0004). Comparison of mean healing time in the pimecrolimus versus placebo group, demonstrated a significant acceleration

both in intention-to-treat analysis (10.7 vs. 20.7 days, F = 17.466, P < 0.0001) and treatment-completed analysis (8.3 vs. 20.7 days, F = 29.289, P < 0.0001). Conclusion:  Pimecrolimus is safe and efficient in the treatment of BD genital ulcers, by accelerating the healing process. "
“Systemic lupus erythematosus (SLE) is an autoimmune disease in which organs undergo damage. Hypoparathyroidism Selleckchem Bioactive Compound Library is a rare disease, which presents in two forms: hereditary and acquired. Cases of hypoparathyroidism and SLE rarely co-exist. Only six cases have been reported; five of them first presented with lupus and then hypoparathyroidism or simultaneously. We present here developing lupus disease in a woman who had idiopathic hypoparathyroidism. FDA approved Drug Library chemical structure According to increasing data about the autoimmune origin of idiopathic hypoparathyroidism, these case reports suggest that there may be an autoimmune process linking these diseases. “
“Systemic lupus erythematosus (SLE) is a chronic autoimmune disease with unknown etiology. Genetic and environmental factors play important roles in the pathogenesis of SLE. The primary objective of this study was to investigate the possible association of eNOS gene intron 4b/a, Glu298Asp and T-786C polymorphisms with SLE in southeast Iran populations. This was a case-control study comparing eNOS polymorphisms in 106 SLE patients

and 196 age- and sex-matched healthy controls. The 4b/a, Glu298Asp and T-786C polymorphisms were analyzed using polymerase chain reaction and restriction fragment length polymorphism.

Our findings indicated that the 4b/a polymorphism was associated with SLE, and the risk of SLE was 3.5- and 1.75-fold higher in patients with aa and ba genotypes than in patients with bb genotype. No association was observed between Glu298Asp and T-786C polymorphisms and SLE. There were no differences in eNOS gene polymorphisms between the Balouch and Fars population. Statistically significant differences were observed in genotypes and allele frequencies of 4b/a polymorphism between patients with SLE and healthy controls in southeast Iran. “
“Behcet’s disease PJ34 HCl (BD) is a rare disease mostly seen along the Silk Road. The prevalence has been reported as 0.12 (USA) to 370 (in a single village, northern Turkey) for 100 000 inhabitants.[1] During the past four decades, due to immigrations, the prevalence in Europe and North America has gradually increased.[2] It is now 4.2 in Germany, 7.2 in France and 8.6 in the US. Behcet’s disease is classified among the vasculitides and the pathophysiology is thought to be autoimmune, although some propose it as an autoinflammatory disease. Human leukocyte antigen (HLA)-B51 is recognized as a genetic factor. Hyperfunction of neutrophils, reactive oxygen species production, T-cell abnormalities, heat shock proteins (microbial and viral) are all involved in the ethiopathogenesis.

Foods that could be regarded as functional foods are subject to regulations drawn up for other food groups. The US Food and Drug Administration (FDA) defined four food categories:

conventional foods, constituting the largest category and including articles of food and drink that do not fall into the other three categories such as foods for special dietary use; medical foods; and dietary supplements. According to Berner & O’Donnell (1998), it is possible to envision ‘functional foods’ in any of the categories of foods and supplements mentioned above. From a legislative standpoint, click here probiotic-containing foods could fit into several of the four categories of foods described by the FDA; however, there is no explicit recognition of any health benefits of probiotic-, prebiotic-, or culture-added dairy foods in the United States. Government regulations regarding safety assessment differ among countries, and the status of probiotics as a component in food is currently not established on an international basis. For the most part, probiotics come under food and dietary supplements because most are delivered by mouth as foods and, as such, are allowed to make only general health claims. The factors that must be addressed in the evaluation of safety of probiotics include pathogenicity, infectivity, and virulence factors comprising toxicity, metabolic activity, and the

