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1.
Rev. argent. microbiol ; 51(3): 268-277, set. 2019. graf, tab
Article in English | LILACS | ID: biblio-1041836

ABSTRACT

Phytophthora parasitica is an important oomycete that causes disease in a variety of plants, dimethomorph fungicides being specific for oomycetes. The aim of this study was to use RNA-seq to rapidly discover the mechanism by which dimethomorph acts in the treatment of P. parasitica. We found that the expression of 832 genes changed significantly after the dimethomorph treatment, including 365 up-regulated genes and 467 down-regulated genes. According to the Gene Ontology (GO) enrichment analysis, pathway enrichment and verification test results, the following conclusions are obtained: (i) the treatment of P. parasitica with dimethomorph causes changes in the expression levels of genes associated with the cell wall and cell wall synthesis; (ii) dimethomorph treatment results in reduced permeability of the cell membrane and changes in the expression of certain transport-related proteins; (iii) dimethomorph treatment increased reactive oxygen species and reduced the expression of genes related to the control of oxidative stress.


Phytophthora parasitica es un importante oomiceto que origina enfermedades en una variedad de plantas; el fungicida dimetomorf es específico contra oomicetos. El objetivo de este estudio fue utilizar la tecnología de RNA-seq para descubrir rápidamente el mecanismo por el que el dimetomorf actúa en el tratamiento de P. parasitica. Descubrimos que la expresión de 832 genes se modificaba significativamente tras el tratamiento con dimetomorf, incluyendo 365 genes que son sobrerregulados y 467 genes que son subrregulados. El análisis de enriquecimiento de ontología de genes (GO), análisis de enriquecimiento de las vías y pruebas de verificación permitieron extraer las conclusiones siguientes: 1) el tratamiento de P. parasitica con dimetomorf origina cambios en los niveles de expresión de los genes relacionados con la pared celular y su síntesis; 2) el tratamiento con dimetomorf origina una reducción de la permeabilidad de la membrana celular, así como cambios en la expresión de ciertas proteínas relacionadas con el transporte, y 3) el tratamiento con dimetomorf incrementó las especies reactivas del oxígeno y redujo la expresión de los genes relacionados con el control del estrés oxidativo.


Subject(s)
Phytophthora/drug effects , RNA, Messenger/biosynthesis , Morpholines/pharmacology , Fungicides, Industrial/pharmacology , RNA-Seq , Phytophthora/genetics , Plant Diseases/parasitology , RNA, Messenger/genetics , Carrier Proteins/biosynthesis , Carrier Proteins/genetics , Cell Membrane Permeability/drug effects , Cell Membrane Permeability/genetics , Cell Wall/metabolism , Gene Expression Regulation/drug effects , Sequence Alignment , Reactive Oxygen Species , Oxidative Stress/genetics , beta-Glucans/analysis , Real-Time Polymerase Chain Reaction , Gene Ontology
2.
Journal of Zhejiang University. Science. B ; (12): 84-94, 2019.
Article in English | WPRIM | ID: wpr-1010445

ABSTRACT

Peach brown rot, caused by Monilinia fructicola, is one of the most serious peach diseases. A strain belonging to the Actinomycetales, named Streptomyces blastmyceticus JZB130180, was found to have a strong inhibitory effect on M. fructicola in confrontation culture. Following the inoculation of peaches in vitro, it was revealed that the fermentation broth of S. blastmyceticus JZB130180 had a significant inhibitory effect on disease development by M. fructicola. The fermentation broth of S. blastmyceticus JZB130180 had an EC50 (concentration for 50% of maximal effect) of 38.3 µg/mL against M. fructicola, as determined in an indoor toxicity test. Analysis of the physicochemical properties of the fermentation broth revealed that it was tolerant of acid and alkaline conditions, temperature, and ultraviolet radiation. In addition, chitinase, cellulase, and protease were also found to be secreted by the strain. The results of this study suggest that S. blastmyceticus JZB130180 may be used for the biocontrol of peach brown rot.


