Complete Inhibition Of Infection Research Articles

Paraphysoderma sedebokerense GlnS III Is Essential for the Infection of Its Host Haematococcus lacustris.

Glutamine synthetase (GlnS) is a key enzyme in nitrogen metabolism. We investigated the effect of the GlnS inhibitor glufosinate on the infection of H. lacustris by the blastocladialean fungus P. sedebokerense, assuming that interfering with the host nitrogen metabolism will affect the success of the parasite. Complete inhibition of infection, which could be bypassed by the GlnS product glutamine, was observed at millimolar concentrations of glufosinate. However, this effect of glufosinate was attributed to its direct interaction with the blastoclad and not the host, which results in development and growth inhibition of the blastoclad. In our P. sedebokerense draft genome, we found that the sequence of GlnS is related to another fungal GlnS, type III, found in many poor known phyla of fungi, including Blastocladiomycota and Chytridiomycota, and absent in the main subkingdom of fungi, the Dikarya. We further tested the ability of the blastoclad to utilize nitrate and ammonia as inorganic nitrogen sources and glutamine for growth. We found that P. sedebokerense equally use ammonia and glutamine and use also nitrate, but with less efficiency. Altogether, our results show that GlnS type III is mandatory for the development and growth of P. sedebokerense and could be an efficient target to develop strategies for the control of the fungal parasite of H. lacustris.

Control of postharvest fungal rots on citrus fruit and sweet cherries using a pomegranate peel extract

A pomegranate peel extract (PGE) was evaluated as a natural antifungal preparation for the control of postharvest rots. In vitro trials revealed a strong fungicidal activity against germination of conidia of Botrytis cinerea, Penicillium digitatum and Penicillium expansum. Almost complete inhibition of all fungal spore germination was achieved after 20h of incubation with PGE. PGE was very effective in inhibiting decay following artificial inoculations of lemons by P. digitatum and Penicillium italicum, grapefruits by P. italicum and apples by P. expansum . At concentrations of 1.2 and 12g/l complete inhibition of infection was achieved in the majority of host pathogen combinations. Furthermore, it was also effective in reducing natural rots under semi-commercial conditions on both sweet cherries and lemons: on cherries Monilinia laxa and B. cinerea rots were reduced by 61.0% (cv. Bigarreau Moreau) and 95.6% (cv. Giorgia), respectively, and on lemons 87.8% reduction of total rot was achieved. PGE treatment showed residual effect as it was effective in inhibiting infections made at 6, 12, and 24h after the application of the extract in fruit wounds. Additionally, PGE exhibited curative activity and reduced the incidence of rots when it was applied 6 and 24h after inoculation. Considering that PGE was extracted and stabilized using safe chemicals (food grade ethanol and citric acid) and that it did not have any apparent phytotoxic effect on treated fruit, PGE proved to be effectively eco-friendly and safe control mean for postharvest rots of fruit.

Erythromycin Ethosomal Systems: Physicochemical Characterization and Enhanced Antibacterial Activity

The rationale behind this work was that a permeation enhancing carrier could facilitate the transport of antibacterial molecules through the two biological barriers: stratum corneum of the skin and bacterial membrane/cell wall. To this end, erythromycin ethosomes (EE) were designed and characterized, and their antibacterial efficiency was evaluated in vitro and in vivo. TEM, CLSM, DLS, DSC and ultracentrifugation tests indicate that EE are small unilamellar soft vesicles encapsulating 78.6% erythromycin. The compositions were stable for at least one year at room temperature. In live/dead viability/cytotoxicity tests, EE systems were nontoxic to cultured 3T3 dermal fibroblasts. Susceptibility studies conducted on three bacterial strains (B. subtilis ATCC 6633, S. aureus ATCC 29213 and S. aureus clinically resistant to erythromycin) showed significantly larger inhibition zones for EE as compared to erythromycin in hydroethanolic solutions. Moreover, EE reduced erythromycin MIC as compared to control solution: from 2.5 to 1.25 microg/ml for S. aureus ATCC 29213 and from 12.5 to 5.0 microg/ml for clinically isolated resistant S. aureus strain. Ethosomal erythromycin applied to the skin of ICR mice inoculated with 10(7)cfu S. aurues ATCC 29213 resulted in complete inhibition of infection. On the contrary, when hydroethanolic solution of erythromycin was applied, deep dermal and subcutaneous abscesses developed within five days after challenge. On day seven, a similar number of S. aureus colonies (1.06x10(7) vs. 0.90x10(7) cfu/g tissue) were isolated from the untreated wounds or treated with hydroethanolic erythromycin. For these animals, histopathological examination showed necrosis, destroyed skin structures and dense infiltrates of neutrophils and macrophages. These findings show that ethosomes are efficient carriers for erythromycin delivery to bacteria localized within the deep skin strata for eradication of staphylococcal infections.

