Effect of Lipopeptide-Loaded Chitosan Nanoparticles on Candida albicans Adhesion and on the Growth of Leishmania major.

Cyclic lipopeptides produced by Bacillus species exhibit interesting therapeutic potential. However, their clinical use remains limited due to their low stability, undesirable interactions with host macromolecules, and their potential toxicity to mammalian cells. The present work aims to develop suitable lipopeptide-loaded chitosan nanoparticles with improved biological properties and reduced toxicity. Surfactin and bacillomycin D lipopeptides produced by Bacillus amyloliquefaciens B84 strain were loaded onto chitosan nanoparticles by ionotropic gelation process. Nanoformulated lipopeptides exhibit an average size of 569 nm, a zeta potential range of 38.8 mV, and encapsulation efficiency (EE) of 85.58%. Treatment of Candida (C.) albicans cells with encapsulated lipopeptides induced anti-adhesive activity of 81.17% and decreased cell surface hydrophobicity (CSH) by 25.53% at 2000 µg/mL. Nanoformulated lipopeptides also induced antileishmanial activity against Leishmania (L.) major promastigote and amastigote forms at respective IC50 values of 14.37 µg/mL and 22.45 µg/mL. Nanoencapsulated lipopeptides exerted low cytotoxicity towards human erythrocytes and Raw 264.7 macrophage cell line with respective HC50 and LC50 values of 770 µg/mL and 234.56 µg/mL. Nanoencapsulated lipopeptides could be used as a potential delivery system of lipopeptides to improve their anti-adhesive effect against C. albicans cells colonizing medical devices and their anti-infectious activity against leishmania.

Potential of Exopolysaccharide from Porphyridium marinum to Contend with Bacterial Proliferation, Biofilm Formation, and Breast Cancer.

Exopolysaccharide (EPS) from marine microalgae are promising sources of a new generation of drugs. However, lot of them remain to be discovered and tested. In this study, EPS produced by Porphyridium marinum and its oligomers prepared by High Pressure Homogenizer have been tested for different biological activities, i.e., antibacterial, anti-fungal and antibiofilm activities on Candida albicans, as well as for their effects on the viability of murine breast cancer cells. Results have shown that all EPS samples present some biological activity. For antibacterial and antibiofilm activities, the native EPS exhibited a better efficiency with Minimum Inhibitory Concentration (MIC) from 62.5 µg/mL to 1000 µg/mL depending on the bacterial strain. For Candida albicans, the biofilm formation was reduced by about 90% by using only a 31.3 µg/mL concentration. Concerning breast cancer cells, lower molar masses fractions appeared to be more efficient, with a reduction of viability of up to 55%. Finally, analyses of polymers composition and viscosity measurements were conducted on all samples, in order to propose hypotheses involving the activities caused by the intrinsic properties of polymers.

Biological control of the soft rot bacterium Pectobacterium carotovorum by Bacillus amyloliquefaciens strain Ar10 producing glycolipid-like compounds.

Four hundred and fifty bacteria were evaluated for antagonistic activity against bacterial soft rot of potato caused by Pectobacterium carotovorum sp strain II16. A strain Ar10 exhibiting potent antagonist activity has been identified as Bacillus amyloliquefaciens on the basis of biochemical and molecular characterization. Cell free supernatant showed a broad spectrum of antibacterial activity against human and phytopathogenic bacteria in the range of 10-60 AU/mL. Incubation of P. carotovorum cells with increasing concentrations of the antibacterial compound showed a killing rate of 94.8 and 96% at MIC and 2xMIC respectively. In addition, the antibacterial agent did not exert haemolytic activity at the active concentration and has been preliminary characterized by TLC and GC-MS as a glycolipid compound. Treatment of potato tubers with strain Ar10 for 72 h significantly reduced the severity of disease symptoms (100 and 85.05% reduction of necrosis deep / area and weight loss respectively). The same levels in disease symptoms severity was also recorded following treatment of potato tubers with cell free supernatant for 1 h. Data suggest that protection against potato soft rot disease may be related to glycolipid production by strain Ar10. The present study affords new alternatives for anti-Pectobacterium carotovorum bioactive compounds against the soft rot disease of potato.

