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1.
Sci Adv ; 6(2): eaaw6284, 2020 01.
Article in English | MEDLINE | ID: mdl-31950075

ABSTRACT

Zika virus (ZIKV) infection during pregnancy is associated with a spectrum of developmental impairments known as congenital Zika syndrome (CZS). The prevalence of this syndrome varies across ZIKV endemic regions, suggesting that its occurrence could depend on cofactors. Here, we evaluate the relevance of protein malnutrition for the emergence of CZS. Epidemiological data from the ZIKV outbreak in the Americas suggest a relationship between undernutrition and cases of microcephaly. To experimentally examine this relationship, we use immunocompetent pregnant mice, which were subjected to protein malnutrition and infected with a Brazilian ZIKV strain. We found that the combination of protein restriction and ZIKV infection leads to severe alterations of placental structure and embryonic body growth, with offspring displaying a reduction in neurogenesis and postnatal brain size. RNA-seq analysis reveals gene expression deregulation required for brain development in infected low-protein progeny. These results suggest that maternal protein malnutrition increases susceptibility to CZS.


Subject(s)
Malnutrition/complications , Zika Virus Infection/congenital , Zika Virus Infection/complications , Animals , Animals, Newborn , Body Weight , Brain/enzymology , Brain/pathology , Brazil/epidemiology , Diet, Protein-Restricted , Disease Outbreaks , Embryo, Mammalian/pathology , Female , Gene Expression Regulation, Developmental , Malnutrition/virology , Mice, Inbred C57BL , Microcephaly/complications , Microcephaly/virology , Neurogenesis , Organ Size , Pregnancy , Syndrome , Viral Load , Zika Virus Infection/virology
2.
Sci Rep ; 8(1): 12774, 2018 08 24.
Article in English | MEDLINE | ID: mdl-30143723

ABSTRACT

Zika virus (ZIKV) is associated with brain development abnormalities such as primary microcephaly, a severe reduction in brain growth. Here we demonstrated in vivo the impact of congenital ZIKV infection in blood vessel development, a crucial step in organogenesis. ZIKV was injected intravenously in the pregnant type 2 interferon (IFN)-deficient mouse at embryonic day (E) 12.5. The embryos were collected at E15.5 and postnatal day (P)2. Immunohistochemistry for cortical progenitors and neuronal markers at E15.5 showed the reduction of both populations as a result of ZIKV infection. Using confocal 3D imaging, we found that ZIKV infected brain sections displayed a reduction in the vasculature density and vessel branching compared to mocks at E15.5; altogether, cortical vessels presented a comparatively immature pattern in the infected tissue. These impaired vascular patterns were also apparent in the placenta and retina. Moreover, proteomic analysis has shown that angiogenesis proteins are deregulated in the infected brains compared to controls. At P2, the cortical size and brain weight were reduced in comparison to mock-infected animals. In sum, our results indicate that ZIKV impairs angiogenesis in addition to neurogenesis during development. The vasculature defects represent a limitation for general brain growth but also could regulate neurogenesis directly.


Subject(s)
Neovascularization, Physiologic , Zika Virus Infection/congenital , Zika Virus/physiology , Animals , Blood Vessels/pathology , Brain/blood supply , Brain/pathology , Brain/virology , Disease Models, Animal , Embryo, Mammalian/pathology , Embryo, Mammalian/virology , Endothelial Cells/pathology , Endothelial Cells/virology , Female , Mice, Inbred C57BL , Neurogenesis , Organ Size , Zika Virus Infection/pathology , Zika Virus Infection/virology
3.
J Med Microbiol ; 65(11): 1311-1321, 2016 Nov.
Article in English | MEDLINE | ID: mdl-27902402

ABSTRACT

Corynebacterium diphtheriae is typically recognized as a colonizer of the upper respiratory tract (respiratory diphtheria) and the skin (cutaneous diphtheria). However, different strains of Corynebacteriumdiphtheriae can also cause invasive infections. In this study, the characterization of a non-toxigenic Corynebacteriumdiphtheriae strain (designated BR-INCA5015) isolated from osteomyelitis in the frontal bone of a patient with adenoid cystic carcinoma was performed. Pathogenic properties of the strain BR-INCA5015 were tested in a Caenorhabditis elegans survival assay showing strong colonization and killing by this strain. Survival rates of 3.8±2.7 %, 33.6±7.3 % and 0 % were observed for strains ATCC 27010T, ATCC 27012 and BR-INCA5015, respectively, at day 7. BR-INCA5015 was able to colonize epithelial cells, showing elevated capacity to adhere to and survive within HeLa cells compared to other Corynebacteriumdiphtheriae isolates. Intracellular survival in macrophages (THP-1 and RAW 264.7) was significantly higher compared to control strains ATCC 27010T (non-toxigenic) and ATCC 27012 (toxigenic). Furthermore, the ability of BR-INCA5015 to induce osteomyelitis was confirmed by in vivo assay using Swiss Webster mice.