Subject(s)
Ascomycota/pathogenicity , Bacterial Proteins/metabolism , Cell Wall/metabolism , Cellulase/metabolism , Chitinases/metabolism , Fermentation , Fruit/microbiology , Pest Control, Biological/methods , Phylogeny , Plant Diseases/prevention & control , Prunus persica/microbiology , Siderophores/metabolism , Streptomyces/physiology
3.
Biol. Res ; 51: 49, 2018. tab, graf
Article in English | LILACS | ID: biblio-1011393

ABSTRACT

BACKGROUND: Antarctic bryophytes (mosses and liverworts) are resilient to physiologically extreme environmental conditions including elevated levels of ultraviolet (UV) radiation due to depletion of stratospheric ozone. Many Antarctic bryophytes synthesise UV-B-absorbing compounds (UVAC) that are localised in their cells and cell walls, a location that is rarely investigated for UVAC in plants. This study compares the concentrations and localisation of intracellular and cell wall UVAC in Antarctic Ceratodon purpureus, Bryum pseudotriquetrum and Schistidium antarctici from the Windmill Islands, East Antarctica. RESULTS: Multiple stresses, including desiccation and naturally high UV and visible light, seemed to enhance the incorporation of total UVAC including red pigments in the cell walls of all three Antarctic species analysed. The red growth form of C. purpureus had significantly higher levels of cell wall bound and lower intracellular UVAC concentrations than its nearby green form. Microscopic and spectroscopic analyses showed that the red colouration in this species was associated with the cell wall and that these red cell walls contained less pectin and phenolic esters than the green form. All three moss species showed a natural increase in cell wall UVAC content during the growing season and a decline in these compounds in new tissue grown under less stressful conditions in the laboratory. CONCLUSIONS: UVAC and red pigments are tightly bound to the cell wall and likely have a long-term protective role in Antarctic bryophytes. Although the identity of these red pigments remains unknown, our study demonstrates the importance of investigating cell wall UVAC in plants and contributes to our current understanding of UV-protective strategies employed by particular Antarctic bryophytes. Studies such as these provide clues to how these plants survive in such extreme habitats and are helpful in predicting future survival of the species studied.


Subject(s)
Pigments, Biological/radiation effects , Pigments, Biological/metabolism , Ultraviolet Rays , Cell Wall/radiation effects , Cell Wall/metabolism , Bryophyta/radiation effects , Bryophyta/metabolism , Seasons , Time Factors , Pigmentation/radiation effects , Analysis of Variance , Chromatography, High Pressure Liquid , Spectroscopy, Fourier Transform Infrared/methods , Plant Leaves/radiation effects , Plant Leaves/metabolism , Microscopy, Confocal , Bryophyta/cytology , Antarctic Regions
4.
Electron. j. biotechnol ; 26: 12-19, Mar. 2017. ilus, graf, tab
Article in English | LILACS | ID: biblio-1009164

ABSTRACT

Background: Expansins play an important role in cell wall metabolism and fruit softening. Determination of expansin activity is a challenging problem since it depends on measuring cell wall properties by using ad hoc extensometers, a fact that has strongly restricted its study. Then, the objective of the work was to adapt a methodology to measure cell wall creep and expansin activity using a commercial texture meter, equipped with miniature tensile grips and an ad hoc cuvette of easy construction. Results: It was possible to measure hypocotyls acid growth and expansin activity in a reliable and reproducible way, using a commercial texture meter, common equipment found in laboratories of food science or postharvest technology. Expansin activity was detected in protein extracts from cucumber hypocotyls, tomato and strawberry fruits, and statistical differences in expansin activity were found in both fruit models at different ripening stages. Conclusions: The possibility of measuring expansin activity following this adapted protocol with a commercial texture meter could contribute to ease and increase the analysis of expansin in different systems, leading to a better understanding of the properties of these proteins under different experimental conditions.


Subject(s)
Plant Proteins/metabolism , Solanum lycopersicum/metabolism , Cucumis sativus/metabolism , Fragaria/metabolism , Plant Proteins/analysis , Cell Wall/metabolism , Hypocotyl/growth & development , Elasticity , Fruit/metabolism
5.
Rev. Soc. Bras. Med. Trop ; 50(1): 9-18, Jan.-Feb. 2017. tab, graf
Article in English | LILACS | ID: biblio-842818

ABSTRACT

Abstract: The lipid-rich cell wall of Mycobacterium tuberculosis is a dynamic structure that is involved in the regulation of the transport of nutrients, toxic host-cell effector molecules, and anti-tuberculosis drugs. It is therefore postulated to contribute to the long-term bacterial survival in an infected human host. Accumulating evidence suggests that M. tuberculosis remodels the lipid composition of the cell wall as an adaptive mechanism against host-imposed stress. Some of these lipid species (trehalose dimycolate, diacylated sulphoglycolipid, and mannan-based lipoglycans) trigger an immunopathologic response, whereas others (phthiocerol dimycocerosate, mycolic acids, sulpholipid-1, and di-and polyacyltrehalose) appear to dampen the immune responses. These lipids appear to be coordinately expressed in the cell wall of M. tuberculosis during different phases of infection, ultimately determining the clinical fate of the infection. This review summarizes the current state of knowledge on the metabolism, transport, and homeostatic or immunostatic regulation of the cell wall lipids, and their orchestrated interaction with host immune responses that results in bacterial clearance, persistence, or tuberculosis.


