Publications by authors named "Carlos Garrido"

31 Publications

Impact of Sequential Inoculation with the Non- and Combined with Strains on Chemicals and Sensory Profile of Rosé Wines.

J Agric Food Chem 2021 Feb 28;69(5):1598-1609. Epub 2021 Jan 28.

Department of Food Science and Health, Andalusian Institute of Agricultural and Fisheries Research and Training (IFAPA), Alameda del Obispo Avda. Menéndez Pidal, s/n., 14004 Córdoba, Spain.

Controlled inoculations of non- yeasts are becoming increasingly used to produce high-quality wines due to their enological potential. In this study, we evaluated the impact of sequential inoculation with the commercial non- yeasts ( and ) in combination with on the chemical and sensory profile of rosé wines. Sequential inoculation with produced wines with an overall reduction in esters, mainly explained by the lower concentrations of ethyl esters of medium-chain fatty acids and isoamyl acetate. The lower ester concentrations of these wines were related to a reduction in fruity descriptors. An increase was observed, however, in other minor esters such as cinnamates and ethyl esters of branched acids. Zinc, ethyl isobutyrate, and ethyl dihydrocinnamate were selected as potential markers for this fermentation strategy. Sequential inoculation with resulted in rosé wines with an enhanced ester profile, reduced acetaldehyde, and increased anthocyans and tannins. Compared to the control wines fermented with , the changes observed in these wines were far subtler, especially for the volatile profile, sensory characteristics, and color parameters, with isobutyl hexanoate and isoamyl butyrate being selected as potential markers.
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http://dx.doi.org/10.1021/acs.jafc.0c06970DOI Listing
February 2021

Identification of the Sesquiterpene Cyclase Involved in the Biosynthesis of (+)-4-Epi-eremophil-9-en-11-ol Derivatives Isolated from .

ACS Chem Biol 2020 10 1;15(10):2775-2782. Epub 2020 Oct 1.

Facultad de Ciencias, Departamento de Química Orgánica, Universidad de Cádiz, Campus Universitario Río San Pedro s/n, Torre sur, 4a planta, Puerto Real, 11510 Cádiz, Spain.

Cultivation of the phytopathogenic fungus using sublethal amounts of copper sulfate yielded a cryptic sesquiterpenoids family, which displayed the basic chemical structure of (+)-4-epi-eremophil-9-ene. The biosynthesis pathway was established, and the route involved the likely transformation of the diphosphate of farnesyl (FDP), to give a -fused eudesmane cation, through ()-hedycaryol, finally yielding the (+)-4-epi-eremophil-9-enol derivatives. An expression study of genes that code for the sesquiterpene cyclases (STC), including the recently reported gene present in the genome, was performed in order to establish the STC involved in this biosynthesis. The results showed a higher expression level for the gene with respect to the other genes in both wild-type strains, B05.10 and UCA992. Deletion of the gene eliminated (+)-4-epi-eremophilenol biosynthesis, which could be re-established by complementing the null mutant with the gene. Chemical analysis suggested that STC7 is the principal enzyme responsible for the key step of cyclization of FDP to eremophil-9-en-11-ols. Furthermore, a thorough study of the two wild-types and the complemented mutant revealed four new eremophilenol derivatives whose structures are reported here.
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http://dx.doi.org/10.1021/acschembio.0c00561DOI Listing
October 2020

Adsorption of bovine serum albumin on gold nanoprisms: interaction and effect of NIR irradiation on protein corona.

J Mater Chem B 2020 09;8(37):8644-8657

Advanced Center for Chronic Diseases (ACCDiS), Santos Dumont 964, Independencia, Santiago, Chile and Departamento de Ciencias Químicas, Facultad de Ciencias Exactas, Universidad Andres Bello, Av. Republica 275, Santiago, Chile.

Because of their photothermal properties, gold nanoparticles (AuNPs) have gained attention regarding their use in drug delivery and therapeutic applications. In this sense, it is interesting to consider their interactions with biologically available proteins, such as serum albumin, as well as the effects of irradiation and photothermal conversion on the protein structure that can lead to a loss of function or generate an immune response. Gold nanoprisms (AuNPrs) have gained interest due to their low toxicity, ease of synthesis, and excellent stability, promoting their use in bioapplications such as surface-enhanced Raman spectroscopy (SERS), drug delivery, and photothermal therapy. The interaction between AuNPrs, with plasmon bands centred in the near-infrared region (NIR), and bovine serum albumin (BSA) has not been explored yet. UV-Vis spectroscopy, dynamic light scattering (DLS) and fluorescence spectroscopy were used to study the interaction between AuNPrs and BSA in addition to estimation of the adsorption rate and kinetic and thermodynamic parameters (K, ΔH°, ΔG°, ΔS°, and Ea) using adsorption isotherms and Langmuir and Freundlich models. The results suggest spontaneous cooperative binding in multilayer adsorption, achieved by the chemisorption of BSA on the AuNPr surface through the S-Au interaction, as confirmed by Raman spectroscopy. On the other hand, the photothermal conversion efficiency (PE) of the coated nanoparticles after NIR irradiation was assessed, resulting in a slight decrease in the PE of BSA coated on AuNPrs in comparison with that of noncapped nanoparticles. The effect of the irradiation on the protein conformation of capped nanoparticles was also assessed; circular dichroism showed BSA unfolding upon interaction with AuNPrs, with a decrease in the α-helix and β-sheet contents, as well as an increase in random coil conformations. Changes in the Raman spectrum suggest a modification of the disposition of the protein residues exposed to the gold surface after NIR irradiation; but at the secondary structure level, no relevant changes were observed. This provides possibilities for the use of NPs-BSA for bioapplications based on the photothermal effect promoted by laser irradiation, since the biological identity of the protein is preserved after NIR irradiation.
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http://dx.doi.org/10.1039/d0tb01246gDOI Listing
September 2020

Use of cervical cancer preventive services among US women aged 21-29: an assessment of the 2010 Affordable Care Act rollout through 2018.

Cancer Causes Control 2020 Sep 29;31(9):839-850. Epub 2020 Jun 29.

Arizona Cancer Center, University of Arizona, 1515 N Campbell Ave, Tucson, AZ, 85719, USA.

