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1.
Biofouling ; 31(8): 639-49, 2015.
Article in English | MEDLINE | ID: mdl-26362127

ABSTRACT

This study assessed the effect of temperature on the release of essential oil components incorporated by melt compounding into polymeric films. Specifically, polyethylene-co-vinylacetate (EVA) films containing carvacrol (CAR) and cinnamaldehyde (ALD), alone and in combination, were prepared and their surface and mechanical properties and antibacterial and anti-biofilm activity against Escherichia coli and Staphylococcus aureus were evaluated. The addition of ALD and CAR did not provoke variation in the surface morphology of EVA and allowed their delivery. At 37°C, films containing CAR, ALD or their combination (25+75%) were found to have the strongest bactericidal effect, whereas at lower temperatures a lower killing rate was observed. There was no clear evidence of the influence of temperature on the anti-biofilm activity of the essential oil component-based polymeric films. The biomass formed on EVA containing ALD, CAR or their combination (25+75) was significantly lower (60-80% reduction) than that formed on the EVA control at both 37° and 22°C.


Subject(s)
Anti-Bacterial Agents/pharmacology , Biofilms/drug effects , Escherichia coli/drug effects , Oils, Volatile/chemistry , Staphylococcus aureus/drug effects , Temperature , Acrolein/analogs & derivatives , Acrolein/chemistry , Acrolein/pharmacology , Anti-Bacterial Agents/chemistry , Biofilms/growth & development , Cymenes , Escherichia coli/growth & development , Monoterpenes/chemistry , Monoterpenes/pharmacology , Polyethylenes/chemistry , Polyethylenes/pharmacology , Polyvinyls/chemistry , Polyvinyls/pharmacology , Staphylococcus aureus/growth & development
2.
Appl Microbiol Biotechnol ; 97(21): 9515-23, 2013 Nov.
Article in English | MEDLINE | ID: mdl-23989976

ABSTRACT

The development of new polymeric materials aimed to control the bacterial biofilm appears to be an important practical approach. The goal of the present study was to prepare and characterize poly(ethylene-co-vinyl acetate) copolymer (EVA) films containing citronellol, eugenol, and linalool and evaluate their efficiency on growth and biofilm formation of Listeria monocytogenes, Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Pseudomonas aeruginosa in monospecies and dual species. The results showed that the addition of oil components influenced the elastic modulus (15 % decrease), the tensile stress (30 % decrease), the elongation at break (10 % increase), and the contact angle values (10-20° decrease) while leaving the homogeneity of the surface unaltered. Among the polymeric films, EVA + citronellol and EVA + eugenol at 7 wt% had the best inhibitory effect. After 24-48 h of incubation, EVA + citronellol was more effective against the growth (30-60 % reduction) than EVA + eugenol (15-30 % inhibition). However, this inhibition decreased after 240 h of incubation. On the contrary, the biofilm evaluation revealed a strong inhibition trend also after prolonged incubation time: the amount of biomass per square centimeter formed on copolymer with oil components was significantly less (40-70 % decrease) than that on pure copolymer control for L. monocytogenes, S. aureus, and E. coli. When polymeric materials were simultaneously inoculated with combinations of S. aureus and E. coli, the biomass accumulated was higher for EVA + citronellol and lower for EVA + eugenol than that in monoculture biofilm. The findings were similar to the results obtained by 2,3-bis[2-methyloxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide assay that measures the metabolic activity of viable cells.


Subject(s)
Anti-Bacterial Agents/metabolism , Bacteria/drug effects , Bacteria/growth & development , Biofilms/drug effects , Biofilms/growth & development , Oils, Volatile/metabolism , Polyvinyls/metabolism , Anti-Bacterial Agents/pharmacology , Bacterial Physiological Phenomena , Oils, Volatile/pharmacology , Polyvinyls/pharmacology
3.
Appl Microbiol Biotechnol ; 96(4): 1029-38, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22555914

ABSTRACT

Polyethylene-co-vinylacetate (EVA) films with different concentrations (3.5 wt% and 7 wt%) of essential oil constituents, carvacrol or cinnamaldehyde, were prepared and characterized by mechanical, antibacterial and antibiofilm properties. The incorporation of the compounds into copolymer films affected their elastic modulus, tensile stress and elongation at break. Carvacrol and cinnamaldehyde act as plasticizers which reduce the intermolecular forces of polymer chains, thus improving the flexibility and extensibility of the film. The analysis of the surface characteristics demonstrated that essential oil constituents lowered the contact angle values without causing any remarkable variation of the surface roughness. The films allowed progressive diffusion of the bioactive molecules and the kinetic of release was correlated with the damaging effect on bacterial growth. The kill curves proved that the film with essential oil constituents (7 wt%) had a significant bactericidal effect (reduction of 4 and 2 log CFU) against Staphylococcus aureus and Escherichia coli and a bacteriostatic effect against Staphylococcus epidermidis and Listeria monocytogenes (reduction of about 1 log CFU). With regard to biofilm formation the biomass formed on polymeric films surface was significantly reduced if compared with the pure copolymer control. The results were confirmed by fluorescence microscopy images by Live/dead staining. The reduction in the surface tension coupled to an inherent bactericidal property of carvacrol and cinnamaldehyde could in turn affect the initial attachment phase of bacteria and compromise the normal biofilm development.


