Effect of inoculation of symbiotic fungi on the growth and antioxidant enzymes’ activities in the presence of <Emphasis Type="Italic">Fusarium subglutinans</Emphasis> f. sp. <Emphasis Type="Italic">ananas</Emphasis> in pineapple plantlets

The inoculation with symbiotic fungi, Arbuscular mycorrhizal fungi (AMF) and/or Piriformospora indica on the growth, nutrient absorption, and induction of antioxidant enzyme activities in plantlets of pineapple ‘Imperial’ (fusariosis-resistant) and ‘Pérola’ (fusariosis-susceptible) in the presence of Fusarium subglutinans f. sp. ananas was investigated. The experiment was comprised by two cultivars, with or without fungal inoculation (Claroideoglomus etunicatum, Rhizophagus clarus, and P. indica, a mixture of all the fungi, and the control—absence of fungal inoculation), with or without applying Fusarium conidia, and with four replicates. In both cultivars, nutrient absorption was higher in the AMF plantlets compared to those inoculated with P. indica or the control ones, although it was more efficient in ‘Imperial’ than in ‘Pérola’. Inoculation with AMF and/or P. indica as well as the pathogen influenced differently the activities of superoxide dismutase, catalase, glutathione reductase, peroxidase, and polyphenol oxidase, in the shoots or roots of pineapple plantlets in both cultivars. Inoculated plantlets with mixture of all the fungi also exhibited a better growth and nutrient absorption, and generally, the ‘Imperial’ responded better than ‘Pérola’. In addition, these plantlets developed better than the control even in the presence of pathogen, indicating that inoculation with AMF and/or P. indica may contribute to the production of more resistant propagative material. Increased antioxidant enzyme activity is a potential strategy for managing this plant for explore biological control as an alternative to reduce environmental and health impacts by reducing the use of fungicides.     

Experimental Evidence that 3-Methylglutaric Acid Disturbs Mitochondrial Function and Induced Oxidative Stress in Rat Brain Synaptosomes: New Converging Mechanisms

3-Methylglutaric acid (3MGA) is an organic acid that accumulates in various organic acidemias whose patients present neurodegeneration events in children coursing with metabolic acidurias. Limited evidence describes the toxic mechanisms elicited by 3MGA in the brain. Herein, we explored the effects of 3MGA on different toxic endpoints in synaptosomal and mitochondrial-enriched fractions of adult rat brains to provide novel information on early mechanisms evoked by this metabolite. At 1 and 5 mM concentration, 3MGA increased lipid peroxidation, but decreased mitochondrial function only at 5 mM concentration. Despite less intense effects were obtained at 1 mM concentration, its co-administration with the kynurenine pathway (KP) metabolite and N-methyl-d-aspartate receptor (NMDAr) agonist, quinolinic acid (QUIN, 50 and 100 µM), produced toxic synergism on markers of oxidative stress and mitochondrial function. The toxicity of 3MGA per se (5 mM) was prevented by the cannabinoid receptor agonist WIN55,212-2 and the NMDAr antagonist kynurenic acid (KYNA), suggesting cannabinoid and glutamatergic components in the 3MGA pattern of toxicity. The synergic model (3MGA?+?QUIN) was also sensitive to KYNA and the antioxidant S-allylcysteine, but not to the nitric oxide synthase inhibitor l-nitroarginine methyl ester. These findings suggest various underlying mechanisms involved in the neurotoxicity of 3MGA that may possibly contribute to the neurodegeneration observed in acidemias.     

Inhibitory Effect of 5-Aminoimidazole-4-Carbohydrazonamides Derivatives Against Candida spp. Biofilm on Nanohydroxyapatite Substrate

Mycopathologia - Candida can adhere and form biofilm on biomaterials commonly used in medical devices which is a key attribute that enhances its ability to cause infections in humans. Furthermore,...     

Daily rhythms of locomotor activity and the influence of a light and dark cycle on gut microbiota species in tambaqui (Colossoma macropomum)

This study aimed to assess the daily rhythms of locomotor activity and to quantify bacteria populations in the tambaqui digestive tract during light and dark periods. Photoelectric cells and a recording computational system were used to quantify the locomotor activity, and the existing Gram-positive and Gram-negative bacteria populations were evaluating during the daytime and night-time periods. The results showed a typical pattern for nocturnal locomotor activity (91%), and lower percentages of Gram-positive 6?×?10³, 24% and Gram-negative cocci (approximately 38%) were observed during the same periods compared with the diurnal period (76 and 62%, respectively). Additionally, lower percentages of Gram-positive (42%) and Gram-negative (44%) Bacillus were observed during the same periods compared with the diurnal period (29?×?10⁶ CFU/g, 58% and 23?×?10⁶ CFU/g (56%), respectively). These data should be considered when animals are subjected to high stress loads for more appropriate management strategies and collaborating with future studies of what is the best schedule to vaccinate or provide probiotics to fish.     