Purpose: The 2010 Affordable Care Act (ACA) provided millions of people with health insurance coverage and facilitated routine cancer screening by requiring insurers to cover preventive services without cost-sharing. Despite greater access to affordable cancer screening, Pap testing has declined over time. The aim of this study is to assess participation in Pap test and HPV vaccination, and adherence to guidelines as outlined by the American Cancer Society (ACS) from the 2010 ACA provision eliminating cost-sharing for preventive services.

Methods: Using multi-year responses from the Behavioral Risk Factor Surveillance System, we examined the association between the ACA and participation in and adherence to Pap testing and HPV vaccination behaviors as set by the ACS. The sample included women aged 21-29 who completed the survey between 2008 and 2018 (every other year) and who live in 24 US States (N = 37,893).

Results: Results showed significant decreases in Pap testing rates but increases in the uptake of the HPV vaccine series for all age groups and across all demographics. Post-ACA year significantly predicted increases in HPV + Pap co-testing participation and adherence. Women with health insurance coverage were more likely to engage in both behaviors.

Conclusion: Findings raise concerns around declines in the proportion of women receiving and adhering to Pap testing guidelines. A need exists for research to examine the role of increases in HPV vaccination uptake on decreases in Pap testing. Moreover, effective strategies should target increases in cervical cancer screening uptake among women vaccinated against HPV.
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http://dx.doi.org/10.1007/s10552-020-01325-wDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7433930PMC
September 2020

Antimony as a tracer of non-exhaust traffic emissions in air pollution in Granada (S Spain) using lichen bioindicators.

Environ Pollut 2020 Aug 3;263(Pt A):114482. Epub 2020 Apr 3.

Instituto Andaluz de Ciencias de la Tierra (IACT), CSIC-UGR, Avda. de las Palmeras 4, E-18100, Armilla, Granada, Spain. Electronic address:

We have studied the metal air pollution trends in a medium-sized Spanish city suffering from traffic emission using in-situ lichen Xanthoria parietina as a bioindicator. The large scale sampling included 97 samples from urban, metropolitan and remote control areas of Granada that were analyzed by Inductively Coupled Plasma-Mass Spectrometry. Enrichment factor of Sb exhibited severe anthropogenic enrichment, whereas Cu and Sb showed significantly higher median values in the urban areas with respect to metropolitan areas. Additionally, bioaccumulation ratios of V, Cr, Ni, Cu, Zn, Cd, Sb, and Pb -associated to exhaust and non-exhaust traffic emissions- enabled us to delineate hot spots of metal(loid) accumulation in the main accesses to the city, characterized by dense traffic and copious traffic jams. To distinguish non-exhaust emissions, we studied the spatial distribution of the Cu:Sb ratio -a tracer of brake wear- highlighting the surroundings of the highway and the main traffic accesses to the city likely due to sudden hard braking and acceleration during frequent traffic jams. Our study shows that the metal(loid) contents in lichens are excellent proxies for non-exhaust traffic emissions and that their contribution to the metal(loid) air pollution in Granada is more significant than previously thought.
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http://dx.doi.org/10.1016/j.envpol.2020.114482DOI Listing
August 2020

Biodegradation and toxicity reduction of nonylphenol, 4-tert-octylphenol and 2,4-dichlorophenol by the ascomycetous fungus Thielavia sp HJ22: Identification of fungal metabolites and proposal of a putative pathway.

Sci Total Environ 2020 Mar 21;708:135129. Epub 2019 Nov 21.

Laboratory of Biochemistry and Enzymatic Engineering of Lipases, National School of Engineers of Sfax, University of Sfax, BP «1173», 3038 Sfax, Tunisia.

Research on the biodegradation of emerging pollutants is gained great focus regarding their detrimental effects on the environment and humans. The objective of the present study was to evaluate the ability of the ascomycetes Thielavia sp HJ22 to remove the phenolic xenobiotics nonylphenol (NP), 4-tert-octylphenol (4-tert-OP) and 2,4-dichlorophenol (2,4-DCP). The strain showed efficient degradation of NP and 4-tert-OP with 95% and 100% removal within 8 h of incubation, respectively. A removal rate of 80% was observed with 2,4-DCP within the same time. Under experimental conditions, the degradation of the tested pollutants concomitantly increased with the laccase production and cytochrome P450 monooxygenases inhibition. This study showed the involvement of laccase in pollutants removal together with biosorption mechanisms. Additionally, results demonstrated the participation of cytochrome P450 monooxygenase in the elimination of 2,4-DCP. Liquid chromatography-mass spectrometry analysis revealed several intermediates, mainly hydroxylated and oxidized compounds with less harmful effects compared to the parent compounds. A decrease in the toxicity of the identified metabolites was observed using Aliivibrio fischeri as bioindicator. The metabolic pathways of degradation were proposed based on the identified metabolites. The results point out the potential of Thielavia strains in the degradation and detoxification of phenolic xenobiotics.
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http://dx.doi.org/10.1016/j.scitotenv.2019.135129DOI Listing
March 2020

Effects of redox oscillations on the phosphogypsum waste in an estuarine salt-marsh system.

Chemosphere 2020 Mar 23;242:125174. Epub 2019 Oct 23.

University Grenoble Alpes, CNRS, IRD, IFSTTAR, ISTerre, 38000, Grenoble, France.

Salt marshes are natural deposits of heavy metals in estuarine systems, where sulphide precipitation associated with redox changes often results in a natural attenuation of contamination. In the present study, we focus on the effects of variable redox conditions imposed to a highly-polluted phosphogypsum stack that is directly piled over the salt marsh soil in the Tinto River estuary (Huelva, Spain). The behaviour of contaminants is evaluated in the phosphogypsum waste and in the marsh basement, separately, in controlled, experimentally-induced oscillating redox conditions. The results revealed that Fe, and to a lesser extent S, control most precipitation/dissolution processes. Ferric iron precipitates in the form of phosphates and oxyhydroxides, while metal sulphide precipitation is insignificant and appears to be prevented by the abundant formation of Fe phosphates. An antagonistic evolution with changing redox conditions was observed for the remaining contaminants such as Zn, As, Cd and U, which remained mobile in solution during most of experimental run. Therefore, these findings revealed that high concentrations of phosphates inhibit the typical processes of immobilisation of pollutants in salt-marshes which highlights the elevated contaminant potential of phosphogypsum wastes on coastal environments.
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http://dx.doi.org/10.1016/j.chemosphere.2019.125174DOI Listing
March 2020

NLRP3 inflammasome suppression improves longevity and prevents cardiac aging in male mice.

