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1.
Materials (Basel) ; 11(8)2018 Jul 30.
Article in English | MEDLINE | ID: mdl-30061545

ABSTRACT

Ultrasonic molding is a new technology for processing small and micro polymeric components with reasonable cost and energy savings when small and medium batch sizes are required. However, when microcomponents are manufactured, the replicability of different micro features has to be guaranteed. The aim is to investigate the capability of ultrasonic molding technology for processing thin-wall plates of polystyrene with a microchannel, analyzing the filling behavior, the optical transparency, and the dimensional accuracy of the thin plate. The replicability of the manufactured microchannel is studied according to dimension and shape. The results reveal that plunger velocity influences transparency and filling cavity, whereas the vibration amplitude has less effect in both cases. The thickness deviation achieved on the final part is below 7% and the replication of the microchannel is better in depth than width, obtaining an average deviation of 4% and 11%, respectively. This replication also depends on the orientation of the microchannels and the distance from the injection gate. The replicability and repeatability for processing thin-wall plates with microchannel in polystyrene polymer are proved in this paper.

2.
Cereb Cortex ; 28(10): 3429-3444, 2018 10 01.
Article in English | MEDLINE | ID: mdl-29194486

ABSTRACT

Characteristic preterm EEG patterns of "Delta-brushes" (DBs) have been reported in the temporal cortex following auditory stimuli, but their spatio-temporal dynamics remains elusive. Using 32-electrode EEG recordings and co-registration of electrodes' position to 3D-MRI of age-matched neonates, we explored the cortical auditory-evoked responses (AERs) after 'click' stimuli in 30 healthy neonates aged 30-38 post-menstrual weeks (PMW). (1) We visually identified auditory-evoked DBs within AERs in all the babies between 30 and 33 PMW and a decreasing response rate afterwards. (2) The AERs showed an increase in EEG power from delta to gamma frequency bands over the middle and posterior temporal regions with higher values in quiet sleep and on the right. (3) Time-frequency and averaging analyses showed that the delta component of DBs, which negatively peaked around 550 and 750 ms over the middle and posterior temporal regions, respectively, was superimposed with fast (alpha-gamma) oscillations and corresponded to the late part of the cortical auditory-evoked potential (CAEP), a feature missed when using classical CAEP processing. As evoked DBs rate and AERs delta to alpha frequency power decreased until full term, auditory-evoked DBs are thus associated with the prenatal development of auditory processing and may suggest an early emerging hemispheric specialization.


Subject(s)
Audiometry, Evoked Response , Cerebral Cortex/physiology , Infant, Premature/physiology , Acoustic Stimulation , Alpha Rhythm/physiology , Delta Rhythm/physiology , Electroencephalography , Evoked Potentials, Auditory/physiology , Female , Gamma Rhythm , Humans , Infant, Newborn , Magnetic Resonance Imaging , Male , Sleep/physiology
3.
Colloids Surf B Biointerfaces ; 141: 19-27, 2016 May 01.
Article in English | MEDLINE | ID: mdl-26820106

ABSTRACT

Cationic double chain surfactants have attracted much interest because they can give rise to cationic vesicles that can be used in biomedical applications. Using a simple and economical synthetic approach, we have synthesized four double-chain surfactants with different alkyl chain lengths (LANHCx). The critical aggregation concentration of the double chain surfactants is at least one order of magnitude lower than the CMC of their corresponding single-chain LAM and the solutions prepared with the LANHCx contain stable cationic vesicles. Encouragingly, these new arginine derivatives show very low haemolytic activity and weaker cytotoxic effects than conventional dialkyl dimethyl ammonium surfactants. In addition, the surfactant with the shortest alkyl chain exhibits good antimicrobial activity against Gram-positive bacteria. The results show that a rational design applied to cationic double chain surfactants might serve as a promising strategy for the development of safe cationic vesicular systems.


Subject(s)
Anti-Infective Agents/chemistry , Arginine/chemistry , Cations/chemistry , Surface-Active Agents/chemistry , 3T3 Cells , Animals , Anti-Infective Agents/pharmacology , Cell Line, Transformed , Cell Survival/drug effects , Gram-Negative Bacteria/drug effects , Gram-Positive Bacteria/drug effects , HeLa Cells , Humans , Hydrophobic and Hydrophilic Interactions , Inhibitory Concentration 50 , Mice , Micelles , Microbial Sensitivity Tests , Models, Chemical , Molecular Structure , Surface Tension , Surface-Active Agents/pharmacology , Temperature
4.
J Appl Microbiol ; 120(1): 99-111, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26519210