Subject(s)
Humans , Cell Wall/metabolism , Lipids/physiology , Mycobacterium tuberculosis/physiology , Membrane Transport Proteins , Cell Wall/physiology , Lipid Metabolism , Immunity, Innate , Membrane Lipids/physiology , Mycobacterium tuberculosis/metabolism
6.
Braz. j. microbiol ; 47(1): 25-32, Jan.-Mar. 2016. tab, graf
Article in English | LILACS | ID: lil-775113

ABSTRACT

Abstract Twelve isolates of Trichoderma spp. isolated from tobacco rhizosphere were evaluated for their ability to produce chitinase and β-1,3-glucanase extracellular hydrolytic enzymes. Isolates ThJt1 and TvHt2, out of 12 isolates, produced maximum activities of chitinase and β-1,3-glucanase, respectively. In vitro production of chitinase and β-1,3-glucanase by isolates ThJt1 and TvHt2 was tested under different cultural conditions. The enzyme activities were significantly influenced by acidic pH and the optimum temperature was 30 °C. The chitin and cell walls of Sclerotium rolfsii, as carbon sources, supported the maximum and significantly higher chitinase activity by both isolates. The chitinase activity of isolate ThJt1 was suppressed significantly by fructose (80.28%), followed by glucose (77.42%), whereas the β-1,3-glucanase activity of ThJt1 and both enzymes of isolate TvHt2 were significantly suppressed by fructose, followed by sucrose. Ammonium nitrate as nitrogen source supported the maximum activity of chitinase in both isolates, whereas urea was a poor nitrogen source. Production of both enzymes by the isolates was significantly influenced by the cultural conditions. Thus, the isolates ThJt1 and TvHt2 showed higher levels of chitinase and β-1,3-glucanase activities and were capable of hydrolyzing the mycelium of S. rolfsii infecting tobacco. These organisms can be used therefore for assessment of their synergism in biomass production and biocontrol efficacy and for their field biocontrol ability against S. rolfsii and Pythium aphanidermatum infecting tobacco.


Subject(s)
Chitinases , Soil Microbiology , Trichoderma/enzymology , Trichoderma/growth & development , Basidiomycota/metabolism , Carbon/metabolism , Cell Wall/metabolism , Chitin/metabolism , Culture Media/chemistry , Hydrogen-Ion Concentration , Nitrogen/metabolism , Rhizosphere , Temperature , Nicotiana , Trichoderma/isolation & purification
7.
Gac. méd. Caracas ; 120(2): 107-114, abr.-jun. 2012. tab
Article in Spanish | LILACS | ID: lil-679022

ABSTRACT

La fibra dietética o fibra alimentaria, forma parte de lo que se considera una dieta equilibrada o saludable. A pesar que la fibra alimentaria no se considera un nutriente, es un componente importante de la dieta diaria. La razón principal de su importancia, es que pasa por el sistema digestivo sin ser absorbida y este hecho fisiológico, trae beneficios a la salud. La fibra alimentaria, se considera como el material alimenticio particularmente de origen vegetal que no es hidrolizado por las enzimas secretadas por el tracto digestivo humano, pero, que puede ser fermentada en el intestino grueso por la microflora colónica. El grupo de enfermedades crónicas no transmisibles, incluye: obesidad, diabetes, enfermedades cardiovasculares y cerebrovasculares, la hipertensión arterial, el cáncer y problemas articulares. Estudios epidemiológicos, muestran el efecto beneficioso de la fibra en el tratamiento terapéutico en algunas de estas enfermedades crónicas


Dietary fiber or alimentary fiber, is part of what is considered a balanced or healthy diet. Although the alimentary fiber is not considered a nutrient, is an important component of the diet. The main reason for its importance is passing through the digestive system without being absorbed and this physiological fact is beneficial to health. The alimentary fiber is considered as particularly the food material of plant origin that is not hydrolyzed by enzymes secreted by the human digestive tract, but that can be fermented in the large intestine by colonic microflora. The group of chronic non-communicable diseases, include: obesity, diabetes, cardiovascular and cerebrovascular diseases, hypertension, cancer and joint problems. Epidemiological studies show the beneficial effect of fiber in the therapeutic treatment in some of these chronic diseases


Subject(s)
Humans , Male , Female , Diabetes Mellitus/metabolism , Chronic Disease/therapy , Cardiovascular Diseases/metabolism , Dietary Fiber/therapeutic use , Obesity/etiology , Cell Wall/metabolism , Organic Chemicals/administration & dosage , Health Behavior , Indigenous Peoples , Digestive System/physiopathology
8.
The Korean Journal of Parasitology ; : 361-364, 2012.
Article in English | WPRIM | ID: wpr-69773