Aging Cell 2020 01 18;19(1):e13050. Epub 2019 Oct 18.

Institute of Nutrition and Food Technology "José Mataix Verdú", Department of Physiology, Biomedical Research Center, University of Granada, Granada, Spain.

While NLRP3-inflammasome has been implicated in cardiovascular diseases, its role in physiological cardiac aging is largely unknown. During aging, many alterations occur in the organism, which are associated with progressive impairment of metabolic pathways related to insulin resistance, autophagy dysfunction, and inflammation. Here, we investigated the molecular mechanisms through which NLRP3 inhibition may attenuate cardiac aging. Ablation of NLRP3-inflammasome protected mice from age-related increased insulin sensitivity, reduced IGF-1 and leptin/adiponectin ratio levels, and reduced cardiac damage with protection of the prolongation of the age-dependent PR interval, which is associated with atrial fibrillation by cardiovascular aging and reduced telomere shortening. Furthermore, old NLRP3 KO mice showed an inhibition of the PI3K/AKT/mTOR pathway and autophagy improvement, compared with old wild mice and preserved Nampt-mediated NAD levels with increased SIRT1 protein expression. These findings suggest that suppression of NLRP3 prevented many age-associated changes in the heart, preserved cardiac function of aged mice and increased lifespan.
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http://dx.doi.org/10.1111/acel.13050DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6974709PMC
January 2020

Lichens as a spatial record of metal air pollution in the industrialized city of Huelva (SW Spain).

Environ Pollut 2019 Oct 17;253:918-929. Epub 2019 Jul 17.

Instituto Andaluz de Ciencias de la Tierra (IACT), CSIC-UGR, Avda. de las Palmeras 4, E-18100 Armilla, Granada, Spain. Electronic address:

Huelva is a highly industrialized city in SW Spain hosting, among others, a Cu smelter, a phosphate fertilizer plant, a power plant, and oil refineries. This study aims to evaluate metal concentrations in lichens as bioindicators of atmospheric pollution in the impacted urban areas. Xanthoria parietina species from Huelva and nearby villages, as well as reference samples from remote, non-contaminated urban areas, were analyzed for trace elements (V, Cr, Mn, Co, Ni, Cu, Zn, Sr, As, Cd, Sb, Cs, Ba, La, Ce, Pr, Nd, Sm, Er, Tm, Yb, Lu, Pb, Th, U) using Inductively Coupled Plasma-Mass Spectrometry; and for major elements (Ca, K, Mg, P, and S) by Inductively Coupled Plasma-Optical Emission Spectrometry after acid digestion. The metal composition of X. parietina exhibits spatial distribution patterns with extremely elevated concentrations (Co, Ni, Cu, Zn, As, Cd, Sb, Ba, Pb, U, and S) in the surroundings of the industrial estates to <1 km distance. Mean concentrations were significantly lower in the urban areas >1 km from the pollution sources. However, air pollution persists in the urban areas up to 4 km away, as the mean concentrations of Cu, Zn, As, Cd, Sb and S remained considerably elevated in comparison to the reference samples. Though rigorous source apportionment analysis was not the aim of this study, a good positive correlation of our results with metal abundances in ambient particulate matter and in pollution sources points to the Cu smelter as the main source of pollution. Hence, the severe air pollution affecting Huelva and nearby urban areas may be considered a serious health risk to local residents.
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http://dx.doi.org/10.1016/j.envpol.2019.07.086DOI Listing
October 2019

Vibrational and electronic spectroscopic detection and quantification of carminic acid in candies.

Food Chem 2019 Jun 14;283:164-169. Epub 2019 Jan 14.

Laboratory of Vibrational Spectroscopy, Faculty of Sciences, Department of Chemistry, Universidad de Chile, Santiago, Chile.

Carminic acid (CA) contained in one kind of gummy candy was detected and quantified by using vibrational and electronic spectroscopy, respectively; the proposed methodology in solution is simple and rapid without sample pretreatment as usually used and reported. The identification of CA in candies was performed through the Raman and surface enhanced Raman scattering (SERS) spectra along with the equivalent spectral data from the natural CA dye. A modified silver colloidal solution was used in order to obtain SERS activity of CA at very low concentration in colloidal aqueous solution. Theoretical calculations allow infer about both the CA/silver surface interaction nature and on the orientation of CA on the surface. The electronic spectroscopic (UV-Vis) data allowed quantify CA in candies; the amount resulted nearly identical to that determined from HPLC traditional measurements. The present results should contribute to the health of children consumers.
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http://dx.doi.org/10.1016/j.foodchem.2018.12.123DOI Listing
June 2019

Unraveling the impact of chronic exposure to metal pollution through human gallstones.

Sci Total Environ 2018 May 27;624:1031-1040. Epub 2017 Dec 27.

Riotinto Hospital, Avda. La Esquila 5, E-21660 Minas de Riotinto, Huelva, Spain.

This study aims to explore the impact of chronic metal exposure derived from persistent pollution from mining activity using human gallstones as proxies. The samples were obtained from patients residing in geologically and environmentally contrasting areas in the Province of Huelva, SW Spain, allowing for the evaluation of the regional effect of metal pollution. The study group resides in the Iberian Pyrite Belt characterized by natural and anthropogenic metal pollution from mining activities, whereas the control group resides in the Ossa Morena Zone famous for its natural parks. A total of 68 gallstones were first classified based on their phase composition and structure and subsequently their chemical composition was studied using solution Inductively Coupled Plasma-Mass Spectrometry. The metal concentrations increased in the cholesterol-rich gallstones from pure, to mixed and composite cholesterol stones along with the increasing amount of minor phases, such as bilirubinate, carbonate, and phosphate. These cholesterol stones did not show an evident enrichment tendency. On the contrary, pigment stones, composed of bilirubinate, carbonate, and phosphate phases, were rich in a variety of elements and the regional comparison showed that the pigment stones from the study area were enriched in sulfide-associated metal(loid)s, Mn, Fe, Cu, Zn, Sr, As, Ag, Sb, and Pb with respect to the control group. Inhalation of polluted airborne particulate matter is considered as one of the main exposure routes among the residents of the study area. Additionally, consumption of local water and locally produced food products such as fruit and vegetables and dermal contact may be possible sources of exposure, but no direct connection was observed.
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http://dx.doi.org/10.1016/j.scitotenv.2017.12.224DOI Listing
May 2018

Stable isotope insights into the weathering processes of a phosphogypsum disposal area.

