Malakoplakia after renal transplantation in the current era of immunosuppressive therapy: case report and literature review.

Malakoplakia is a rare chronic granulomatous disease of unknown cause. It is thought to be caused by an acquired bactericidal defect of macrophages. Malakoplakia is associated with chronic infections and immunosuppression. Although it occurs mainly in the urinary tract, it has already been reported in almost every organ system. The isolation of bacteria, especially Escherichia coli, is common in malakoplakia patients. Here, we present a case of primary cutaneous malakoplakia in a kidney transplant recipient who had been taking prednisone, tacrolimus, and mycophenolate. Culture of a lesion grew Burkholderia cepacia complex. Treatment with high doses of trimethoprim-sulfamethoxazole was successful. We also present a systematic review of the literature, identifying 4 previously reported cases of malakoplakia after renal transplantation under similar immunosuppressive therapy, most occurring in the urinary tract or perineum and following benign courses to cure. Data in the literature suggest that malakoplakia has become even rarer since changes were made in the immunosuppressive therapy employed after kidney transplantation.

Outcome measurement instruments in Rett syndrome: A systematic review.

OBJECTIVE: The aim of the study was to identify and characterize outcome measures for objective and subjective assessment in persons with Rett syndrome (RS). METHODS: A systematic review was conducted consulting the EBSCO, Cochrane, Web of Science, Scielo, MEDLINE and PsycINFO databases for published studies describing the use of patient-reported outcome measures (PROMs) and other outcome measures in persons with RS. Validation studies and observational studies were included. The PROMs were first described, and then the measurement properties were evaluated using predefined criteria according to the COnsensus-based Standards for the selection of health Measurement Instruments (COSMIN). The outcome measures were then grouped according to the International Classification of Functioning, Disability and Health (ICF) to establish a relationship between outcome measures and ICF domains. RESULTS: Twenty out of 2327 articles were appraised, and seventeen different outcome measures were identified and described. Ten outcome measures corresponded to evaluation questionnaires, while the remaining seven assessed functional outcomes: walking distance, physical activity level and ability to interact visually. A relation between these outcome measures that assess RS and the ICF allows understanding that most of the instruments (fifteen) include the assessment of activity limitations. CONCLUSIONS: The findings of this study seem to be promising for their use by clinicians and researchers, although they have methodological limitations. The accuracy and quality of these individual outcome measures should continue to be assessed in an attempt to gather a consensus on the best tools used in RS.

Cdc42p controls yeast-cell shape and virulence of Paracoccidioides brasiliensis.

Paracoccidioides brasiliensis is characterized by a multiple budding phenotype and a polymorphic cell growth, leading to the formation of cells with extreme variations in shape and size. Since Cdc42 is a pivotal molecule in establishing and maintaining polarized growth for diverse cell types, as well as during pathogenesis of certain fungi, we evaluated its role during cell growth and virulence of the yeast-form of P. brasiliensis. We used antisense technology to knock-down PbCDC42's expression in P. brasiliensis yeast cells, promoting a decrease in cell size and more homogenous cell growth, altering the typical polymorphism of wild-type cells. Reduced expression levels also lead to increased phagocytosis and decreased virulence in a mouse model of infection. We provide genetic evidences underlying Pbcdc42p as an important protein during host-pathogen interaction and the relevance of the polymorphic nature and cell size in the pathogenesis of P. brasiliensis.

Vascular permeability, neutrophil migration and edematogenic effects induced by the latex of Cryptostegia grandiflora.

Inflammatory responses have been described as occurring after exposure to some latex materials. In this study pro-inflammatory activity in the latex of Cryptostegia grandiflora was investigated. The soluble proteins of the latex (CgLP) were isolated from the whole latex and evaluated by in vivo assays. CgLP induced strong inflammatory activity mediated by neutrophil migration, enlarging vascular permeability and increasing myeloperoxidase activity locally in rats. CgLP-induced inflammation was observed in peritonitis, paw edema and air push models. In addition, CgLP caused hyperemia in a healing model. The peritonitis effect was lost when CgLP was previously boiled suggesting the involvement of pro-inflammatory proteins. Thioglycollate increased the neutrophil migration induced by CgLP, but not by fMLP. Mast cell depletion provoked by 40/80 compound did not modify the course of inflammation triggered by CgLP, being similar to fMLP, which suggested that neutrophil migration was induced by direct mechanism mediated by macrophages. Neutrophil migration stimulated by CgLP was strongly inhibited by Dexamethasone and to a lesser extent by Thalidomide, indicating the involvement of cytokines in mediating neutrophil infiltration. Celecoxib and Indomethacin were inhibitory suggesting the involvement of prostaglandins. Cimetidine was effective only in the initial phase of edema. PCA 4248 was ineffective. It is concluded that the latex of C. grandiflora is a potent inflammatory fluid, and also that laticifer proteins may be implicated in this process.