Enhanced B-phycoerythrin production by the red microalga Porphyridium marinum: A powerful agent in industrial applications.

The production of B-phycoerythrin (B-PE) from the red microalga Porphyridium marinum was optimized before to purify it and subsequently study its antioxidant activities. NaNO3, K2HPO4 and metal traces concentrations of the culture medium, and luminosity parameters were chosen, according to the Plackett-Burman design, as the most influent factors on the B-PE production by P. marinum. The optimization of these factors according to the Box-Behnken plan gave a maximum of B-PE production equal to 40 mg/g dry weight under the following conditions: NaNO3 = 3.4 g/L; K2HPO4 = 0 g/L; light intensity = 70 µmol photons/m2/s and metal solution = 1.5 mL/L. The B-PE also showed an interesting capacity to chelate Fe3+ (IC50 = 13.91 ±â€¯0.21 µg/mL) and a significant reducing power (OD700nm = 0.485 ±â€¯0.011 at 100 µg/mL). The present study reports the antioxidant potential of purified B-PE from P. marinum that could be potentially used as a source of bioactive protein for a wide range of cosmetic and pharmaceutical applications.

Antifungal mechanism of the combination of Cinnamomum verum and Pelargonium graveolens essential oils with fluconazole against pathogenic Candida strains.

The present study aimed to investigate the anti-Candida activity of ten essential oils (EOs) and to evaluate their potential synergism with conventional drugs. The effect on secreted aspartic protease (SAP) activity and the mechanism of action were also explored. The antifungal properties of essential oils were investigated using standard micro-broth dilution assay. Only Cinnamomum verum, Thymus capitatus, Syzygium aromaticum, and Pelargonium graveolens exhibited a broad spectrum of activity against a variety of pathogenic Candida strains. Chemical composition of active essential oils was performed by gas chromatography-mass spectrometry (GC-MS). Synergistic effect was observed with the combinations C. verum/fluconazole and P. graveolens/fluconazole, with FIC value 0.37. Investigation of the mechanism of action revealed that C. verum EO reduced the quantity of ergosterol to 83%. A total inhibition was observed for the combination C. verum/fluconazole. However, P. graveolens EO may disturb the permeability barrier of the fungal cell wall. An increase of MIC values of P. graveolens EO and the combination with fluconazole was observed with osmoprotectants (sorbitol and PEG6000). Furthermore, the combination with fluconazole may affect ergosterol biosynthesis and disturb fatty acid homeostasis in C. albicans cells as the quantity of ergosterol and oleic acid was reduced to 52.33 and 72%, respectively. The combination of P. graveolens and C. verum EOs with fluconazole inhibited 78.31 and 64.72% SAP activity, respectively. To our knowledge, this is the first report underlying the mechanism of action and the inhibitory effect of SAP activity of essential oils in synergy with fluconazole. Naturally occurring phytochemicals C. verum and P. graveolens could be effective candidate to enhance the efficacy of fluconazole-based therapy of C. albicans infections.

Production and identification of iturin A lipopeptide from Bacillus methyltrophicus TEB1 for control of Phoma tracheiphila.

A lipopeptide-producing endophytic Bacillus methyltrophicus TEB1 strain exhibited potent antifungal activity against Phoma tracheiphila. Lipopeptide production started at the early growth phase plateaued after 36 h of culture where it reduced the mycelium growth by 80%. The crude lipopeptide extract harvested at the stationary phase efficiently inhibited the growth of P. tracheiphila mycelium and MIC values displaying 50 and 90% inhibition of conidia germination were around 47.5 and 100 µg ml(-1) , respectively. Increasing lipopeptide extract till 3 mg ml(-1) induced 10% swelling and 3% crumbling of P. tracheiphila conidia whereas 5 mg ml(-1) induced 40% swelling and 20% crumbling. Mass spectrometry analysis of the lipopeptide extract indicated that surfactin production took place from 12 to 20 h, iturin A from 16 to 72 h, and fengycin from 12 to 72 h and that the main active compound against P. tracheiphila was identified as C15 iturin A lipopeptide. Iturin A appeared as a potential biological control agent able to substitute the currently used chemical pesticides in agriculture.