Subject(s)
Acrolein/analogs & derivatives , Biofilms/drug effects , Food Packaging/instrumentation , Monoterpenes/chemistry , Polymers/chemistry , Acrolein/chemistry , Acrolein/pharmacology , Anti-Bacterial Agents/chemical synthesis , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Cymenes , Escherichia coli O157/drug effects , Escherichia coli O157/physiology , Kinetics , Listeria monocytogenes/drug effects , Listeria monocytogenes/physiology , Monoterpenes/pharmacology , Polymers/chemical synthesis , Polymers/pharmacology , Staphylococcus aureus/drug effects , Staphylococcus aureus/physiology
4.
Clin Nutr ; 30(2): 221-33, 2011 Apr.
Article in English | MEDLINE | ID: mdl-20864228

ABSTRACT

BACKGROUND & AIMS: Functional deficits following spinal cord injury (SCI) arise from both mechanical injury and from secondary tissue reactions involving inflammation. Natural almond skins (NS) were tested to evaluate anti-inflammatory effects on an animal model of SCI. METHODS: SCI was induced by the application of vascular clips to the dura via a four-level T5-T8 laminectomy. In the present study, to elucidate whether the protective effects of NS are related to the total phenolic content, we also investigated the effect of a blanched (BS) almond skins (industrially obtained by removing bran from the nut) in SCI. NS and BS (30 mg/kg respectively) were administered per os, 1 h and 6 h, after SCI. RESULTS: SCI in mice resulted in severe injury characterized by edema, tissue damage, production of inflammatory mediators and apoptosis (measured by Bax, Bcl-2 and Tunel assay). NS treatment, 1 and 6 h after SCI, reduced all parameters of inflammation as neutrophil infiltration, NF-κB activation, PAR formation, iNOS expression and apoptosis. However, treatment with BS did not exert any protective effect. CONCLUSIONS: Our results suggest that NS treatment, reducing the development of inflammation and tissue injury, may be useful in the treatment of SCI.


Subject(s)
Anti-Inflammatory Agents/pharmacology , Neuroprotective Agents/pharmacology , Prunus/chemistry , Spinal Cord Injuries/drug therapy , Spinal Cord Injuries/pathology , Analysis of Variance , Animals , Apoptosis/drug effects , Disease Models, Animal , Inflammation/complications , Inflammation/drug therapy , Lipid Peroxidation/drug effects , Male , Mice , NF-kappa B/metabolism , Neutrophil Infiltration/drug effects , Plant Extracts/pharmacology , Plant Structures/chemistry , Spinal Cord Injuries/complications , Tyrosine/analogs & derivatives , Tyrosine/metabolism
5.
Osteoporos Int ; 22(11): 2869-77, 2011 Nov.
Article in English | MEDLINE | ID: mdl-21116815

ABSTRACT

UNLABELLED: This study evaluates cytokines production in bone and bone marrow of patients with an osteoporotic fracture or with osteoarthritis by real time PCR, Western blot and immunohistochemistry. We demonstrate that the cytokine pattern is shifted towards osteoclast activation and osteoblast inhibition in patients with osteoporotic fractures. INTRODUCTION: Fragility fractures are the resultant of low bone mass and poor bone architecture typical of osteoporosis. Cytokines involved in the control of bone cell maturation and function are produced by both bone itself and bone marrow cells, but the roles of these two sources in its control and the amounts they produce are not clear. This study compares their production in patients with an osteoporotic fracture and those with osteoarthritis. METHODS: We evaluated 52 femoral heads from women subjected to hip-joint replacement surgery for femoral neck fractures due to low-energy trauma (37), or for osteoarthritis (15). Total RNA was extracted from both bone and bone marrow, and quantitative PCR was used to identify the receptor activator of nuclear factor kB Ligand (RANKL), osteoprotegerin (OPG), macrophage colony stimulating factor (M-CSF), transforming growth factor ß (TGFß), Dickoppf-1 (DKK-1) and sclerostin (SOST) expression. Immunohistochemistry and Western blot were performed in order to quantify and localize in bone and bone marrow the cytokines. RESULTS: We found an increase of RANKL/OPG ratio, M-CSF, SOST and DKK-1 in fractured patients, whereas TGFß was increased in osteoarthritic bone. Bone marrow produced greater amounts of RANKL, M-CSF and TGFß compared to bone, whereas the production of DKK-1 and SOST was higher in bone. CONCLUSIONS: We show that bone marrow cells produced the greater amount of pro-osteoclastogenic cytokines, whereas bone cells produced higher amount of osteoblast inhibitors in patients with fragility fracture, thus the cytokine pattern is shifted towards osteoclast activation and osteoblast inhibition in these patients.