Publisher Correction: Characterization of nitrogen doped graphene bilayers synthesized by fast, low temperature microwave plasma-enhanced chemical vapour deposition.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Characterization of nitrogen doped grapheme bilayers synthesized by fast, low temperature microwave plasma-enhanced chemical vapour deposition.

New techniques to manipulate the electronic properties of few layer 2D materials, unveiling new physical phenomena as well as possibilities for new device applications have brought renewed interest to these systems. Therefore, the quest for reproducible methods for the large scale synthesis, as well as the manipulation, characterization and deeper understanding of these structures is a very active field of research. We here report the production of nitrogen doped bilayer graphene in a fast single step (2.5 minutes), at reduced temperatures (760 °C) using microwave plasma-enhanced chemical vapor deposition (MW-PECVD). Raman spectroscopy confirmed that nitrogen-doped bilayer structures were produced by this method. XPS analysis showed that we achieved control of the concentration of nitrogen dopants incorporated into the final samples. We have performed state of the art parameter-free simulations to investigate the cause of an unexpected splitting of the XPS signal as the concentration of nitrogen defects increased. We show that this splitting is due to the formation of interlayer bonds mediated by nitrogen defects on the layers of the material. The occurrence of these bonds may result in very specific electronic and mechanical properties of the bilayer structures.

Utilization of short-chain monocarboxylic acids by the yeast Torulaspora delbrueckii: specificity of the transport systems and their regulation.

Cells of Torulaspora delbrueckii IGC 4478 grown in a medium with DL-lactic acid (0.5% v/v, at pH 5.0) exhibited Michaelis-Menten kinetics for labelled L-lactic acid transport with the following parameters at pH 5.0: Vmax, 0.38 nmol of total L-lactic acid s-1 per mg dry weight of cells and Km, 0.05 mM total L-lactic acid. Furthermore, evidence was available indicating that a proton symport for the charged form of the acid was involved. D-lactic, acetic, propionic, pyruvic and formic acids were competitive inhibitors of labelled L-lactic acid transport, suggesting that these acids used the same transport system. The ability of T. delbrueckii IGC 4478 to grow with acetic acid as the carbon source was dependent on the acid concentration and on the pH of the culture medium. When the cells were grown in 0.5% (v/v) acetic acid (pH 6.0), the transport of labelled acetic acid followed a Michaelis-Menten kinetics with the following parameters at pH 5.0: Vmax, 2.93 nmol of total acetic acid s-1 per mg dry weight of cells and and Km, 0.55 mM total acetic acid. The system also displayed a behavior consistent with a proton symport mechanism. However, the specificity of this carrier was distinct from that observed for the monocarboxylate transport in DL-lactic acid grown cells. While propionic and formic acids were competitive inhibitors of the labelled acetic acid transport, DL-lactic and pyruvic acids did not exhibit any inhibitory effects on that transport. Moreover, under the same conditions, no uptake was observed when the transport was measured with labelled L-lactic acid. Both systems were inducible and subjected to repression by glucose, fructose or sucrose. Accordingly, diauxic growth was observed in a medium containing a mixture of any of these sugars plus lactic pyruvic or acetic acid. While the induction of the acetate proton-symport appeared to be exclusively associated with acetic acid, the lactate proton-symport could be induced by either lactic or pyruvic acid but not by acetic acid. Besides, glucose repressed cells were still permeable to the undissociated form of the acids which entered the cells by simple diffusion. Furthermore, the activities of the lactate proton-symport and of the acetate proton-symport appeared not to be associated with the activity of the L-lactate (cytochrome) dehydrogenase.

Effects of ethanol and other alkanols on passive proton influx in the yeast Saccharomyces cerevisiae.