intrinsic properties of the microorganisms. Donohue & Salminen (1996) provided some methods for assessing the safety of lactic acid bacteria through the use of ITF2357 price in vitro studies, animal studies, and human clinical studies and indicated that some current probiotic strains are reported to fulfill the required safety standards. Salminen & Marteau (1997) also proposed studies on intrinsic properties, pharmacokinetics, and interactions between the host and probiotics as means to assess the

safety of probiotics. It was recognized that there is a need to accurately enumerate the probiotic bacteria in food products to include them on a label and that proper manufacture and handling procedures be employed to ensure the maintenance of viability and probiotic Selleckchem CHIR 99021 activity through processing, handling, and storage of probiotic foods, including powdered milk products. Good evidence exists that specific strains of probiotics are safe for human use and able to confer some health benefits on the host, but such benefits cannot be extrapolated to other strains without experimentation. As there has been an increased influx of probiotic products in the Indian market during the last decade, therefore an initiative was taken by the Indian Council of Medical Research and Department of Biotechnology, Government of India, to formulate guidelines for the regulation of probiotic products in the country (Ganguly et al., 2011), defining a set of parameters required for a product/strain to be termed as ‘probiotic’.

It is particularly important to prevent activation of enzymes that modify proteins, lipids and nucleic acids, due to hypoxia and cellular stress. Likewise, preservation of membranes is essential to prevent dispersion of soluble proteins out of cells and organelles. Hypoxia can also dramatically increase exocytosis, in particular from presynaptic transmitter vesicles. For biochemical and neurochemical analyses, rapid dissection of the tissue of interest and

cooling on ice, followed by homogenisation in the presence of enzyme inhibitors, is usually sufficient for yielding high-quality protein Cabozantinib cell line and nucleic acid preparations. For immunohistochemistry, chemical fixation, most commonly with aldehydes, is necessary to ensure preservation of histological sections throughout the staining procedure. We, and others, have shown extensively that chemical fixation markedly reduces antigenicity and/or accessibility of synaptic proteins, thereby impairing or preventing their characterisation by immunohistochemistry (Nusser et al., 1995; Fritschy et al., 1998; Watanabe et al., 1998; Sassoè-Pognetto et al., 2000; Lorincz

& Nusser, 2008). Several antigen retrieval procedures have been proposed to circumvent these limitations. In particular, minimizing exposure to fixatives is a key factor for detecting synaptic proteins in brain tissue. Thus, using perfusion-fixation with low concentration of paraformaldehyde (1–2%) and skipping post-fixation also allows highly sensitive detection of pre- and post-synaptic proteins (Eyre et al., 2012); alternatively, we have shown that Org 27569 immersion-fixation of Volasertib solubility dmso living tissue slices allows detection of both transmembrane synaptic proteins and soluble neuronal markers, in particular eGFP (Schneider Gasser et al., 2006, 2007). Here, we show that it is possible, via a brief perfusion with ice-cold, oxygenated and glucose-supplemented ACSF, to keep brain tissue alive and in optimal conditions, suitable for both homogenisation for biochemical analysis and immersion-fixation for immunohistochemistry. The possibility to combine multiple analytical methods (qPCR, Western blotting, immunofluorescence/immunoperoxidase

staining, immunoelectron microscopy) on brain tissue from the same animal represents a major advantage for correlative studies. In addition, it allows a marked reduction of the number of animals needed for studies requiring a combination of analytical methods. Although we did not attempt here to perform electrophysiology on slices prepared from ACSF-perfused mice, it is a routine procedure, in particular for preparing tissue for patch-clamp recordings. Therefore, we expect that this protocol is also suitable for concurrent (or sequential) functional and immunohistochemical/biochemical analysis of tissue from the same animal. A further benefit of immersion-fixation over perfusion-fixation is to minimise human exposure to aldehyde vapors, especially in laboratories devoid of a ventilated cabinet.