ABSTRACT

AIMS: Apply response surface methodology (RSM) to develop and optimize an economical medium for lichenysin production, which is a surfactant produced by Bacillus licheniformis and evaluate the application of lichenysin in the prevention and disruption of pathogenic micro-organism biofilm that creates health problems in the food industry and hospitals. RESULTS: An economical medium containing molasses was optimized to enhance lichenysin production by RSM. A production of 3·2 g l(-1) of lichenysin was achieved with an optimum medium containing 107·82 g l(-1) of molasses, 6·47 g l(-1) of NaNO3 and 9·7 g l(-1) of K2 HPO4 /KH2 PO4 , in which molasses and phosphate salts had a significant effect on biosurfactant production. Lichenysin was effectively applied in a surface pre-treatment to avoid microbial biofilm development of methicillin-resistant Staphylococcus aureus (MRSA) (68·73%) and Candida albicans (74·35%), with ED50 values of 8·3 and 17·2 µg ml(-1) respectively. It was also very efficient in a surface post-treatment to remove biofilms of MRSA (55·74%) and Yersinia enterocolitica (51·51%), with an ED50 of 2·79 and 4·09 µg ml(-1) respectively. CONCLUSIONS: Lichenysin was found to have notable anti-adhesion activity, being able to prevent and eliminate the biofilm formation by pathogenic strains associated with foodborne illness. This new medium resulted in a four-fold increase in production compared with the nonoptimized medium. SIGNIFICANCE AND IMPACT OF THE STUDY: Molasses can be regarded as a useful resource for biotechnological applications, such as the production of lichenysin. The use of agro-industrial substrates has an important role in the sustainable and competitive development of several industrial sectors, as well as in industrial residues management. Additionally, lichenysin is particularly effective in preventing biofilm formation by strains problematic for the food industry and in the hospital environment. Lichenysin also efficiently disrupts biofilm.


Subject(s)
Bacillus/metabolism , Biofilms/drug effects , Candida albicans/drug effects , Methicillin-Resistant Staphylococcus aureus/drug effects , Surface-Active Agents/metabolism , Surface-Active Agents/pharmacology , Bacillus/chemistry , Candida albicans/physiology , Culture Media/metabolism , Methicillin-Resistant Staphylococcus aureus/physiology
5.
Biochim Biophys Acta ; 1838(3): 776-83, 2014 Mar.
Article in English | MEDLINE | ID: mdl-24239913

ABSTRACT

Rhamnolipids (RL) are one of the most important classes of biosurfactants produced by microorganisms using a wide range of carbon sources, from a simple carbon source like glucose to complex wastes such as the used cooking oils used in this work. The objective of this work was to learn about the rhamnolipid-phospholipid dipalmitoyl phosphatidyl choline (DPPC) molecular interactions through the behaviour observed in the neat products and four RL/DPPC mixtures. Size and z-potential were used to characterize the size and the charge of the vesicles, and small angle X-ray scattering (SAXS) was used to measure the vesicle bilayer characteristics, and the release of carboxyfluorescein to study the bilayer disrupting effect promoted by rhamnolipids. The results show that rhamnolipids are disposed in ordered bilayers with long repeating distances, which are stabilized by the charging of the bilayer and also by a strong fluidity of the bilayers. The ability of rhamnolipids to increase the fluidity of DPPC bilayers may be related with the strong haemolytic power of these molecules.


Subject(s)
1,2-Dipalmitoylphosphatidylcholine/metabolism , Erythrocytes/metabolism , Glycolipids/metabolism , Lipid Bilayers/metabolism , Water/metabolism , 1,2-Dipalmitoylphosphatidylcholine/chemistry , Calorimetry, Differential Scanning , Glycolipids/chemistry , Hemolysis , Humans , Lipid Bilayers/chemistry , Liposomes , Scattering, Small Angle , Water/chemistry , X-Ray Diffraction
6.
J Colloid Interface Sci ; 394: 368-79, 2013 Mar 15.
Article in English | MEDLINE | ID: mdl-23332937

ABSTRACT

The commercial application of a new biosurfactant such as the one produced by Sphingobacteriumdetergens needs a cost-effective process and knowledge of its properties. In the present study, a specific medium and a downstream process have been developed to enhance biosurfactant production. Optimal concentrations of nutrients in MCA medium were (g/L) the following: KH(2)PO(4), 1; K(2)HPO(4), 2; CO(NH(2))(2) 0.88; CaCl(2) 0.01; FeSO(4)·7H(2)O, 0.01; MgSO(4)·7H(2)O 0.5; KCl, 1.0; trace elements 0.05 mL. Biosurfactant production in the MCA medium required a bacterial co-metabolism of glucose and an n-alkane. A fed-batch culture with supernatant lyophilization prior to organic extraction produced 466 mg/L of organic extract, which represents a 6.9-fold increase in production. The newly obtained biosurfactant was a complex mixture of molecules. The three characterized fractions consisted of the complete fraction and two second-level purification fractions with apolar and polar characteristics. The complete and apolar fractions have been shown to self-aggregate in the form of lamellar liquid crystals at a high concentration and bilayers at lower concentrations. Negatively charged particles were identified, which were neutralized at a low pH with a concomitant increase in size. The pH affected the surface tension of the solutions congruently with phosphate headgroups.