ABSTRACT

The mature cyst of Acanthamoeba is highly resistant to various antibiotics and therapeutic agents. Cyst wall of Acanthamoeba are composed of cellulose, acid-resistant proteins, lipids, and unidentified materials. Because cellulose is one of the primary components of the inner cyst wall, cellulose synthesis is essential to the process of cyst formation in Acanthamoeba. In this study, we hypothesized the key and short-step process in synthesis of cellulose from glycogen in encysting Acanthamoeba castellanii, and confirmed it by comparing the expression pattern of enzymes involving glycogenolysis and cellulose synthesis. The genes of 3 enzymes, glycogen phosphorylase, UDP-glucose pyrophosphorylase, and cellulose synthase, which are involved in the cellulose synthesis, were expressed high at the 1st and 2nd day of encystation. However, the phosphoglucomutase that facilitates the interconversion of glucose 1-phosphate and glucose 6-phosphate expressed low during encystation. This report identified the short-cut pathway of cellulose synthesis required for construction of the cyst wall during the encystation process in Acanthamoeba. This study provides important information to understand cyst wall formation in encysting Acanthamoeba.


Subject(s)
Acanthamoeba castellanii/enzymology , Amebiasis/parasitology , Cell Wall/metabolism , Cellulose/biosynthesis , Glucosyltransferases/genetics , Glycogen Phosphorylase/genetics , Protozoan Proteins/genetics , UTP-Glucose-1-Phosphate Uridylyltransferase/genetics
9.
Indian J Biochem Biophys ; 2011 Oct; 48(5): 341-345
Article in English | IMSEAR | ID: sea-135338

ABSTRACT

Stripe rust (Puccinia striiformis f.sp. tritici) is the most devastating disease of wheat (Triticum aestivum L.) accounting huge economical losses to the industry worldwide. HD 2329 was a widely grown wheat cultivar which had become highly susceptible to stripe rust and was used to understand the biochemical aspects of the host pathogen interaction through characterization of superoxide dismutase (SOD). In the present study, two types of SOD, ionically or covalently bound to the particulate fraction were found in the stripe rust infected and uninfected wheat leaves of susceptible cultivar HD 2329. Cell walls of leaves contained a high level of SOD, of which 41-44% was extractable by 2 M NaCl and 10-13% by 0.5% EDTA in infected and uninfected leaves. The NaCl-released SOD constituted the predominant fraction. It exhibited maximum activity at pH 9.0, had a Km value of 1.82-2.51 for uninfected and 1.77-2.37 mM for infected, respectively with pyrogallol as the substrate, and a Vmax of 9.55-21.4 and 12.4-24.1 A min-1g-1FW. A temperature optimum of 20oC was observed for SOD of both uninfected and infected leaves. SOD showed differential response to metal ions, suggesting their distinctive nature. Inhibition of wall bound SOD by iodine and its partial regeneration of activity by mercaptoethanol suggested the involvement of cysteine in active site of the enzyme. These two forms showed greater differences with respect to thermodynamic properties like energy of activation (Ea) and enthalpy change (H), while entropy change (S) and free energy change (G) were similar. The results further showed that pathogen infection of the leaves of susceptible wheat cultivar induced a decrease in the SOD activity and kinetics which might be critical during the response of plant cells to the infection.


Subject(s)
Basidiomycota/metabolism , Basidiomycota/pathogenicity , Cell Wall/chemistry , Cell Wall/enzymology , Cell Wall/metabolism , Enzyme Inhibitors/chemistry , Hydrogen-Ion Concentration , Kinetics , Metals/chemistry , Plant Cells/enzymology , Plant Diseases/microbiology , Plant Leaves/enzymology , Superoxide Dismutase/chemistry , Superoxide Dismutase/pharmacokinetics , Temperature , Triticum/enzymology
10.
Indian J Biochem Biophys ; 2010 Aug; 47(4): 249-253
Article in English | IMSEAR | ID: sea-135273