An audit of more recent perinatal transmissions occurring in the UK commenced in 2012 and is

expected to report in 2014 [6]. In 2009 the National Screening Committee considered the introduction of a routine repeat screening test in the third trimester to identify seroconversions Fluorouracil mw in pregnancy, but concluded that a universal re-offer should not be introduced at that time. However, it was reiterated that women who declined the initial offer should be re-offered screening at around 28 weeks’ gestation, and that repeat tests could be offered to any woman who was thought to be at continuing risk of infection, and to any woman who requested a second or subsequent test [13]. It is the responsibility of clinicians caring for women with HIV and their children to report them prospectively to the NSHPC. Aggregated data tables from the UK and Ireland of antiretroviral exposure and congenital malformations are regularly sent to the Antiretroviral Pregnancy Registry (APR). Individual prospective reports should also be made to the APR antenatally with post-natal follow-up. Antiretroviral

Pregnancy Registry Research Park, 1011 Ashes Drive, Wilmington, NC 28405, USA In UK call Tel: 0800 5913 1359; Fax: 0800 5812 1658; For forms visit: www.apregistry.com This is the UK and Ireland’s surveillance system for obstetric and paediatric HIV, based at the UCL Institute www.selleckchem.com/products/Bleomycin-sulfate.html of Child Health, London. HIV-infected children and children born to HIV-infected women are reported through the British Paediatric Surveillance Unit of the Royal College of Paediatrics and Child Health, or in the case of some units with large caseloads direct to the NSHPC. Diagnosed pregnant women are reported prospectively through a parallel

reporting scheme run under the auspices of the Royal College of Obstetricians and Gynaecologists. Longer-term data on infected children are subsequently collected through the Collaborative HIV Paediatric Study (CHIPS). For further information see the NSHPC website (www.ucl.ac.uk/nshpc), the CHIPS website (www.chipscohort.ac.uk), or email ([email protected]). 4.1.1 Sexual health screening is recommended for pregnant women newly diagnosed with HIV. Grading: 1B 4.1.2 O-methylated flavonoid For HIV-positive women already engaged in HIV care who become pregnant sexual health screening is suggested. Grading: 2C 4.1.3 Genital tract infections should be treated according to BASHH guidelines. Grading: 1B There are few data regarding the prevalence of genital infections in HIV-positive women in the UK [15]. At present, the majority of pregnant HIV-positive women in the UK come from, and mostly acquired HIV in, sub-Saharan Africa where the prevalence of genital infections, particularly in the HIV-positive population, can be high [16].

circinelloides could cause a series of pathological

changes in host tissues and was disseminated in different organs. Vessel thrombosis and tissue necrosis are two major hallmarks of mucormycosis in human infection (Chkhotua et al., 2001). We found tissue necrosis and vessel congestion were present in tissue sections from infected fish. Moreover, Wang et al. (2005) and Yang et al. (2006) previously reported Mucor mycelium could invade blood vessels and cause embolism in humans and fish, respectively. From the immersion infection and analysis of the human literature (Henderson et al., 2001; Lenane click here et al., 2003; Brown, 2005), we speculated that infection of healthy fish with M. circinelloides would be difficult. Results suggest that M. circinelloides may first attach to the surface selleck chemicals llc of healthy fish and upon injury or stress invade the hypodermis. Broad hyphae or spores can penetrate the mucous membrane to invade ground substance and cause focal necrosis. Fungal incursions into the vessels result in circulatory disturbance and infarction. According to Yang et al. (2006), these

acidic necrotic tissues accelerate the Mucor infection. The infected fish may perhaps have consequently died from dyspnea. New studies are expected to further the understanding of the pathogenic mechanism of mucormycosis in fish. This work was supported by the National Basic Research Program (also called 973 Program) under Grant 2009 CB118700, National Key Technology R&D Program under Grant 2007BAD37B03 and Research Program of Institute of Hydrobiology, heptaminol Chinese Academy of Sciences under Grant 095A03. “
“Initial ecosystems are characterized by a low availability of nutrients and a low soil organic matter content. Interactions of plants and microorganisms in such environments, particularly in relation to litter decomposition,