A new troglomorphic species of Harmonicon (Araneae,Mygalomorphae, Dipluridae) from Pará, Brazil,with notes on the genus

A new species of Harmonicon F. O. Pickard-Cambridge, 1896 (Araneae, Dipluridae) is described, from a medium-sized lateritic cave in Parauapebas, Pará, Brazil. The male holotype and only specimen known of H. cerberus sp. n. was found near the entrance of Pequiá cave. This taxon is the fourth species described and the southernmost record for the genus. The new species displays some troglomorphic characteristics, such as reduction and merging of the posterior median and both pairs of lateral eyes and pale yellow to light brown coloration. Both characters are diagnostic when compared to the normal separated eyes and reddish to dark brown of other Harmonicon species. Other diagnostic characteristics are isolated, long, rigid setae distal to the lyra and the shape of the copulatory bulb. This is the second troglomorphic mygalomorph species from Brazil and the first from the Amazonian region.     

Affinity purified anti-citrullinated protein/peptide antibodies target antigens expressed in the rheumatoid joint

Introduction

A major subset of patients with rheumatoid arthritis (RA) is characterized by the presence of circulating autoantibodies directed to citrullinated proteins/peptides (ACPAs). These autoantibodies, which are commonly detected by using an enzyme-linked immunosorbent assay (ELISA) based on synthetic cyclic citrullinated peptides (CCPs), predict clinical onset and a destructive disease course. In the present study, we have used plasma and synovial fluids from patients with RA, for the affinity purification and characterization of anti-CCP2 reactive antibodies, with an aim to generate molecular tools that can be used in vitro and in vivo for future investigations into the pathobiology of the ACPA response. Specifically, this study aims to demonstrate that the surrogate marker CCP2 can capture ACPAs that bind to autoantigens expressed in vivo in the major inflammatory lesions of RA (that is, in the rheumatoid joint).

Methods

Plasma (n = 16) and synovial fluid (n = 26) samples were collected from RA patients with anti-CCP2 IgG levels of above 300 AU/mL. Total IgG was isolated on Protein G columns and subsequently applied to CCP2 affinity columns. Purified anti-CCP2 IgG was analyzed for reactivity and specificity by using the CCPlus® ELISA, in-house peptide ELISAs, Western blot, and immunohisto-/immunocytochemistry.

Results

Approximately 2% of the total IgG pool in both plasma and synovial fluid was CCP2-reactive. Purified anti-CCP2 reactive antibodies from different patients showed differences in binding to CCP2 and differences in binding to citrullinated peptides from α-enolase, vimentin, fibrinogen, and collagen type II, illustrating different ACPA fine-specificity profiles. Furthermore, the purified ACPA bound not only in vitro citrullinated proteins but, more importantly, in vivo-generated epitopes on synovial fluid cells and synovial tissues from patients with RA.

Conclusions

We have isolated ACPAs from plasma and synovial fluid and demonstrated that the CCP2 peptides, frequently used in diagnostic ELISAs, de facto act as surrogate antigens for at least four different, well-characterized, largely non-cross-reactive, ACPA fine specificities. Moreover, we have determined the concentration and proportion of CCP2-reactive IgG molecules in rheumatoid plasma and synovial fluid, and we have shown that the purified ACPAs can be used to detect both in vitro- and in vivo-generated citrullinated epitopes by various techniques. We anticipate that these antibodies will provide us with new opportunities to investigate the potential pathogenic effects of human ACPAs.     

Evaluation of adenosine deaminase seric activity in the diagnosis of bovine tuberculosis

Determination of seric levels of adenosine deaminase (ADA), an enzyme produced by monocytes/macrophages and lymphocytes, has been used in the diagnosis of human tuberculosis (TB). In the present study, ADA seric activity was evaluated comparatively to the comparative tuberculin test in the diagnosis of bovine tuberculosis. Two hundred fifty-six cattle were classified by origin and by the comparative tuberculin test as TB-positive animals (n = 52, from herds where the Mycobacterium bovis had previously been isolated), and TB-negative animals (n = 204, TB-free herds). The mean ADA seric value from the TB-positive group (4.45 +/- 2.33 U/L) was significantly lower (p = 0.008) than that observed in sera from the TB-negative group (6.12 +/- 4.47 U/L). When animals from a herd with clinical cases of enzootic bovine leukosis of TB-negative group were withdrawn from analysis, the mean ADA seric values of TB-negative group (5.12 +/- 3.75 U/L) was not significantly different anymore from that of the TB-positive group (p = 0.28). There was no agreement in the diagnosis of bovine TB between comparative tuberculin test and determination of ADA seric values, using two different cutoff points, being 6.12 U/L and 15.0 U/L, (kappa = -0.086 and kappa = -0.082, respectively). In conclusion, the determination of ADA seric activity was not a good auxiliary test for bovine TB, because it was not able to distinguish between TB-positive and TB-negative animals.     

