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    Full-text · Article · Feb 2016
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    ABSTRACT: Objectives the aim of this study was to evaluate the effect of SLPI on the growth and biological processes of Candida albicans. Methods Two C. albicans strains were used in this study, a clinical isolate resistant to fluconazole (PRI) and a reference strain ATCC 24433. The minimal inhibitory concentration (MIC) was determined according to the CLSI methodology. The influence of SLPI on secreted serine proteinase activities (SSP) was measured by the cleavage of specific substrate, and surface hydrophobicity was determined by the aqueous-hydrocarbon biphasic separation method. Flow cytometry was performed to investigate receptors for SLPI and variations in the cell wall mannoprotein expression. Interaction between yeast and epithelium was assessed using the MA-104 cells lineage. Ultrastructure was analyzed by transmission electron microscopy (TEM). Results MIC values were calculated as 18 and 18.9 μM for the PRI and ATCC 24433, respectively. SSP activity was reduced by 48.8% by 18 μM of SLPI and cell surface hydrophobicity increased by 11.1%. Flow cytometry suggest the existence of SLPI binding sites on the surface of the yeast. Results showed a reduction in the expression of mannoproteins in 20.8% by the cells treated with 80 μM of SLPI, and 18 μM reduced the adhesion of yeasts to mammalian cells in 60.1%. TEM revealed ultrastructural changes in cells treated with 80 μM of SLPI, such as the presence of membrane-like structures within the cytoplasm. Conclusions SLPI exerts a significant influence on C. albicans viability and biological processes. Considering its constitutive and physiologic features, SLPI may become a promising tool for the development of new methodologies for the treatment and control of candidiasis.
    Preview · Article · Sep 2014 · Archives of Oral Biology
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    ABSTRACT: 4) H&N Homeopatia-Farmácias HNCristiano; (5) Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil; (6) Universita' degli studi di Verona, Verona, Italy Zincum metallicum (ZM) is a homeopathic medicine whose material medica is defined by diverse behavioral and mental symptoms, including depression. Moreover, as a microelement, zinc itself is involved in several functions of Central Nervous System, including development and cell maturity during the intra-uterus life. Herein, the putative transgenerational effects of different homeopathic potencies of ZM upon behavioral parameters in P and F1 generations were evaluated. Since mice and the tail suspension test (TST) are references for evaluating antidepressant agent activity, the TST together with the open field test (OPT) and the elevated plus maze test (EPM) were used to analyze offspring behavioral parameters. All animal procedures were in agreement with the Brazilian ethical research practices and were approved by the institutional ethical committee (CEUA-UNIP) under the protocol 156/2013. Four groups of seven females Balb/C mice were exposed to 0,1mL of ZM 5cH, 30cH, 200cH and lactose 5cH, diluted in 250mL of drinking water, during pregnancy and post partum period, in a total of 31 days. The flasks were coded before the remedies administration and all experimental procedures, including statistical analysis were done in blind. The parents were previously distributed in a Completely Randomized Design for the mates, according to the TST previous results. Mothers were re-evaluated for TST after weaning and mice of F1 generation were evaluated for TST, EPM and OPT when they reached two months old. According to the time of immobilization in TST, animals were classified as healthy (h), intermediate (i) and depressed (d) (< 116; 117-180 and >180 seconds of immobilization, respectively). No significant changes were seen among the groups regarding to the number of newborns, sex proportions, TST, OPT and EPM behavioral parameters, besides the fact that the treatment with ZM 200cH was associated to the majority of healthy F1 mice (male: n=8: 7h+1i+0d; female: n=8: 8h+0i+0d), in relation to the number of delivery per group (Fisher test, p ≤ 0.01). Treatment with ZM 5cH, instead, produced reduced number of pups with no male mouse among F1 generation. We conclude that the treatment of pregnant females with ZM 200cH produced the best results in F1 generation regarding reproduction and behavioral parameters and the treatment with ZM 5cH reduced births in relation to control. The involved mechanisms have to be elucidated in the next steps of the study.