Subject(s)
Industrial Microbiology/methods , Sphingobacterium/metabolism , Surface-Active Agents/chemistry , Surface-Active Agents/metabolism , Batch Cell Culture Techniques , Sphingobacterium/chemistry , Surface Tension , Surface-Active Agents/isolation & purification
7.
J Colloid Interface Sci ; 361(1): 195-204, 2011 Sep 01.
Article in English | MEDLINE | ID: mdl-21684551

ABSTRACT

Strain 6.2S, isolated from soil and identified as a Sphingobacterium sp., is the first strain in this genus to be reported as a biosurfactant producer, being able to reduce the surface tension of its culture supernatant to 32 mN/m. In this work, biosurfactants from the culture supernatant were purified and partially characterized. The crude extract (10 g/L) was very effective in reducing surface tension (22 mN/m). Thin layer chromatography (TLC) indicated that a mixture of various biosurfactants was present in the 6.2S crude extract. After purification, Fraction A, a phospholipid mixture, reduced surface tension to 33 mN/m. Fraction B was a mixture of lipopetides and at least one glycolipid. The surface tension-concentration curve showed two plateaux, the first of which can be attributed to a critical aggregation concentration of the biosurfactant with a protein (2.7 g/L) and the second to the true cmc in water (6.3g/L).


Subject(s)
Soil Microbiology , Sphingobacterium/chemistry , Surface-Active Agents/isolation & purification , Chromatography, Thin Layer , Glycolipids/isolation & purification , Lipopeptides/isolation & purification , Phospholipids/isolation & purification , Surface Tension
8.
Rev. calid. asist ; 26(2): 83-89, mar.-abr. 2011. tab
Article in Spanish | IBECS | ID: ibc-87982

ABSTRACT

Introducción. Las auditorías clínicas son el análisis crítico y sistemático de la calidad de la asistencia médica. La artroplastia total de cadera (ATC) primaria es un proceso habitual y coste-efectivo, aunque se dispone de poca información sobre la calidad asistencial de ella. Objetivo. Evaluar el impacto de un ciclo de auditorías clínicas en la calidad asistencial en el procedimiento de la ATC primaria por enfermedad no traumática. Pacientes y métodos. Se realizó un ciclo de 2 auditorías (primera auditoría en el 2005 y segunda auditoría en el 2007). Se incluyó a pacientes de ambos sexos con ATC primaria por enfermedad no traumática y un seguimiento de 6 meses. Se adoptaron como indicadores de gestión: el tiempo (días) de estancia ingresado en el hospital y el índice (porcentaje) de reingresos, y como indicadores de la práctica clínica: el índice (porcentaje) de luxación y el índice (porcentaje) de infección. Se compararon estos índices entre ambas auditorías. Resultados. Se estudió a un total de 160 pacientes: 79 y 81, primera y segunda auditoría, respectivamente. Indicadores de gestión: la mediana (rango) de los días de ingreso fue 8 (7-78) y 7 (6-16), p<0,001; el porcentaje de reingresos, el 5% (4/79) y 0 (0/81), p=0,057. Indicadores de la práctica clínica: el porcentaje de luxación fue el 8% (6/79) y 0 (0/81), p=0,013; y el porcentaje de infección el 1% (1/79) y el 1% (1/81), p=1. En un análisis multivariante no se encontraron otros factores relacionados con estos indicadores. Conclusiones. La implementación de un ciclo de auditorías clínicas ha mejorado la calidad asistencial del procedimiento de ATC primaria por enfermedad no traumática(AU)


Background. Clinical audits are critical and systematic quality analysis of medical care. Total hip arthroplasty (THA) is a routine practice and cost-effective, although there is little information on the quality of care of it. Objective. To evaluate the impact of a clinical audit cycle in the quality of care in the primary THA procedures for non-traumatic cause. Patients and methods. A series of two audits (first audit in 2005 and second one in 2007) were performed. Patients of both sexes with non-traumatic primary THA and with a follow-up of 6 months were included. Time (days) in hospital stay and the rate (percentage) of readmissions were used as indicators of management; and as indicators of clinical practice: the index (percentage) of dislocation and the rate (percentage) of infection. Both audits were compared with respect to these indicators. Results. A total of 160 patients (79 and 81, first and second audit respectively) were analysed. Management indicators: median (range) of hospital stay was 8 (7-78) and 7 (6-16), p<0.001, and the percentage of readmissions 5% (4/79) and 0 (0/81), p=0.057. Indicators of clinical practice: the rate of dislocation was 8% (6/79) and 0 (0/81), p=0.013, and the rate of infection 1% (1/79) and 1% (1/81), p=1. A multivariate analysis did not find other factors related to these indicators. Conclusions. The implementation of a clinical audit cycle has improved the quality of care of primary THA procedures for non-traumatic cause(AU)