Water Res 2018 09 28;140:344-353. Epub 2018 Apr 28.

Department of Earth Sciences & Research Center on Natural Resources, Health and the Environment, University of Huelva, Campus 'El Carmen', E-21071 Huelva, Spain. Electronic address:

Highly acidic phosphogypsum wastes with elevated potential for contaminant leaching are stack-piled near coastal areas worldwide, threatening the adjacent environment. Huge phosphogypsum stacks were disposed directly on the marshes of the Estuary of Huelva (SW Spain) without any impermeable barrier to prevent leaching and thus, contributing to the total contamination of the estuarine environment. According to the previous weathering model, the process water ponded on the surface of the stack, initially used to carry the waste, was thought to be the main washing agent through its infiltration and subsequently the main component of the leachates emerging as the edge outflows. Preliminary restorations have been applied to the site and similar ones are planned for the future considering process water as the only pollution agent. Further investigation to validate the pollution pathway was necessary, thus an evaluation of the relationship between leachates and weathering agents of the stack was carried out using stable isotopes (δO, δH, and δS) as geochemical tracers. Quantification of the contribution of all possible end-members to the phosphogypsum leachates was also conducted using ternary mixing via the stable isotopic tracers. The results ruled out ponded process water as main vector of edge outflow pollution and unveiled a continuous infiltration of estuarine waters to the stack implying that is subjected to an open weathering system. The isotopic tracers revealed a progressive contribution downstream from fluvial to marine signatures in the composition of the edge outflows, depending on the location of each disposal zone within the different estuarine morphodynamic domains. Thus, the current study suggests that the access of intertidal water inside the phosphogypsum stack, for instance through secondary tidal channels, is the main responsible for the weathering of the waste in depth, underlying the necessity for new, more effective restorations plans.
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http://dx.doi.org/10.1016/j.watres.2018.04.060DOI Listing
September 2018

Biosynthesis of abscisic acid in fungi: identification of a sesquiterpene cyclase as the key enzyme in Botrytis cinerea.

Environ Microbiol 2018 07 26;20(7):2469-2482. Epub 2018 Jul 26.

UMR BIOGER, INRA, AgroParisTech, Université Paris-Saclay, 78850 Thiverval-Grignon, France.

While abscisic acid (ABA) is known as a hormone produced by plants through the carotenoid pathway, a small number of phytopathogenic fungi are also able to produce this sesquiterpene but they use a distinct pathway that starts with the cyclization of farnesyl diphosphate (FPP) into 2Z,4E-α-ionylideneethane which is then subjected to several oxidation steps. To identify the sesquiterpene cyclase (STC) responsible for the biosynthesis of ABA in fungi, we conducted a genomic approach in Botrytis cinerea. The genome of the ABA-overproducing strain ATCC58025 was fully sequenced and five STC-coding genes were identified. Among them, Bcstc5 exhibits an expression profile concomitant with ABA production. Gene inactivation, complementation and chemical analysis demonstrated that BcStc5/BcAba5 is the key enzyme responsible for the key step of ABA biosynthesis in fungi. Unlike what is observed for most of the fungal secondary metabolism genes, the key enzyme-coding gene Bcstc5/Bcaba5 is not clustered with the other biosynthetic genes, i.e., Bcaba1 to Bcaba4 that are responsible for the oxidative transformation of 2Z,4E-α-ionylideneethane. Finally, our study revealed that the presence of the Bcaba genes among Botrytis species is rare and that the majority of them do not possess the ability to produce ABA.
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http://dx.doi.org/10.1111/1462-2920.14258DOI Listing
July 2018

Effects of seawater mixing on the mobility of trace elements in acid phosphogypsum leachates.

Mar Pollut Bull 2018 Feb 5;127:695-703. Epub 2018 Jan 5.

Instituto Andaluz de Ciencias de la Tierra, CSIC & UGR, Avenida de las Palmeras 4, 18100 Armilla, Granada, Spain. Electronic address:

This research reports the effects of pH increase on contaminant mobility in phosphogypsum leachates by seawater mixing, as occurs with dumpings on marine environments. Acid leachates from a phosphogypsum stack located in the Estuary of Huelva (Spain) were mixed with seawater to achieve gradually pH7. Concentrations of Al, Fe, Cr, Pb and U in mixed solutions significantly decreased with increasing pH by sorption and/or precipitation processes. Nevertheless, this study provides insight into the high contribution of the phosphogypsum stack to the release of other toxic elements (Co, Ni, Cu, Zn, As, Cd and Sb) to the coastal areas, as 80-100% of their initial concentrations behaved conservatively in mixing solutions with no participation in sorption processes. Stable isotopes ruled out connexion between different phosphogypsum-related wastewaters and unveiled possible weathering inputs of estuarine waters to the stack. The urgency of adopting effective restoration measures in the study area is also stressed.
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http://dx.doi.org/10.1016/j.marpolbul.2018.01.001DOI Listing
February 2018

The F-actin capping protein is required for hyphal growth and full virulence but is dispensable for septum formation in Botrytis cinerea.

Fungal Biol 2016 10 22;120(10):1225-35. Epub 2016 Jul 22.