are very important for further ecosystem development. In a litter decomposition study using an initial substrate from a former mining area, we applied the litter of two contrasting pioneer plant species (legume vs. pasture plants), Lotus corniculatus and Calamagrostis epigejos, which are commonly observed in the study area. Litter decomposition was investigated and carbon (C) translocation from litter into soil microorganisms was described by following 13C from labelled plant litter materials into the fraction of phospholipid fatty acids. Labile C compounds of both plant litter types were easily degraded during the first 4 weeks of litter decomposition. In contrast to climax ecosystems, where the importance of fungi for litter degradation has been shown in many studies, in our experiment, data clearly indicate an outcompetition of fungi by Gram-positive bacteria as soon as available nitrogen is limited in the detritusphere. During the development of soil ecosystems, both above- and belowground biodiversity is changing and determines the structures and processes related to pedogenesis (Schaaf et al., 2011).

, 1999). In contrast to LipA, LipC is expressed only in very low amounts and a physiological function has not yet been assigned to this enzyme. Transcription of the lipC gene is repressed

by the products of two pilus biogenesis genes, pilX and pilY1 (Alm et al., 1996; Martinez et al., 1999), suggesting that LipC function may Selleck MK 2206 affect either type IV pilus biogenesis or pilus-dependent cellular functions. The strains and plasmids used are listed in Table 1. For mutant construction, the lipC gene was isolated by PCR amplification using Pfu-DNA-polymerase (Fermentas) and the LipCup1 (5′-GTAATGGCGTGCGCCGGGAGCC CAAC-3′) and LipCdwn1 (5′-CGCGAACAGGAGGTGA TATCCAGGTGCCATTAG-3′) primers. The resulting 1.1-kb fragment

was ligated into the EcoRV-digested cloning vector pUCPSK. The resulting plasmid pUCPLipC carrying the lipC gene under Plac control was digested with BamHI and HindIII, and the resulting lipC fragment was cloned into BamHI/HindIII-digested pSUP202 (Simon et al., 1983), yielding pSUPLipC. GSK2118436 mw The lipC gene was disrupted by exchange of an internal SalI fragment of lipC in pSUPLipC by a Gmr cassette taken from SalI-digested pWKR202 ligated into SalI sites of pSUPLipC. For gene replacement by homologous recombination, the resulting plasmid pSUPLipCGM was transferred to P. aeruginosa PAO1 by diparental mating using Escherichia coli S17.1 (Simon et al., 1983) and transconjugants were selected on Gm-containing Luria–Bertani (LB) agar. Resulting clones were tested by contra-selection on Cm-containing agar to ensure Farnesyltransferase the loss of vector sequences. The integration of the disrupted lipC allele

was confirmed by Southern blot and PCR. For Southern blot analysis, SmaI-digested chromosomal DNA of the resulting clones was probed with the Gm cassette and the lipC PCR fragment. For PCR analysis, internal primers of the Gm cassette and the lipC flanking primers LipCup1 and LipCdwn1 were used and the resulting products were analysed in terms of their length on agarose gels (data not shown). For complementation, the lipC gene was PCR-amplified using Pfu-DNA-polymerase and primers LipCup2 and LipCdwn2 (5′-GGAGTCTCGCATATGAACAAGAA CAAGACG-3′; 5′-GTAGGATCCAGGTGATATCCAGGTGC CATTAG-3′), which were designed to add an NdeI site overlapping the lipC translational startcodon and a BamHI site downstream of lipC. The resulting 1.1-kb fragment was digested with the respective enzymes and ligated to the NdeI-/BamHI-sites of pET22b (Novagen). The resulting plasmid pET22bLipC was digested with BglII and BamHI and a fragment containing the pET22b ribosomal-binding site and the T7 promoter preceding the lipC gene was ligated to BamHI-digested pBBL7, which is a derivative of pBBR1MCS containing the lipA/lipH operon of P.