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Clinical Audit , Arthroplasty, Replacement, Hip/economics , Arthroplasty, Replacement, Hip/methods , Medical Assistance/organization & administration , Medical Assistance/standards , Medical Assistance , Health Status Indicators , Comorbidity , Clinical Audit/methods , Clinical Audit/trends , Medical Assistance/statistics & numerical data , Medical Assistance/trends , Indicators of Health Services/standards , Multivariate Analysis , 28599 , Analysis of Variance
9.
Rev Calid Asist ; 26(2): 83-9, 2011.
Article in Spanish | MEDLINE | ID: mdl-21339078

ABSTRACT

BACKGROUND: Clinical audits are critical and systematic quality analysis of medical care. Total hip arthroplasty (THA) is a routine practice and cost-effective, although there is little information on the quality of care of it. OBJECTIVE: To evaluate the impact of a clinical audit cycle in the quality of care in the primary THA procedures for non-traumatic cause. PATIENTS AND METHODS: A series of two audits (first audit in 2005 and second one in 2007) were performed. Patients of both sexes with non-traumatic primary THA and with a follow-up of 6 months were included. Time (days) in hospital stay and the rate (percentage) of readmissions were used as indicators of management; and as indicators of clinical practice: the index (percentage) of dislocation and the rate (percentage) of infection. Both audits were compared with respect to these indicators. RESULTS: A total of 160 patients (79 and 81, first and second audit respectively) were analysed. Management indicators: median (range) of hospital stay was 8 (7-78) and 7 (6-16), p<0.001, and the percentage of readmissions 5% (4/79) and 0 (0/81), p=0.057. Indicators of clinical practice: the rate of dislocation was 8% (6/79) and 0 (0/81), p=0.013, and the rate of infection 1% (1/79) and 1% (1/81), p=1. A multivariate analysis did not find other factors related to these indicators. CONCLUSIONS: The implementation of a clinical audit cycle has improved the quality of care of primary THA procedures for non-traumatic cause.


Subject(s)
Arthroplasty, Replacement, Hip/statistics & numerical data , Medical Audit , Quality Improvement , Aged , Aged, 80 and over , Arthroplasty, Replacement, Hip/standards , Awards and Prizes , Comorbidity , Female , Hip Dislocation/epidemiology , Humans , Length of Stay/statistics & numerical data , Male , Osteoarthritis, Hip/surgery , Patient Readmission/statistics & numerical data , Postoperative Complications/epidemiology , Prosthesis-Related Infections/epidemiology , Quality Indicators, Health Care/statistics & numerical data , Spain/epidemiology , Surgical Wound Infection/epidemiology
10.
J Appl Microbiol ; 109(4): 1139-49, 2010 Oct.
Article in English | MEDLINE | ID: mdl-20456525

ABSTRACT

AIMS: To study cellular damage induced by Cinnamomum verum essential oil in Pseudomonas aeruginosa ATCC 27853 and Staphylococcus aureus ATCC 29213. METHODS AND RESULTS: The effect of cinnamon bark essential oil on these two strains was evaluated by plate counts, potassium leakage, flow cytometry and transmission electron microscopy (TEM). Exposure to this oil induced alterations in the bacterial membrane of Ps. aeruginosa, which led to the collapse of membrane potential, as demonstrated by bis-oxonol staining, and loss of membrane-selective permeability, as indicated by efflux of K(+) and propidium iodide accumulation. Thus, respiratory activity was inhibited, leading to cell death. In Staph. aureus, cells treated with the oil entered a viable but noncultivable (VNC) state. The oil initially caused a considerable decrease in the metabolic activity and in the replication capacity of these bacterial cells. The loss of membrane integrity appeared later, as indicated by bis-oxonol and Propidium iodide (PI) staining. Data provided by TEM showed various structural effects in response to cinnamon essential oil. In Ps. aeruginosa cells, coagulated cytoplasmic material was observed, and intracellular material was seen in the surrounding environment, while oil-treated Staph. aureus showed fibres extending from the cell surface. CONCLUSIONS: Cinnamon essential oil damages the cellular membrane of Ps. aeruginosa, which leads to cell death. There is evidence of VNC Staph. aureus after exposure to the oil. SIGNIFICANCE AND IMPACT OF THE STUDY: Cinnamon essential oil shows effective antimicrobial activity and health benefits and is therefore considered a potential food additive. To use this oil as a natural food preservative, especially in combination with other preservation methods, a thorough understanding of the mechanism through which this oil exerts its antibacterial action is required.