Ethanol, isopropanol, propanol and butanol enhanced the passive influx of protons into deenergized cells of Saccharomyces cerevisiae. The influx followed first-order kinetics with a rate constant that increased exponentially with the alkanol concentration. The exponential enhancement constants increased with the lipid solubility of the alkanols, which indicated hydrophobic membrane regions as the target sites. While the enhancement constants were independent of pH over the range tested (3.3-5.0), the rate constants decreased linearly with increasing extracellular proton concentration, indicating the presence of an additional surface barrier against proton penetration, the effectiveness of which increased with protonation. The alkanols affected the acidification curves of energized yeast suspensions in such a way that the final pH values were linear functions of the alkanol concentrations. These results were consistent with a balance between active and passive proton movements at the final pH, the exponential enhancement constants calculated from the slopes being nearly identical with those obtained with deenergized cells. It was concluded that passive proton influx contributes to the kinetics of acidification in S. cerevisiae and that uncoupling contributes to the overall kinetics of alkanol-inhibited secondary active transport across the yeast plasma membrane.

Quantitative analysis of proton movements associated with the uptake of weak carboxylic acids. The yeast Candida utilis as a model.

A quantitative analysis of the proton movements associated with the initial uptake rate of weak short-chain carboxylic acids was developed in order to estimate proton/carboxylate symports stoichiometries. The yeast Candida utilis was used as a biological model and the deduced equations were applied on the elucidation of the proton/carboxylate symports stoichiometries of lactate, succinate and citrate in a strain of that yeast species at different pH values. At pH 5.0, the proton/lactate and the proton/succinate symport stoichiometry was 1:1. In the cases of the proton/lactate and proton/citrate symports it appears that the stoichiometry ratio increased with increasing extracellular pH.

Relationship between protein kinase C and derepression of different enzymes.

The PKC1 gene in the yeast Saccharomyces cerevisiae encodes for protein kinase C which is known to control a MAP kinase cascade consisting of different kinases: Bck1, Mkk1 and Mkk2, and Mpk1. This cascade affects the cell wall integrity but the phenotype of pkc1Delta mutants suggests additional targets that have not yet been identified [Heinisch et al., Mol. Microbiol. 32 (1999) 671-680]. The pkc1Delta mutant, as opposed to other mutants in the MAP kinase cascade, displays defects in the control of carbon metabolism. One of them occurs in the derepression of SUC2 gene after exhaustion of glucose from the medium, suggesting an involvement of Pkc1p in the derepression process that is not shared by the downstream MAP kinase cascade. In this work, we demonstrate that Pkc1p is required for the increase of the activity of enzymatic systems during the derepression process. We observed that Pkc1p is involved in the derepression of invertase and alcohol dehydrogenase activities. On the other hand, it seems not to be necessary for the derepression of the enzymes of the GAL system. Our results suggest that Pkc1p is acting through the main glucose repression pathway, since introduction of an additional mutation in the PKC1 gene in yeast strains already presenting mutations in the HXKII or MIG1 genes does not interfere with the typical derepressed phenotype observed in these single mutants. Moreover, our data indicate that Pkc1p participates in this process through the control of the cellular localization of the Mig1 transcriptional factor.

Sequential inactivation of ammonium and glucose transport in Saccharomyces cerevisiae during fermentation.

Ethanol at concentrations above 12% (v/v) in mineral medium with glucose and with ammonium as the only nitrogen source induced rapid inactivation of the ammonium transport system in the strain IGC 3507 of Saccharomyces cerevisiae terminating protein synthesis. Subsequently, when glucose was present, the glucose transport system was irreversibly inactivated. This two-step mechanism may play a decisive role when ethanol stops fermentation by S. cerevisiae, before all the fermentable sugar has been consumed.

Effects of ethanol and acetic acid on the transport of malic acid and glucose in the yeast Schizosaccharomyces pombe: implications in wine deacidification.

Ethanol and acetic acid, at concentrations which may occur during wine-making, inhibited the transport of L-malic acid in Schizosaccharomyces pombe. The inhibition was non-competitive, the decrease of the maximum initial velocity following exponential kinetics. Glucose transport was not significantly affected either by ethanol (up to 13%, w/v) or by acetic acid (up to 1.5%, w/v). The uptake of labelled acetic acid followed simple diffusion kinetics, indicating that a carrier was not involved in its transport. Therefore, the undissociated acid appears to be the only form that enters the cells and is probably responsible for the toxic effects. Accordingly, deacidification by Ss. pombe during wine fermentation should take place before, rather than after, the main alcoholic fermentation by Saccharomyces cerevisiae.