Institut für Biologie und Biotechnologie der Pflanzen (IBBP), Westfälische Wilhelms-Universität Münster, Schlossplatz 8, 48143 Münster, Germany. Electronic address:

Filamentous (F-) actin is an integral part of the cytoskeleton allowing for cell growth, intracellular motility, and cytokinesis of eukaryotic cells. Its assembly from G-actin monomers and its disassembly are tightly regulated processes involving a number of actin-binding proteins (ABPs) such as F-actin nucleators and cross-linking proteins. F-actin capping protein (CP) is an alpha/beta heterodimer known from yeast and higher eukaryotes to bind to the fast growing ends of the actin filaments stabilizing them. In this study, we identified the orthologs of the two CP subunits, named BcCPA1 and BcCPB1, in the plant pathogenic fungus Botrytis cinerea and showed that the two proteins physically interact in a yeast two-hybrid approach. GFP-BcCPA1 fusion proteins were functional and localized to the assumed sites of F-actin accumulation, i.e. to the hyphal tips and the sites of actin ring formation. Deletion of bccpa1 had a profound effect on hyphal growth, morphogenesis, and virulence indicating the importance of F-actin capping for an intact actin cytoskeleton. As polarized growth - unlike septum formation - is impaired in the mutants, it can be concluded that the organization and/or localization of actin patches and cables are disturbed rather than the functionality of the actin rings.
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http://dx.doi.org/10.1016/j.funbio.2016.07.007DOI Listing
October 2016

Rapid and not culture-dependent assay based on multiplex PCR-SSR analysis for monitoring inoculated yeast strains in industrial wine fermentations.

Arch Microbiol 2017 Jan 8;199(1):135-143. Epub 2016 Sep 8.

Laboratorio de Microbiología, Facultad de Ciencias del Mar y Ambientales, Universidad de Cádiz, 11510, Puerto Real, Cádiz, Spain.

Wine industry needs a simple method for rapid diagnosis of the dominance of inoculated strains that could be performed routinely during the fermentation process. We present a suitable, high-throughput, and low-cost method to monitor rapidly the dominance of inoculated yeast strains in industrial fermentations of red and white wines using an activated carbon cleaning pretreatment, and a rapid DNA extraction method plus multiplex PCR-SSR analysis. We apply this technique directly to samples of fermenting wines without previously isolating yeast colonies. Results are obtained in a maximum time of 4.5 h.
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http://dx.doi.org/10.1007/s00203-016-1287-4DOI Listing
January 2017

Combined microstructural and mineralogical phase characterization of gallstones in a patient-based study in SW Spain - Implications for environmental contamination in their formation.

Sci Total Environ 2016 Dec 27;573:433-443. Epub 2016 Aug 27.

Riotinto Hospital, Avda. La Esquila 5, E-21660 Minas de Riotinto, Huelva, Spain. Electronic address:

This study explores the environmental impact of metal exposure on humans through detailed phase and structural characterization of gallstones from two environmentally contrasting populations in Huelva Province (SW Spain). A total of 42 gallstone samples, obtained after surgical intervention at the Riotinto Hospital, were studied by powder X-ray diffraction (XRD), Fourier Transformed Infra-Red spectroscopy (FTIR), FTIR-μ-ATR (Attenuated Total Reflection) coupled with an optical microscope, and by Environmental Scanning Electron Microscope with Energy Dispersive X-ray Spectroscopy (ESEM-EDS), and subsequently classified according to their phase composition and structure. Additionally, the patients were enquired for their living habits in order to analyze the source of possible exposure to metal contamination. The gallstones were classified into pure, mixed and composite cholesterol stones, black and brown pigment stones, and carbonate stones. The patients from the study group residing in a region with acknowledged metal contamination of both natural and anthropogenic origin have a higher risk of metal exposure through contaminated soil, particle matter in the air, and consumption of local water and food products. According to our findings, the metal exposure is related to a higher tendency of forming black pigment stones in the study group in comparison to the control group residing in a natural park with nearly pristine environmental conditions. Moreover, the gallstones from the study group showed to contain more abundant metal components, such as Cu, Fe, Ni, and Zn, than those from the control group. To our knowledge this is the first study to examine the regional environmental impact of metal exposure on human gallstones.
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http://dx.doi.org/10.1016/j.scitotenv.2016.08.110DOI Listing
December 2016

What Facial Appearance Reveals Over Time: When Perceived Expressions in Neutral Faces Reveal Stable Emotion Dispositions.

Front Psychol 2016 30;7:986. Epub 2016 Jun 30.

Department of Psychological and Brain Sciences, Dartmouth College, Hanover NH, USA.

It might seem a reasonable assumption that when we are not actively using our faces to express ourselves (i.e., when we display nonexpressive, or neutral faces), those around us will not be able to read our emotions. Herein, using a variety of expression-related ratings, we examined whether age-related changes in the face can accurately reveal one's innermost affective dispositions. In each study, we found that expressive ratings of neutral facial displays predicted self-reported positive/negative dispositional affect, but only for elderly women, and only for positive affect. These findings meaningfully replicate and extend earlier work examining age-related emotion cues in the face of elderly women (Malatesta et al., 1987a). We discuss these findings in light of evidence that women are expected to, and do, smile more than men, and that the quality of their smiles predicts their life satisfaction. Although ratings of old male faces did not significantly predict self-reported affective dispositions, the trend was similar to that found for old female faces. A plausible explanation for this gender difference is that in the process of attenuating emotional expressions over their lifetimes, old men reveal less evidence of their total emotional experiences in their faces than do old women.
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http://dx.doi.org/10.3389/fpsyg.2016.00986DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4927908PMC
July 2016

Chemically Induced Cryptic Sesquiterpenoids and Expression of Sesquiterpene Cyclases in Botrytis cinerea Revealed New Sporogenic (+)-4-Epieremophil-9-en-11-ols.

ACS Chem Biol 2016 05 8;11(5):1391-400. Epub 2016 Mar 8.

Departamento de Química Orgánica, Facultad de Ciencias, Universidad de Cádiz , Campus Universitario Río San Pedro s/n, Torre sur, 4° planta, 11510, Puerto Real, Cádiz, Spain.