Subject(s)
Bone Marrow/metabolism , Cytokines/metabolism , Femur Head/metabolism , Osteoarthritis/metabolism , Osteoporotic Fractures/metabolism , Adaptor Proteins, Signal Transducing , Aged , Aged, 80 and over , Blotting, Western , Bone Morphogenetic Proteins/metabolism , Female , Genetic Markers , Humans , Intercellular Signaling Peptides and Proteins/metabolism , Macrophage Colony-Stimulating Factor/metabolism , Middle Aged , Osteoblasts/metabolism , Osteoclasts/metabolism , Osteoprotegerin/metabolism , RANK Ligand/metabolism , Real-Time Polymerase Chain Reaction , Transforming Growth Factor beta/metabolism
6.
Antiviral Res ; 89(1): 83-8, 2011 Jan.
Article in English | MEDLINE | ID: mdl-21095205

ABSTRACT

Our previous study demonstrated that Melaleuca alternifolia (tea tree) oil (TTO) had an interesting antiviral activity against Influenza A in MDCK cells. In fact, when we tested TTO and some of its components, we found that TTO had an inhibitory effect on influenza virus replication at doses below the cytotoxic dose; terpinen-4-ol, terpinolene, and alfa-terpineol were the main active components. The aim of this study was to investigate the mechanism of action of TTO and its active components against Influenza A/PR/8 virus subtype H1N1 in MDCK cells. None of the test compounds showed virucidal activity nor any protective action for the MDCK cells. Thus, the effect of TTO and its active components on different steps of the replicative cycle of influenza virus was studied by adding the test compounds at various times after infection. These experiments revealed that viral replication was significantly inhibited if TTO was added within 2h of infection, indicating an interference with an early step of the viral replicative cycle of influenza virus. The influence of the compound on the virus adsorption step, studied by the infective center assay, indicated that TTO did not interfere with cellular attachment of the virus. TTO did not inhibit influenza virus neuraminidase activity, as shown by the experiment measuring the amount of 4-methylumbelliferone, cleaved by the influenza virus neuraminidase from the fluorogenic substrate 2'-O-(4-methylumbelliferyl)-N-acetylneuraminic acid. The effect of TTO on acidification of cellular lysosomes was studied by vital staining with acridine orange using bafilomycin A1 as positive control. The treatment of cells with 0.01% (v/v) of TTO at 37°C for 4h before staining inhibited the acridine orange accumulation in acid cytoplasmic vesicles, indicating that TTO could inhibit viral uncoating by an interference with acidification of intralysosomal compartment.


Subject(s)
Antiviral Agents/pharmacology , Influenza A Virus, H1N1 Subtype/drug effects , Melaleuca/chemistry , Tea Tree Oil/pharmacology , Animals , Antiviral Agents/isolation & purification , Cell Line , Dogs , Hydrogen-Ion Concentration , Influenza A Virus, H1N1 Subtype/physiology , Lysosomes/chemistry , Lysosomes/drug effects , Lysosomes/virology , Tea Tree Oil/isolation & purification , Virus Internalization/drug effects , Virus Replication/drug effects
7.
Phytomedicine ; 17(5): 317-22, 2010 Apr.
Article in English | MEDLINE | ID: mdl-19699074

ABSTRACT

The application of antimicrobial combinations may address the rising resistance to established classes of both systemic and topical agents and their clinical relevance is related to the presence of a significant postantibiotic effect (PAE). We investigated the effectiveness in vitro of the association between tobramycin and tea tree oil (TTO) against Gram-positive and Gram-negative bacteria. The minimal inhibitory concentrations, the bacterial killing and the PAE of tobramycin and TTO were determined both singly and in combination against Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 29213. A synergistic interaction was observed against both strains tested: the mean PAEs were 1.3 and 1.7h for tobramycin against E. coli and S. aureus respectively, 10.8h for tobramycin and TTO (0.05%) against E. coli, 10.4h and 17.4h against S. aureus for tobramycin and TTO (0.25 and 0.50%, respectively). Longer PASMEs were observed with S. aureus after TTO/tobramycin exposure. In vitro interactions can improve the antimicrobial effectiveness of the antibiotic and may contribute for the development of novel topical agents for the treatment of skin lesions including conjunctiva and respiratory infections by inhalation.