1% Coomassie brilliant blue R250 (CBB), 40% (v/v) methanol, and 1% glacial acetic acid solution, followed by destaining in 50% (v/v) methanol. Total proteins of the cells were solubilized for 2 h at 4 °C in a buffer containing 50 mM Tris–HCl (pH 7.4), 0.15 M NaCl, 2.5% (w/v) sodium deoxycholate, 2% (v/v) NP-40, 1 mM N, N, N′, N′-ethylenediaminetetraacetic acid

(EDTA), protease inhibitors (1 mM PMSF, 1 μg mL−1 aprotinin, 1 μg mL−1 leupeptin, and 1 μg mL−1 pepstatin), and phosphatase inhibitors (1 mM Na3VO4 and 1 mM NaF). The phosphoproteins were trapped on Phos-tag agarose phosphate-affinity beads and isolated according to the methods reported by Saracatinib manufacturer Kinoshita-Kikuta et al. (2009). For LC-MS/MS analysis, the blots used in Phos-tag detection assays were incubated in 1 N aqueous NH3 for 15 min three times to remove the biotinylated Phos-tag complex (Nakanishi et al., 2007). Prior to LC-MS/MS analysis, the protein bands of interest on the blotting membranes were cut out and digested with 10−7 M lysyl endopeptidase (Wako) for 1 h at 37 °C. Peptides

produced by protease digestion were separated by a 0–40% linear acetonitrile gradient for 60 min, followed by analyses with a Waters UPLC Xevo Qtof. Obtained data were processed with ProteinLynx Global Server 2.4 and searched against Alveolata protein entries in the Uniprot Knowledgebase (UniprotKB) Selleckchem PLX4032 and Alveolata protein records in Entrez. When cells of C. cucullus cultured for 1–2 days were stimulated to encyst by

overpopulation in the presence of 0.1 mM Ca2+, the phosphorylation level of several proteins was enhanced within 1 h after onset of encystment induction (Fig. 1a, ‘P-tag’ right lane). In this case, the integrated optical density between in two lanes of CBB-stained blots (Fig. 1a, ‘CBB’) determined by NIH Image analysis was almost equivalent, indicating that an elevation of the Phos-tag signal (Fig. 1a, ‘P-tag’) should not be attributed to a difference in amounts of proteins between the lanes, but rather to a real enhancement of the protein phosphorylation level. In most of these cases, the increased phosphorylation old level was maintained for at least 12 h (data not shown). In the preparation of the protein samples shown in Fig. 1a, the sample buffer for SDS-PAGE did not contain inhibitors of proteases or phosphatases. Therefore, the phosphorylated proteins detected in this assay may have contained protein fragments digested by endogenous proteases, and some of the proteins may have been dephosphorylated by endogenous phosphatases during sample preparation. Therefore, as shown in Fig. 1b, the phosphorylation assay of the encystment-induced Colpoda proteins was re-examined in the presence of protease and phosphatase inhibitors (PPI).

EMSA in the presence of IPA or its analogous substrates demonstrated that IPA had the ability to inhibit the binding of IphR to this operator region. In conclusion, the iph operon is negatively

autoregulated by the binding of IphR to the operator region, and this repression is released by the presence of IPA. Phthalate isomers: phthalate, terephthalate (TPA), and isophthalate (IPA), broadly used as plasticizers, are potential starting compounds for the production of an intermediate metabolite of the protocatechuate (PCA) 4,5-cleavage pathway, 2-pyrone-4,6-dicarboxylic acid (PDC). This metabolite is a useful chemical building block for the synthesis of biodegradable and highly functional polymers (Michinobu et al., 2008, 2009; selleck antibody Hasegawa et al., 2009). To establish an efficient bioprocess for the production of PDC from inexpensive