Characterization of endophytic Bacillus strains from tomato plants (Lycopersicon esculentum) displaying antifungal activity against Botrytis cinerea Pers.

Eighty endophytic bacteria were isolated from healthy tissues of roots, stems, leaves and fruits of tomato plants (Lycopersicon esculentum). Four strains, named BL1, BT5, BR8 and BF11 were selected for their antagonism against Botrytis cinerea, a phytopathogenic fungus responsible of gray mold in several important crops, with growth inhibitory activity ranging from 27 to 53%. Morphological, biochemical, and molecular parameters as 16S rDNA sequencing demonstrated that the selected bacterial strains were related to Bacillus species which are known to produce and secrete a lot of lipopeptides with strong inhibitory effect against pathogen mycelial growth. Electrospray mass spectrometry analysis showed that these strains produced heterogeneous mixture of antibiotics belonging to fengycin and surfactin for BL1 and BT5, to iturin and surfactin for BR8, to bacillomycin D, fengycin and surfactin for BF11. Furthermore, these bacteria exhibited biocontrol potential by reducing the disease severity when tested on detached leaflets. Based on their antifungal activity against Botrytis cinerea, these strains could be used for biological control of plant diseases.

Metabolic interactions between hyperhomocysteinemia and endothelin-1 among Tunisian patients with acute coronary diseases.

BACKGROUND: Acute coronary syndromes (ACS) are complex and polygenic diseases which are a real problem of public health. These syndromes require multidisciplinary studies to understand the pathogenesis mechanisms and metabolic interactions between different risk factors.This study aimed to explore the variation of two coronary risk parameters not mentioned by Framingham cohorts, hyperhomocysteinemia and endothelin-1 (ET-1) in Tunisian coronary and the study of the variation of these parameters based on various cardiac risk factors and metabolic relationship between them.To 157 coronary and 142 healthy subjects, the concentration of homocysteine was quantified by fluorescence polarization immunoassay; the concentration of ET-1 was measured by an analytical technique, the High Performance Liquid Chromatography (HPLC) coupled with mass spectrometry. RESULTS: Our study showed that homocysteine and ET-1 were significantly higher in patients compared to healthy subjects (24.40 ± 12.5 µmol/L vs 7.44 ± 2.5 µmol/L p <0.00001) for homocysteine and (15.2 ± 5.3 nmol/L vs 7.1 ± 2.7 nmol/L, p <0.00001) for ET-1. On the other hand, homocysteine varies according to tobacco and diabetes while ET-1 depends on the sex, hypertension, smoking, obesity and dyslipidemia and a statistically negative correlation was shown between homocysteine and ET-1 in coronary patients (r = -0.66 p <0.00001). CONCLUSION: The study of the variation of these two parameters in coronary patients and metabolic exploration of the relationship between homocysteine and ET-1 according to various risk factors and the interactions between themselves facilitates the decision of therapeutic treatment.

Metabolic interactions between hyperhomocysteinemia and endothelin-1 among Tunisian patients with acute coronary diseases

BACKGROUND: Acute coronary syndromes (ACS) are complex and polygenic diseases which are a real problem of public health. These syndromes require multidisciplinary studies to understand the pathogenesis mechanisms and metabolic interactions between different risk factors.This study aimed to explore the variation of two coronary risk parameters not mentioned by Framingham cohorts, hyperhomocysteinemia and endothelin-1 (ET-1) in Tunisian coronary and the study of the variation of these parameters based on various cardiac risk factors and metabolic relationship between them.To 157 coronary and 142 healthy subjects, the concentration of homocysteine was quantified by fluorescence polarization immunoassay; the concentration of ET-1 was measured by an analytical technique, the High Performance Liquid Chromatography (HPLC) coupled with mass spectrometry. RESULTS: Our study showed that homocysteine and ET-1 were significantly higher in patients compared to healthy subjects (24.40 ± 12.5 µmol/L vs 7.44 ± 2.5 µmol/L p <0.00001) for homocysteine and (15.2 ± 5.3 nmol/L vs 7.1 ± 2.7 nmol/L, p <0.00001) for ET-1. On the other hand, homocysteine varies according to tobacco and diabetes while ET-1 depends on the sex, hypertension, smoking, obesity and dyslipidemia and a statistically negative correlation was shown between homocysteine and ET-1 in coronary patients (r = -0.66 p <0.00001. CONCLUSION: The study of the variation of these two parameters in coronary patients and metabolic exploration of the relationship between homocysteine and ET-1 according to various risk factors and the interactions between themselves facilitates the decision of therapeutic treatment.