Subject(s)
Corynebacterium diphtheriae/isolation & purification , Corynebacterium diphtheriae/pathogenicity , Osteomyelitis/microbiology , Adult , Animals , Caenorhabditis elegans , Corynebacterium diphtheriae/classification , Corynebacterium diphtheriae/genetics , Epithelial Cells/microbiology , Female , Humans , Macrophages/microbiology , Male , Mice , RAW 264.7 Cells , Virulence
4.
Talanta ; 155: 107-15, 2016 08 01.
Article in English | MEDLINE | ID: mdl-27216663

ABSTRACT

In this work, a non-commercial triaxial geometry energy dispersive X-ray Fluorescence (EDXRF) setup and a benchtop µ-XRF system were used to identify postmortem contamination in buried bones. For two of the individuals, unusually high concentrations of Cu and Pb, but also Zn (in one individual) were observed. The pigments of the burial shroud coverings have been identified as the source of contamination. Accurate and precise quantitative results were obtained by nondestructive process using fundamental parameters method taking into account the matrix absorption effects. A total of 30 bones from 13 individuals, buried between the mid-XVIIIth to early XIXth centuries, were analyzed to study the elemental composition and elemental distribution. The bones were collected from a church in Almada (Portugal), called Ermida do Espírito Santo, located near the Tagus River and at the sea neighbourhood. The triaxial geometry setup was used to quantify Ca, Fe, Cu, Zn, Br, Sr and Pb of powder pressed bone pellets (n=9 for each bone). Cluster analysis was performed considering the elemental concentrations for the different bones. There was a clear association between some bones regarding Fe, Cu, Zn, Br and Pb content but not a categorization between cortical and trabecular bones. The elemental distribution of Cu, Zn and Pb were assessed by the benchtop µ-analysis, the M4 Tornado, based on a polycapillary system which provides multi-elemental 2D maps. The results showed that contamination was mostly on the surface of the bone confirming that it was related to the burial shroud covering the individuals.


Subject(s)
Bone and Bones/chemistry , Bone and Bones/diagnostic imaging , Molecular Imaging , Spectrometry, X-Ray Emission , Archaeology , Autopsy , Humans , Limit of Detection
6.
Oncogene ; 34(10): 1270-9, 2015 Mar 05.
Article in English | MEDLINE | ID: mdl-24662834

ABSTRACT

Intratumoral heterogeneity (ITH) represents an obstacle for cancer diagnosis and treatment, but little is known about its functional role in cancer progression. The A Desintegrin And Metalloproteinase 23 (ADAM23) gene is epigenetically silenced in different types of tumors, and silencing is often associated with advanced disease and metastasis. Here, we show that invasive breast tumors exhibit significant ADAM23-ITH and that this heterogeneity is critical for tumor growth and metastasis. We demonstrate that while loss of ADAM23 expression enhances invasion, it causes a severe proliferative deficiency and is not itself sufficient to trigger metastasis. Rather, we observed that, in ADAM23-heterotypic environments, ADAM23-negative cells promote tumor growth and metastasis by enhancing the proliferation and invasion of adjacent A23-positive cells through the production of LGI4 (Leucine-rich Glioma Inactivated 4) and nitric oxide (NO). Ablation of LGI4 and NO in A23-negative cells significantly attenuates A23-positive cell proliferation and invasion. Our work denotes a driving role of ADAM23-ITH during disease progression, shifting the malignant phenotype from the cellular to the tissue level. Our findings also provide insights for therapeutic intervention, enforcing the need to ascertain ITH to improve cancer diagnosis and therapy.