    Full-text · Conference Paper · Jun 2014
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    Garcia Sheila · PINHEIRO · Mariana Santos · AGUIAR · P. F. · HOLANDINO · GARCIA

    Full-text · Article · Jan 2014
  • Anderson de Oliveira Ferreira · Carla Holandino
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    ABSTRACT: Enteric coated capsules are characterized by their resistance to dissolution in low pH environments, such as the stomach, and by their rapid disintegration in a higher pH environment, such as the intestine. The surface of hard gelatin capsules is usually smooth and nonporous, which limits their coating efficiency. We developed a simple, quick, and easily reproducible compounding preparation method for enteric-release hard gelatin capsules. Twenty-two batches of 60 diclofenac sodium capsules each were prepared and then divided into three groups. Each group was submitted to a different coating process using a small-scale enteric coating machine and a coating process based on the atomization (spraying) of organic solutions of polymers. The results of dissolution testing were compared statistically within the groups and also with a reference drug control (Voltaren DR). Some of the batches in group I and all the batches in groups II and III met the pharmacopeial requirements for enteric release, in both acid and pH 6.8 phosphate buffer stages. Dissolution of capsules coated with Eudragit L100 was more efficient than that of other tested systems (P less than 0.05). The dissolution assay results for group III in the acid stage were superior to the results for the reference drug, but no statistical difference was noticed between these two in the phosphate buffer stage. Diclofenac sodium hard gelatin capsules coated with cellulose acetate Phthalate or Eudragit L100 presented dissolution performance that met pharmacopeial requirements. Furthermore, results obtained with Eudragit-coated capsules were comparable to those obtained with the reference drug.
    No preview · Article · Aug 2013 · International Journal of Pharmaceutical Compounding
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    ABSTRACT: Electrochemical treatment is an alternative modality for tumor treatment based on the application of a low intensity direct electric current to the tumor tissue through two or more platinum electrodes placed within the tumor zone or in the surrounding areas. This treatment is noted for its great effectiveness, minimal invasiveness and local effect. Several studies have been conducted worldwide to evaluate the antitumoral effect of this therapy. In all these studies a variety of biochemical and physiological responses of tumors to the applied treatment have been obtained. By this reason, researchers have suggested various mechanisms to explain how direct electric current destroys tumor cells. Although, it is generally accepted this treatment induces electrolysis, electroosmosis and electroporation in tumoral tissues. However, action mechanism of this alternative modality on the tumor tissue is not well understood. Although the principle of Electrochemical treatment is simple, a standardized method is not yet available. The mechanism by which Electrochemical treatment affects tumor growth and survival may represent more complex process. The present work analyzes the latest and most important research done on the electrochemical treatment of tumors. We conclude with our point of view about the destruction mechanism features of this alternative therapy. Also, we suggest some mechanisms and strategies from the thermodynamic point of view for this therapy. In the area of Electrochemical treatment of cancer this tool has been exploited very little and much work remains to be done. Electrochemical treatment constitutes a good therapeutic option for patients that have failed the conventional oncology methods.
    Full-text · Article · Apr 2013 · Chinese Journal of Cancer Research
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    ABSTRACT: Influenza viruses cause highly contagious acute respiratory illnesses with significant mortality, especially among young children, elderly people, and individuals with serious medical conditions. This encourages the development of new treatments for human flu. Biotherapies are diluted solutions prepared from biological products compounded following homeopathic procedures. To develop a biotherapy prepared from the infectious influenza A virus (A/Aichi/2/68 H3N2) and to verify its in vitro response. The ultradiluted influenza virus solution was prepared in the homeopathic dilution 30dH, it was termed Influenzinum RC. The cellular alterations induced by this preparation were analyzed by optical and electron microscopy, MTT and neutral red assays. Glycolytic metabolism (PFK-1) was studied by spectrophotometric assay. Additionally, the production of tumor necrosis factor-α (TNF-α) by J774.G8 macrophage cells was quantified by ELISA before and after infection with H3N2 influenza virus and treatment. Influenzinum RC did not cause cytotoxic effects but induced morphological alterations in Madin-Darby canine kidney (MDCK) cells. After 30 days, a significant increase (p < 0.05) in mitosis rate was detected compared to control. MDCK mitochondrial activity was changed after treatment for 10 and 30 days. Treatment significantly diminished (p < 0.05) PFK-1 activity. TNF-α in biotherapy-stimulated J774.G8 macrophages indicated a significant (p < 0.05) increase in this cytokine when the cell supernatant was analyzed. Influenzinum RC altered cellular and biochemical features of MDCK and J774G8 cells.