Protective effect of grape seed and skin extract against high-fat diet-induced liver steatosis and zinc depletion in rat.

BACKGROUND: Obesity is a tremendous public health problem, characterized by ectopic deposition of fat into non-adipose tissues as liver generating an oxidative stress that could lead to steato-hepatitis. Grape seed and skin extract (GSSE) is a complex mixture of polyphenolics exhibiting robust antioxidative properties. AIM: We hypothesize that GSSE could protect the liver from fat-induced lipotoxicity and have a beneficial effect on liver function. METHODS: Hepatoprotective effect of GSSE was measured by using an experimental model of fat-induced rat liver steatosis. Male rats were fed a standard diet or a high-fat diet (HFD) during 6 weeks and treated or not with 500 mg/kg bw GSSE. Lipid deposition into the liver was assessed by triglyceride, cholesterol and phospholipid measurements. Fat-induced lipoperoxidation, carbonylation, depletion of glutathione and of antioxidant enzyme activities were used as oxidative stress markers with a special emphasis on transition metal distribution. RESULTS: HFD induced liver hypertrophy and inflammation as assessed by high liver transaminases. HFD also induced an oxidative stress characterized by increased lipid and protein oxidation, a drop in glutathione and antioxidant enzyme activities as glutathione peroxidase and superoxide dismutase and a drastic depletion in liver zinc. Importantly, GSSE prevented all the deleterious effects of HFD treatment. CONCLUSIONS: Data suggest that GSSE could be used as a safe preventive agent against fat-induced liver lipotoxicity which could also have potential applications in other non-alcoholic liver diseases.

Grape seed and skin extract alleviates high-fat diet-induced renal lipotoxicity and prevents copper depletion in rat.

Obesity is a public health problem that contributes to morbidity and mortality from diabetes, heart disease, stroke, and cancers. The purpose of this investigation was to analyse the link between obesity-induced oxidative stress, renal steatosis, and kidney dysfunction, as well as the protective effect of grape seed and skin extract. Rats were fed a standard diet or a high-fat diet for 6 weeks and were either treated or not treated with grape seed and skin extract. Fat-induced oxidative stress was evaluated in the kidney with a special emphasis on transition metals. High-fat diet induced triglyceride deposition and disturbances in kidney function parameters, which are linked to an oxidative stress status and depletion of copper from the kidney. Grape seed and skin extract abrogated almost all fat-induced kidney disturbances. Grape seed and skin extract exerted potential protection against fat-induced kidney lipotoxicity and should find potential application in other kidney-related diseases.

Resveratrol Provides Cardioprotection after Ischemia/reperfusion Injury via Modulation of Antioxidant Enzyme Activities.

In this study, we investigated the cardioprotective effects of resveratrol. Rats were intraperitoneally administered with resveratrol (25 mg/kg bw) or vehicle (ethanol 10%) for 7 days and their heart subjected to ischemia/reperfusion injury. Isolated hearts were langendorff perfused, left ventricular functions as heart rate and developed pressure, as well as, heart antioxidant status were determined. Data showed that resveratrol improved recovery of post-ischemic ventricular functions when compared to control. Resveratrol also improved myocardial lipoperoxidation, free iron and antioxidant enzyme activities. Resveratrol decreased significantly catalase while it increased peroxidase and superoxide dismutase activities. In this later case, native PAGE analysis of superoxide dismutase isoforms revealed that resveratrol up regulated iron and manganese isoforms. Resveratrol exerted potential cardioprotection partly by its antioxidant properties.

Grape seed and skin extract prevents high-fat diet-induced brain lipotoxicity in rat.