ABSTRACT

Plant cell wall expresses monoamine oxidases (MAOs) that catalyze oxidation of secreted amines and produce H2O2 in the process. The H2O2, so produced is used by cell wall peroxidases for lignification of cell wall or for plant defense. The natural substrates for these MAOs are elusive, but polyamines and certain catecholamines have been proposed as candidates. Reactive oxygen species are also known to act as signaling molecules controlling plant metabolism. Mungbean (Vigna radiata) has long served as the plant model of choice while studying molecular programs followed during germination and seed development. In this study, we tested the effect of externally added MAO substrates epinephrine and H2O2 on storage protein mobilization in germinating seeds of Vigna radiata. The seeds were imbibed in the presence of 50 M epinephrine and 10 M H2O2. These low concentrations of the two compounds were used to exclude direct effects on proteolysis and were arrived at after testing a range of the two and choosing the most effective concentration. These seeds showed 11% and 7% decrease in fresh weight respectively, indicating greater storage mobilization and a corresponding 19% and 46% increase in axis length as compared to untreated seeds. Soluble protein in seeds treated with epinephrine and H2O2 decreased significantly by 34% and 33% as compared to untreated seeds. Electrophoretic analysis of seed proteins revealed a startling and selective depletion of storage proteins in treated seeds. The results indicated a clear involvement of H2O2 in storage protein mobilization in the cotyledons. We propose that H2O2 generated within cell walls of seeds serves as a signaling molecule guiding germination events, including protein reserve mobilization.


Subject(s)
Cell Wall/enzymology , Cell Wall/metabolism , Densitometry/methods , Electrophoresis, Polyacrylamide Gel/methods , Epinephrine/chemistry , Epinephrine/pharmacology , Fabaceae/enzymology , Germination/drug effects , Germination/physiology , Hydrogen Peroxide/chemistry , Hydrogen Peroxide/pharmacology , Lignin/chemistry , Monoamine Oxidase/chemistry , Plant Proteins/chemistry , Reactive Oxygen Species , Seeds/chemistry , Signal Transduction
11.
Article in English | IMSEAR | ID: sea-22213

ABSTRACT

BACKGROUND & OBJECTIVE: Slime is a major determinant of Staphylococcus epidermidis adherence.The established methods of laboratory detection of slime production by this organism i.e., Christensen's tube method and congo red agar plate method, can both yield inconclusive and/or intermediate results. We, therefore tried to find out electronmicroscopically the localization of slime in relation to the bacterial cell wall and look for the effect, if any of the slime location on the staphylococcal adherence as well as on the quantum of slime production. METHODS: A total of 132 coagulase negative staphylococci from cases of infectious keratitis were identified as S. epidermidis following the recommended protocol. Slime was detected both by Christensen's tube method and congo red agar plate method. Antibiotic sensitivity testing was performed by standardized disc diffusion method. Adherence of the organisms to artificial surfaces was determined by a quantitative method and transmission electron microscopy was carried out by the conventional techniques. RESULTS: Of the total 132 isolates, 57 (43.2%) were slime positive and 75 (56.8%) were slime negative.Twenty seven (47.4%) of the 57 slime producing organisms were multi drug resistant as compared to only 12 (16%) of 75 nonslime-producing organisms (P<0.001). A majority i.e., 45 (78.9%) of 57 adherent organisms were slime producers as against 12 (16%) of 75 nonadherent organisms. Electron microscopic study revealed a thick viscid layer of slime anchoring to the bacterial cell wall, especially in adherent organisms and those yielding positive slime test. Some of the organisms showed loose nonadherent slime and those were mostly nonadherent to artificial surfaces. INTERPRETATION & CONCLUSION: Slime and multi drug resistance were the important virulence factors of S. epidermidis in bacterial keratitis. It was the adherent slime (i.e., slime in intimate association with the bacterial cell wall as shown by electron microscopy) only, which was responsible for resistance to multiple antibiotics and for the adhesion phenomenon observed in the quantitative slime test.


Subject(s)
Agar/chemistry , Animals , Anti-Bacterial Agents/chemistry , Bacterial Adhesion , Cell Wall/metabolism , Congo Red/pharmacology , Humans , Keratitis/microbiology , Microbial Sensitivity Tests , Microscopy, Electron, Transmission , Staphylococcus epidermidis/metabolism , Virulence Factors
12.
Indian J Exp Biol ; 2005 Oct; 43(10): 897-901
Article in English | IMSEAR | ID: sea-62236

ABSTRACT

Saccharomyces cerevisiae cells when grown on synthetic medium plates containing 10 mM of 4-aminopyridine (4-AP) undergo cell lysis. Using an ethylmethane sulfonate mutagenesis (EMS) screen, 4-AP resistant mutants (apr) were isolated which could grow on inhibitory concentration of 4-AP. Eighty mutants were obtained that were recessive, monogenic and formed two complementation groups. To identify genes, whose products might be interacting with the apr loci, extragenic suppressors were isolated, which reverted 4-AP resistance phenotype of apr mutants. The suppressors, when genetically characterized, were found to be recessive and represented two loci with overlapping functions. Representative alleles from apr mutants were analyzed for cell wall composition. They were found to have a higher amount of alkali-insoluble glucan signifying the role of alkali-insoluble glucan in cell wall maintenance.