Subject(s)
ADAM Proteins/metabolism , Breast Neoplasms/metabolism , Breast Neoplasms/pathology , Extracellular Matrix Proteins/metabolism , Nitric Oxide/metabolism , ADAM Proteins/genetics , Breast Neoplasms/genetics , Cell Line, Tumor , Cell Proliferation , DNA Methylation , Epigenesis, Genetic , Extracellular Matrix Proteins/genetics , Female , Gene Silencing , Humans , Neoplasm Metastasis , Nerve Tissue Proteins , Tumor Burden , Tumor Microenvironment
7.
Cell Death Dis ; 2: e224, 2011 Nov 03.
Article in English | MEDLINE | ID: mdl-22048167

ABSTRACT

Wilms' tumors (WTs) originate from metanephric blastema cells that are unable to complete differentiation, resulting in triphasic tumors composed of epithelial, stromal and blastemal cells, with the latter harboring molecular characteristics similar to those of the earliest kidney development stages. Precise regulation of Wnt and related signaling pathways has been shown to be crucial for correct kidney differentiation. In this study, the gene expression profile of Wnt and related pathways was assessed in laser-microdissected blastemal cells in WTs and differentiated kidneys, in human and in four temporal kidney differentiation stages (i.e. E15.5, E17.5, P1.5 and P7.5) in mice, using an orthologous cDNA microarray platform. A signaling pathway-based gene signature was shared between cells of WT and of earliest kidney differentiation stages, revealing genes involved in the interruption of blastemal cell differentiation in WT. Reverse transcription-quantitative PCR showed high robustness of the microarray data demonstrating 75 and 56% agreement in the initial and independent sample sets, respectively. The protein expression of CRABP2, IGF2, GRK7, TESK1, HDGF, WNT5B, FZD2 and TIMP3 was characterized in WTs and in a panel of human fetal kidneys displaying remarkable aspects of differentiation, which was recapitulated in the tumor. Taken together, this study reveals new genes candidate for triggering WT onset and for therapeutic treatment targets.


Subject(s)
Gene Expression Regulation, Neoplastic , Genes, Wilms Tumor , Kidney Neoplasms/genetics , Kidney/physiology , Wilms Tumor/genetics , Animals , DNA, Complementary/genetics , HEK293 Cells , Humans , Kidney/embryology , Kidney/metabolism , Kidney/pathology , Kidney Neoplasms/pathology , Mice , Nucleic Acid Hybridization , Signal Transduction , Wilms Tumor/pathology , Wnt Proteins/biosynthesis , Wnt Proteins/genetics
8.
Vet Microbiol ; 153(3-4): 323-31, 2011 Dec 15.
Article in English | MEDLINE | ID: mdl-21742447

ABSTRACT

During the last decade the majority of diphtheria cases in Europe had Corynebacterium ulcerans as the etiologic agent with dogs and cats as the reservoir hosts. However, little has been documented about the virulence factors of this zoonotic pathogen. To set up an in vivo experimental C. ulcerans infection model, conventional Swiss Webster mice were intravenously infected with different doses (from 1 × 10(7) to 5 × 10(9) bacteria per mouse) of C. ulcerans strains, namely 809 (from human lower respiratory tract), BR-AD22 (from asymptomatic dog nares) and CDC-KC279. Mortality rates were demonstrated by LD(50) values ranging from 1.9 × 10(8) to 1.3 × 10(9). Viable bacteria were recovered from blood, kidneys, liver, spleen and joints. For CDC-KC279 and 809 strains (2 × 10(8)mL(-1)) approximately 85% and 72% of animals with articular lesions were observed, respectively; BR-AD22-infected mice showed no signs of arthritis. CDC-KC279 and 809 strains exhibited higher arthritogenic potential when compared to the homologous toxigenic (ATCC27012) and non-toxigenic (ATCC27010) strains of Corynebacterium diphtheriae. A high number of affected joints and arthritis index in addition to the histopathological features, including subcutaneous edema, inflammatory infiltrate, damage to bone tissue and synoviocyte hypertrophy, indicated a strain-dependent ability of C. ulcerans strains to cause severe polyarthritis. A correlation between the arthritis index and systemic levels of IL-6 and TNF-α was observed for C. ulcerans strains, with the exception of the non-arthritogenic BR-AD22 strain. In conclusion, C. ulcerans revealed a strain-dependent arthritogenic potential independent of DNAse, PLD and diphtheria toxin production.


