Eraldo Batista - Academia.edu (original) (raw)

Papers by Eraldo Batista

Research paper thumbnail of parte I) Amado

Research paper thumbnail of Apical Periodontium Response to Enamel Matrix Derivative as an Intracanal Medication in Rat Immature Teeth with Pulp Necrosis: Radiographic and Histologic Findings

Journal of Endodontics, 2012

Introduction: The aim of this study was to evaluate the enamel matrix derivative (EMD) biomateria... more Introduction: The aim of this study was to evaluate the enamel matrix derivative (EMD) biomaterial in nonvital immature teeth. Methods: To arrest root development, pulpectomies were performed in the lower first molars of 36 4-week-old rats; the cavities were left exposed to the oral environment for 3 weeks. Then, chemical disinfection was performed, and triple antibiotic paste (TAP) or EMD was applied in the root canals. A control group did not receive any treatment. Radiographic and histological data were evaluated after 3 and 6 weeks. Results: At 3 weeks, TAP promoted a milder inflammatory response and increased root lengths compared with the control group. At 6 weeks, root development and reduced periapical lesions could be observed in both test groups, mainly because of the deposition of a cementum-like tissue. EMD promoted narrower canals compared with TAP (P < .05). Conclusions: EMD deserves attention as a potential tool in the treatment of nonvital immature teeth. The ingrowth of cementum-like tissues into canal spaces favored dental wall thickness and may contribute to tooth resistance and support. (J Endod 2012;38:449-453)

Research paper thumbnail of Response to Intracanal Medication in Immature Teeth with Pulp Necrosis: An Experimental Model in Rat Molars

Journal of Endodontics, 2011

Introduction: The present study aimed at developing an experimental model in rat molars for evalu... more Introduction: The present study aimed at developing an experimental model in rat molars for evaluating treatment strategies in necrotic immature teeth. Methods: To define the periods to be adopted in the experimental procedures and to confirm induction of periapical lesions and interruption of root embryogenesis, the left lower first molars of 4-weeks-old Wistar rats underwent pulpectomy and were left open to the oral environment. Comparisons with the right lower first molars (vital teeth) were performed in animals with ages of 7, 10, 13, and 16 weeks. In another group of animals the teeth were left open for 3 weeks, and then interventions for disinfection including the use of an antibiotic paste were carried out. Root formation was then assessed after 3 and 6 weeks on the basis of radiographic and histologic evaluation. Results: Vital teeth showed increase of root length and hard tissue thickness throughout the experimental periods. On the other hand, induction of necrosis arrested root formation. Teeth subjected to disinfection with sodium hypochlorite associated with the triple antibiotic paste showed significant reduction of periapical lesions, gain in root length, and increased wall thickness compared with the control (P < .05). Conclusions: The root canal disinfection protocol used was able to reduce periapical lesion size and improve root development. The experimental model presented should contribute to studies that aim at improving therapeutic strategies for necrotic immature teeth by using a rat model. (J Endod 2011;37:1069-1073

Research paper thumbnail of Moesin-induced signaling in response to lipopolysaccharide in macrophages: Moesin-mediated signaling in macrophages