Subject(s)
Anti-Bacterial Agents/pharmacology , Escherichia coli/drug effects , Melaleuca/chemistry , Plant Extracts/pharmacology , Staphylococcus aureus/drug effects , Tea Tree Oil/pharmacology , Tobramycin/pharmacology , Drug Resistance , Drug Synergism , Drug Therapy, Combination , Microbial Sensitivity Tests , Plant Extracts/chemistry
8.
Lett Appl Microbiol ; 49(6): 806-8, 2009 Dec.
Article in English | MEDLINE | ID: mdl-19843207

ABSTRACT

AIMS: To investigate the in vitro antiviral activity of Melaleuca alternifolia essential oil (TTO) and its main components, terpinen-4-ol, alpha-terpinene, gamma-terpinene, p-cymene, terpinolene and alpha-terpineol. METHODS AND RESULTS: The antiviral activity of tested compounds was evaluated against polio type 1, ECHO 9, Coxsackie B1, adeno type 2, herpes simplex (HSV) type 1 and 2 viruses by 50% plaque reduction assay. The anti-influenza virus assay was based on the inhibition of the virus-induced cytopathogenicity. Results obtained from our screening demonstrated that the TTO and some of its components (the terpinen-4-ol, the terpinolene, the alpha-terpineol) have an inhibitory effect on influenza A/PR/8 virus subtype H1N1 replication at doses below the cytotoxic dose. The ID(50) value of the TTO was found to be 0.0006% (v/v) and was much lower than its CD(50) (0.025% v/v). All the compounds were ineffective against polio 1, adeno 2, ECHO 9, Coxsackie B1, HSV-1 and HSV-2. None of the tested compounds showed virucidal activity. Only a slight virucidal effect was observed for TTO (0.125% v/v) against HSV-1 and HSV-2. CONCLUSIONS: These data show that TTO has an antiviral activity against influenza A/PR/8 virus subtype H1N1 and that antiviral activity has been principally attributed to terpinen-4-ol, the main active component. SIGNIFICANCE AND IMPACT OF THE STUDY: TTO should be a promising drug in the treatment of influenza virus infection.


Subject(s)
Antiviral Agents/pharmacology , Influenza A Virus, H1N1 Subtype/drug effects , Monoterpenes/pharmacology , Tea Tree Oil/pharmacology , Drug Resistance, Viral/drug effects , Enterovirus/drug effects , Inhibitory Concentration 50 , Insecticides/pharmacology , Microbial Sensitivity Tests , Simplexvirus/drug effects
9.
Appl Environ Microbiol ; 74(14): 4264-70, 2008 Jul.
Article in English | MEDLINE | ID: mdl-18502914

ABSTRACT

Almonds are known to have a number of nutritional benefits, including cholesterol-lowering effects and protection against diabetes. They are also a good source of minerals and vitamin E, associated with promoting health and reducing the risk for chronic disease. For this study we investigated the potential prebiotic effect of almond seeds in vitro by using mixed fecal bacterial cultures. Two almond products, finely ground almonds (FG) and defatted finely ground almonds (DG), were subjected to a combined model of the gastrointestinal tract which included in vitro gastric and duodenal digestion, and the resulting fractions were subsequently used as substrates for the colonic model to assess their influence on the composition and metabolic activity of gut bacteria populations. FG significantly increased the populations of bifidobacteria and Eubacterium rectale, resulting in a higher prebiotic index (4.43) than was found for the commercial prebiotic fructooligosaccharides (4.08) at 24 h of incubation. No significant differences in the proportions of gut bacteria groups were detected in response to DG. The increase in the numbers of Eubacterium rectale during fermentation of FG correlated with increased butyrate production. In conclusion, we have shown that the addition of FG altered the composition of gut bacteria by stimulating the growth of bifidobacteria and Eubacterium rectale.


Subject(s)
Bifidobacterium/growth & development , Eubacterium/growth & development , Nuts/chemistry , Prunus/chemistry , Butyrates/analysis , Colony Count, Microbial , Digestion , Duodenum/microbiology , Feces/microbiology , Fermentation , Food Analysis , Humans , Oligosaccharides/analysis
10.
Bioresour Technol ; 99(11): 5130-3, 2008 Jul.
Article in English | MEDLINE | ID: mdl-17951057

ABSTRACT

Feruloyl esterase (FAE) and xylanase activities were detected in culture supernatants from Humicola grisea var. thermoidea and Talaromyces stipitatus grown on brewers' spent grain (BSG) and wheat bran (WB), two agro-industrial by-products. Maximum activities were detected from cultures of H. grisea grown at 150 rpm, with 16.9 U/ml and 9.1 U/ml of xylanase activity on BSG and WB, respectively. Maximum FAE activity was 0.47 U/ml and 0.33 U/ml on BSG and WB, respectively. Analysis of residual cell wall material after microbial growth shows the preferential solubilisation of arabinoxylan and cellulose, two main polysaccharides present in BSG and WB. The production of low-cost cell-wall-deconstructing enzymes on agro-industrial by-products could lead to the production of low-cost enzymes for use in the valorisation of food processing wastes.