    Full-text · Article · Jan 2013 · Homeopathy: the journal of the Faculty of Homeopathy
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    ABSTRACT: This work aimed to produce poly(methyl methacrylate) nanoparticles for use in drug encapsulation. The polymer nanoparticles were produced using miniemulsion polymerization technique. Monomer miniemulsion showed moderate stability and polymer average particle size was about 90 nm. PMMA nanoparticles were tested for toxicity in human leukemic cell strain K562 and they did not show any adverse effect on cell viability. Therefore, poly(methyl methacrylate) nanoparticles are suitable to encapsulate antitumor agents.
    Full-text · Article · Sep 2012 · Macromolecular Symposia
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    ABSTRACT: The literature shows that the effects of direct electric currents on biological material are numerous, including bactericidal, fungicidal, parasiticidal, and anti-tumoral, among others. Non-pathogenic trypanosomatids, such as Herpetomonas samuelpessoai, have emerged as important models for the study of basic biological processes performed by a eukaryotic cell. The present study reports a dose-dependent anti-protozoan effect of direct electric treatment with both cathodic and anodic current flows on H. samuelpessoai cells. The damaging effects can be attributable to the electrolysis products generated during electric stimulation. The pH of the cell suspension was progressively augmented from 7.4 to 10.5 after the cathodic treatment. In contrast, the anodic treatment caused a pH decrease varying from 7.4 to 6.5. Transmission electron microscopy analyses revealed profound alterations in vital cellular structures (e.g., mitochondrion, kinetoplast, flagellum, flagellar pocket, nucleus, and plasma membrane) after exposure to both cathodic and anodic current flows. Specifically, cathodic current flow treatment induced the appearance of autophagic-like structures on parasite cells, while those submitted to an anodic current flow presented marked disorganization of plasma membrane and necrotic appearance. However, parasites treated in the intermediary chamber (without contact with the electrodes) did not present significant changes in viability or morphology, and no pH variation was detected in this system. The use of H. samuelpessoai as a biological model and the direct electric current experimental approach used in our study provide important information for understanding the mechanisms involved in the cytotoxic effects of this physical agent. Bioelectromagnetics. © 2011 Wiley Periodicals, Inc.
    Full-text · Article · May 2012 · Bioelectromagnetics
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    ABSTRACT: Staphylococcus lugdunensis is an unusually virulent coagulase-negative species, which causes serious infection similar to S. aureus. We evaluated the expression of virulence factors such as S. lugdunensis synergistic haemolysin (SLUSH), fibrinogen-binding protein (Fbl), biofilm production and biofilm-production-related genes in 23 S. lugdunensis clinical isolates and one type strain that had been previously characterized for their genotypes. In addition, the biofilm composition and the ability of isolates to adhere to and invade human epithelial lung cells were also investigated. The PCR method used detected the presence of slush and intercellular adhesin (ica) virulence genes in all isolates. All isolates produced the Fbl protein and, with the exception of the type strain, all isolates produced the SLUSH haemolysin. Fourteen (60.9 %) isolates produced biofilms. The detachment assay, using sodium metaperiodate or proteolytic enzymes to analyse the biofilm composition, showed protein-mediated biofilms in two representative isolates, one for each colony type (rough and smooth). All strongly biofilm-producing isolates, including three with rough colony morphology, had the same prevalent PFGE pattern. However, among the representative strains tested, only the S. lugdunensis isolate that formed rough colonies was able to adhere to and invade A549 cell monolayers in the same quantities as those observed with S. aureus isolates (P = 1.000). No significant adhesion or invasion was observed for the other isolates in comparison with the S. aureus isolate, independent of biofilm production or clonality. Our results could explain the incredible ability of this pathogen to cause infections that are as aggressive as S. aureus. In addition, the ability of S. lugdunensis to adhere to and invade eukaryotic cells was also noticed for isolates with rough colony morphology, reinforcing the increased virulence in this species.
