Yersinia enterocolitica YopP inhibits MAP kinase-mediated antigen uptake in dendritic cells

Yersinia enterocolitica (Ye) targets mouse dendritic cells (DCs) and inhibits their ability to trigger T cell activation. Here we have investigated whether Ye might interfere with antigen presentation in DCs. Infection of DCs with the Ye wild-type strain reduced OVA uptake by DCs as demonstrated by flow cytometry and confocal laser scan microscopy. In contrast, DCs infected with Yersinia outer protein P (YopP)-deficient mutant strain rapidly internalized OVA. Furthermore, transfection of DCs with YopP, but not with a cysteine protease deficient YopP-C172A mutant, reduced uptake of OVA. This finding suggests that YopP, a virulence factor of Ye, inhibits OVA uptake by DCs. By the use of MAPK inhibitors we provide evidence that YopP mediates reduction of OVA uptake by its ability to block MAPK signalling pathways in host cells. Using transferrin (Tf) as specific marker for clathrin-mediated endocytosis (CME) and lucifer yellow (LY) as specific marker for macropinocytosis (MP) we could show that YopP inhibits CME, whereas other Yops inhibit MP. In keeping with these data, activation and proliferation of OVA-specific T cells was reduced when DCs were treated with MAPK inhibitors. Together, our data demonstrate that (i) MAPK play an important role in antigen uptake by CME in DCs, and (ii) that YopP inhibits this pathway of antigen uptake in DCs, which might contribute to evasion of adaptive immunity.     

L-Tryptophan-dependent biosynthesis of indole-3-acetic acid (IAA) improves plant growth and colonization of maize by <Emphasis Type="BoldItalic">Burkholderia phytofirmans</Emphasis> PsJN

Burkholderia phytofirmans PsJN is a well-known plant growth-promoting bacterium that establishes rhizospheric and endophytic colonization in different plants. PsJN inoculation promotes growth of different horticultural crops. L-Tryptophan (L-TRP) application may further improve its effectiveness, due to substrate (L-TRP)-dependent inoculum (PsJN)-derived auxins in the rhizosphere. In the present study, the substrate (L-TRP)-dependent response of PsJN inoculation to maize growth and auxin biosynthesis was evaluated under pot conditions. In vitro auxin biosynthesis by PsJN was determined in the absence and presence of L-TRP, a physiological precursor of auxins. Surface-disinfected seeds were treated with peat-based inoculum and L-TRP solutions (10^?4 and 10^?5 M). Results revealed that L-TRP application and PsJN inoculation, when applied separately, significantly increased the growth parameters of maize compared to untreated control. However, PsJN inoculation supplemented with L-TRP (10^?5 M) gave the most promising results and significantly increased plant height, photosynthesis, chlorophyll content, root biomass and shoot biomass up to 18, 16, 45, 62 and 55 %, respectively, compared to the uninoculated control. Similarly, higher values of N, P and IAA content were observed with precursor (L-TRP)–inoculum (PsJN) interaction. The inoculant strain efficiently colonized maize seedlings and was recovered from the rhizosphere, root and shoot of plants. The results imply that substrate (L-TRP)-derived IAA biosynthesis in the rhizosphere by PsJN inoculation could be a useful approach for improving the growth, photosynthesis and nutrient content of maize plants.     

Evaluation of antileishmanial activity of South Indian medicinal plants against Leishmania donovani

Infections due to protozoa of the genus Leishmania are a major worldwide health problem, with high endemicity in developing countries. The aim of this study was to evaluate the in vitro antileishmanial activity of the acetone and methanol leaf extracts of Anisomeles malabarica, flower of Gloriosa superba, leaf of Ocimum basilicum, leaf and seed of Ricinus communis against promastigotes form of Leishmania donovani. Antiparasitic evaluations of different plant crude extracts were performed on 96 well plates at 37°C for 24-48h. Out of the 10 experimental plant extracts tested, the leaf methanol extracts of A. malabarica, and R. communis showed good antileishmanial activity (IC(50)=126±19.70 and 184±39.33μg/mL), respectively against promastigotes. Effective antileishmanial activity was observed making these plants as good candidates for isolation of antiprotozoal compounds which could serve as new lead structures for drug development.     