Subject(s)
Anti-Bacterial Agents/pharmacology , Oils, Volatile/pharmacology , Pseudomonas aeruginosa/drug effects , Staphylococcus aureus/drug effects , Cell Membrane Permeability/drug effects , Cinnamomum/chemistry , Membrane Potentials/drug effects , Potassium/metabolism , Pseudomonas aeruginosa/metabolism , Pseudomonas aeruginosa/ultrastructure , Staphylococcus aureus/metabolism , Staphylococcus aureus/ultrastructure , Thiobarbiturates/analysis
11.
Chem Phys Lipids ; 163(4-5): 341-6, 2010 May.
Article in English | MEDLINE | ID: mdl-20188718

ABSTRACT

In microorganisms hydroxy fatty acids are produced from the biotransformation of unsaturated fatty acids. Such compounds belong to a class of oxylipins which are reported to perform a variety of biological functions such as anti-inflammatory or cytotoxic activity. These compounds have been found in rice and timothy plants after being infected by specific fungus. When grown in submerged culture with linoleic acid, Pseudomonas 42A2 accumulated in the supernatant several hydroxy fatty acids. In this work LC-MS/MS has been used to elucidate the structure of the components form the organic extract: 9-hydroxy-10,12-octadecadienoic acid; 13-hydroxy-9,11-octadecadienoic acid; 7,10-dihydroxy-8E-octadecenoic acid; 9,10,13-trihydroxy-11-octadecenoic acid and 9,12,13-trihydroxy-10-octadecenoic acid. Antimicrobial activity against several pathogenic fungal strains is presented: MIC (microg/mL) Verticillium dhaliae, 32; Macrophonia phaesolina, 32; Arthroderma uncinatum, 32; Trycophyton mentagrophytes, 64.


Subject(s)
Antifungal Agents/administration & dosage , Antifungal Agents/chemistry , Chromatography, Liquid/methods , Fungi/drug effects , Mass Spectrometry/methods , Oxylipins/administration & dosage , Oxylipins/chemistry , Pseudomonas/metabolism , Antifungal Agents/isolation & purification , Cell Survival/drug effects , Cell-Free System/chemistry , Fungi/cytology , Oxylipins/isolation & purification
12.
Chem Phys Lipids ; 158(2): 110-7, 2009 Apr.
Article in English | MEDLINE | ID: mdl-19428355

ABSTRACT

This study analyzed the chemical and physical properties of a biosurfactant synthesized by Rhodococcus sp. 51T7. The biosurfactant was a trehalose tetraester (THL) consisting of six components: one major and five minor. The hydrophobic moieties ranged in size from 9 to 11 carbons. The critical micelle concentration (CMC) was 0.037g L(-1) and the interfacial tension against hexadecane was 5mN m(-1). At pH 7.4 the glycolipid CMC/critical aggregation concentration (CAC) was 0.05g L(-1) and at pH 4 it was 0.034g L(-1). A phase diagram revealed effective emulsification with water and paraffin or isopropyl myristate. A composition of 11.3-7.5-81.8 (isopropyl myristate-THL-W) was stable for at least 3 months. The HLB was 11 and the phase behaviour of the glycolipid revealed the formation of lamellar and hexagonal liquid-crystalline textures.


Subject(s)
Cell Survival/drug effects , Glycolipids/analysis , Glycolipids/toxicity , Rhodococcus/chemistry , Trehalose/analysis , Trehalose/toxicity , Animals , Cell Line , Emulsions/chemistry , Fibroblasts/cytology , Fibroblasts/drug effects , Genes, Bacterial , Genes, rRNA , Glycolipids/isolation & purification , Humans , Hydrogen-Ion Concentration , Keratinocytes/cytology , Keratinocytes/drug effects , Mice , Micelles , Osmolar Concentration , Phase Transition , Rhodococcus/genetics , Trehalose/isolation & purification
13.
J Appl Microbiol ; 106(5): 1558-68, 2009 May.
Article in English | MEDLINE | ID: mdl-19226402

ABSTRACT

AIMS: Evaluation of the cellular effects of Origanum compactum essential oil on Pseudomonas aeruginosa ATCC 27853 and Staphylococcus aureus ATCC 29213. METHODS AND RESULTS: The damage induced by O. compactum essential oil on these two strains has been studied using different techniques: plate count, potassium leakage, flow cytometry (FC) and transmission electron microscopy (TEM). The results showed that oil treatment led to reduction of cells viability and dissipated potassium ion gradients. Flow cytometric analysis showed that oil treatment promoted the accumulation of bis-oxonol and the membrane-impermeable nucleic acid stain propidium iodide (PI), indicating the loss of membrane potential and permeability. The ability to reduce 5-cyano-2,3-ditolyl tetrazolium chloride was inhibited. Unlike in Ps. aeruginosa, membrane potential and membrane permeability in Staph. aureus cells were affected by oil concentration and contact time. Finally, TEM showed various structural effects. Mesosome-like structures were seen in oil-treated Staph. aureus cells whereas in Ps. aeruginosa, coagulated cytoplasmic material and liberation of membrane vesicles were observed, and intracellular material was seen in the surrounding environment. Both FC and TEM revealed that the effects in Ps. aeruginosa were greater than in Staph. aureus. CONCLUSIONS: Oregano essential oil induces membrane damage showed by the leakage of potassium and uptake of PI and bis-oxonol. Ultrastructural alterations and the loss of cell viability were observed. SIGNIFICANCE AND IMPACT OF THE STUDY: Understanding the mode of antibacterial effect of the oil studied is of a great interest in it further application as natural preservative in food or pharmaceutical industries.