The sequencing of the genomes of the B05.10 and T4 strains of the fungus Botrytis cinerea revealed an abundance of novel biosynthetic gene clusters, the majority of which were unexpected on the basis of the previous analyses of the fermentation of these and closely related species. By systematic alteration of easy accessible cultivation parameters, using chemical induction with copper sulfate, we have found a cryptic sesquiterpenoid family with new structures related to eremophil-9-ene, which had the basic structure of the sesquiterpene (+)-5-epiaristolochene ((+)-4-epieremophil-9-ene). An expression study of the sesquiterpene cyclase genes present in the Botrytis cinerea genome, under culture conditions, is reported. In general, a 3 day delay and a higher BcSTC genes expression were observed when copper (5 ppm) was fed to the fermentation broth. In addition, to the observed effect on the BcBOT2 (BcSTC1) gene, involved in the biosynthesis of the botrydial toxin, a higher expression level for BcSTC3 and BcSTC4 was observed with respect to the control in the strain B05.10. Interestingly, under copper conditions, the BcSTC4 gene was the most expressed gene in the Botrytis cinerea UCA992 strain. In vitro evaluation of the biological role of these metabolites indicates that they contributed to the conidial development in B. cinerea and appear to be involved in self-regulation of the production of asexual spores. Furthermore, they promoted the formation of complex appressoria or infection cushions.
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http://dx.doi.org/10.1021/acschembio.5b00931DOI Listing
May 2016

On the neural implausibility of the modular mind: Evidence for distributed construction dissolves boundaries between perception, cognition, and emotion.

Behav Brain Sci 2016 Jan;39:e246

Department of Psychology,Northeastern University,Boston,MA

Firestone & Scholl (F&S) rely on three problematic assumptions about the mind (modularity, reflexiveness, and context-insensitivity) to argue cognition does not fundamentally influence perception. We highlight evidence indicating that perception, cognition, and emotion are constructed through overlapping, distributed brain networks characterized by top-down activity and context-sensitivity. This evidence undermines F&S's ability to generalize from case studies to the nature of perception.
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http://dx.doi.org/10.1017/S0140525X15002770DOI Listing
January 2016

Proteomic profiling of Botrytis cinerea conidial germination.

Arch Microbiol 2015 Mar 21;197(2):117-33. Epub 2014 Aug 21.

Laboratory of Microbiology, Marine and Environmental Sciences Faculty, Andalusian Center for Grape and Grapevine Research, CeiA3 International Campus of Excellence in Agrifood, University of Cádiz, Pol. Río San Pedro s/n, 11510, Puerto Real, Cádiz, Spain.

Botrytis cinerea is one of the most relevant plant pathogenic fungi. The first step during its infection process is the germination of the conidia. Here, we report on the first proteome analysis during the germination of B. cinerea conidia, where 204 spots showed significant differences in their accumulation between ungerminated and germinated conidia by two-dimensional polyacrylamide gel electrophoresis and qPCR. The identified proteins were grouped by gene ontology revealing that the infective tools are mainly preformed inside the ungerminated conidia allowing a quick fungal development at the early stages of conidial germination. From 118 identified spots, several virulence factors have been identified while proteins, such as mannitol-1-phosphate dehydrogenase, 6,7-dimethyl-8-ribityllumazine synthase or uracil phosphoribosyltransferase, have been disclosed as a new potential virulence factors in botrytis whose role in pathogenicity needs to be studied to gain new insights about the role of these proteins as therapeutic targets and virulence factors.
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http://dx.doi.org/10.1007/s00203-014-1029-4DOI Listing
March 2015

PROSPECTIVE STUDY ON SEVENTY-SIX CASES OF FRACTURED ACETABULUM WITH SURGICAL TREATMENT.

Rev Bras Ortop 2011 Sep-Oct;46(5):520-5. Epub 2015 Dec 6.

Resident Physician in Orthopedics and Traumatology, Hospital São Bento Cardioclínica S/A, Belo Horizonte, MG, Brazil.

Objective: To conduct a prospective study on unstable and incongruent fractures of the acetabulum, in comparison with the literature, covering the type of access, fixation materials, degree of reduction, type of fracture and results after surgery.

Methods: The authors evaluated 76 patients with unstable and incongruent fracture of the acetabulum between January 1999 and December 2009. The Marvin Tile classification was used, and all cases were treated surgically using the technique recommended by the AO-ASIF group.

Results: The radiological evaluation was done in accordance with the Ruesch criteria, and good, excellent or perfect results were obtained for 64 patients (84.2%). The clinical evaluation was done in accordance with the Harris criteria, and good or excellent results were obtained for 62 patients (81.6%). The complications were assessed and, in comparison with the literature, were shown to be compatible with previous reports even after the statistical analysis.

Conclusion: It was concluded that anatomical reduction and stable fixation are important for good results.
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http://dx.doi.org/10.1016/S2255-4971(15)30406-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4799313PMC
March 2016

Biphasic calcium phosphate bioceramics for orthopaedic reconstructions: clinical outcomes.

Int J Biomater 2011 28;2011:129727. Epub 2011 Jun 28.

Department of Orthopaedic Surgery, Hospital São Bento Cardioclinica Ltda., Rua Crucis 50, 30.360-290 Belo Horizonte, Minas Gerais, Brazil.

BCP are considered the most promising biomaterials for bone reconstruction. This study aims at analyzing the outcomes of patients who received BCP as bone substitutes in orthopaedic surgeries. Sixty-six patients were categorized according to the etiology and morphology of the bone defects and received scores after clinical and radiographic evaluations. The final results corresponded to the combination of both parameters and varied from 5 (excellent result) to 2 or lower (poor result). Most of the patients who presented cavitary defects or bone losses due to prosthesis placement or revision, osteotomies, or arthrodesis showed good results, and some of them excellent results. However, patients with segmental defects equal or larger than 3 cm in length were classified as moderate results. This study established clinical parameters where the BCP alone can successfully support the osteogenic process and where the association with other tissue engineering strategies may be considered.
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http://dx.doi.org/10.1155/2011/129727DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3132521PMC
November 2011

COMPARATIVE STUDY BETWEEN RADIOLOGICAL CLASSIFICATION AND MACRO AND MICROSCOPIC ANALYSIS ON OSTEOARTHRITIS LESIONS OF THE KNEE.

Rev Bras Ortop 2011 Mar-Apr;46(2):155-9. Epub 2015 Dec 6.