Journal of Periodontal Research, 2010

Zawawi KH, Kantarci A, Schulze-Späte U, Fujita T, Batista EL Jr, Amar S, Van Dyke TE. Moesin-indu... more Zawawi KH, Kantarci A, Schulze-Späte U, Fujita T, Batista EL Jr, Amar S, Van Dyke TE. Moesin-induced signaling in response to lipopolysaccharide in macrophages. J Periodont Res 2010; 45: 589–601.©2010 John Wiley & Sons A/SBackground and Objective: Many physiological and pathophysiological conditions are attributable in part to cytoskeletal regulation of cellular responses to signals. Moesin (membrane-organizing extension spike protein), an ERM (ezrin, radixin and moesin) family member, is involved in lipopolysaccharide (LPS)-mediated events in mononuclear phagocytes; however, its role in signaling is not fully understood. The aim of this study was to investigate the LPS-induced moesin signaling pathways in macrophages.Material and Methods: Macrophages were stimulated with 500 ng/mL LPS in macrophage serum-free medium. For blocking experiments, cells were pre-incubated with anti-moesin antibody. Moesin total protein and phosphorylation were studied with western blotting. Moesin mRNA was assessed using quantitative real-time PCR. To explore binding of moesin to LPS, native polyacrylamide gel electrophoresis (PAGE) gel shift assay was performed. Moesin immunoprecipitation with CD14, MD-2 and Toll-like receptor 4 (TLR4) and co-immunoprecipitation of MyD88–interleukin-1 receptor-associated kinase (IRAK) and IRAK–tumor necrosis factor receptor-activated factor 6 (TRAF6) were analyzed. Phosphorylation of IRAK and activities of MAPK, nuclear factor κB (NF-κB) and IκBα were studied. Tumor necrosis factor α, interleukin-1β and interferon β were measured by ELISA.Results: Moesin was identified as part of a protein cluster that facilitates LPS recognition and results in the expression of proinflammatory cytokines. Lipopolysaccharide stimulates moesin expression and phosphorylation by binding directly to the moesin carboxyl-terminus. Moesin is temporally associated with TLR4 and MD-2 after LPS stimulation, while CD14 is continuously bound to moesin. Lipopolysaccharide-induced signaling is transferred downstream to p38, p44/42 MAPK and NF-κB activation. Blockage of moesin function interrupts the LPS response through an inhibition of MyD88, IRAK and TRAF6, negatively affecting subsequent activation of the MAP kinases (p38 and ERK), NF-κB activation and translocation to the nucleus.Conclusion: These results suggest an important role for moesin in the innate immune response and TLR4-mediated pattern recognition in periodontal disease.

Research paper thumbnail of The role of CXCR2 chemokine receptors in the oral squamous cell carcinoma

Investigational New Drugs, 2000

This study evaluated the relevance of CXCR2 chemokine receptors in oral squamous cell carcinoma, ... more This study evaluated the relevance of CXCR2 chemokine receptors in oral squamous cell carcinoma, by means of in vitro and in vivo approaches. The in vitro incubation of the selective and non-peptide CXCR2 receptor antagonist N-(2-hydroxy-4-nitrophenyl)-N9-(2-bromophenyl) Urea (SB225002; 25 to 800 nM) produced a time- and concentration-dependent inhibition of SCC158 (rat) and HN30 (human) cell lines viability. Conversely, this antagonist did not significantly affect the viability of the immortalized keratinocyte lineage, HaCaT. Additionally, the incubation of human IL-8 and rat CINC-1 CXCR2 agonists produced a concentration-related increase on HN30 and SCC158 proliferation. The submucosal injection of SCC158 cells (5 × 106 cells) into the tongue of Fischer 344 rats induced tumor development, which displayed typical clinical features. Immunohistochemical analysis of rat tongue biopsies revealed a marked increase of CXCR2 receptor immunoreactivity, which was accompanied by augumented expression of VEGF and caspase-3. Our data suggests an important role for CXCR2 receptors in oral squamous cell carcinoma.

Research paper thumbnail of Response to Intracanal Medication in Immature Teeth with Pulp Necrosis: An Experimental Model in Rat Molars

Journal of Endodontics, 2011

The present study aimed at developing an experimental model in rat molars for evaluating treatmen... more The present study aimed at developing an experimental model in rat molars for evaluating treatment strategies in necrotic immature teeth.To define the periods to be adopted in the experimental procedures and to confirm induction of periapical lesions and interruption of root embryogenesis, the left lower first molars of 4-weeks-old Wistar rats underwent pulpectomy and were left open to the oral environment. Comparisons with the right lower first molars (vital teeth) were performed in animals with ages of 7, 10, 13, and 16 weeks. In another group of animals the teeth were left open for 3 weeks, and then interventions for disinfection including the use of an antibiotic paste were carried out. Root formation was then assessed after 3 and 6 weeks on the basis of radiographic and histologic evaluation.Vital teeth showed increase of root length and hard tissue thickness throughout the experimental periods. On the other hand, induction of necrosis arrested root formation. Teeth subjected to disinfection with sodium hypochlorite associated with the triple antibiotic paste showed significant reduction of periapical lesions, gain in root length, and increased wall thickness compared with the control (P < .05).The root canal disinfection protocol used was able to reduce periapical lesion size and improve root development. The experimental model presented should contribute to studies that aim at improving therapeutic strategies for necrotic immature teeth by using a rat model.