Transport of acetate in mutants of Saccharomyces cerevisiae defective in monocarboxylate permeases.

The strain Saccharomyces cerevisiae W303-1a, able to grow in a medium containing acetic acid as the sole carbon and energy source, was subjected to mutagenesis in order to obtain mutants deficient in monocarboxylate permeases. Two mutant clones exhibiting growth in ethanol, but unable to grow in a medium with acetic acid as the sole carbon and energy source, were isolated (mutants Ace12 and Ace8). In both mutants, the activity for the acetate carrier was strongly affected. The mutant Ace8 revealed not to be affected in the transport of lactate, while the mutant Ace12 did not display activity for that carrier. These results reinforced those previously found in the strain IGC 4072, where two distinct transport systems for monocarboxylates have been described, depending on the growth carbon source. It is tempting to postulate that the Ace8 mutant seems to be affected in the gene coding for an acetate permease. In contrast, the absence of activity for both monocarboxylate permeases in mutant Ace12 could be attributed to a mutation in a gene coding for a regulatory protein not detected before.

Lack of lactate-proton symport activity in pck1 mutants of Saccharomyces cerevisiae.

Mutants of Saccharomyces cerevisiae without phosphoenolpyruvate carboxykinase activity showed no measurable lactate proton symport, while mutants without fructose-1,6-bisphosphatase had normal transport activity. Incubation of a pck1 mutant, under derepression conditions in the presence of glycerol, restored the activity of the lactate-proton symport, with identical kinetic characteristics to that in the wild-type. For efficient lactate-proton symport activity, not only is an external inducer such as lactic acid needed, but also a molecule derived from the acid metabolism may be necessary.

Utilization and transport of acetic acid in Dekkera anomala and their implications on the survival of the yeast in acidic environments.

The yeast Dekkera anomala IGC 5153 exhibited a restricted ability to use weak acids as the only carbon and energy sources. Of the monocarboxylic, dicarboxylic, and tricarboxylic acids tested, only acetic acid was used in such a way. The cells were able to grow at acetic acid concentrations of 0.1 to 3% (vol/vol) over a pH range of 3.5 to 5.5, and the specific growth rates decreased exponentially with the increase of the undissociated acetic acid concentration in the culture medium. Transport assays carried out in cells that exhibited higher specific growth rates showed the presence of an acetate-proton symport associated with a simple diffusion component of the undissociated acetic acid, the weight of the latter increasing with the undissociated acid concentration in the culture media. The acetate carrier was shared by propionic, formic, and sorbic acids and was inducible and repressed by glucose and concentrations of undissociated acetic acid in the culture medium above 0.3% (vol/vol). In undissociated acetic acid repression conditions, the lowest values for the yeast specific growth rates were obtained, and the simple diffusion of the undissociated acid was the only mechanism involved in the acetic acid uptake by the cells. The results will be discussed in terms of the high tolerance of D. anomala to the acidic stress conditions present in wine.

A differential medium for the enumeration of the spoilage yeast Zygosaccharomyces bailii in wine.

A collection of yeasts, isolated mostly from spoiled wines, was used in order to develop a differential medium for Zygosaccharomyces bailii. The 118 selected strains of 21 species differed in their origin and resistance to preservatives and belonged to the genera Pichia, Torulaspora, Dekkera, Debaryomyces, Saccharomycodes, Issatchenkia, Kluyveromyces, Kloeckera, Lodderomyces, Schizosaccharomyces, Rhodotorula, Saccharomyces, and Zygosaccharomyces. The design of the culture medium was based on the different ability of the various yeast species to grow in a mineral medium with glucose and formic acid (mixed-substrate medium) as the only carbon and energy sources and supplemented with an acid-base indicator. By manipulating the concentration of the acid and the sugar it was possible to select conditions where only Z. bailii strains gave rise to alkalinization, associated with a color change of the medium (positive response). The final composition of the mixed medium was adjusted as a compromise between the percentage of recovery and selectivity for Z. bailii. This was accomplished by the use of pure or mixed cultures of the yeast strains and applying the membrane filtration methodology. The microbiological analysis of two samples of contaminated Vinho Verde showed that the new medium can be considered as a differential medium to distinguish Z. bailii from other contaminating yeasts, having potential application in the microbiological control of wines and probably other beverages and foods.