Orthopedic Doctor and Specialist in Knee Surgery at the Hospital São Bento Cardioclínica S/A - Belo Horizonte, MG.

Objectives: To compare the modified Ahlbäck radiological classification with macroscopic analysis of knee injuries and locate a chondral lesion in the tibial plateau, and to correlate this with integrity or lack of integrity of the anterior cruciate ligament.

Material And Methods: Between July and December 2009, 40 patients of mean age 67.1 years with an indication for total knee arthroplasty were selected. The modified Ahlbäck radiological classification was used. The International Cartilage Repair Society classification was used for macroscopic analysis of the lesions. Chondral injuries were correlated with the integrity or lack of integrity of the anterior cruciate ligament.

Results: Regarding the radiological classification of the knees, three (7.5%) were classified as grade 1, two (5%) as grade 2, 17 (42.5%) as grade 3, 16 (40%) as grade 4 and two (5%) as grade 5. The macroscopic analysis of the knee showed that 25 patients (62.5%) had very severe injury and 15 (37.5%), severe. In eight knees (20%) with ruptured ACL, the lesion extended to the posterior region of the medial tibial plateau. When the ACL was intact, the lesion was located in the anterior-central region.

Conclusion: Knee osteoarthritis of grades 4 and 5 in the radiological classification, showed agreement with the macroscopic analysis, i.e. very severe chondral injury. However, grades 1, 2 and 3 were discordant. In the cases of osteoarthritis with intact ACL, the lesion was located in the anterior-central region of the medial tibial plateau, and those with ruptured ACL had the lesion extending to the posterior region of the plateau.
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http://dx.doi.org/10.1016/S2255-4971(15)30232-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4799151PMC
March 2016

Development of proteomics-based fungicides: new strategies for environmentally friendly control of fungal plant diseases.

Int J Mol Sci 2011 Jan 21;12(1):795-816. Epub 2011 Jan 21.

Laboratory of Microbiology, Faculty of Marine and Environmental Sciences, University of Cádiz, Pol. Río San Pedro s/n, 11510 Puerto Real, Spain.

Proteomics has become one of the most relevant high-throughput technologies. Several approaches have been used for studying, for example, tumor development, biomarker discovery, or microbiology. In this "post-genomic" era, the relevance of these studies has been highlighted as the phenotypes determined by the proteins and not by the genotypes encoding them that is responsible for the final phenotypes. One of the most interesting outcomes of these technologies is the design of new drugs, due to the discovery of new disease factors that may be candidates for new therapeutic targets. To our knowledge, no commercial fungicides have been developed from targeted molecular research, this review will shed some light on future prospects. We will summarize previous research efforts and discuss future innovations, focused on the fight against one of the main agents causing a devastating crops disease, fungal phytopathogens.
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http://dx.doi.org/10.3390/ijms12010795DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3039980PMC
January 2011

USE OF BIOCERAMICS IN FILLING BONE DEFECTS.

Rev Bras Ortop 2010 Jul-Aug;45(4):433-8. Epub 2015 Nov 17.

MSc in Rehabilitation from Unifesp; PhD in General Pathology from UFMG. Resident Physician in Orthopedics and Traumatology, Hospital Sao Bento Cardioclínica S/A.

Objective: To present the results from using biological ceramics for filling bone defects resulting from post-traumatic or orthopedic injuries.

Methods: Thirty-six patients with bone defects caused by trauma or orthopedic injury were evaluated. Nineteen patients were male (52.8%) and 17 were female (47.2%). Their ages ranged from 19 to 84 years, with a mean of 45.7 years and median of 37 years. Only patients with defects that required at least five grams of biological ceramic were included. Eighteen cases were classified as orthopedic: bone defects were observed in 11 cases of total hip arthroplasty; one case of primary total hip arthroplasty, due to coxarthrosis; five cases of femoral or tibial open wedge osteotomy; and one case of tarsal arthrodesis. There were 18 cases of trauma-related defects; uninfected pseudarthrosis, eight cases; recent fractures of the tibial plateau with compression of the spongy bone, three cases; and exposed fractures treated with external fixators, seven cases. The surgical technique consisted of curetting and debriding the injury until bone suitable for grafting was found. Biological ceramic was then used to fill the defect and some kind of fixation was applied.

Results: Among the 36 patients evaluated, it was seen that 35 (97.2%) presented integration of the biological ceramic, while one case of open fracture treated with external fixation had poor integration of the biological ceramic.

Conclusion: Treatment of bone defects of orthopedic or post-traumatic etiology using a phosphocalcium ceramic composed of hydroxyapatite was shown to be a practical, effective and safe method.
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http://dx.doi.org/10.1016/S2255-4971(15)30393-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4799113PMC
March 2016

Altered blood glucose concentration is associated with risk of death among patients with community-acquired Gram-negative rod bacteremia.

BMC Infect Dis 2010 Jun 22;10:181. Epub 2010 Jun 22.

Instituto de Investigación y Formación Marques de Valdecilla, 5 Planta de la Escuela Universitaria de Enfermería, Avda de Valdecilla s/n, 39008, Santander, Spain.

Background: Altered blood glucose concentration is commonly observed in patients with sepsis, even among those without hypoglycemic treatments or history of diabetes mellitus. These alterations in blood glucose are potentially detrimental, although the precise relationship with outcome in patients with bacteremia has not been yet determined.

Methods: A retrospective cohort study design for analyzing patients with Gram negative rod bacteremia was employed, with the main outcome measure being in-hospital mortality. Patients were stratified in quintiles accordingly deviation of the blood glucose concentration from a central value with lowest mortality. Cox proportional-hazards regression model was used for determining the relationship of same day of bacteremia blood glucose and death.