Subject(s)
4-Aminopyridine/pharmacology , Cell Wall/metabolism , Drug Resistance , Ethyl Methanesulfonate/pharmacology , Genetic Complementation Test , Glucan 1,3-beta-Glucosidase/metabolism , Glucans/chemistry , Mutagens , Mutation , Phenotype , Potassium/pharmacokinetics , Protein Binding , Saccharomyces cerevisiae/metabolism , beta-Glucans/chemistry
13.
Indian J Exp Biol ; 2005 Oct; 43(10): 892-6
Article in English | IMSEAR | ID: sea-60601

ABSTRACT

Fungal cell wall degrading chitinases and glucanases attained significance in agriculture, medicine, and environment management. The present study was conducted to describe the optimum conditions required for the production of beta-1,4-N-acetyl glucosaminidase (NAGase) and beta-1,3-glucanase by a biocontrol strain of Bacillus subtilis AF 1. B. subtilis AF 1 was grown in minimal medium with colloidal chitin (3.0%) and yeast extract (0.3% YE ) and incubated at pH 7.0 and 30 degrees C on constant shaker at 180 rpm for 6 days produced highest amounts of NAGase. Presence of 0.5 mM of phenyl methyl sulfonyl fluoride (PMSF) and 0.04% of Tween 20 further improved the enzyme production. B. subtilis AF 1 grown in minimal medium with laminarin (1%) and yeast extract (0.3%) for 3 days produced maximum amount of beta-1,3-glucanase. These conditions can be further scaled-up for large-scale production of NAGase and beta-1,3-glucanase by B. subtilis AF 1.


Subject(s)
Acetylglucosaminidase/metabolism , Bacillus subtilis/enzymology , Carbon/chemistry , Cell Wall/metabolism , Culture Media , Detergents/pharmacology , Dose-Response Relationship, Drug , Glucan 1,3-beta-Glucosidase/metabolism , Hydrogen-Ion Concentration , Polysaccharides/pharmacology , Polysorbates/pharmacology , Temperature , Time Factors , Tosyl Compounds/pharmacology
14.
Genet. mol. res. (Online) ; 4(2): 430-449, 30 jun. 2005. tab, ilus
Article in English | LILACS | ID: lil-445278

ABSTRACT

The rise in antifungal resistance, observed as a result of the increasing numbers of immunocompromised patients, has made the discovery of new targets for drug therapy imperative. The description of the Paracoccidioides brasiliensis transcriptome has allowed us to find alternatives to refine current therapy against paracoccidioidomycosis. We used comparative analysis of expressed sequence tags to find promising drug targets that have been addressed in other pathogens. We divided the analysis into six different categories, based on the involvement of the targeted mechanisms in the cell: i) cell wall construction, ii) plasma membrane composition, iii) cellular machinery, iv) cellular metabolism, v) signaling pathways, and vi) other essential processes. Through this approach, it has been possible to infer strategies to develop alternative drugs against this pathogen.


Subject(s)
Humans , Antifungal Agents/pharmacology , Drug Design , Expressed Sequence Tags , Paracoccidioides/genetics , Transcription, Genetic/genetics , Paracoccidioides/drug effects , Paracoccidioides/metabolism , Cell Wall/drug effects , Cell Wall/enzymology , Cell Wall/metabolism
15.
Genet. mol. res. (Online) ; 4(2): 309-325, 30 jun. 2005. tab, ilus
Article in English | LILACS | ID: lil-445285

ABSTRACT

The cell wall of a human pathogenic fungus is in contact with the host, serves as a barrier against host defense mechanisms and harbors most fungal antigens. In addition, cell wall biosynthesis pathways have been recognized as essential to viability and as specific drug targets. Paracoccidioides brasiliensis is a dimorphic fungus that presents mycelium morphology in the free environment and causes infection in a yeast form. The morphogenetic conversion is correlated with changes in the cell wall composition, organization and structure. Based on transcriptome analysis, the enzymes involved in the biosynthesis and remodeling of cell wall polysaccharides, as well as several cell wall-associated molecules of P. brasiliensis, were identified and addressed in further detail.