    Preview · Article · Nov 2011 · Journal of Medical Microbiology
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    ABSTRACT: In this study, zinc phthalocyanine (ZnPc) was loaded onto poly-ɛ-caprolactone (PCL) nanoparticles (NPs) using a solvent emulsification-evaporation method. The process yield and encapsulation efficiency were 74.2% ± 1.2% and 67.1% ± 0.9%, respectively. The NPs had a mean diameter of 187.4 ± 2.1 nm, narrow distribution size with a polydispersity index of 0.096 ± 0.004, zeta potential of -4.85 ± 0.21 mV, and spherical shape. ZnPc has sustained release, following Higuchi's kinetics. The photobiological activity of the ZnPc-loaded NPs was evaluated on human lung adenocarcinoma A549 cells. Cells were incubated with free ZnPc or ZnPc-loaded NPs for 4 h and then washed with phosphate-buffered saline. Culture medium was added to the wells containing the cells. Finally, the cells were exposed to red light (660 nm) with a light dose of 100 J/cm(2). The cellular viability was determined after 24 h of incubation. ZnPc-loaded NPs and free photosensitizer eliminated about 95.9% ± 1.8% and 28.7% ± 2.2% of A549 cells, respectively. The phototoxicity was time dependent up to 4 h and concentration dependent at 0-5 μg ZnPc. The cells viability decreased with the increase of the light dose in the range of 10-100 J/cm(2). Intense lysis was observed in the cells incubated with the ZnPcloaded NPs and irradiated with red light. ZnPc-loaded PCL NPs are the release systems that promise photodynamic therapy use.
    Full-text · Article · Jan 2011 · International Journal of Nanomedicine
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    ABSTRACT: Background: Candida spp is naturally found in humans' flora of skin, gastrointestinal and genitourinary tracts and, in general, up to 75% of the population does not have any symptom [1]. However, oral candidiasis is very common among HIV patients and patients undergoing chemotherapy. The treatment of oral candidiasis is necessary once the disease causes discomfort and dysphagia, resulting in poor nutrition, slow recovery, and prolonged hospital stay [2,3]. Preliminary results obtained by our group with a new biotherapic prepared from Candida albicans (Candida 30x) showed a great potential to reduce the candida yeast adhesion rate when the epithelial cells were pre-treated. This study is currently being developed with the evaluation of mutagenic and genotoxic potentials of several homeopathic solutions. Aims: The goal of this study was to assess the genotoxic and mutagenic potentials of different homeopathic potencies of C. albicans. Methodology: One part of C. albicans yeast obtained from Brazilian patient's blood [4] was diluted in 9 parts of sterile water. This sample was submitted to 100 mechanical succussions (approximately 3 Hz), using Autic® Brazilian machine, originating the first dilution (1x). Then, 1 ml of this solution was diluted in 9 ml of solvent, submitted to 100 succussions, obtaining 2x potency. This procedure was successively repeated to obtain 30x potency, according to Brazilian Homeopathic Pharmacopoeia [5]. By the same technique, water vehicle was prepared until 30x to be used as control. All samples were prepared in sterile and aseptic conditions, using laminar flow cabinet, class II and were stored in the refrigerator (8°C). The samples 1x, 6x, 12x, 18x, 24x and 30x of C. albicans and water 30x (vehicle control) were analysed by: the Inductest, which assesses the ability of physical or chemical agents to promote lysogenic induction as a reflection of damage in DNA molecules in lysogenic bacteria, and the Ames test, which uses indicator strains of Salmonella typhimurium, sensitive to substances that can induce different types of mutation. Results: In the Inductest no decrease in survival fraction of bacteria and no increase in the formation of lysogenic induction were detected independently of the homeopathic potency employed. The same profile was obtained after the Ames test, with similar results to negative control. Conclusion: Afterwards, we can conclude that these samples are not able to induce DNA damage in the cells tested. So, the use of this medicine does not present any side effects related to mutagenesis and genotoxicity.
