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PubMed1992

[Paracoccidioidomycosis assessed with 4 isotopic studies].

Paracoccidioidomycosis (south-american blastomycosis) is a consumptive granulomatous disease that is found in all Latin-America. The ethiological agent is a fungus (Paracoccidiosis brasiliensis) and mechanisms of disease transmission are not clear. Affection is frequent in the lungs, liver, spleen, ganglia, oropharynx and brain. Disease's response to therapy (Amphotericin, Ketoconazole) is good but recurrence is frequent. We have studied 12 patients with gallium-67, bone scintigraphy, bone marrow scintigraphy and lymph-scintigraphy. Six of these patients were re-studied after variable time of therapy. Gallium-67 accumulated in all patients's lungs. Other abnormal areas included ganglia, liver, spleen and bone. Bone scintigraphy was abnormal in all bone sites that were abnormal at the gallium study. Bone marrow scintigraphy showed expansion to the periphery in 6/12 cases and lymph nodes accumulated the radiopharmaceutical in 5 patients. Patients re-studied after the beginning of therapy presented decreased degrees or normalization of the radio-pharmaceutical's uptake (gallium and MDP), a regression of the bone marrow periphery expansion (bone marrow scintigraphy) and normalization of the lymph node aspects. These four radioisotopical studies may be useful for staging and for therapy follow-up. The sensitivity of the method is greater than the one noted for the conventional radiological studies.

Bone and Bones↗
PubMed2005

Loss of pectin is an early event during infection of cocoyam roots by Pythium myriotylum.

Cocoyam (Xanthosoma sagittifolium) is an important tuber crop in most tropical zones of Africa and America. In Cameroon, its cultivation is hampered by a soil-borne fungus Pythium myriotylum which is responsible for root rot disease. The mechanism of root colonisation by the fungus has yet to be elucidated. In this study, using microscopical and immunocytochemical methods, we provide a new evidence regarding the mode of action of the fungus and we describe the reaction of the plant to the early stages of fungal invasion. We show that the fungal attack begins with the colonisation of the peripheral and epidermal cells of the root apex. These cells are rapidly lost upon infection, while cortical and stele cells are not. Labelling with the cationic gold, which binds to negatively charged wall polymers such as pectins, is absent in cortical cells and in the interfacial zone of the infected roots while it is abundant in the cell walls of stele cells. A similar pattern of labelling is also found when using the anti-pectin monoclonal antibody JIM5, but not with anti-xyloglucan antibodies. This suggests that early during infection, the fungus causes a significant loss of pectin probably via degradation by hydrolytic enzymes that diffuse and act away from the site of attack. Additional support for pectin loss is the demonstration, via sugar analysis, that a significant decrease in galacturonic acid content occurred in infected root cell walls. In addition, we demonstrate that one of the early reactions of X. sagittifolium to the fungal invasion is the formation of wall appositions that are rich in callose and cellulose.

Cell Wall↗
PubMed2005

Root response to Fusarium solani f. sp . glycines: temporal accumulation of transcripts in partially resistant and susceptible soybean.

Sudden death syndrome (SDS) of soybean is a complex of root rot disease caused by the semi-biotrophic fungus Fusarium solani f. sp. glycines (Fsg) and a leaf scorch disease caused by toxins produced by the pathogen in the roots. Development of partial rate-reducing resistance in roots to SDS was studied. The recombinant inbred line 23 (RIL23) that carried resistance conferred by six quantitative trait loci (QTL) derived from cultivars 'Essex' x 'Forrest' was compared to the susceptible cultivar Essex. Roots of RIL23 and its susceptible parent Essex were inoculated with Fsg. Transcript abundance (TA) of 191 ESTs was studied at five time points after inoculation. For most of the genes, there was an initial decrease in TA in the inoculated roots of both genotypes. By days 7 and 10 the inoculated roots of Essex failed to increase expression of the transcripts of defense-related genes. In RIL23 inoculated roots, the TA of 81 genes was increased by at least two-fold at day 3 (P=0.004), 88 genes at day 7 (P=0.0023) and 129 genes at day 10 (P=0.0026). A set of 35 genes maintained at least a two-fold higher abundance at all three time points. The increase in TA in RIL23 was in contrast to that observed in Essex where most of the ESTs showed either no change or a decreased TA. The ESTs with an increased TA had homology to the genes involved in resistance (analogs), signal transduction, plant defense, cell wall synthesis and transport of metabolites. Pathways that responded included the protein phosphorylation cascade, the phospholipase cascade and the phenolic natural products pathways, including isoflavone and cell wall synthesis.