Research paper thumbnail of Increased Tissue Factor Expression and Poor Nephroblastoma Prognosis

Journal of Urology, 2009

There is potential interaction between malignant cell growth and the coagulation pathway. Recent ... more There is potential interaction between malignant cell growth and the coagulation pathway. Recent studies suggest that tissue factor, a primary initiator of the extrinsic coagulation pathway, is expressed in various solid tumors in association with increased angiogenesis. To our knowledge we report for the first time the detection of tissue factor expression by immunohistochemistry in Wilms tumors and its correlation with clinical outcomes.Tissue factor expression detected by immunohistochemistry was assessed in 41 formalin fixed, paraffin embedded Wilms tumor cases treated at university hospitals. We correlated findings with tumor recurrence and cancer specific survival.Positive immunohistochemistry detection of tissue factor was observed in 88.3% of the tumors analyzed. Tissue factor on immunohistochemistry was associated with tumor recurrence and survival (p = 0.01 and 0.02, respectively). Increased immunohistochemical detection of tissue factor was the most important risk factor for recurrence and mortality in our population on bivariate and multivariate analysis.Tissue factor is a promising research subject as a prognostic factor for Wilms tumor. More studies are needed to clarify the mechanisms by which tissue factor affects cancer progression and outcome, and its potential role as a therapeutic target.

Research paper thumbnail of Sequential Chemotactic and Phagocytic Activation of Human Polymorphonuclear Neutrophils

Infection and Immunity, 2007

Human polymorphonuclear neutrophils (PMN) chemotax to a foreign entity. When the chemoattractants... more Human polymorphonuclear neutrophils (PMN) chemotax to a foreign entity. When the chemoattractants' origins are reached, specific receptors bind to the invader's surface, initiating phagocytosis, phagosome formation, and fusion with granule membranes, generating the bactericidal oxidative burst, and releasing lytic enzymes, specific peptides, and proteins. We explored the initial signaling involved in these functions by observing naïve, unprimed PMN in suspension using fluorescent indicators of cytoplasmic signals (⌬[Ca 2؉ ] i and ⌬pH i ) and of bactericidal entities (oxidative species and elastase) exposed to N-formyl-methionyl-leucylphenylalanine (fMLP) and/or multivalent immune complexes (IC). fMLP and IC each initiate a rapid transient rise in [Ca 2؉ ] i , mostly from intracellular stores, simultaneously with a drop in pH i ; these are followed by a drop in [Ca 2؉ ] i and a rise in pH i , with the latter being due to a Na ؉ /H ؉ antiport. The impact of a second stimulation depends on the order in which stimuli are applied, on their dose, and on their nature. Provided that [Ca 2؉ ] i is restored, 10 ؊7 M fMLP, previously shown to elicit maximal ⌬[Ca 2؉ ] i but no bactericidal functions, did not prevent the cells' responses with ⌬[Ca 2؉ ] i to a subsequent high dose of fMLP or IC; conversely, cells first exposed to 120 g/ml IC, previously shown to elicit maximal ⌬[Ca 2؉ ] i and bactericidal functions, exhibited no subsequent ⌬[Ca 2؉ ] i or ⌬pH i to either stimulus. While exposure to 10 ؊7 M fMLP, which saturates the PMN high-affinity receptor, did not elicit bactericidal release from these naïve unprimed PMN in suspension, 10 ؊5 M fMLP did, presumably via the low-affinity receptor, using a different Ca 2؉ source. on February 9, 2015 by guest http://iai.asm.org/ Downloaded from FIG. 4. Sequential stimulation of PMN by saturating doses (120 g/ml) of IC followed 5 min later by 10 Ϫ7 M fMLP. (a) Control; (b) 5 mM EGTA added 15 s before IC injection; (c) 5 mM EGTA added before fMLP injection; (d) release of elastase (shown as F 460 ) and of oxidative products (shown as F 530 ). Each figure is representative of five independent experiments. VOL. 75, 2007 SEQUENTIAL CHEMOTACTIC AND IC ACTION ON PMN 3995 on February 9, 2015 by guest