Subject(s)
Arthritis, Infectious/microbiology , Corynebacterium Infections/microbiology , Corynebacterium Infections/pathology , Corynebacterium/physiology , Animals , Arthritis, Infectious/pathology , Bacterial Load , Corynebacterium/immunology , Corynebacterium Infections/immunology , Corynebacterium diphtheriae/physiology , Cytokines/metabolism , Disease Models, Animal , Female , Male , Mice , Species Specificity , Time Factors
9.
Biomacromolecules ; 12(9): 3243-51, 2011 Sep 12.
Article in English | MEDLINE | ID: mdl-21786745

ABSTRACT

To scrutinize materials for specific biomedical applications, we need sensitive and selective analytical methods that can give more insight into the process of their biodegradation. In the present study, the enzymatic degradation of multiblock poly(ester amide) based on natural amino acids, such as lysine and leucine, was performed with serine proteases (α-chymotrypsin (α-CT) and proteinase K (PK)) in phosphate-buffered saline solution at 37 °C for 4 weeks. Fully and partially degraded water-soluble products were analyzed by liquid chromatography hyphenated with time-of-flight mass spectrometry using an electrospray interface (LC-ESI-ToF-MS). Tracking the release of monomeric and oligomeric products into the enzyme media during the course of enzymatic degradation revealed the preferences of α-CT and PK toward ester and amide bonds: both α-CT and PK showed esterase and amidase activity. Although within the experimental time frame up to 30 and 15% weight loss was observed in case of α-CT and PK, respectively, analysis by size exclusion chromatography showed no change in the characteristic molecular-weight averages of the remaining polymer. This suggests that the enzymatic degradation occurs at the surface of this biomaterial. A sustained and linear degradation over a period of 4 weeks supports the potential of this class of poly(ester amide)s for drug delivery applications.


Subject(s)
Biocompatible Materials/metabolism , Chymotrypsin/metabolism , Endopeptidase K/metabolism , Polyesters/metabolism , Amides/chemistry , Animals , Biocompatible Materials/chemistry , Biodegradation, Environmental , Cattle , Chromatography, Liquid , Drug Delivery Systems/methods , Fungi , Leucine/chemistry , Lysine/chemistry , Molecular Weight , Polyesters/chemistry , Spectrometry, Mass, Electrospray Ionization
10.
J Phys Chem A ; 115(30): 8447-57, 2011 Aug 04.
Article in English | MEDLINE | ID: mdl-21707051

ABSTRACT

Methyl 2-azidopropionate (N(3)CH(3)CHCOOCH(3), M2AP) has been synthesized and characterized by different spectroscopic methods, and the thermal decomposition of this molecule has been investigated by matrix isolation infrared (IR) spectroscopy and ultraviolet photoelectron spectroscopy (UVPES). Computational methods have been employed in the spectral simulation of both UVPES and matrix IR spectra and in the rationalization of the thermal decomposition results. M2AP presents a HOMO vertical ionization energy (VIE) of 9.60 ± 0.03 eV and contributions from all four lowest-energy conformations of this molecule are detected in the gas phase. Its thermal decomposition starts at ca. 400 °C and is complete at ca. 650 °C, yielding N(2), CO, CO(2), CH(3)CN, and CH(3)OH as the final decomposition products. Methyl formate (MF) and CH(4) are also found during the pyrolysis process. Analysis of the potential energy surface of the decomposition of M2AP indicates that M2AP decomposes preferentially into the corresponding imine (M2IP), through a 1,2-H shift synchronous with the N(2) elimination (Type 1 mechanism), requiring an activation energy of 160.8 kJ/mol. The imine further decomposes via two competitive routes: one accounting for CO, CH(3)OH, and CH(3)CN (ΔE(G3) = 260.2 kJ/mol) and another leading to CO(2), CH(4), and CH(3)CN (ΔE(G3) = 268.6 kJ/mol). A heterocyclic intermediate (Type 2 mechanism)-4-Me-5-oxazolidone-can also be formed from M2AP via H transfer from the remote O-CH(3) group, together with the N(2) elimination (ΔE(G3) = 260.2 kJ/mol). Finally, a third pathway which accounts for the formation of MF through an M2AP isomer is envisioned.


Subject(s)
Azides/chemistry , Heterocyclic Compounds/chemistry , Imines/chemical synthesis , Propionates/chemistry , Thermodynamics , Ultraviolet Rays , Azides/chemical synthesis , Imines/chemistry , Models, Molecular , Photoelectron Spectroscopy , Propionates/chemical synthesis , Quantum Theory , Spectrophotometry, Infrared
11.
J Chromatogr A ; 1218(3): 449-58, 2011 Jan 21.
Article in English | MEDLINE | ID: mdl-21167489