Subject(s)
Oils, Volatile/pharmacology , Origanum/chemistry , Plant Oils/pharmacology , Pseudomonas aeruginosa/drug effects , Staphylococcus aureus/drug effects , Anti-Bacterial Agents/pharmacology , Cell Membrane Permeability/drug effects , Chlorhexidine/pharmacology , Membrane Potentials/drug effects , Microbial Sensitivity Tests , Microscopy, Electron, Transmission , Oils, Volatile/chemistry , Plant Oils/chemistry , Polymyxin B/pharmacology , Potassium/metabolism , Pseudomonas aeruginosa/cytology , Pseudomonas aeruginosa/ultrastructure , Staphylococcus aureus/cytology , Staphylococcus aureus/ultrastructure
14.
Appl Microbiol Biotechnol ; 78(4): 587-96, 2008 Mar.
Article in English | MEDLINE | ID: mdl-18224316

ABSTRACT

Pseudomonas aeruginosa 42A2 produces a polyunsaturated polyhydroxyalkanoates (PHA-L) when grown on linseed oil as a substrate. Its high unsaturation content (36.5%) provides highly reactive PHA-L, generating a cross-linked biopolymer after ultraviolet (UV) irradiation. Both PHAs (PHA-L and uvPHA-L) were characterized by nuclear magnetic resonance, Fourier transform infrared spectroscopy, gel permeation chromatography, gas chromatography-mass spectrometry and differential scanning calorimetry-thermogravimetric analysis. The structural analysis of the new polymer revealed a dramatic decrease in unsaturated monomer content (8.5%), due to the complete disappearance of the polyunsaturated monomers (C(12:2), C(14:2), and C(14:3)). The cross-linking reaction was also confirmed by atomic force microscopy (AFM) and transmission electron microscopy. AFM showed morphological changes in bacteria cells with and without PHA granules. The microscope techniques provided us with micrographs of the native and cross-linked polymers, showing the formation of a reticular structure as the consequence of the cross-linking reaction.


Subject(s)
Inclusion Bodies/chemistry , Microscopy, Atomic Force , Microscopy, Electron, Transmission , Polyhydroxyalkanoates/chemistry , Pseudomonas aeruginosa/chemistry , Inclusion Bodies/metabolism , Inclusion Bodies/ultrastructure , Magnetic Resonance Spectroscopy , Polyhydroxyalkanoates/isolation & purification , Polyhydroxyalkanoates/metabolism , Polymers/chemistry , Polymers/metabolism , Pseudomonas aeruginosa/metabolism , Pseudomonas aeruginosa/ultrastructure , Ultraviolet Rays
15.
J Appl Microbiol ; 104(4): 1075-81, 2008 Apr.
Article in English | MEDLINE | ID: mdl-18005346

ABSTRACT

AIMS: To study the growth inhibitory properties of a series of phytosphingosine (PHS) and phytoceramide (PHC) analogues. METHODS AND RESULTS: A panel of two yeast (Candida albicans and Saccharomyces cerevisiae) and six moulds (Aspergillus repens, Aspergillus niger, Penicillium chrysogenum, Cladosporium cladosporioides, Arthroderma uncinatum and Penicillium funiculosum) has been used in this study. A series of new PHS and PHC analogues differing at the sphingoid backbone and the functional group at C1 position were synthesized. CONCLUSIONS: Among PHS analogues, 1-azido derivative 1c, bearing the natural D-ribo stereochemistry, showed a promising growth inhibitory profile. Among PHC analogues, compound 12, with a bulky N-pivaloyl group and a Z double bond at C3 position of the sphingoid chain, was the most active growth inhibitor. Minimal inhibitory concentration values were in the range of 23-48 micromol l(-1) for 1c and 44-87 micromol l(-1) for 12. SIGNIFICANCE AND IMPACT OF THE STUDY: Only scattered data on the antifungal activity of phytosphingolipids have been reported in the literature. This is the first time that a series of analogues of this kind are tested and compared to discern their structural requirements for antifungal activity.