Induction of systemic and mucosal immune response and decrease in Streptococcus pneumoniae colonization by nasal inoculation of mice with recombinant lactic acid bacteria expressing pneumococcal surface antigen A

Mucosal epithelia constitute the first barriers to be overcome by pathogens during infection. The induction of protective IgA in this location is important for the prevention of infection and can be achieved through different mucosal immunization strategies. Lactic acid bacteria have been tested in the last few years as live vectors for the delivery of antigens at mucosal sites, with promising results. In this work, Streptococcus pneumoniae PsaA antigen was expressed in different species of lactic acid bacteria, such as Lactococcus lactis, Lactobacillus casei, Lactobacillus plantarum, and Lactobacillus helveticus. After nasal inoculation of C57Bl/6 mice, their ability to induce both systemic (IgG in serum) and mucosal (IgA in saliva, nasal and bronchial washes) anti-PsaA antibodies was determined. Immunization with L. lactis MG1363 induced very low levels of IgA and IgG, possibly by the low amount of PsaA expressed in this strain and its short persistence in the nasal mucosa. All three lactobacilli persisted in the nasal mucosa for 3 days and produced a similar amount of PsaA protein (150-250 ng per 10(9) CFU). However, L. plantarum NCDO1193 and L. helveticus ATCC15009 elicited the highest antibody response (IgA and IgG). Vaccination with recombinant lactobacilli but not with recombinant L. lactis led to a decrease in S. pneumoniae recovery from nasal mucosa upon a colonization challenge. Our results confirm that certain Lactobacillus strains have intrinsic properties that make them suitable candidates for mucosal vaccination experiments.     

Statistical correlation of spectroscopic analysis and enzymatic hydrolysis of poplar samples

Spectroscopic characterization of poplar wood samples with different crystallinity indices, lignin contents, and acetyl contents was performed to determine changes in the biomass spectra and the effects of these changes on the hydrolysis yield. The spectroscopic methods used were X-ray diffraction for determining cellulose crystallinity (CrI), diffuse reflectance infrared (DRIFT) for changes in C-C and C-O bonds, and fluorescence to determine lignin content. Raman spectroscopy was also used to determine its effectiveness in the determination of crystallinity and C-C and C-O bond changes in the biomass as a complement to better-known methods. Changes in spectral characteristics and crystallinity were statistically correlated with enzymatic hydrolysis results to identify and better understand the fundamental features of biomass that influence enzymatic conversion to monomeric sugars. In addition, the different spectroscopic methods were evaluated separately to determine the minimum amount of spectroscopic data needed to obtain accurate predictions. The principal component regression (PCR) model with only the DRIFT data gives the best correlation and prediction for both initial rate of hydrolysis and also the 72-h hydrolysis yield. The factor that most affects both the initial rate and the 72-h conversion is the O-H bond content of the sample, which directly relates to the breakage of structural carbohydrates into smaller molecules.     

Bioinformatics describes novel Loci for high resolution discrimination of leptospira isolates

Background

Leptospirosis is one of the most widespread zoonoses in the world and with over 260 pathogenic serovars there is an urgent need for a molecular system of classification. The development of multilocus sequence typing (MLST) schemes for Leptospira spp. is addressing this issue. The aim of this study was to identify loci with potential to enhance Leptospira strain discrimination by sequencing-based methods.

Methodology and Principal Findings

We used bioinformatics to evaluate pre-existing loci with the potential to increase the discrimination of outbreak strains. Previously deposited sequence data were evaluated by phylogenetic analyses using either single or concatenated sequences. We identified and evaluated the applicability of the ligB, secY, rpoB and lipL41 loci, individually and in combination, to discriminate between 38 pathogenic Leptospira strains and to cluster them according to the species they belonged to. Pairwise identity among the loci ranged from 82.0–92.0%, while interspecies identity was 97.7–98.5%. Using the ligB-secY-rpoB-lipL41 superlocus it was possible to discriminate 34/38 strains, which belong to six pathogenic Leptospira species. In addition, the sequences were concatenated with the superloci from 16 sequence types from a previous MLST scheme employed to study the association of a leptospiral clone with an outbreak of human leptospirosis in Thailand. Their use enhanced the discriminative power of the existing scheme. The lipL41 and rpoB loci raised the resolution from 81.0–100%, but the enhanced scheme still remains limited to the L. interrogans and L. kirschneri species.

Conclusions

As the first aim of our study, the ligB-secY-rpoB-lipL41 superlocus demonstrated a satisfactory level of discrimination among the strains evaluated. Second, the inclusion of the rpoB and lipL41 loci to a MLST scheme provided high resolution for discrimination of strains within L. interrogans and L. kirschneri and might be useful in future epidemiological studies.