    No preview · Article · Jan 2011 · International Journal of High Dilution Research
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    ABSTRACT: Introduction: Strains of macrophages, such as murine J774.G8 macrophages, are susceptible to influenza A infection [1]. One of the responses to viral infection involves the production of various types of immunostimulatory cytokines by infected cells [2]. Methods: In the present study, the macrophage strain J774.G8, maintained in RPMI medium, was submitted to treatment with 10% V/V of two different biotherapics prepared from influenza H3N2, both at 30x. Additionally, two control groups were analyzed: macrophages stimulated with water 30x and macrophages without any treatment. Biotherapics were prepared from intact H3N2 influenza virus and H3N2 inactivated by alcohol 70%. The compounding of both biotherapics followed this procedure: one part of viral particles was diluted in 9 parts of sterile distilled water. The 1:10 sample was submitted to 100 mechanical succussions using Autic® Brazilian machine, originating the first dilution, named decimal (1x). 1 ml of this solution was diluted in 9 ml of solvent and was submitted to 100 succussions, generating biotherapic 2x. This procedure was successively repeated, according to Brazilian Homeopathic Pharmacopoeia, to obtain the biotherapic 30x. By the same technique, water vehicle was prepared in the potency of 30x to be used as control. All samples were prepared under sterile and aseptic conditions, using laminar flow cabinet, class II, and were stored in the refrigerator (8°C), to avoid microbiological contamination. J774.G8 macrophages were stimulated for 2 days, in a total of six stimuli. Immediately before infection with 25 μl of H3N2 influenza virus, the supernatants were collected and frozen at -20 °C for later analysis. Next, 24 hours after the virus infection, the supernatants were aliquoted and frozen under the same conditions. Three independent experiments were done in triplicate. Analysis of supernatants was performed by flow cytometry using the Mouse Inflammation Kit. The cytokines detected in this experiment were IL-10, IL 12, TNF-α and MCP1. Results: In all cases, there were no significant differences compared to control groups. However, the production of TNF-α detected in macrophages treated by intact and inactivated biotherapics presented a tendency to increase after infection. In fact, similar results were previously detected in other experiments conducted only with the intact biotherapic [3]. The release of the cytokine MCP1 in all experimental situations presented a tendency to decrease after the viral infection when compared to untreated macrophages. No statistically significant difference was detected in the production of IL 12 and IL 10. These experiments will be repeated to confirm the data obtained.
    No preview · Article · Jan 2011 · International Journal of High Dilution Research
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    ABSTRACT: Background: Oral candidiasis is an opportunist fungal infection in humans, mainly caused by Candida albicans. It occurs when the host presents an imbalance in the immune system and Candida spp., normally found in human flora, become able to develop the infection [1]. This disease is very common in HIV patients, and in all individuals that present immunossupression, such as patients treated with chemotherapy. Considering this scenario, the development of new medicines to treat oral candidiasis is mandatory. Aims: The aim of this study was to evaluate citotoxicity, morphology and quantify the adhesion rates of C. albicans to biotherapic-treated Ma104 cells. Methodology: The biotherapic was prepared following the Roberto Costa technique and Brazilian Homeopathic Pharmacopeia protocol [2]. Briefly, biotherapic 1X was prepared with 1 mL of aqueous solution containing 108 yeasts of living Candida albicans plus 9 ml of sterile distilled water. This solution was submmited to 100 mechanical succussions. Biotherapic 2X was obtained after addition of 1 ml of 1X solution in 9 ml of sterile distilled water and it was also submitted to 100 mechanical succussions. This procedure was repeated until biotherapic 30X was obtained. As a control, sterile dynamized water (30X) was used. The inhibition of fungal growth induced by biotherapic was evaluated by MTT method after 24 hours of treatment. The morphological aspects of Ma104-biotherapic-treated cells were analyzed by Giemsa staining after 5, 10 and 60 days, and compared with control groups (water 30X and untreated cells). Additionally, Ma104 cells were treated during 5 and 30 days with biotherapic in parallel with respective controls, and the index adhesion of yeast cells was quantified. Results: The biotherapic was not able to reduce the viability of treated C. albicans when compared with controls. On the other hand, Ma104 treated cells presented important morphological alterations after 60 days, such as: cytoplasmic vacuoles, halos around the nucleolus and elongation of the plasmatic membrane. These changes were not observed in,untreated cells nor in ones treated with water 30X. The adhesion index to Ma104 cells was reduced around 27% after 5 and 30 days of treatment when compared to controls. Conclusion: These results showed that the biotherapic did not present any citotoxicity, but was able to modify the morphological aspects of Ma-104 cells. Additionally, the interaction between host cells and ethilogic agent is directly influenced by biotherapic treatment, suggesting a promising antifungal potential of this medicine.