ABSTRACT

Synthetic biomaterials have evoked extensive interest for applications in the field of health care. Prior to administration to the body a quantitative study is necessary to evaluate their composition. An in vitro method was developed for the quick hydrolytic degradation of poly-2-hydroxyethyl methacrylate (pHEMA), poly(lactide-co-glycolide50/50)1550-diol (PLGA(50:50)(1550)-diol), PLGA(50:50)(1550)-diol(HEMA)(2) and PLGA(50:50)(1550)-diol(etLDI-HEMA)(2) containing ethyl ester lysine diisocyanate (etLDI) linkers using a microwave instrument. Hydrolysis time and temperature were optimized while monitoring the degree of hydrolysis by (1)H NMR spectroscopy. Complete hydrolytic degradation was achieved at 120°C and 3 bar pressure after 24 h. Chemical structure elucidations of the degradation products were carried out using (1)H and (13)C NMR spectroscopy. The molecular weight (MW) of the polymethacrylic backbone was estimated via size-exclusion chromatography coupled to refractive index detection (SEC-dRI). A bimodal MW distribution was found experimentally, also in the pHEMA starting material. The number average molecular weights (M(n)) of the PLGA-links (PLGA(50:50)(1550)-diol) were calculated by high pressure liquid chromatography-time-of-flight mass spectrometry (HPLC-TOF-MS) and (1)H NMR. The amounts of the high and low MW degradation products were determined by SEC-dRI and, HPLC-TOF-MS, respectively. The main hydrolysis products poly (methacrylic acid) (PMAA), ethylene glycol (EG), diethylene glycol (DEG), lactic acid (LA), glycolic acid (GA) and lysine were recovered almost quantitatively. The current method leads to the complete hydrolytic degradation of these materials and will be helpful to study the degradation behavior of these novel cross-linked polymeric biomaterials.


Subject(s)
Lactic Acid/chemistry , Polyamines/chemistry , Polyglycolic Acid/chemistry , Polyhydroxyethyl Methacrylate/analogs & derivatives , Chromatography, Gel , Chromatography, High Pressure Liquid , Hydrolysis , Materials Testing , Nuclear Magnetic Resonance, Biomolecular , Polyhydroxyethyl Methacrylate/chemistry , Polylactic Acid-Polyglycolic Acid Copolymer , Spectrometry, Mass, Electrospray Ionization
12.
Braz J Med Biol Res ; 43(5): 460-6, 2010 May.
Article in English | MEDLINE | ID: mdl-20490433

ABSTRACT

The construction of a hexahistidine-tagged version of the B fragment of diphtheria toxin (DTB) represents an important step in the study of the biological properties of DTB because it will permit the production of pure recombinant DTB (rDTB) in less time and with higher yields than currently available. In the present study, the genomic DNA of the Corynebacterium diphtheriae Park Williams 8 (PW8) vaccine strain was used as a template for PCR amplification of the dtb gene. After amplification, the dtb gene was cloned and expressed in competent Escherichia coli M15 cells using the expression vector pQE-30. The lysate obtained from transformed E. coli cells containing the rDTB PW8 was clarified by centrifugation and purified by affinity chromatography. The homogeneity of the purified rDTB PW8 was confirmed by immunoblotting using mouse polyclonal anti-diphtheria toxoid antibodies and the immune response induced in animals with rDTB PW8 was evaluated by ELISA and dermonecrotic neutralization assays. The main result of the present study was an alternative and accessible method for the expression and purification of immunogenically reactive rDTB PW8 using commercially available systems. Data also provided preliminary evidence that rabbits immunized with rDTB PW8 are able to mount a neutralizing response against the challenge with toxigenic C. diphtheriae.


Subject(s)
Corynebacterium diphtheriae/genetics , Diphtheria Toxin/genetics , Gene Expression Regulation, Bacterial/genetics , Animals , Corynebacterium diphtheriae/classification , DNA, Bacterial , Male , Mice , Polymerase Chain Reaction , Rabbits , Sequence Analysis, DNA
13.
Braz. j. med. biol. res ; 43(5): 460-466, May 2010. ilus
Article in English | LILACS | ID: lil-546334

ABSTRACT

The construction of a hexahistidine-tagged version of the B fragment of diphtheria toxin (DTB) represents an important step in the study of the biological properties of DTB because it will permit the production of pure recombinant DTB (rDTB) in less time and with higher yields than currently available. In the present study, the genomic DNA of the Corynebacterium diphtheriae Park Williams 8 (PW8) vaccine strain was used as a template for PCR amplification of the dtb gene. After amplification, the dtb gene was cloned and expressed in competent Escherichia coli M15™ cells using the expression vector pQE-30™. The lysate obtained from transformed E. coli cells containing the rDTB PW8 was clarified by centrifugation and purified by affinity chromatography. The homogeneity of the purified rDTB PW8 was confirmed by immunoblotting using mouse polyclonal anti-diphtheria toxoid antibodies and the immune response induced in animals with rDTB PW8 was evaluated by ELISA and dermonecrotic neutralization assays. The main result of the present study was an alternative and accessible method for the expression and purification of immunogenically reactive rDTB PW8 using commercially available systems. Data also provided preliminary evidence that rabbits immunized with rDTB PW8 are able to mount a neutralizing response against the challenge with toxigenic C. diphtheriae.


