-5 curcumin analogues was carried out. The new compounds demonstrated good cytotoxicity against chronic myeloid leukemia (KBM5) and colon cancer (HCT116) cell lines. Further, these compounds were found to have better potential to inhibit TNF-α-induced NF-κB activation in comparison to curcumin, which show their potential to act as anti-inflammatory agents. Some compounds were found to show higher cytotoxicity against cancer cell lines in comparison to curcumin used as standard.

Antimicrobial Potential and Chemical Characterization of Serbian Liverwort (Porella arboris-vitae): SEM and TEM Observations

The chemical composition of Porella arboris-vitae extracts was determined by solid phase microextraction, gas chromatography-mass spectrometry (SPME GC-MS), and 66 constituents were identified. The dominant compounds in methanol extract of P. arboris-vitae were β-caryophyllene (14.7%), α-gurjunene (10.9%), α-selinene (10.8%), β-elemene (5.6%), γ-muurolene (4.6%), and allo-aromadendrene (4.3%) and in ethanol extract, β-caryophyllene (11.8%), α-selinene (9.6%), α-gurjunene (9.4%), isopentyl alcohol (8.8%), 2-hexanol (3.7%), β-elemene (3.7%), allo-aromadendrene (3.7%), and γ-muurolene (3.3%) were the major components. In ethyl acetate extract of P. arboris-vitae, undecane (11.3%), β-caryophyllene (8.4%), dodecane (6.4%), α-gurjunene (6%), 2-methyldecane (5.1%), hemimellitene (4.9%), and D-limonene (3.9%) were major components. The antimicrobial activity of different P. arboris-vitae extracts was evaluated against selected food spoilage microorganisms using microbroth dilution method. The Minimal Inhibitory Concentration (MIC) varied from 0.5 to 1.5u2009mg/mL and 1.25 to 2u2009mg/mL for yeast and bacterial strains, respectively. Significant morphological and ultrastructural alterations due to the effect of methanolic and ethanolic P. arboris-vitae extracts on S. Enteritidis have also been observed by scanning electron microscope and transmission electron microscope, respectively. The results provide the evidence of antimicrobial potential of P. arboris-vitae extracts and suggest its potential as natural antimicrobial agents for food preservation.

Extraction and characterization of volatile compounds and fatty acids from red and green macroalgae from the Romanian Black Sea in order to obtain valuable bioadditives and biopreservatives

Three species of macroalgae, Ceramium virgatum (Rhodophyta), Ulva intestinalis, and Cladophora vagabunda (Chlorophyta), harvested from the Romanian Black Sea coast, were studied as sources of valuable compounds that could be used as additives and biopreservatives. Volatile compounds including hexanal (11.2xa0%), octane (9.8xa0%), nonanal (7.0xa0%), octanal (6.7xa0%), 2,5,5-trimethyl-2-hexene (4.7xa0%), 3-hexen-2-one (4xa0%), and o-cymene (3.6xa0%) were identified as the major components in the biomass extract of C. vagabunda. In C. virgatum, the major volatile components were 3-hexen-2-one (27.9xa0%), acetone (12.4xa0%), hexanal (3.4xa0%), and o-cymene (2.7xa0%). The major volatile compounds of U. intestinalis were hexanal (14.6xa0%), trichloromethane (7.3xa0%), nonanal (5.6xa0%), 3-hexen-2-one (5.3xa0%), and octanal (3.1xa0%). Some of these compounds have industrial applications as additives in the food, pharmaceutical, or cosmetics industries. The U. intestinalis extract had a greater content of mono- and polyunsaturated fatty acids around 46.0xa0% as compared with 42.0xa0% for C. vagabunda and 31.9xa0% for C. virgatum. The most abundant fatty acids were palmitic acid (C16:0), arachidonic acid (C20:4n-6), and oleic acid (C18:1ω-9cis). The antimicrobial effect of fatty acid extracts was tested against four pathogenic bacteria. The minimum inhibitory concentrations of C. vagabunda, C. virgatum, and U. intestinalis fatty acids extracts were 1.8, 3.8, and 3.8xa0mgxa0mL−1, respectively, for all bacterial strains. This study can help the efforts of finding new, value-added uses for natural marine resources.