Subject(s)
Ascomycota/enzymology , Carboxylic Ester Hydrolases/biosynthesis , Endo-1,4-beta Xylanases/biosynthesis , Food-Processing Industry , Industrial Waste , Talaromyces/enzymology , Ascomycota/growth & development , Cell Wall/metabolism , Edible Grain/metabolism , Talaromyces/growth & development
11.
Phytother Res ; 22(3): 313-7, 2008 Mar.
Article in English | MEDLINE | ID: mdl-18058988

ABSTRACT

Herpes simplex virus type 1 (HSV-1) and herpes simplex virus type 2 (HSV-2) are common human pathogens that in particular cases can also cause severe problems especially in immunodeficient patients. The present paper reports the antiviral and immunomodulatory properties of a methanolic extract of C. spinosa buds (CAP), rich in flavonoids, including several quercetin and kaempferol glycosides. In particular we have investigated whether the in vitro exposure of human peripheral blood mononuclear cells (PBMCs) to CAP might inhibit the replication of HSV-2 and modulate the induction kinetics of IL-12, TNF-alpha IFN-gamma. Our findings have shown that CAP treatment interferes with HSV-2 replication in PBMCs inhibiting the extracellular virus release upregulating their production of IL-12, IFN-gamma and TNF-alpha. One could speculate that CAP may contribute in improving immune surveillance of PBMCs toward virus infection by up-regulating expression of peculiar proinflammatory cytokines; it should thus be successfully employed for treatment of HSV-2 infections in immunocompromised hosts.


Subject(s)
Antiviral Agents/pharmacology , Capparis/chemistry , Immunologic Factors/pharmacology , Leukocytes, Mononuclear/drug effects , Up-Regulation/drug effects , Cells, Cultured , Cytokines/analysis , Cytokines/biosynthesis , Flowers/chemistry , Freeze Drying , Herpesvirus 2, Human/growth & development , Leukocytes, Mononuclear/immunology , Leukocytes, Mononuclear/virology , Methanol/chemistry , Virus Replication/drug effects
12.
J Appl Microbiol ; 103(6): 2056-64, 2007 Dec.
Article in English | MEDLINE | ID: mdl-18045389

ABSTRACT

AIMS: To evaluate the antimicrobial properties of flavonoid-rich fractions derived from bergamot peel, a byproduct from the Citrus fruit processing industry and the influence of enzymatic deglycosylation on their activity against different bacteria and yeast. METHODS AND RESULTS: Bergamot ethanolic fractions were tested against Gram-negative bacteria (Escherichia coli, Pseudomonas putida, Salmonella enterica), Gram-positive bacteria (Listeria innocua, Bacillus subtilis, Staphylococcus aureus, Lactococcus lactis) and the yeast Saccharomyces cerevisiae. Bergamot fractions were found to be active against all the Gram-negative bacteria tested, and their antimicrobial potency increased after enzymatic deglycosylation. The minimum inhibitory concentrations of the fractions and the pure flavonoids, neohesperidin, hesperetin (aglycone), neoeriocitrin, eriodictyol (aglycone), naringin and naringenin (aglycone), were found to be in the range 200 to 800 microg ml(-1). The interactions between three bergamot flavonoids were also evaluated. CONCLUSION: The enzyme preparation Pectinase 62L efficiently converted common glycosides into their aglycones from bergamot extracts, and this deglycosylation increased the antimicrobial potency of Citrus flavonoids. Pairwise combinations of eriodictyol, naringenin and hesperetin showed both synergistic and indifferent interactions that were dependent on the test indicator organism. SIGNIFICANCE AND IMPACT OF THE STUDY: Bergamot peel is a potential source of natural antimicrobials that are active against Gram-negative bacteria.