phthalate isomers, we isolated and characterized the genes involved in the catabolism of TPA and IPA from a phthalate isomers-degrading bacterium, Comamonas I-BET-762 order sp. strain E6 (Sasoh et al., 2006; Fukuhara et al., 2008, 2010; Kasai et al., 2010). To date, the IPA catabolic genes have been reported for E6 and Comamonas testosteroni YZW-D (Wang et al., 1995), but the details of their actual functions have been reported only for the E6 genes (Fukuhara et al., 2010). The IPA degradation genes, iphACBDR code for an oxygenase component of IPA dioxygenase (IPADO), a periplasmic IPA binding receptor, a 1,2-dihydroxy-3,5-cyclohexadiene-1,5-dicarboxlylate

dehydrogenase, a reductase component of IPADO, and an IclR-type transcriptional regulator (IphR), respectively. Reverse transcription (RT)-PCR analysis revealed that the iph genes constitute an operon, and transcription of the iph operon was induced during the growth of E6 on IPA. Disruption of iphR suggested that IphR acts as a repressor for the iph operon (Fukuhara et al., 2010). IclR-type transcriptional regulators are proteins with around 250 amino acid residues, which have a helix-turn-helix DNA-binding motif in the N-terminal domain and regions involved Ribonuclease T1 in subunit multimerization and cofactor binding in the C-terminal domain (Tropel & van der Meer, 2004; Molina-Henares et al., 2006). Proteins in this family are known to act as activators, repressors, and regulators with both functions. Among the IclR-type transcriptional regulators of catabolic pathway genes for aromatic compounds, activators such as PcaU of Acinetobacter baylyi ADP1 (Gerischer et al., 1998; Trautwein & Gerischer, 2001; Popp et al., 2002) and PcaR of Pseudomonas putida PRS2000 (Parales & Harwood, 1993; Romero-Steiner et al., 1994; Guo & Houghton, 1999), which positively regulate the pca genes for the catabolism of PCA, have been well documented.

The PCR mixtures for the MT Ivan mutants contained 6 mM Tris-HCl, pH 8.5, 2.5 mM KCl, 1 mM 2-mercaptoethanol, 0.01% Triton X-100, 1.5 mM Nutlin 3a MgCl2, 100 ng DNA, 25 pmol of each primer, 200 μM dNTPs and 2.5 U Taq polymerase in a final volume of 25 μL. After an initial denaturation step of 2.5 min at 96 °C, 35 cycles of 1 min at 96 °C, 30 s at 55 °C (MT Ivan) or 50 °C (MT Iver) and 1 min at 72 °C were performed. After the last cycle, a final elongation step of 15 min at 72 °C was carried out. The purified fragments were used as templates in the second PCR step. The PCR mixtures for the second step were the same as those used for the first PCR, but without adding DNA or primers. After the addition of 50 ng of each fragment, the reaction was started with a first denaturation step of 2.5 min at 96 °C, which was followed by 20 cycles of 30 s at 96 °C and 20 s at 72 °C. Thereafter, the primers were Selleckchem Etoposide added (MT Ivan primers 1 and 5, MT Iver primers 3 and 5, see Table 1). Following a first denaturation step of 30 s at 96 °C, 35 cycles of 20 s at 96 °C, 30 s at 55 °C (MT Ivan)

or 50 °C (MT Iver) and 2 min at 72 °C were performed. In a last step, 20 cycles of 30 s at 96 °C and 1 min at 72 °C are followed by a last elongation step of 15 min at 72 °C. The mutated genes and the vector pET11a were digested with NdeI and BamHI according to the manufacturer’s protocol (Fermentas, St. Leon-Rot, Germany) and ligated in 1 × ligation buffer and 1 U T4 ligase (1 h, 22 °C). Escherichia coli TB 1 (New England Biolabs, Frankfurt/Main, Germany) was transformed with the plasmids using heat shock. For the detection of clones that contain the insert, the plasmids were isolated using the GeneJET™ Plasmid Miniprep