Subject(s)
Humans , Expressed Sequence Tags/metabolism , Mycelium/cytology , Paracoccidioides/cytology , Cell Wall/metabolism , Transcription, Genetic/genetics , Sequence Alignment , Genes, Fungal , Mycelium/enzymology , Mycelium/genetics , Paracoccidioides/enzymology , Paracoccidioides/genetics , Cell Wall/chemistry , Cell Wall/genetics , Gene Expression Profiling
16.
Indian J Exp Biol ; 2004 Jun; 42(6): 636-8
Article in English | IMSEAR | ID: sea-57183

ABSTRACT

Ruminal fungal isolates (Orpinomyces sp.; C-14, Piromyces sp.; C-15, Orpinomyces sp.; B-13 and Anaeromyces sp.; B-6), were evaluated under anoxic conditions for their effect on in vitro dry matter digestibility, neutral detergent fibre, acid detergent fibre and acid detergent lignin using rice and wheat straw as substrate. There was no significant effect of the fungal isolates on the disappearance of the substrates along with rumen liquor when compared to control. The doses of 10(6) cfu/ml of the isolate were found to have maximum degradation of straws in comparison to the doses of 10(3) cfu/ml.


Subject(s)
Cell Wall/metabolism , Edible Grain/microbiology , Detergents/pharmacology , Neocallimastigales/metabolism , Oryza/microbiology , Temperature , Time Factors , Triticum/microbiology
17.
Biocell ; 27(2): 173-179, Aug. 2003.
Article in English | LILACS | ID: lil-384246

ABSTRACT

Vicilins (7S storage proteins) found in various legume seeds have been previously shown to interfere with the germination of spores or conidia of phytopathogenic fungi and inhibit yeast growth and glucose stimulated acidification of the medium by yeast cells. In the present work vicilins from cowpea (Vigna unguiculata) seeds were added to the growth medium of Saccharomyces cerevisiae cells and Fusarium oxysporum conidia. Helix pomatia lectin, wheat germ agglutinin and Ulex europaeus lectin were used to identify differences in the binding of the vicilins to the surface of cells of S. cerevisiae and F. oxysporum treated with this protein. After the growth period, the material in suspension (yeast cells) was centrifuged and the final pellet was also treated with different sugar (glucose, sucrose, glucosamine, N-acetyl-glucosamine) concentrations and 0.1 M HCl for extraction of vicilins associated to chitinous structures present in yeast cells. Our results showed that vicilin sub-units were present in the different sugar extracts of yeast cells pretreated with the vicilins and these proteins were eluted by 0.5 M solutions of sugars in the following order of efficiency of elution: N-acetyl-glucosamine, sucrose/glucose and glucosamine.


Subject(s)
Carbohydrates/pharmacology , Binding, Competitive/drug effects , Cell Membrane/drug effects , Cell Wall/drug effects , Plant Proteins/pharmacology , Acetylglucosamine/pharmacology , Fungi/drug effects , Fungi/growth & development , Fungi/ultrastructure , Fusarium/drug effects , Fusarium/growth & development , Fusarium/ultrastructure , Glucosamine/pharmacology , Glucose/pharmacology , Binding, Competitive/physiology , Microscopy, Electron , Cell Membrane/metabolism , Cell Membrane/ultrastructure , Cell Wall/metabolism , Cell Wall/ultrastructure , Sucrose/pharmacology , Saccharomyces cerevisiae/drug effects , Saccharomyces cerevisiae/growth & development , Saccharomyces cerevisiae/ultrastructure , Binding Sites/drug effects , Binding Sites/physiology
18.
Biocell ; 27(1): 11-18, Apr. 2003.
Article in English | LILACS | ID: lil-384255

ABSTRACT

Cell wall pectins are some of the most complex biopolymers known, and yet their functions remain largely mysterious. The aim of this paper was to deepen the study of the spatial pattern of pectin distribution in the aperture of Oenothera hookeri.velans ster/+ster fertile pollen. We used "in situ" immunocytochemical techniques at electron microscopy, involving monoclonal antibodies JIM5 and JIM7 directed against pectin epitopes in fertile pollen grains of Oenothera hookeri.velans ster/+ster. The same region was also analyzed by classical cytochemistry for polysaccharide detection. Immunogold labelling at the JIM7 epitope showed only in mature pollen labelling mainly located at the intine endo-aperture region. Cytoplasmic structures near the plasma membrane of the vegetative cell showed no labelling gold grains. In the same pollen stge the labelling at the JIM5 epitope was mostly confined to a layer located in the limit between the endexine and the ektexine at the level of the border of the oncus. Some tubuli at the base of the ektexine showed also an accumulation of gold particles. No JIM5 label was demonstrated in the aperture chamber and either in any cytoplasmic structure of the pollen grains. The immunocytochemical technique, when compared with the traditional methods for non-cellulose polysaccharide cytochemistry is fare more sensitive and allows the univocal determination of temporal and spatial location of pectins recognized by the JIM7 and JIM5 MAbs.