Obesity is related to an elevated risk of dementia and the physiologic mechanisms whereby fat adversely affects the brain are poorly understood. The present investigation analyzed the effect of a high fat diet (HFD) on brain steatosis and oxidative stress and the intracellular mediators involved in signal transduction, as well as the protection offered by grape seed and skin extract (GSSE). HFD induced ectopic deposition of cholesterol and phospholipid but not triglyceride. Moreover brain lipotoxicity is linked to an oxidative stress characterized by increased lipoperoxidation and carbonylation, inhibition of glutathione peroxidase and superoxide dismutase activities, depletion of manganese and a concomitant increase in ionizable calcium and acetylcholinesterase activity. Importantly GSSE alleviated all the deleterious effects of HFD treatment. Altogether our data indicated that HFD could find some potential application in the treatment of manganism and that GSSE should be used as a safe anti-lipotoxic agent in the prevention and treatment of fat-induced brain injury.

Anti-candida effect of bacillomycin D-like lipopeptides from Bacillus subtilis B38.

Bacillus subtilis B38, isolated from soil, showed antimicrobial activity against human pathogenic Candida albicans species. Specific PCR primers revealed the presence of the bamC gene, which is involved in the biosynthesis of bacillomycin D. Three anti-Candida compounds designated a(1) , a(2) and a(3) were purified from culture supernatant and identified using matrix-assisted laser desorption/ionization time-of-flight MS as analogues of bacillomycin D-like lipopeptides of 14, 15 and 16 carbon fatty acid long chains, respectively. The compound a(3) displayed the strongest fungicidal activity against pathogenic C. albicans strains. It was even more active than amphotericin B with a lethal concentration of 59.07 vs. 135.26 µM of the antimycotic drug against the pathogenic strain C. albicans sp. 311 isolated from finger nail. Only moderate or weak anti-Candida activity was recorded for a(1) and a(2) compounds. Furthermore, a(3) showed the highest hemolytic activity, reaching 50% hemolysis at 22.14 µM, whereas a(1) and a(2) displayed a limited hemolysis at 68.26 and 37.41 µM, respectively. These findings suggest that the acyl chain length of bacillomycin D-like lipopeptides plays a major role in hemolytic and antifungal activities.

Triggering of the antibacterial activity of Bacillus subtilis B38 strain against methicillin-resistant Staphylococcus aureus.

When cultured in minimal growth medium, the B38 strain of Bacillus subtilis did not exhibit any antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA) clinical isolate. Coculturing B38 strain with viable MRSA cells weakly increased antibacterial activity production (20 AU/ml). Addition of dead MRSA cells in a B38 culture, increased by 8-fold the B. subtilis strain antibacterial activity reaching 160 AU/ml against MRSA strain. This antibacterial activity recovered from cell-free supernatants was stimulated by an autoinducing compound which is sensitive to the action of proteinase K suggesting a proteinaceous nature. This compound was heat-stable till 80 °C and showed a molecular mass around 20 kDa as determined by SDS-PAGE. These results suggest that the production of antibacterial compounds by B38 strain is dependent on the amount of the autoinducing compound.

A new antibacterial and antioxidant S07-2 compound produced by Bacillus subtilis B38.

An antibacterial compound, S07-2, was purified to homogeneity by hydrophobic interaction, anion exchange, C18 reverse-phase and HS PEG HPLC. The molecular mass of S07-2 was 905.6 Da as determined by MS. The S07-2 compound was resistant to high temperatures (up to 100 degrees C) and could withstand a wide range of pH from 3 to 10. In addition, its antibacterial activity was preserved after treatment with proteases. Biochemical characterization revealed its cyclic peptide structure. This compound showed a bactericidal effect against important food-spoilage bacteria and food-borne pathogens including Listeria monocytogenes and Enterococcus faecalis with lethal concentration values of 62.5 microg mL(-1) and against Salmonella enteritidis at a concentration of 31.25 microg mL(-1). However, no cytotoxic effect against human erythrocytes was recorded. Furthermore, the S07-2 compound displayed a remarkable Fe(2+)-chelating activity (EC(50)=9.76 microg mL(-1)) and 1-diphenyl-2-picrylhydrazyl-scavenging capacity (IC(50)=65 microg mL(-1)). All these chemical and biological features make S07-2 a useful compound in the food industry as a natural preservative.