    No preview · Article · Jan 2011 · International Journal of High Dilution Research
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    ABSTRACT: Background: The influenza virus has been responsible for contagious respiratory diseases with high mortality rates [1]. Some drugs have been used to treat human influenza. However, these drugs cause many common side effects and induce the appearance of resistant viral strains [2]. The impact caused by the influenza virus has motivated the development of new approaches for the prevention and control of influenza [3]. Therefore, a new homeopathic medicine was developed using, as a starting point, the infectious influenza virus [4]. This belongs to a group called living nosodes [5]. However, its mutagenic and genotoxic potentials, especially when used in low dilutions, has not yet been evaluated and it is important because this biotherapic is prepared from living microorganisms. Different methods can be used to detect mutagenic and genotoxicic effects. Aims: This study aims to evaluate the genotoxic and mutagenic potentials of influenza A living nosode at different homeopathic potencies.
    Full-text · Article · Jan 2011 · International Journal of High Dilution Research
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    ABSTRACT: Inhibition of tumor growth induced by treatment with direct electric current (DC) has been reported in several models. One of the mechanisms responsible for the antitumoral activity of DC is the generation of oxidative species, known as chloramines. With the aim of increasing chloramine production in the electrolytic medium and optimizing the antitumoral effects of DC, poly(ɛ-caprolactone) (PCL) nanoparticles (NPs) loaded with the amino acid tyrosine were obtained. The physical-chemical characterization showed that the NPs presented size in nanometric range and monomodal distribution. A slightly negative electrokinetic potential was also found in both blank NPs and L-tyrosine-loaded PCL NPs. The yield of the loading process was approximately 50%. Within 3 h of dissolution assay, a burst release of about 80% L-tyrosine was obtained. The in vitro cytotoxicity of DC was significantly increased when associated with L-tyrosine-loaded NPs, using a murine multidrug-resistant melanoma cell line model. This study showed that the use of the combination of nanotechnology and DC has a promising antineoplastic potential and opens a new perspective in cancer therapy.
    Full-text · Article · Nov 2010 · International Journal of Nanomedicine
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    ABSTRACT:   There are several kinds of oral soft tissue lesions that are common manifestations observed in human immunodeficiency virus (HIV)-infected children; for example, linear gingival erythema (LGE) that is a distinctive fiery red band along the margin of the gingivae. The etiology and pathogenesis of LGE are questionable, but a candidal origin has been suggested. Proteases are key virulence attributes produced by a variety of pathogenic fungi, including Candida. The objective of the present study is to identify the protease production in Candida species including, C. albicans (n=5), C. dubliniensis (n=1) and C. tropicalis (n=1), isolated directly from typical LGE lesions observed in six HIV-positive children, and also to test the effect of a serine protease inhibitor on the interaction of Candida spp. and epithelial cells in vitro. The ability of Candida strains to release proteases in the culture supernatant fluids was visualized by gelatin-SDS-PAGE. Gel strips containing 30-fold concentrated supernatant (1.5×10(8) yeasts) were incubated at 37°C for 48 h in 50 mM sodium phosphate buffer, pH 5.5. The concentrated supernatants were also incubated with fibronectin, laminin, immunoglobulin G, bovine serum albumin and human serum albumin. The effect of serine protease inhibitor on the interaction of Candida spp. and epithelial cells (MA 104) was measured after pre-treatment of fungi with the inhibitor (phenylmethylsulphonyl fluoride, PMSF). All the extracellular proteases were completely inhibited by PMSF, identifying these activities as serine-type proteases. Interestingly, a common 62-kDa serine protease was observed in all Candida strains. The culture supernatants, rich in serine protease activities, cleaved several soluble proteinaceous substrates. Additionally, we demonstrated that pre-treatment of C. albicans, C. dubliniensis and C. tropicalis with PMSF diminished the interaction with epithelial cells. Collectively, our results show that Candida spp. isolated from LGE lesions produced and secreted serine proteases and these enzymes may be involved in the initial colonization events.