Crosses, Genetic↗
PubMed1999

THE DARK SIDE OF THE MYCELIUM: Melanins of Phytopathogenic Fungi.

Melanins are darkly pigmented polymers that protect organisms against environmental stress. Even when not directly involved in pathogenesis, fungal melanin is likely required by melanizing phytopathogens for survival in the environment. However, some phytopathogenic fungi that produce melanized appressoria for host invasion require appressorial melanogenesis for pathogenicity. Much less is known about the role melanins play in pathogenesis during infection by other phytopathogens that do not rely on appressoria for host penetration. Here we focus on one such phytopathogenic fungus, Gaeumannomyces graminis var. tritici, the etiologic agent of the devastating root disease of cereals, take-all. This fungus is lightly pigmented in culture, but requires melanin biosynthesis for pathogenesis, perhaps to produce melanized, ectotrophic macrohyphae on roots. However, the constitutively melanized, asexual Phialophora anamorph of G. graminis var. tritici is nonpathogenic. In addition, melanization of G. graminis var. graminis is not required to produce root disease on its rice host. Explanations for these apparent contradictions are suggested, as are other functions for the melanins of phytopathogenic fungi.

Journal Article↗
PubMed2006

Molecular cloning and expression analysis of rice OsTVLP1, encoding a protein with similarity to TGF-beta receptor interacting proteins and vacuolar assembly Vam6p/Vps39p proteins.

We describe the cloning and identification of a rice cDNA, OsTVLP1, encoding a protein with similarity to TGF-beta receptor interacting proteins and vacuolar assembly Vam6p/Vps39p proteins. OsTVLP1 has an open reading frame of 2955 bp, which encodes a 984 amino acid protein, containing a citron homology (CNH) domain at its N-terminal and a clathrin heavy-chain repeat homology (CLH) domain at its C-terminal. The expression of OsTVLP1 was induced by treatments with benzothiadiazole (BTH), a chemical activator of plant disease resistance responses, and by infection of the blast fungus, Magnaporthe grisea. Importantly, the expression of OsTVLP1 was activated specifically in disease resistance response induced by BTH and in an incompatible interaction between rice and the blast fungus. Our observations suggest that OsTVLP1 may play a role in rice disease resistance response against pathogen infection.

Amino Acid Sequence↗
PubMed1999

Alterations in bovine serum biochemistry profiles associated with prolonged consumption of endophyte-infected tall fescue.

Tall fescue (Festuca arundinacea) is a forage grass that is widely used in pastures in the eastern US for cattle, sheep and horses. The endophytic fungus Neotyphodium coenophialum is endemic in tall fescue pastures in the US. The turfgrass industry intentionally infects fescue cultivars with strains of the fungus to impart desirable growth and disease tolerance qualities to the plants. In contrast, ergot and pyrrolizidine alkaloid toxins produced by fungus-infected plants have been incriminated causally in bovine tall fescue toxicosis, a poorly defined syndrome of morbidity that occurs in cattle that consume endophyte-infected tall fescue (E+TF). We compared the serum biochemistry profiles from cattle that grazed E+TF with those from control cattle that grazed on endophyte-free tall fescue (E-TF). Cattle were bled on 7 dates from April 1 to August 30, 1996 and on 5 dates from May 1 to July 30, 1997. Cattle that grazed E+TF retained rough winter haircoats and had lesser weight gains, typical of tall fescue toxicosis, compared to those grazing E-TF. They had decreased activities of alkaline phosphatase and alanine aminotransferase. Compared to controls, they had lower values for serum prolactin and globulin concentrations. The concentration of creatinine and the albumin/globulin ratio were increased in the cattle grazing E+TF. Isozyme determination of alkaline phosphatase indicated that the decrease in serum activity of cattle grazing E+TF was due to decreases in both intestinal and bone isozymes. Serum protein electrophoresis indicated that the decrease in serum globulin concentration was due to decreases in both alpha and gamma globulin fractions of this protein. The data collected in these experiments add to our understanding of the alterations that occur in the serum chemistry profiles when cattle consume E+TF for prolonged periods of time.