Results: Of 869 patients identified 63 (7.4%) died. Same day of bacteremia blood glucose concentration had a U-shaped relationship with in-hospital mortality. The lowest mortality (2%) was detected in the range of blood glucose concentration from 150 to 160 mg/dL. Greater deviation of blood glucose concentration from the central value of this range (155 mg/dL, reference value) was directly associated with higher risk of death (p = 0.002, chi for trend). The low-risk group (quintile 1) had a mortality of 3.3%, intermediate-risk group (quintiles 2, 3 and 4) a mortality of 7.1%, and the high-risk group (quintile 5) a mortality of 12.05%. In a multivariable Cox regression model, the hazard ratio for death among patients in the intermediate-risk group as compared with that in the low risk group was 2.88 (95% confidence interval, 1.01 to 8.18; P = 0.048), and for the high risk group it was 4.26 (95% confidence interval, 1.41 to 12.94; P = 0.01).

Conclusions: Same day of bacteremia blood glucose concentration is related with outcome of patients with Gram-negative rod bacteremia. Lowest mortality is detected in patients with blood glucose concentration in an interval of 150-160 mg/dL. Deviations from these values are associated with an increased risk of death.
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http://dx.doi.org/10.1186/1471-2334-10-181DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2912309PMC
June 2010

Diabetes does not affect outcome in patients with Enterobacteriaceae bacteremia.

BMC Infect Dis 2009 Jun 13;9:94. Epub 2009 Jun 13.

Instituto de Formación e Investigación Marqués de Valdecilla, 39008 Santander, Spain.

Background: There is limited information about the effect of diabetes on the prognosis of patients with bacterial infections. We performed a retrospective cohort study to investigate possible correlations between diabetes and prognosis in patients with Enterobacteriaceae bacteremia.

Methods: We reviewed the medical charts of 1112 patients who were treated at a community teaching hospital for Enterobacteriaceae bacteremia from January 1997 through June 2007. Factors associated with in-hospital mortality were analyzed by logistic regression analysis.

Results: Among the 1112 patients with Enterobacteriaceae bacteremia, 181 (16.3%) were diabetic patients; 90 patients (8.1%) died while in the hospital. Compared to non-diabetic patients, diabetic patients were older (75.4 +/- 11.9 years vs. 70 +/- 16.6 years, p < 0.001) and had more comorbidities. However, mortality among diabetic and non-diabetic patients was not different [7.2% vs. 8.2%, RR 1.13; 95% CI (0.67-1.9); p = 0.39]. In a multivariate analysis, the variables associated with in-hospital mortality were age, the origin of the bacteremia, and the presence of immunosuppression. Diabetes was not associated with outcome.

Conclusion: In this cohort of patients with Enterobacteriaceae bacteremia, diabetes was not associated with a poorer prognosis.
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http://dx.doi.org/10.1186/1471-2334-9-94DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2711956PMC
June 2009

Impact of antibiotic resistance and of adequate empirical antibiotic treatment in the prognosis of patients with Escherichia coli bacteraemia.

J Antimicrob Chemother 2007 Oct 20;60(4):855-63. Epub 2007 Jul 20.

Internal Medicine Service, Sierrallana Hospital, Barrio de Ganzo s/n, Torrelavega, Cantabria, Spain.

Background: Escherichia coli is the most frequent Gram-negative organism causing bacteraemia. There are few data about prognostic factors of bloodstream infections due to E. coli. In particular, the consequences of antibiotic resistance and of adequate empirical antibiotic treatment on outcome remain broadly unknown.

Methods: We conducted a retrospective cohort study of patients with E. coli bacteraemia between January 1997 and June 2005 to identify any association between antibiotic resistance, adequacy of empirical antibiotic therapy and mortality.

Results: Of 663 patients with E. coli bacteraemia, 36 (5.4%) died. Patients with multidrug-resistant (MDR) E. coli bacteraemia had a significantly lower frequency of correct empirical antibiotic treatment than patients with non-MDR E. coli bacteraemia [relative risk (RR) 0.53; 95% confidence interval (CI) 0.48-0.67], and also had a significantly higher mortality (RR 3.31; 95% CI 1.72-6.36). An association between the number of antibiotics to which E. coli was resistant with adequacy of empirical antibiotic (P < 0.001) and with mortality (P < 0.001) was detected. After adjustment for other significant risk factors and confounders, the inadequacy of empirical antibiotic treatment was associated with an increased mortality (adjusted OR 2.98; 95% CI 1.25-7.11). When the adequacy of empirical treatment was excluded from the model, the presence of MDR E. coli in blood cultures was also associated with the prognosis (adjusted OR 3.11; 95% CI 1.3-7.44). In multivariate analysis, other variables associated with the outcome were age, the presence of severe sepsis or shock, Charlson index score and a non-urinary origin of the bacteraemia.

Conclusions: Adequacy of empirical antibiotic treatment is an independent risk factor for mortality in patients with E. coli bacteraemia. MDR E. coli bacteraemia had a worse prognosis due, at least in part, to a lower frequency of correct empirical treatment.
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http://dx.doi.org/10.1093/jac/dkm279DOI Listing
October 2007

Proteomic analysis of phytopathogenic fungus Botrytis cinerea as a potential tool for identifying pathogenicity factors, therapeutic targets and for basic research.

Arch Microbiol 2007 Mar 24;187(3):207-15. Epub 2006 Nov 24.

Laboratory of Microbiology, Marine and Environmental Sciences Faculty, University of Cádiz, Pol. Río San Pedro, 11510 Puerto Real, Cádiz, Spain.

Botrytis cinerea is a phytopathogenic fungus causing disease in a substantial number of economically important crops. In an attempt to identify putative fungal virulence factors, the two-dimensional gel electrophoresis (2-DE) protein profile from two B. cinerea strains differing in virulence and toxin production were compared. Protein extracts from fungal mycelium obtained by tissue homogenization were analyzed. The mycelial 2-DE protein profile revealed the existence of qualitative and quantitative differences between the analyzed strains. The lack of genomic data from B. cinerea required the use of peptide fragmentation data from MALDI-TOF/TOF and ESI ion trap for protein identification, resulting in the identification of 27 protein spots. A significant number of spots were identified as malate dehydrogenase (MDH) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The different expression patterns revealed by some of the identified proteins could be ascribed to differences in virulence between strains. Our results indicate that proteomic analysis are becoming an important tool to be used as a starting point for identifying new pathogenicity factors, therapeutic targets and for basic research on this plant pathogen in the postgenomic era.
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http://dx.doi.org/10.1007/s00203-006-0188-3DOI Listing
March 2007