Subject(s)
Animals , Male , Mice , Rabbits , Corynebacterium diphtheriae/genetics , Diphtheria Toxin/genetics , Gene Expression Regulation, Bacterial/genetics , Corynebacterium diphtheriae/classification , DNA, Bacterial , Polymerase Chain Reaction , Sequence Analysis, DNA
14.
Toxicon ; 51(4): 502-14, 2008 Mar 15.
Article in English | MEDLINE | ID: mdl-18155119

ABSTRACT

Envenoming from snakebites is an important public health issue in Brazil. In 2005, 28,597 cases were notified (15 cases/100,000 inhabitants), 87.5% due to Bothrops and 9.2% to Crotalus genus. Antivenoms available in Brazil are liquid preparations containing purified equine Fab'2. Since 1987, the National Institute for Quality Control in Health (INCQS/FIOCRUZ) has been testing all lots prior to batch release. Between 2000 and 2006, 619 lots of antivenoms were tested, comprising 2,513,690 ampoules. The potency assay was performed only for bothropic and crotalic antivenoms (485 lots corresponding to 1,866,726 ampoules) due to the unavailability of the other reference venoms. This paper aims to report the last 7-year activities of INCQS on the quality control, batch release and potency evaluation of antivenoms.


Subject(s)
Antivenins/pharmacology , Laboratories , Animals , Brazil , Female , Male , Quality Control , Reference Standards , Reproducibility of Results , Time Factors
15.
Lett Appl Microbiol ; 45(3): 270-5, 2007 Sep.
Article in English | MEDLINE | ID: mdl-17718838

ABSTRACT

AIMS: To investigate different autochthonous isolates of wood-rotting fungi for the removal of both colour and phenolic compounds from olive mill wastewaters (OMW). METHODS AND RESULTS: The isolates Bjerkandera adusta Ba-100, Fomes fomentarius Ff-106, Ganoderma applanatum Ga-20, Irpex lacteus Il-3, Trametes versicolor Tv-101 and Tv-103 were preliminarily screened for their OMW-decolourizing potential on potato dextrose agar supplemented with different OMW concentrations. A further screening of batch cultures under different agitation speeds, to test the effect of shear stress, resulted in the selection of isolate G. applanatum Ga-20. Batch cultures grown in OMW-based medium exhibited strong laccase induction and significant decrease in the values of phenols, colour and chemical oxygen demand. Concomitant onset of laccase activity and colour removal was observed, and apart from laccase, neither lignin peroxidase nor manganese-dependent peroxidase activities were detected. Moreover, the depletion of aromatic compounds with high and low apparent molecular mass was observed by chromatographic analysis. CONCLUSIONS: Isolate G. applanatum Ga-20 exhibited interesting properties for its use in bioremediation of OMW, namely high removal of recalcitrant phenolic compounds and strong colour abatement. SIGNIFICANCE AND IMPACT OF THE STUDY: For the first time, the white-rot fungus G. applanatum proves to be effective for the decolourization and dephenolization of OMW.


Subject(s)
Ganoderma/enzymology , Industrial Microbiology , Industrial Waste , Phenol/metabolism , Plant Oils/chemistry , Waste Disposal, Fluid , Biodegradation, Environmental , Color , Ganoderma/isolation & purification , Laccase/metabolism , Olive Oil , Phenol/chemistry , Water Pollutants/metabolism
16.
J Chromatogr A ; 1156(1-2): 111-23, 2007 Jul 13.
Article in English | MEDLINE | ID: mdl-17113590