Alkaline Phosphatase↗
PubMed1994

Spinal cord degeneration in adult dairy cows associated with the feeding of sorghum beer residues.

A neurological syndrome in a herd of Friesland cows (n = 100) on a diet including sorghum beer residues is described. Over a period of 8 months, 76 cows developed weakness of the hindquarters, progressively worsening ataxia and, eventually, paralysis and permanent recumbency. The course of the disease varied from 2 to 8 weeks. The lesions were significant in the 2 cows necropsied and included extensive, bilaterally symmetrical dilation of myelin sheaths, axonal swelling and loss with Wallerian degeneration, and depletion of myelin in both ascending and descending tracts at all levels of the spinal cord. Focal neuronal degeneration in the spinal cord grey matter and dorsal root ganglia, and focal loss of axons and ovoid formation in the spinal nerve roots and ischiatic nerves were also evident. The pathogenesis of the spinal lesions appears to involve a primary axonopathy with secondary myelin loss. The epidemiology, clinical signs and pathology suggest that the disease was associated with the fungus Aspergillus clavatus.

Animal Feed↗
PubMed1996

Inhibition of the alveolar macrophage oxidative burst by a diffusible component from the surface of the spores of the fungus Aspergillus fumigatus.

BACKGROUND: Aspergillus fumigatus is a fungus that grows on dead and decaying organic matter in the environment and whose spores are present ubiquitously in the air. The fungus causes a range of diseases in the human lung. A study was undertaken to demonstrate and partially characterise an inhibitor of the macrophage respiratory burst from the surface of A fumigatus spores that could be an important factor in allowing the fungus to colonise the lung. METHODS: The spore-derived inhibitor of the respiratory burst of rat alveolar macrophages, as measured by generation of superoxide anion, was demonstrated in Hank's balanced salt solution extracts of four clinical isolates and an environmental isolate of A fumigatus. The time course of the release of the inhibitor into aqueous solution was assessed and the cytotoxic potential of the spore-derived inhibitor towards macrophages was tested using the propidium iodide method. An oxygen electrode was used to confirm the superoxide anion measurements. Molecular weight cutoff filters were used to determine the size of the inhibitor as assessed in the respiratory burst assay and also by its ability to inhibit macrophage spreading on glass. The crude diffusate from the spore surface was fractionated by reversed phase high pressure liquid chromatography (HPLC) and the fractions analysed for inhibitory activity, protein, and carbohydrate content. RESULTS: A small molecular weight (< 10 kD) heat stable toxin was released from the spores of clinical and environmental isolates of A fumigatus within minutes of deposition in aqueous solution. The key effect of the toxin demonstrated here was its ability to inhibit the oxidative burst of macrophages as measured by superoxide anion release. The inhibition was not due to cell death or detectable loss of membrane integrity as measured by permeability to propidium iodide. The toxin was not a scavenger of superoxide anion. Oxygen electrode studies suggested indirectly that the inhibitor acted to inhibit the assembly of the macrophage NADPH-oxidase complex. Fractions of < 10 kD also inhibited the spreading of alveolar macrophages, confirming that the toxin had an additional effect on macrophages that leads to loss of adherence or impairment of cytoskeletal function. In reversed phase HPLC fractions the inhibitory activity eluted with an associated carbohydrate, although the exact chemical nature of the toxin remains to be elucidated. CONCLUSIONS: This spore toxin may, through its ability to diffuse rapidly into lung lining fluid, diminish the macrophage respiratory burst and play a part in allowing A fumigatus to persist in the lung and manifest its well known pathogenic effects. Future research will be focused on further molecular characterisation of the toxin and elaboration of the effect of the toxin on intracellular signalling pathways involved in the activation of alveolar macrophages.