Survey of hydrogen sulphide production by wine yeasts.

Twenty-one strains of commercial wine yeasts and 17 non-Saccharomyces species of different provenance were surveyed for their ability to produce hydrogen sulphide in synthetic grape juice medium indicator agar with different nitrogen sources, as well as in natural grape juice. Bacto Biggy agar, a commercially available bismuth-containing agar, was used to compare our results with others previously reported in the literature. Under identical physiological conditions, the strains used in this study displayed similar growth patterns but varied in colony color intensity in all media, suggesting significant differences in sulphite reductase activity. Sulphite reductase activity was absent for only one strain of Saccharomyces cerevisiae. All other strains produced an off-odor to different extents, depending significantly (P <0.05) on medium composition. Within the same species of some non-Saccharomyces yeasts, strain variation existed as it did for Saccharomyces. In natural musts, strains fell into three major groups: (i) nonproducers, (ii) must-composition-dependent producers, and (iii) invariable producers. In synthetic media, the formation of sulphide by strains of S. cerevisiae results from the reduction of sulphate. Therefore, this rapid screening methodology promises to be a very useful tool for winemakers for determining the risk of hydrogen sulphide formation by a given yeast strain in a specific grape juice.

Activity of essential oils from Mediterranean Lamiaceae species against food spoilage yeasts.

The essential oils from aerial parts of Melissa officinalis, Lavandula angustifolia, Salvia officinalis, and Mentha piperita were analyzed by gas chromatography and gas chromatography-mass spectrometry. Their antimicrobial activities were evaluated against five food spoilage yeasts, Torulaspora delbrueckii, Zygosaccharomyces bailii, Pichia membranifaciens, Dekkera anomala, and Yarrowia lipolytica. Saccharomyces cerevisiae was also used as a reference. The oils were preliminarily screened by a disc diffusion technique, with the most active being the oil from M. officinalis. MICs were determined by the broth dilution method, and the main components of the oils were also tested by this method. The essential oil of M. officinalis at 500 microg/ml completely inhibited the growth of all yeast species. The main component of the oil of M. officinalis is citral (neral plus geranial) (58.3%), which showed a marked fungitoxic effect, contributing to its high activity.

Renal effect of gentamicin in diabetic rats.

To evaluate the protective effect of diabetes mellitus (DM) on renal function of rats treated with gentamicin (GM), male Wistar-EPM rats (250-350 g) were treated with streptozotocin (SZ; 45 mg/kg) and starting 10 days after induction of diabetes, GM was given for ten consecutive days at a daily dose of 40 mg/kg. In the GM-treated group (G), a significant fall in inulin and sodium-p-amino-hippurate clearance was obtained (3.57 +/- 0.16 and 12.59 +/- 0.61 ml min-1 kg-1 vs 6.43 +/- 0.21 and 17.98 +/- 0.47 ml min-1 kg-1 in control rats (C), respectively) while in the animals previously injected with SZ (diabetic+gentamicin, DG group) these changes were not observed. The diabetic (D), G and DG groups showed a significant rise in urinary flow compared to C (0.165 +/- 0.009, 0.145 +/- 0.007 and 0.173 +/- 0.009 ml min-1 kg-1 vs 0.109 +/- 0.003 ml min-1 kg-1, respectively); however, only in G was the U/P inulin ratio significantly decreased when compared to C. The fractional excretion (FE) of sodium and potassium was significantly augmented in G when compared to C, D and DG. Thus, diabetes protected against gentamicin nephrotoxicity at both the glomerular and tubular level, although it did not promote a reduction in urinary flow.

Yeasts as a model for assessing the toxicity of the fungicides Penconazol, Cymoxanil and Dichlofluanid.

In the present work the sensitivity of yeast strains of Kluyveromyces marxianus, Pichia anomala, Candida utilis, Schizosaccharomyces pombe and Saccharomyces cerevisiae, to the fungicides cymoxanil, penconazol, and dichlofluanid, was evaluated. Dichlofluanid induced the most negative effects, whereas penconazol in general was not very toxic. Overall, our results show that the parameters IC50 for specific respiration rates of C. utilis and S. cerevisiae and C(D) for cell viability of S. cerevisiae can be applied to quantify the toxicity level of the above compounds in yeast. Hence, could be explored as an alternative or at least as a complementary test in toxicity studies and, therefore, its potential for inclusion in a tier testing toxicity test battery merits further research.