IgG from acutely infected cats blocks mucosal feline immunodeficiency virus infection

We have previously shown an absence of detectable systemic or local infection in cats exposed to an infectious (100 TCID 50) feline immunodeficiency virus (FIV) plasma inoculum via either the rectal or vaginal mucosa (Burkhard, M.J., Obert, L.A., O’Neil, L.L., Diehl, L.J., Hoover, E.A., 1997. Mucosal transmission of cell-associated and cell-free feline immunodeficiency virus. AIDS Res. Hum. Retroviruses 13, 347–355). In contrast, this same plasma inoculum was infectious via parenteral inoculation. Moreover an equivalent dose of cell-free tissue culture-origin virus inoculum infected 100% of cats by either the rectal or vaginal exposure route. To evaluate this phenomena, we used a tissue culture system to identify a heat-stable factor in the plasma of cats acutely (3 weeks) infected with FIV that blocked infection of naive peripheral blood mononuclear cells (PBMC) by either cell-free or cell-associated FIV in vitro. A single application of as little as a 1:200 dilution of either heparinized or Alsevier's anticoagulated plasma effectively inhibited production of FIV p26 in culture over a 21-day co-culture period. Depletion of antibody using a protein A column abrogated the inhibitory effect of FIV plasma against in vitro FIV infection. Co-inoculation of heat-inactivated plasma with 400 TCID 50 FIV-B-2542 cell-free supernatant virus onto the vaginal mucosa of two cats resulted in complete inhibition of infection in one cat and increased time to infection in the second. Thus, antibody found in the plasma of cats acutely infected with FIV blocks cell-associated and cell-free infection, inhibits virus production in previously infected cells, and reduces mucosal transmission efficiency in vivo. Extrapolation may help explain the relatively inefficient mucosal transmission of human immunodeficiency virus-1 (HIV) and other lentiviruses.

Antimicrobial activity of a porcine myeloperoxidase against plant pathogenic bacteria and fungi

Porcine myeloperoxidase was evaluated for its antimicrobial activity against plant pathogenic bacteria and fungi. The results indicated that the enzyme, in the presence of a small amount of hydrogen peroxide, was effective against a broad spectrum of plant pathogens. The growth of seven bacterial species, including nine pathovars, from the genera Erwinia, Pseudomonas and Xanthomonas, was significantly inhibited by the enzyme at a concentration as low as 0·4 U ml−1, while 4·0 U ml−1 was lethal to all plant pathogenic bacteria examined. Myeloperoxidase, at 40 U ml−1, was lethal to germinating spores from three isolates of the fungal plant pathogen Fusarium solani and two isolates from each of Colletotrichum gloeosporioides and C. malvarum. The enzyme’s antifungal effects on the rice blast pathogen Magnaporthe grisea were studied both in vitro and on host plants. The enzyme significantly inhibited spore germination of two isolates of M. grisea races IC17 and IB49 at concentrations over 16 U ml−1, and disintegration of fungal spore walls was caused by 80 U ml−1. The enzyme was even more effective in reducing disease incidence of blast on young rice plants treated with 0·5 U ml−1, while 2·5 U ml−1 resulted in complete inhibition of infection. These results support and further extend the suggestion that myeloperoxidase could be used as a broad-spectrum biocontrol agent or as a transgenically expressed protein to combat diseases caused by plant pathogenic bacteria and fungi.

Effects of granulocyte-macrophage colony-stimulating factor and tumor necrosis factor alpha on Trypanosoma cruzi trypomastigotes.

We have previously shown that the addition of exogenous granulocyte-macrophage colony-stimulating factor (GM-CSF) to nonactivated mouse peritoneal macrophages (MPM) limits Trypanosoma cruzi infections in vitro (E. Olivares Fontt and B. Vray, Parasite Immunol. 17:135-141, 1995). Lower levels of infection were correlated with a higher level of production of tumor necrosis factor alpha (TNF-alpha) in the absence of nitric oxide (NO) release. These data suggested that GM-CSF and/or TNF-alpha might have a direct parasitocidal effect on T. cruzi trypomastigotes, independently of NO release. To address this question, T. cruzi trypomastigotes were treated with recombinant murine GM-CSF (rmGM-CSF), recombinant murine TNF-alpha (rmTNF-alpha), or both cytokines in a cell-free system. Treatment with rmGM-CSF but not rmTNF-alpha caused morphological changes in the parasites, and most became spherical after 7 h of incubation. Both cytokines exerted a cytolytic activity on the trypomastigotes, yet the trypanolytic activity of rmTNF-alpha was more effective than that of rmGM-CSF. Viable rmGM-CSF- and rmTNF-alpha-treated parasites were less able to infect MPM than untreated parasites, and this reduction in infectivity was greatest for rmGM-CSF. Treatments with both cytokines resulted in more lysis and almost complete inhibition of infection. The direct parasitocidal activity of rmTNF-alpha was inhibited by carbohydrates and monoclonal antibodies specific for the lectin-like domain of TNF-alpha. Collectively, these results suggest that cytokines such as GM-CSF and TNF-alpha may directly control the level of T. cruzi trypomastigotes at least in vitro and so could determine the outcome of infection in vivo.

Pradimicin A inhibition of human immunodeficiency virus: Attenuation by mannan

The mechanism of action of a novel anti-HIV antibiotic, pradimicin A, has been studied using cell-free (MT-4 cells) and cell to cell (MOLT-4 and MOLT-4/HIV HTLV-3B Cells) HIV infection systems. The data indicate that (1) preincubation of the cells with HIV at 0° for 1 hr, followed by the addition of pradimicin A and further incubation at 37°, resulted in a complete inhibition of infection while preincubation at 37° did not. Similar data were obtained with the coculture between MOLT-4 and MOLT-4/HIV cells. (2) Virus treated with pradimicin A followed by washing did not show any inhibition on subsequent HIV binding to cells. (3) The inhibitory effect of pradimicin A on HIV infection was prevented by mannan but not by other sugars tested. Mannan, however, did not interfere with the anti-HIV activity of other known inhibitors. (4) Use of EGTA suggested that pradimicin A required Ca ions to exert its anti-HIV activity. These data imply that pradimicin A inhibits an early step in HIV infection, probably through its binding to mannose residues of HIV glycoprotein.