Subject(s)
Anti-Bacterial Agents/pharmacology , Citrus , Flavonoids/pharmacology , Gram-Negative Bacteria/drug effects , Oils, Volatile , Plant Extracts/pharmacology , Bacteriological Techniques , Colony Count, Microbial , Flavanones/pharmacology , Gram-Positive Bacteria/drug effects , Hesperidin/pharmacology , Microbial Sensitivity Tests , Polygalacturonase/pharmacology , Saccharomyces cerevisiae/drug effects
13.
Appl Microbiol Biotechnol ; 73(5): 1173-9, 2007 Jan.
Article in English | MEDLINE | ID: mdl-17021882

ABSTRACT

The prebiotic effect of a pectic oligosaccharide-rich extract enzymatically derived from bergamot peel was studied using pure and mixed cultures of human faecal bacteria. This was compared to the prebiotic effect of fructo-oligosaccharides (FOS). Individual species of bifidobacteria and lactobacilli responded positively to the addition of the bergamot extract, which contained oligosaccharides in the range of three to seven. Fermentation studies were also carried out in controlled pH batch mixed human faecal cultures and changes in gut bacterial groups were monitored over 24 h by fluorescent in situ hybridisation, a culture-independent microbial assessment. Addition of the bergamot oligosaccharides (BOS) resulted in a high increase in the number of bifidobacteria and lactobacilli, whereas the clostridial population decreased. A prebiotic index (PI) was calculated for both FOS and BOS after 10 and 24 h incubation. Generally, higher PI scores were obtained after 10 h incubation, with BOS showing a greater value (6.90) than FOS (6.12).


Subject(s)
Bacteria/growth & development , Bacteria/metabolism , Feces/microbiology , Oligosaccharides/metabolism , Plant Extracts/metabolism , Bacteria/classification , Bacteria/isolation & purification , Fermentation , Humans , Hydrogen-Ion Concentration , In Situ Hybridization, Fluorescence
14.
Int J Immunopathol Pharmacol ; 20(4): 707-17, 2007.
Article in English | MEDLINE | ID: mdl-18179743

ABSTRACT

Vaccination with tumor-loaded dendritic cells (DC) is a promising treatment strategy for patients with renal cell carcinoma (RCC). Cells undergoing cell death proved useful as a source of tumor antigen for DC loading. Both apoptotic and necrotic tumor cells have been shown to efficiently load RCC-tumor antigens on DC. However, no direct comparison of these two kinds of death has been attempted in the same RCC. We compared DC pulsed with apoptotic cells, whole cell lysates or their supernatants of the cell line K1, derived from a patient with clear cell RCC, to determine their ability to activate T cells. Monocyte-derived DCs were pulsed with the different sources of tumor antigen, matured and co-cultured with autologouos peripheral blood lymphocytes. After three weekly re-stimulations with DCs, generation of cytotoxic T lymphocytes CTL was assessed by IFN-gamma release in an ELISpot assay in the presence of the sensitizing target. By comparison with lysate, apoptotic tumor cells induced a higher frequency of MHC class I-restricted IFN-gamma releasing lymphocytes. A higher CTL response was induced by pulsing DCs with cell lysate supernatant compared with whole cell lysate. These results indicate that, although necrotic death has been regarded as highly permissive when compared to apoptotic death, the immunogenicity of the death treatment may vary from one tumor to another.


Subject(s)
Antigen-Presenting Cells/physiology , Apoptosis/physiology , Carcinoma, Renal Cell/immunology , Carcinoma, Renal Cell/pathology , Kidney Neoplasms/immunology , Kidney Neoplasms/pathology , Antibodies, Monoclonal/pharmacology , Antigens, Neoplasm/pharmacology , Cell Line, Tumor , Cell Membrane/drug effects , Cell Membrane/pathology , Cell Proliferation/drug effects , Coculture Techniques , Cytokines/pharmacology , Dendritic Cells/drug effects , Enzyme-Linked Immunosorbent Assay , Humans , Interferon-gamma/biosynthesis , Lymphocyte Culture Test, Mixed , Necrosis , Phenotype , T-Lymphocytes/drug effects , T-Lymphocytes/immunology , T-Lymphocytes/metabolism
15.
Phytomedicine ; 13(8): 558-63, 2006 Sep.
Article in English | MEDLINE | ID: mdl-16920510

ABSTRACT

Olea europaea preparations are traditionally employed in a variety of troubles, including skin infections. Olive extracts and some of their pure compounds have shown antimicrobial activity in vitro. The present study deals with the antifungal activity of some aliphatic aldehydes from olive fruit [hexanal, nonanal, (E)-2-hexenal, (E)-2-heptenal, (E)-2-octenal, (E)-2-nonenal] against Tricophyton mentagrophytes (6 strains), Microsporum canis (1 strains) and Candida spp. (7 strains). The capability of these substances to inhibit elastase, a virulence factor essential for the dermatophytes colonization, and their cytotoxicity on cultures of reconstructed human epidermis, are also described. Aldehydes tested, inhibited the growth of T. mentagrophytes and M. canis in the range of concentration between <1.9 and 125 microg/ml; the unsaturated aldehydes showed the most broad spectrum of activity in that inhibited all strains tested. None of the aldehydes exhibited activity against Candida spp. strains. (E)-2-octenal and (E)-2-nonenal inhibited the elastase activity in a concentration-dependent manner; the anti-elastase activity suggests an additional target of the antimicrobial activity of these compounds. Aldehydes were devoid of cytotoxicity on cultures of human reconstructed epidermis. The antifungal activity of the aldehydes from olive fruit here reported, substantiates the use of olive and olive oil in skin diseases and suggests that these natural compounds could be useful agents in the topical treatment of fungal cutaneous infections.