    Full-text · Article · Nov 2010 · Journal of Oral Pathology and Medicine
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    ABSTRACT: This study describes the expression of acidic ectophosphatase activity on twenty isolates of C. albicans from oral cavities of HIV-infected children (HIV+) and compares them with fifteen isolates from HIV-negative children (HIV-), as well as the fungal adhesion to epithelial cells and medical records. The activities were measured in intact cells grown in BHI medium for 48 h at 37 degrees C. Phosphatase activity was assayed at pH 5.5 using 4-methylumbelliferyl phosphate. Yeast adhesion was measured using the MA 104 epithelial cell line. Mean values of ectophosphatase activity were 610.27 +/- 166.36 and 241.25 +/- 78.96 picomoles 4-methylumbelliferone/h/10(7) cells for HIV+ and HIV- group, respectively (P = 0.049). No correlation between C. albicans enzyme activity from HIV children with viral load and CD4 percentual was observed. Yeasts with high enzyme activity, isolated from HIV+ children showed greater adherence than yeasts with basal levels of ectophosphatases from HIV- (Spearman correlation, r = 0.8). Surface phosphatase activity was apparently involved in the adhesion to host cells, as the enhanced attachment of C. albicans to host epithelial cells was reversed by pretreatment of yeast with sodium orthovanadate (1 mM), an acid phosphatase inhibitor. These results show that C. albicans from HIV+ has an ectophosphatase activity significantly higher than the other isolates. Yeasts expressing higher levels of surface phosphatase activity showed greater adhesion to epithelial cells. So, the activity of acidic surface phosphatases on these cells may contribute to the early mechanisms required for disease establishment.
    Full-text · Article · Mar 2010 · Oral Diseases
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    ABSTRACT: Background: although Hahnemann described the fifty-milesimal (LM) method in the 6th edition of the Organon of the Medical Art, very little research has been carried out on the physical chemical properties of these homeopathic preparations. Furthermore, there is still no evidence allowing for the correlation between the alleged physical chemical properties and the biological effects of high dilutions. Aims: to evaluate physical chemical characteristics of LM preparations including electrical conductivity, pH and refraction index, and their effect on biological experimental models. Materials and methods: preparations tested for physical chemical analysis were dilutions 1 lm to 10 lm of Euphorbia tirucalli L. prepared from the latex and the juice of the plant. To rule the seasonal characteristics of this plant, 2 different populations were used, one collected in June 2007 and the other in May 2008. Furthermore, the cytotoxic effect of Euphorbia tirucalli 5 lm was tested on human breast cancer cells (MCF7) through MTT assay. Some differences among the two collections were observed. However, any clear correlation could be observed between physical chemical properties and biological activity.
    Full-text · Article · Jan 2010
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    ABSTRACT: Cadmium is an important toxic environmental heavy metal. Several studies have demonstrated that a major site of cadmium toxicity in humans and in other animals is the proximal tubule of the kidney. A well established model for nefrotoxicity is the use of in vitro technique with proximal tubule epithelial cell lines, as LLC-PK1. Herein, we have the intention to study the possible protective effect of highdiluted CdCl2 solutions. In a blinding way, LLC-PK1 cells were pre-treated with highdiluted cadmium chloride in the potencies 10 cH, 15 cH and 20cH. After 4 days, these cells have received CdCl2 in a pre-determined toxic concentration. The cell viability was assessed by MTT assay. We have identified a protective effect of two CdCl2 highdiluted solutions, 10 cH and 20 cH, when cells were intoxicated by sublethal CdCl2 concentration. The results indicate that probably the highdilutions have an expressive action on cells in sublethal intoxication.
    Full-text · Article · Jan 2010

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