Kit (Fermentas) and the DNA inserts were sequenced (GATC, Konstanz, Germany). Methyltransferase gene expression was performed in the E. coli expression strain BL21 (DE3) (New England Biolabs). The E. coli BL21 (DE3) cultures were grown in Luria–Bertani media containing 0.1 g L−1 Plasmin ampicillin and 0.1% glucose. The induction of MT Ivan C286A and MT Iver C277A was performed at 28 °C for 2 h with 0.25 mM isopropyl-β-d-thiogalactopyranoside (IPTG) as an inducer. All the other mutants were induced at 18 °C for 16 h with 0.5 (MT Ivan D150A, MT Ivan D154A, MT Iver E88A, MT Iver C111A and C128A), 0.25 (MT Iver C210A) or 0.1 mM IPTG (all other mutants). After induction, the cells were harvested by centrifugation for 10 min at 10 000 g and 10 °C and stored at −20 °C. The cells were disrupted under aerobic conditions using a French pressure cell at 137 MPa and the cell debris was removed by centrifugation for 30 min at 10 000 g and 10 °C.

Moreover, it is also well known that the suppression Compound Library of phagocytic function of macrophage occurs by binding of adenosine to A2 receptors (Bours et al., 2006; Haskóet al., 2008; Kumar & Sharma, 2009). Both adenosine receptor types A2A and A2B are expressed in neutrophils, monocytes, macrophages, dendritic cells and T lymphocytes, and its EC50 for adenosine varies at 0.56–0.95 and 16.2–64.1 μM, respectively (Bours et al., 2006). Using adenosine

at the same range, at micromolar concentrations, we observed an inhibition of 50% in the percentage of infected macrophages (Fig. 6a and b). Although 5′-AMP, at the same concentration, did not have an effect in the interaction, 1 mM of 5′-AMP presented similar results to that observed with 100 μM of adenosine. This fact could be explained by the action of C. parapsilosis ecto-5′-nucleotidase activity in generating free adenosine to the medium. At 100 μM on of 5′-AMP, the rate of adenosine released could not achieve the effective concentration of free adenosine necessary to limit macrophage function, whereas at a higher concentration of 5′-AMP, the rate of extracellular adenosine could be more expressive. However, the presence of an ecto-5′-nucleotidase

activity on the external surface of macrophages find more (Edelson & Cohn, 1976a, b), able to hydrolyze 5′-AMP, could indicate that during the interaction assays, macrophages could be also responsible for adenosine generation contributing to reduction in the number of infected macrophages. Recently, our laboratory characterized ecto-ATPase activity on C. parapsilosis. The sequential dephosphorylation of ATP to adenosine was demonstrated by reverse-phase HPLC experiments, suggesting the presence of different enzymatic activities (ecto-ATPase, ecto-ADPase and ecto-5′-nucleotidase) on the surface of C. parapsilosis Inositol oxygenase (Kiffer-Moreira et al., 2010). Ecto-ATPase was also associated with in vitro infectious processes because pretreatment with ATPase inhibitors led to a decrease of C. parapsilosis adhesion to host cells (Kiffer-Moreira et al., 2010). Colonization and infection with C.

parapsilosis are dependent upon the ability of the fungus to adhere to host cells and tissues, particularly mucosal surfaces (Trofa et al., 2008). The specific functions of ecto-ATPases and ecto-5′-nucleotidases are not fully known, but it has been demonstrated that they participate in many relevant biological processes (Zimmermann, 2000; Meyer-Fernandes, 2002). In C. parapsilosis, both enzymes play a role in the control of extracellular nucleotide concentrations and could have a role in limiting inflammation and immune responses from the host, favoring the establishment of infectious processes. The involvement of ecto-5′-nucleotidases and free adenosines during infections has been described for several microorganisms including protozoa (de Almeida Marques-da-Silva et al., 2008), bacteria (Thammavongsa et al.