Simple Pseudo-dipeptides with a P2' Glutamate: A NOVEL INHIBITOR FAMILY OF MATRIX METALLOPROTEASES AND OTHER METZINCINS

A series of pseudo-peptides with general formula X-l-Glu-NH(2) (with X corresponding to an acyl moiety with a long aryl-alkyl side chain) have been synthesized, evaluated as inhibitors of matrix metalloproteases (MMPs), and found to display remarkable nanomolar affinity. The loss in potency associated with a substitution of the P(2)' l-glutamate by a l-glutamine corroborates the importance of a carboxylate at this position. The binding mode of some of these inhibitors was characterized in solution and by x-ray crystallography in complex with various MMPs. The x-ray crystal structures reveal an unusual binding mode with the glutamate side chain chelating the active site zinc ion. Competition experiments between these inhibitors and acetohydroxamic acid, a small zinc-binding molecule, are in accord with the crystallographic results. One of these pseudo-dipeptides displays potency and selectivity toward MMP-12 similar to the best MMP-12 inhibitors reported to date. This novel family of pseudo peptides opens new opportunities to develop potent and selective inhibitors for several metzincins.     

Microbial ACC-Deaminase: Prospects and Applications for Inducing Salt Tolerance in Plants

Salinity is one of the most important stresses that hamper agricultural productivity in nearly every part of the world. Enhanced biosynthesis of ethylene in plants under salinity stress is well established. Higher ethylene concentration inhibits root growth and ultimately affects the overall plant growth. Overcoming this ethylene-induced root inhibition is a prerequisite for successful crop production. Recent studies have shown that ethylene level in plants is regulated by a key enzyme 1-aminocyclopropane-1-carboxylicacid (ACC)-deaminase. This enzyme is present in plant growth-promoting bacteria (PGPR) and lowers the ethylene level by metabolizing its precursor ACC into α-ketobutyrate and ammonia (NH₃). Inoculation of plants under salinity stress with PGPR having ACC-deaminase activity mitigates the inhibitory effects of salinity on root growth by lowering the ethylene concentration in the plant. This in turn results in prolific root growth, which is beneficial for the uptake of nutrients and maintenance of growth under stressful environment. The present review critically discusses the effects of salinity stress on plant growth with special reference to ethylene production and the effects of rhizobacteria containing ACC-deaminase on crop improvement under salinity stress. It also discusses how much progress has been made in producing transgenic lines of different crops over-expressing the gene encoding ACC-deaminase and how far such transformed lines can tolerate salinity stress.     

Synthesis of prostaglandin E2 and prostaglandin F2α by toadfish red blood cells

Saline washed red blood cells of the toadfish convert [1-¹⁴C] arachidonic acid to products that cochromatograph with prostaglandin E₂ and prostaglandin F_2α. This synthesis is inhibited by indomethacin (10 μg/ml). Conversion of arachidonic acid to prostaglandin E₂ was confirmed by mass spectrometry. When saline washed toadfish red blood cells were incubated with a mixture of [1-¹⁴C]-arachidonic acid and [5,6,8,9,11,12,14,15,-³H]-arachidonic acid, comparison of the isotope ratios of the radioactive products indicated that prostaglandin F_2α was produced by reduction of prostaglandin E₂. The capacity of toadfish red blood cells to reduce prostaglandin E₂ to prostaglandin F_2α was confirmed by incubation of the cells with [1-¹⁴C] prostaglandin E₂.     

Kinetics and effects of trace elements and electron complexes on 2-keto-4-methylthiobutyric acid-dependent biosynthesis of ethylene in soil