ABSTRACT

UV-cured networks prepared from mixtures of di-functional (polyethylene-glycol di-acrylate) and mono-functional (2-ethylhexyl acrylate) acrylates were analysed after hydrolysis, by aqueous size-exclusion chromatography coupled to on-line reversed-phase liquid-chromatography. The mean network density and the fraction of dangling chain ends of these networks were varied by changing the concentration of mono-functional acrylate. The amount and the molar-mass distribution of the polyethylene-glycol chains between cross-links (M(XL)) and polyacrylic acid (PAA) backbone chains (the so-called kinetic chain length (kcl)) in the different acrylate networks were determined quantitatively. The molar-mass distribution of kcl revealed an almost linear dependence on the concentration of mono-functional acrylate. Analysis of the starting materials showed a significant concentration of mono-functional polyethylene-glycol acrylate. In combination with the analysis of the extractables of the UV-cured networks (polymers not attached to the network, impurities that originate from the photo-initiator and unreacted monomers), more insight in the total network structure was obtained. It was shown that the UV-cured networks contain only small fractions of residual compounds. With these results, the chemical network structure for the different UV-cured acrylate polymers was expressed in network parameters such as the number of PAA units which are cross-linked, the degree of cross-linking, and the network density, which is the molar concentration of effective network chains between cross-links per volume of the polymers. The mean molar mass of chains between chemical network junctions (M(C)) was calculated and compared with results obtained from solid-state NMR and DMA. The mean molar mass of chains between network junctions as determined by these methods was similar.


Subject(s)
Chromatography, Gel/methods , Polyethylene Glycols/radiation effects , Acrylates/chemistry , Acrylates/radiation effects , Chromatography, Liquid/methods , Hydrolysis , Polyethylene Glycols/chemistry , Ultraviolet Rays
17.
Diabetes Metab ; 31(6): 581-7, 2005 Dec.
Article in English | MEDLINE | ID: mdl-16357807

ABSTRACT

AIM: To evaluate the response to an oral lipid overload, inflammatory markers and carotid intima-media thickness in subjects with impaired glucose tolerance. METHODS: 54 subjects, both sexes, 58 y-old average were submitted to 1) Clinical evaluation 2) Glucose tolerance test with 75 g glucose; classified as normal (2 h plasma glucose<140 mg/dl, n=37) or IGT (2 h G 140-200 mg/dl, n=17), 3) 12 h fasting sample (plasma glucose, lipids, C-reactive protein, fibrinogen and HOMA-IR calculation); 4 and 6 h after the oral lipid overload (1000 kcal, lipids 65 g) glycemia, fibrinogen and triglycerides were reevaluated. Intima-media thickness was calculated by the average of 6 measurements (3 highest of each carotid) evaluated by ultrasonography (7 MHZ transducer). RESULTS: The IGT group had higher (P<0.001) fasting plasma glucose (89.4 +/- 13 vs 104.4 +/- 8 mg/dl), HOMA-IR (1.69 +/- 1.2 vs 2.93 +/- 2.2) and waist (91 +/- 14 vs 101 +/- 9 cm), similar fasting lipids, intima-media thickness (P=0.58) and post-oral lipid overload triglycerides (P=0.74), but higher fibrinogen (284.3 +/- 6 and 305 +/- 10 mg/dl, P=0.05) and C-reactive protein (2.11 +/- 0.33 and 4.19 +/- 0.65 mg/l, P=0.003). C-reactive protein was positively correlated with HOMA-IR (r=0.45, P=0.001), fasting plasma glucose (r=0.43, P=0.002) and waist (r=0.45, P=0.0006), but not with postprandial lipids. CONCLUSION: A higher C-reactive protein in IGT, and its positive correlation with insulin resistance indices, but not with postprandial lipaemia, suggests that the clustering of these factors, characteristic of the metabolic syndrome, occurs earlier than postprandial lipid abnormalities.


Subject(s)
Atherosclerosis/epidemiology , Glucose Intolerance/blood , Glucose Intolerance/physiopathology , Blood Glucose/metabolism , Body Size , C-Reactive Protein/metabolism , Carotid Arteries/pathology , Fasting , Female , Glucose Tolerance Test , Humans , Lipids/blood , Male , Middle Aged , Risk Factors , Tunica Intima/pathology , Tunica Media/pathology
18.
Mem Inst Oswaldo Cruz ; 100 Suppl 1: 25-7, 2005 Mar.
Article in English | MEDLINE | ID: mdl-15962095

ABSTRACT

We have previously reported that in comparison with normal rats, the presence of experimental allergic encephalomyelitis (EAE) leads to decreased endogenous inhibitory activity (EIA) of Ca2+-dependent nitric oxide synthase (NOS) in both brain and serum, and increased expression of protein 3-nitrotyrosine (NT) in brain. In this work we show that animals recovered from the clinical signs of EAE are not different from controls in terms of either brain NOS activity, EIA of NOS, or NT expression. These results suggest that parallel to the reversal of the disease symptoms, a normalization of the production of nitric oxide and related species occurs.