Subject(s)
Antifungal Agents/pharmacology , Sphingolipids/pharmacology , Antifungal Agents/chemical synthesis , Arthrodermataceae/drug effects , Aspergillus/drug effects , Candida albicans/drug effects , Ceramides/chemical synthesis , Ceramides/chemistry , Ceramides/pharmacology , Cladosporium/drug effects , Microbial Sensitivity Tests , Penicillium/drug effects , Penicillium chrysogenum/drug effects , Saccharomyces cerevisiae/drug effects , Sphingolipids/chemical synthesis , Sphingolipids/chemistry , Sphingosine/chemical synthesis , Sphingosine/chemistry , Sphingosine/pharmacology
16.
J Appl Microbiol ; 96(5): 903-12, 2004.
Article in English | MEDLINE | ID: mdl-15078505

ABSTRACT

AIMS: Here we study the effect of monohydrochloride of L-arginine, N(alpha)-lauroyl ethylester (LAE), a cationic preservative derived from lauric acid and arginine, on the cell envelopes of Salmonella typhimurium and Staphylococcus aureus at sub-lethal concentration such as their respective minimal inhibitory concentrations, 32 and 8 microg ml(-1), respectively. METHODS AND RESULTS: Bacterial populations were studied by using transmission electron and fluorescence microscopy (TEM and FM), flow cytometry (FC) and ion-flux across the cellular membrane. Cell integrity was altered mainly in the outer membrane of S. typhimurium, but there was no significant change in the cytoplasm. However, in Staph. aureus, clear zones, abnormal septation and mesosome-like structures were observed in the cytoplasm. Bacterial populations were double-stained with propidium iodide (PI) and SYTO-13 for FC analysis. In S. typhimurium the proportion of damaged cells after 24 h was 97% and in Staph. aureus 56.3%. LAE induced transmembrane ion flux, the increase of potassium leakage after 30 min of contact was 7.7 and 3.34 microg ml(-1) for Staph. aureus and S. typhimurium, respectively. Membrane disruption was detected by measuring the proton flow across the membrane. CONCLUSIONS: Disturbance in membrane potential and structural changes was caused by LAE, although cells were not disrupted. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first time the cellular effects of LAE on bacterial cells were studied.


Subject(s)
Anti-Infective Agents/pharmacology , Arginine/pharmacology , Salmonella typhimurium/drug effects , Staphylococcus aureus/drug effects , Arginine/analogs & derivatives , Cell Membrane/metabolism , Colony Count, Microbial , Flow Cytometry/methods , Microbial Sensitivity Tests/methods , Microscopy, Electron/methods , Microscopy, Fluorescence/methods , Potassium/metabolism , Salmonella typhimurium/ultrastructure , Staphylococcus aureus/ultrastructure
17.
Antonie Van Leeuwenhoek ; 85(2): 129-39, 2004 Feb.
Article in English | MEDLINE | ID: mdl-15028873

ABSTRACT

The isolation of a new lipoxygenase-like (LOX-like) enzyme from Pseudomonas 42A2 and its characterization is described. The enzyme, located in the periplasm of the cell, which contained 0.55 mol of Fe2+ per mol of protein, is monomeric and has a molecular mass of 45 kDa. In the presence of oxygen, the enzyme converts oleic acid into (E)-10-hydroperoxy-8-octadecenoic acid (HPOD), which decomposes to the corresponding (E)-10-hydroxy-8-octadecenoic acid (HOD). The absolute configuration of this acid was determined as S on the basis of exciton-coupled CD data, and specific rotation and NMR analysis of the corresponding p -bromobenzoate derivative. The reaction in vivo leads to the dihydroxy derivative (E)-7,10-dihydroxy-8-octadecenoic acid (DHOD), so that the three hydroxy-fatty acids can be isolated from the culture medium. The activity of the enzyme was optimal between 25 and 30 degrees C and 44% of its activity still remained at 55 degrees C. Its optimal pH is 8.5-9; and the presence of magnesium ions increased LOX activity by 1.5. The activity of the LOX is highest in unsaturated fatty acids containing double bonds in position 9 (oleic, linoleic and linolenic acids), linoleic acid being preferred (100% activity) over linolenic (60.4%) and oleic acids (46%). However, kinetic studies showed that the affinity of the enzyme is similar for the three substrates.