Subject(s)
Antifungal Agents/therapeutic use , Candida/drug effects , Dermatomycoses/drug therapy , Olea/chemistry , Plant Extracts/therapeutic use , Antifungal Agents/toxicity , Candida/enzymology , Enzyme Inhibitors/analysis , Enzyme Inhibitors/therapeutic use , Epidermis/drug effects , Humans , Microbial Sensitivity Tests , Microsporum/drug effects , Microsporum/enzymology , Olea/toxicity , Plant Extracts/pharmacology
16.
Appl Microbiol Biotechnol ; 71(5): 622-9, 2006 Aug.
Article in English | MEDLINE | ID: mdl-16292533

ABSTRACT

The bioconversion of waste residues (by-products) from cereal processing industries requires the cooperation of enzymes able to degrade xylanolytic and cellulosic material. The type A feruloyl esterase from Aspergillus niger, AnFaeA, works synergistically with (1-->4)-beta-D-xylopyranosidases (xylanases) to release monomeric and dimeric ferulic acid (FA) from cereal cell wall-derived material. The esterase was more effective with a family 11 xylanase from Trichoderma viride in releasing FA and with a family 10 xylanase from Thermoascus aurantiacus in releasing the 5,5' form of diferulic acid from arabinoxylan (AX) derived from brewers' spent grain. The converse was found for the release of the phenolic acids from wheat bran-derived AXs. This may be indicative of compositional differences in AXs in cereals.


Subject(s)
Carboxylic Ester Hydrolases/metabolism , Coumaric Acids/metabolism , Edible Grain/metabolism , Endo-1,4-beta Xylanases/metabolism , Xylans/metabolism , Ascomycota/enzymology , Aspergillus niger/enzymology , Drug Synergism , Edible Grain/chemistry , Hydroxybenzoates/metabolism , Industrial Waste , Trichoderma/enzymology
17.
Minerva Pediatr ; 57(4): 173-80, 2005 Aug.
Article in English | MEDLINE | ID: mdl-16172596

ABSTRACT

AIM: Elimination of the offending food is imperative in the management of children with cow-milk allergy/intolerance (CMA/CMI). Herein we report the result of randomized clinical trial carried out to test the efficacy and safety of a new almond-based food (hereinafter named almond milk) in a group of infant with CMI/CMA. METHODS: A group of 52 infants aged 5 to 9 months and with documented CMI/CMA was enrolled and randomized to: almond milk (Group A, n=26); soy-based formula (Group B, n=13); protein hydrolysate-based formula (n=13). The main efficacy outcomes were the improvement in clinical symptoms and the decrease in serum levels of soluble CD30 (a potential marker for atopic disorders; sCD30). RESULTS: Elimination of the offending food and supplementation with a milk protein-free formula produced a considerable improvement of clinical manifestations within 5-12 days in all cases examined (at the onset of the study: 26.4+/-5.4 U/mL and 7.9+/-5.2 U/mL in IgE+ and IgE- infants respectively, after 6 months of supplementation: 16.6+/-4.8 U/mL and 7.1+/-4.5 U/mL in IgE+ and IgE- infants respectively). No difference in growth rate (increment of weight, length and head circumference) was found, during the entire study, between infants given the almond milk and babies given the soy-based formula or the protein hydrolysate-based formula. Supplementation with the soy-based and protein hydrolysate-based formulas caused the development, in some subjects, of a secondary sensitization (23% to soy-based and 15% protein hydrolysate-based formula), whereas supplementation with the almond milk did not. CONCLUSIONS: Though preliminary, the present findings seem to demonstrate that the almond milk may an efficacious substitute of cow milk in infants with CMA/CMI. One could speculate that some active principles contained in the almond milk could contribute to its beneficial effect observed in CMI/CMA-affected infants.