Animals↗
PubMed1998

Role of calcium in signal transduction during the hypersensitive response caused by basidiospore-derived infection of the cowpea rust fungus

The hypersensitive response (HR) of disease-resistant plant cells to fungal invasion is a rapid cell death that has some features in common with programmed cell death (apoptosis) in animals. We investigated the role of cytosolic free calcium ([Ca2+]i) in the HR of cowpea to the cowpea rust fungus. By using confocal laser scanning microscopy in conjunction with a calcium reporter dye, we found a slow, prolonged elevation of [Ca2+]i in epidermal cells of resistant but not susceptible plants as the fungus grew through the cell wall. [Ca2+]i levels declined to normal levels as the fungus entered and grew within the cell lumen. This elevation was related to the stage of fungal growth and not to the speed of initiation of subsequent cell death. Elevated [Ca2+]i levels also represent the first sign of the HR detectable in this cowpea-cowpea rust fungus system. The increase in [Ca2+]i was prevented by calcium channnel inhibitors. This effect was consistent with pharmacological tests in which these inhibitors delayed the HR. The data suggest that elevation of [Ca2+]i is involved in signal transduction leading to the HR during rust fungal infection.

Journal Article↗
PubMed1979

Effect of bacterial polysaccharides on the growth of Gaeumannomyces graminis var. tritici and wheat roots.

Agrobacterium sp. and related species which in the soil and in the rhizosphere of wheat accompany the fungus Gaemannomyces graminis var. tritici and cause take-all of the wheat roots produced polysaccharides in pure cultures (glucans, mannoglucans and galactomannoglucans). These polysaccharides were utilized better by the mycelium of G. graminis than glucose and polysaccharides of plant origin that occurred on the surface of wheat roots (the so-called mucigel). At lower concentrations these bacterial polysaccharides stimulated growth of wheat roots, higher concentrations (more than 0.1%) were inhibitory. Bacteria inoculated on the surface of wheat first inhibited and then stimulated the development of the plants and their growth. Changes in the growth rate of wheat, the rhizosphere of which was colonized by bacteria simultaneously with the fungus G. graminis and also some changes in the course of the disease of wheat roots caused by the fungus can be explained by the inhibitory or stimulatory effect of polysaccharides of accompanying bacteria.

Ascomycota↗
PubMed1991

Detection of an elicitor on infection structures of Puccinia graminis using monoclonal antibodies.

The basidiomycetous fungus Puccinia graminis f. sp. tritici causes the stem rust disease of wheat. Resistance of wheat to the fungus is often associated with the hypersensitive reaction of infected host cells. A glycoprotein isolated from germ tube cell walls of the pathogen elicits a hypersensitive-like response when injected into wheat leaves. Infection structures morphologically identical to those grown on wheat were induced in the absence of the host plant, and indirect immunofluorescence together with specific monoclonal antibodies to the elicitor was employed to locate the antigen at fungal infection structures. No binding occurred to germ tubes or appressoria. The antibodies located the antigen only at that part of the fungal infection structure that develops endophytically in nature and, moreover, only at the youngest part of this structure. In rust-infected wheat leaves, the immunolabel appeared only at haustoria, the structures thought to be involved in specific recognition between host and parasite.

Antibodies, Monoclonal↗
PubMed2005

Biochemical changes during the development of witches' broom: the most important disease of cocoa in Brazil caused by Crinipellis perniciosa.

Witches' broom disease (WBD) is caused by the hemibiotrophic basidiomycete fungus Crinipellis perniciosa, which is one of the most important diseases of cocoa in the western hemisphere. In this study, the contents of soluble sugars, amino acids, alkaloids, ethylene, phenolics, tannins, flavonoids, pigments, malondialdehyde (MDA), glycerol, and fatty acids were analysed in cocoa (Theobroma cacao) shoots during the infection and development of WBD. Alterations were observed in the content of soluble sugars (sucrose, glucose, and fructose), asparagine and alkaloids (caffeine and theobromine), ethylene, and tannins. Ethylene and tannins increased prior to symptom development and declined with the death of the infected tissues. Furthermore, MDA and glycerol concentrations were higher in infected tissue than in the controls, while fatty acid composition changed in the infected tissues. Chlorophylls a and b were lower throughout the development of the disease while carotenoids and xanthophylls dropped in the infected tissue by the time of symptom development. These results show co-ordinated biochemical alterations in the infected tissues, indicating major stress responses with the production of ethylene. Ethylene levels are hypothesized to play a key role in broom development. Some of the other biochemical alterations are directly associated with ethylene synthesis and may be important for the modification of its effect on the infected tissues.

Agaricales↗