AIMS: 2-Keto-4-methylthiobutyric acid (KMBA) is an established intermediate in microbial biosynthesis of ethylene from methionine. This study demonstrates the kinetics and effects of trace elements and electron complexes on substrate (KMBA)-derived C2H4 biosynthesis in soil. METHODS AND RESULTS: We have previously reported KMBA-dependent C2H4 production in soil. We studied the kinetics and effects of various trace elements and electron complexes on KMBA-derived C2H4 biosynthesis in soil by gas chromatography. Kinetic analysis revealed that ethylene forming enzyme (EFE) reaction was linear (R2 = 0.9448) when velocity of reaction (V) was plotted against substrate [S] over the range from 2.5 to 10 mmol l(-1) and thus followed a first order reaction. Application of three linear transformations of the Michaelis-Menten equation indicated high affinity of EFE for the substrate because Km values ranged between 5.4 and 6.67 mmol l(-1) and Vmax of reaction was between 22.4 and 35.7 nmol kg(-1) soil 120 cm(-1). Most of the trace elements exhibited positive effects on KMBA-dependent C2H4 production in soil. Maximum stimulatory effect on C2H4 biosynthesis was observed in response to Co(II) application, while Fe(III) inhibited the biotransformation of KMBA into C2H4. Contrarily, most of the tested electron complexes inhibited KMBA-derived C2H4 biosynthesis in the soil. However, lower concentrations (1.0 mmol l(-1)) of mannitol and hydroquinone were stimulatory to C2H4 production in soil compared with controls (substrate only). Conclusions: The results revealed that both kind and concentration of trace elements and electron complexes affected the substrate-dependent production of C2H4 in soil with different degrees of efficacy. SIGNIFICANCE AND IMPACT OF THE STUDY: The C2H4 in the root environment could be physiologically active even at low concentrations, so knowledge regarding various factors which regulate C2H4 biosynthesis in soil could be of significance for plant growth and development.     

Effects of subinhibitory concentrations of nitroxoline on the surface properties of Escherichia coli

Nitroxoline (5-nitro-8-quinolinol; NIQ) at subinhibitory concentrations (sub-MIC) decreased the adherence of uropathogenicEscherichia coli to catheter surface and significantly enhanced cell surface hydrophobicity. The surface hydrophobicity increased in the presence of sub-MIC of NIQ and also in an excess of Mg²⁺. The effect of NIQ on the cell surface was not related to the bacteriostatic effect of this agent. The increase in nitrogen and decrease in phosphate content in the cell surface was found in the presence of NIQ. NIQ did not inhibit the expression of fimbriae.     

Calreticulin, a potential cell surface receptor involved in cell penetration of anti-DNA antibodies

A 50-kDa protein was purified as a potential receptor, using an affinity matrix containing biotinylated F14.6 or H9.3 anti-DNA mAbs derived from autoimmune (New Zealand Black x New Zealand White)F(1) mouse and membrane extracts from cells. This protein was identified as calreticulin (CRT) by microsequencing. Confocal microscopy and FACS analysis showed that CRT was present on the surface of various cells. CRT protein was recognized by a panel of anti-DNA mAbs in ELISA. The binding of F14.6 to lymphocytes and Chinese hamster ovary cells was inhibited by soluble CRT or SPA-600. Thus, the anti-DNA mAbs used in this study bound to CRT, suggesting that CRT may mediate their penetration into the cells and play an important role in lupus pathogenesis.     

Mitigation of salinity-induced negative impact on the growth and yield of wheat by plant growth-promoting rhizobacteria in naturally saline conditions

Salinity is one of the major environmental threats for successful crop production, hampering plant growth due to the osmotic effect and nutritional and hormonal imbalances. The application of naturally occurring plant growth-promoting rhizobacteria (PGPR) is an emerging technology aimed at ameliorating the negative impact of salinity. However, the results obtained in the laboratory can sometimes not be reproduced in the field. The aim of the study reported here was to evaluate the effect of PGPR inoculation on seed germination in a saline environment under axenic conditions and on enhancement of the growth and yield of wheat under natural salt-affected field conditions. Wheat seeds were inoculated with pre-isolated strains of Pseudomonas putida, Enterobacter cloacae, Serratia ficaria, and Pseudomonas fluorescens and sown at different salinity levels (1, 2, 3, 6, 9, 12, 15 dS m^-1). Inoculation with these strains was found to enhance the germination percentage, germination rate, and index of wheat seeds up to 43, 51, and 123 %, respectively, over the uninoculated control at the highest salinity level. The potential of these PGPR for improving the growth and yield of wheat was also evaluated at two natural salt-affected sites. Inoculation with PGPR resulted a significant increase in the growth and yield parameters of wheat at both sites. The inoculated plants also improved the nutrient status of the wheat plants. The inoculated plants had low sodium and high nitrogen, phosphorus, and potassium contents. Our results show that such rhizobacterial strains may be used as an effective tool for enhancing plant growth under salinity stress and for maximizing the utilization of salt-affected soils.