Subject(s)
Brain/enzymology , Encephalomyelitis, Autoimmune, Experimental/enzymology , Nitric Oxide Synthase Type III/metabolism , Nitric Oxide/metabolism , Animals , Cytoplasmic Dyneins , Dyneins/metabolism , Encephalomyelitis, Autoimmune, Experimental/blood , Male , Nitric Oxide Synthase Type III/antagonists & inhibitors , Rats , Rats, Inbred Lew
19.
Mem. Inst. Oswaldo Cruz ; 100(supl.1): 25-27, Mar. 2005. ilus, tab
Article in English | LILACS | ID: lil-402172

ABSTRACT

We have previously reported that in comparison with normal rats, the presence of experimental allergic encephalomyelitis (EAE) leads to decreased endogenous inhibitory activity (EIA) of Ca2+-dependent nitric oxide synthase (NOS) in both brain and serum, and increased expression of protein 3-nitrotyrosine (NT) in brain. In this work we show that animals recovered from the clinical signs of EAE are not different from controls in terms of either brain NOS activity, EIA of NOS, or NT expression. These results suggest that parallel to the reversal of the disease symptoms, a normalization of the production of nitric oxide and related species occurs.


Subject(s)
Animals , Male , Rats , Brain/enzymology , Encephalomyelitis, Autoimmune, Experimental/enzymology , Nitric Oxide Synthase Type III/metabolism , Nitric Oxide/metabolism , Dyneins/metabolism , Encephalomyelitis, Autoimmune, Experimental/blood , Nitric Oxide Synthase Type III/antagonists & inhibitors , Rats, Inbred Lew
20.
J Leukoc Biol ; 75(4): 649-56, 2004 Apr.
Article in English | MEDLINE | ID: mdl-14726497

ABSTRACT

Pentraxin 3 (PTX3) is a tumor necrosis factor and interleukin-1beta-stimulated gene that encodes a long PTX with proinflammatory activity. Here, we show that peritoneal macrophages derived from PTX3 transgenic (Tg) mice express higher levels of PTX3 mRNA than macrophages from wild-type (WT) mice, at basal level as well as upon stimulation with zymosan (Zy). Macrophages from Tg mice also showed improved opsonin-independent phagocytosis of Zy particles and the yeast form of the fungus Paracoccidioides brasiliensis. In the case of P. brasiliensis, an enhanced microbicidal activity accompanied by higher production of nitric oxide was also observed in macrophages from Tg mice. Using fluorescein-activated cell sorter analysis and reverse transcriptase-polymerase chain reaction, we demonstrated that basal level of Toll-like receptor-6 and Zy-induced dectin-1 expression was slightly but consistently higher in macrophages from Tg mice than in macrophages from WT mice. Recombinant (r)PTX3 protein binds to Zy particles as well as to yeast cells of P. brasiliensis and addition of rPTX3, to a culture of WT-derived macrophages containing Zy leads to an increase in the phagocytic index, which parallels that of Tg-derived macrophages, demonstrating the opsonin-like activity of PTX3. It is important that blockade of dectin-1 receptor inhibited the phagocytosis of Zy particles by WT and PTX3 Tg macrophages, pointing out the relevant role of dectin-1 as the main receptor involved in Zy uptake. Our results provide evidence for a role of PTX3 as an important component of the innate-immune response and as part of the host mechanisms that control fungal recognition and phagocytosis.


Subject(s)
C-Reactive Protein/genetics , Macrophages, Peritoneal/metabolism , Membrane Proteins/metabolism , Nerve Tissue Proteins/metabolism , Opsonin Proteins/genetics , Phagocytosis/genetics , Serum Amyloid P-Component/genetics , Zymosan/immunology , Animals , Binding Sites/drug effects , Binding Sites/genetics , C-Reactive Protein/metabolism , Female , Immunity, Innate/genetics , Lectins, C-Type , Macrophages, Peritoneal/drug effects , Macrophages, Peritoneal/immunology , Male , Membrane Glycoproteins/drug effects , Membrane Glycoproteins/metabolism , Membrane Proteins/antagonists & inhibitors , Mice , Mice, Transgenic , Nerve Tissue Proteins/antagonists & inhibitors , Nitric Oxide/metabolism , Opsonin Proteins/metabolism , Paracoccidioides/immunology , Phagocytosis/drug effects , Phagocytosis/immunology , RNA, Messenger/drug effects , RNA, Messenger/metabolism , Receptors, Cell Surface/drug effects , Receptors, Cell Surface/metabolism , Recombinant Fusion Proteins/metabolism , Serum Amyloid P-Component/metabolism , Toll-Like Receptor 6 , Zymosan/metabolism , Zymosan/pharmacology
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