Subject(s)
Oenothera/metabolism , Pectins/metabolism , Pollen/metabolism , Antibodies, Monoclonal/metabolism , Epitopes/metabolism , Immunohistochemistry , Oenothera/cytology , Cell Wall/chemistry , Cell Wall/metabolism , Cell Wall/ultrastructure , Pollen/ultrastructure , Polysaccharides/chemistry , Polysaccharides/metabolism , Sensitivity and Specificity , Time Factors , Tissue Distribution
19.
P. R. health sci. j ; 17(4): 323-6, Dec. 1998. ilus, graf
Article in English | LILACS | ID: lil-234845

ABSTRACT

OBJECTIVE: To determine whether cell cycle changes can be detected in myosin II-deficient cells using flow cytometry techniques. BACKGROUND: Although the primary role of myosin II (Myo1p) in the yeast Saccharomyces cerevisiae is in cytokinesis we have reported that this conventional myosin also appears to inuence the regulation of cell wall metabolism as indicated by increases in the expression of chitin metabolizing enzymes in a null mutant of the MYO1 gene. The expression of these enzymes is known to be regulated in the cell cycle suggesting that cell cycle changes may alter their expression. METHODS: Flow cytometry was employed to assess the nuclear DNA content of logarithmic yeast cell cultures as a means of determining changes in the cell cycle of Myo1p-deficient cells. RESULTS: Significant changes were observed in the Myo1p-deficient strain suggesting that these cells are arrested in G2/M-phase of the cell cycle. CONCLUSIONS: Based on the results of this preliminary study, we propose a model in which the increased activity of chitin metabolizing enzymes may be explained by a mitotic arrest in these cells.


Subject(s)
Myosin Heavy Chains/metabolism , Yeasts/cytology , Yeasts/metabolism , Cell Culture Techniques , Cell Cycle , Cell Division , Cell Wall/metabolism , Chitin Synthase/genetics , Chitin Synthase/metabolism , Chitin/metabolism , Flow Cytometry , Gene Expression , Haploidy , Mitosis , Myosin Heavy Chains/deficiency , Myosin Heavy Chains/genetics , RNA, Messenger/genetics , Saccharomyces cerevisiae/cytology , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , Yeasts/genetics
20.
Arch. latinoam. nutr ; 48(1): 41-6, mar. 1998. tab
Article in English | LILACS | ID: lil-217536

ABSTRACT

Four samples each of black beans representing two types of vegetative growth were collected from farmers' fields in four locationes in Guatemala. Soon after collection, samples were stored at 4 and 38 degrees Celsius at ambient relative humidity and subsamples were withdrawn at 0,45, 90 and 135 days of strage for determination of water absorption, cooking time and analysis of neutral-and acid detergent fiber, cellulose, hemicellulose and lignin. The fiber fraction analysis were done on samples of 0,45 and 90 days of storage. Water absorption for all 4 samples of the bush type was similar at both storage T, however the samples stored at 38 degrees Celsius and at 135 days absorbed more water than when stored at 4 degrees Celsius. The 4 vine types of beans showed different water absorption rates, with two showing patterns similar to those beans of the bush type and two which did absorbed water at a very slow rate. For both types of beans stored at 4 degrees Celsius, cooking time decreased from 0 to 135 days of storage. On the other hand for all bean samples of the two types cooking time increased when stored at 38 degrees Celsius. Analysis of variance showed highly significant effects due to plant type, days of storage, temperature and locality, and for some interactions. Analysis of variance of the fiber fractions showed high significant differences for days of storage for NDF, ADF, cellulose, hemicellulose, and lignin. Plant type gave significant differences for cellulose and hemicellulose. Highly significant differences for hemicellulose were found for the interactions of type x days, type x temperature, locality x type, and type x days x temperature. The rate of synthesis of the 5 fractions were calculated by simple regression analysis. For the bush type of beans some synthesis occurred at 4 degrees Celsius, but it was enhanced when stored at 38 degrees Celsius. For vine type of beans at 4 degrees Celsius relative high rates of synthesis were observed, which were higher at 38 degrees Celsius for NDF, hemicellulose and lignin. Cooking time and fiber fraction contents were subjected to regression analysis. The correlations at 38 degrees Celsius were higher than at 4 degrees Celsius for all fractions for both types of beans, but statistical significance was obtained only for NDF, ADF and cellulose for vince type of beans. These data show therefore that synthesis of cell wall structure fractions, and not only lignin formation, are responsible...


Subject(s)
Fabaceae , Food Handling/methods , Analysis of Variance , Cell Wall/metabolism , Fabaceae/cytology , Fabaceae/growth & development , Guatemala , Time Factors
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