Subject(s)
Milk Substitutes , Milk/adverse effects , Prunus , Animals , Female , Humans , Immunoglobulin E/blood , Infant , Male , Milk Hypersensitivity
18.
J Ethnopharmacol ; 96(1-2): 227-32, 2005 Jan 04.
Article in English | MEDLINE | ID: mdl-15588675

ABSTRACT

Trichilia emetica Vahl. (Meliaceae) is a tree widely distributed in Tropical Africa. It has been used in Mali folk medicine for the treatment of various illnesses. The aim of this work was to study the hepatoprotective and antibacterial effects of a crude aqueous extract from Trichilia emetica root. An ethyl ether fraction from the aqueous extract was also prepared and studied. We have examined the hepatoprotective activity of the extracts on CCl4-induced damage in rat hepatocytes, their toxicity using the brine shrimp bioassay and their antibacterial activity against clinical isolated bacterial strains, which are commonly responsible for respiratory infections. A preliminary phytochemical analysis showed a high polyphenolic content in the aqueous extract and the presence of limonoids in the ethyl ether fraction. These latter compounds may be considered responsible for the good activity against the bacterial strains tested. Trichilia emetica extracts exerted also a significant (P<0.05) hepatoprotective effect at a dose of 1000 microg/ml both on plasma membrane and mitochondrial function as compared to silymarin used as a positive control. These activities may be a result of the presence of either polyphenols or limonoids. Finally, both the aqueous extract and its ethyl ether fraction did not show toxicity (LC50>1000 microg/ml) in the brine shrimp bioassay.


Subject(s)
Anti-Bacterial Agents/pharmacology , Chemical and Drug Induced Liver Injury/prevention & control , Hepatocytes/drug effects , Meliaceae , Protective Agents/pharmacology , Animals , Anti-Bacterial Agents/toxicity , Carbon Tetrachloride Poisoning/pathology , Carbon Tetrachloride Poisoning/prevention & control , Cell Survival/drug effects , Cells, Cultured , Chemical and Drug Induced Liver Injury/pathology , Haemophilus influenzae/drug effects , Hepatocytes/pathology , Male , Medicine, African Traditional , Microbial Sensitivity Tests , Moraxella catarrhalis/drug effects , Phenols/analysis , Plant Extracts/chemistry , Plant Extracts/pharmacology , Plant Extracts/toxicity , Protective Agents/toxicity , Rats , Rats, Wistar , Staphylococcus aureus/drug effects , Streptococcus pneumoniae/drug effects , Streptococcus pyogenes/drug effects
19.
Appl Microbiol Biotechnol ; 64(5): 644-50, 2004 Jun.
Article in English | MEDLINE | ID: mdl-14730410

ABSTRACT

An enzyme preparation from the thermophilic fungus Humicola insolens, Ultraflo L, was able to solubilise more than half of the biomass of brewer's grain and wheat bran, two agro-industrial co-products. While almost all of the ferulic acid was released in the free form, the majority of diferulates were released still attached to soluble feruloylated oligosaccharides, except for the 8,5' benzofuran form, which remained mostly in the residue. H. insolens also produced an esterase capable of releasing over 50% of p-coumaric acid present in wheat bran, but only 9% from the brewer's grain. The polysaccharide content in the residues after enzyme treatment comprised mostly cellulose and arabinoxylan, which suggests that part of the arabinoxylan in these residues is inaccessible to the xylanases of H. insolens. Differences in the solubilised arabinose-to-xylose ratio coupled to high free ferulate release suggest that the structure of feruloylated arabinoxylan in barley and wheat may differ.


Subject(s)
Ascomycota/metabolism , Carboxylic Ester Hydrolases/metabolism , Coumaric Acids/metabolism , Dietary Fiber/metabolism , Glycoside Hydrolases/metabolism , Hordeum/metabolism , Xylans/metabolism , Ascomycota/enzymology
20.
Inflamm Res ; 53(11): 601-3, 2004 Nov.
Article in English | MEDLINE | ID: mdl-15693608

ABSTRACT

Familial chronic nail candidiasis (FCNC.MIM 607644) is a rare disorder characterized by early onset infections caused by different species of Candida and restricted to the nails; this disorder is genetically associated with low serum concentration of intercellular adhesion molecule 1 (ICAM-1). Herein we report the evidence of high circulating levels of malondialdehyde (MDA) and 4-hydroxy-2,3-nonenal (HNE) in seven patients of a five-generation Italian family affected by FCNC.MIM 607644. The present data evidence, in these patients, an increase in circulating MDA and HNE levels. Only some merely speculative hypotheses may be suggested to explain the mechanisms subserving the oxidative stress condition observed in these genetically ICAM-1 deficient patients; however, one has to point out that a chronic oxidative stress condition could contribute to the development of concurrent pathological alterations in which an overproduction of free radicals may play a central role.


Subject(s)
Aldehydes/blood , Candidiasis, Chronic Mucocutaneous/diagnosis , Malondialdehyde/blood , Nail Diseases/diagnosis , Oxidative Stress , Adolescent , Adult , Biomarkers/blood , Candidiasis, Chronic Mucocutaneous/blood , Candidiasis, Chronic Mucocutaneous/metabolism , Child , Female , Humans , Intercellular Adhesion Molecule-1/metabolism , Italy , Male , Middle Aged , Nail Diseases/blood , Nail Diseases/metabolism
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