Subject(s)
Lipoxygenase/isolation & purification , Lipoxygenase/metabolism , Oleic Acid/metabolism , Pseudomonas/enzymology , Biotransformation , Cell Fractionation , Electrophoresis, Polyacrylamide Gel , Enzyme Stability , Hydrogen-Ion Concentration , Kinetics , Lipoxygenase/chemistry , Oleic Acids/metabolism , Thermodynamics
18.
Antonie Van Leeuwenhoek ; 85(1): 1-8, 2004 Jan.
Article in English | MEDLINE | ID: mdl-15028876

ABSTRACT

Pseudomonas aeruginosa LBI isolated from petroleum-contaminated soil produced rhamnolipids (RL(LBI)) when cultivated on soapstock as the sole carbon source. HPLC-MS analysis of the purified culture supernatant identified 6 RL homologues (%): R(2) C(10) C(10) 28.9; R(2) C(10) C(12:1) 23.0; R(1) C(10) C(10) 23.4; R(2) C(10) C(12) 11.3; R(2) C(10) C(12) 7.9; R(2) C(10) C(12) 5.5. To assess the potential antimicrobial activity of the new rhamnolipid product, RL(LBI), its physicochemical properties were studied. RL(LBI) had a surface tension of 24 mN m(-1) and an interfacial tension of 1.31 mN m(-1); the cmc was 120 mg l(-1). RL(LBI) produced stable emulsions with hydrocarbons and vegetable oils. This product showed good antimicrobial behaviour against bacteria: MIC for Bacillus subtilis, Staphylococcus aureus and Proteus vulgaris was 8 mg l(-1), for Streptococcus faecalis 4 mg l(-1), and for Pseudomonas aeruginosa 32 mg l(-1). RL(LBI) was active against phytopathogenic fungal species, MIC values of 32 mg l(-1) being found against Penicillium, Alternaria, Gliocadium virens and Chaetonium globosum. Due to its physicochemical properties and antimicrobial behaviour, RL(LBI) could be used in bioremediation treatment and in the food, cosmetic and pharmaceutical industries.


Subject(s)
Pseudomonas aeruginosa/metabolism , Surface-Active Agents/metabolism , Surface-Active Agents/pharmacokinetics , Chromatography, High Pressure Liquid , Glycolipids/metabolism , Glycolipids/pharmacology , Kinetics , Microbial Sensitivity Tests , Petroleum , Pseudomonas aeruginosa/drug effects , Pseudomonas aeruginosa/growth & development , Pseudomonas aeruginosa/isolation & purification , Spectrometry, Mass, Electrospray Ionization , Surface-Active Agents/chemistry , Surface-Active Agents/isolation & purification
19.
Biotechnol Bioeng ; 81(3): 316-22, 2003 Feb 05.
Article in English | MEDLINE | ID: mdl-12474254

ABSTRACT

Pseudomonas aeruginosa 47T2, grown in submerged culture with waste frying oil as a carbon source, produced a mixture of rhamnolipids with surface activity. Up to 11 rhamnolipid homologs (Rha-Rha-C(8)-C(10); Rha-C(10)-C(8)/Rha-C(8)-C(10);Rha-Rha-C(8)-C(12:1); Rha-Rha-C(10)-C(10); Rha-Rha-C(10)-C(12:1); Rha-C(10)-C(10); Rha-Rha-C(10)-C(12)/Rha-Rha-C(12)-C(10); Rha-C(10)-C(12:1)/Rha-C(12:1)-C(10); Rha-Rha-C(12:1)-C(12); Rha-Rha-C(10)-C(14:1); Rha-C(10)-C(12)/Rha-C(12)-C(10)) were isolated from cultures of P. aeruginosa 47T2 from waste frying oil and identified by HPLC-MS analysis. This article deals with the production, isolation, and chemical characterization of the rhamnolipid mixture RL(47T2). The physicochemical and biological properties of RL(47T2) as a new product were also studied. Its surface tension decreased to 32.8 mN/m; and the interfacial tension against kerosene to 1 mN/m. The critical micellar concentration for RL(47T2) was 108.8 mg/mL. The product showed excellent antimicrobial properties. Antimicrobial activity was evaluated according to the minimum inhibitory concentration (MIC), the lowest concentration of an antimicrobial agent that inhibits development of visible microbial growth. Low MIC values were found for bacteria Serratia marcescens (4 microg/mL), Enterobacter aerogenes (8 microg/mL), Klebsiella pneumoniae (0.5 microg/mL), Staphylococcus aureus and Staphylococcus epidermidis (32 microg/mL), Bacillus subtilis (16 microg/mL), and phytopathogenic fungal species: Chaetonium globosum (64 microg/mL), Penicillium funiculosum (16 microg/mL), Gliocadium virens (32 microg/mL) and Fusarium solani (75 microg/mL).


Subject(s)
Bacteria/drug effects , Fungi/drug effects , Glycolipids/biosynthesis , Glycolipids/chemistry , Anti-Bacterial Agents , Anti-Infective Agents/pharmacology , Biodegradation, Environmental , Cells, Cultured , Glycolipids/isolation & purification , Glycolipids/pharmacology , Microbial Sensitivity Tests , Olive Oil , Plant Oils/metabolism , Pseudomonas aeruginosa/metabolism , Refuse Disposal/methods , Surface Properties
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