Research paper thumbnail of Identification of mRNAs for the various diacylglycerol kinase isoforms in neutrophils from patients with localized aggressive periodontitis

Journal of Periodontal Research, 2003

Background: Diacylglycerol kinase (DGK) metabolizes diacylglycerol (DAG), an endogenous activato... more Background: Diacylglycerol kinase (DGK) metabolizes diacylglycerol (DAG), an endogenous activator of protein kinase C, to phosphatidic acid. We have previously reported increased DAG in neutrophils from patients with localized aggressive periodontitis (LAP) associated with reduced DGK activity. This reduction could be related to a mutation, post-translational modification, differential expression, or lack of expression of a particular isoform(s).Objective: The aim of this study was to identify the mRNAs for DGK isoforms in normal and LAP neutrophils.Methods: The α-, γ-, and δ-isoforms of DGK were identified by polymerase chain reaction (PCR) using specific oligonucleotide primers for each isoform. The PCR products were isolated and sequenced for comparison to published sequences to confirm the validity of the PCR reaction. Total RNA was isolated from LAP and normal neutrophils, and northern blotting and semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was used to examine the level of mRNA for each isoform.Results: No major differences were found in the isoform pattern between resting normal and LAP neutrophils. However, the levels of mRNA for the α- and γ-isoforms of DGK were increased in normal neutrophils while slightly decreased in LAP cells upon stimulation with N-formyl-methionyl-leucyl-phenylalanine (fMLP).Conclusion: These data suggest that alterations in the mRNAs for the various isoforms of DGK during cell stimulation and the involvement of DGK that is expressed in multiple forms are subject to a variety of regulatory/control mechanisms and these mechanisms may explain the role of the ‘primed’ neutrophil phenotype associated with LAP.

Research paper thumbnail of Differentiation of HL60 cells to granulocytes involves regulation of select diacylglycerol kinases (DGKs)

Journal of Cellular Biochemistry, 2005

AbstractDiacylglycerol Kinases (DGKs) are a family of enzymes that regulate the levels of differe... more AbstractDiacylglycerol Kinases (DGKs) are a family of enzymes that regulate the levels of different pools of diacylglycerol (DAG), affecting DAG-mediated signal transduction. Since DAG is known to play several important regulatory roles in granulocyte physiology, we investigated the expression pattern of DGK isoforms throughout differentiation of HL-60 cells to granulocytes. HL-60 cells were incubated with 1.25% dimethyl-sulfoxide (DMSO) to initiate differentiation and total RNA isolated at different time points. DGK expression was assessed through Northern blot, end-point PCR, and real-time PCR. The non-selective inhibitors R59022 and R59949 were used to block DGK at different time points throughout differentiation. CD11b and GPI-80, reactive oxygen species (ROS) generation, changes in the cell cycle, and apoptosis were used as markers of differentiation. Of the nine isoforms of DGK evaluated (α, δ, ε, γ, ζ, β, θ, ι, η), only five (α, δ, ε, γ, and ζ) were expressed in HL-60 cells. DGKα was virtually absent in non-differentiated cells, but was markedly upregulated throughout differentiation. The other isoforms (δ, ε, γ, and ζ) were expressed in undifferentiated HL-60 cells but were substantially decreased throughout differentiation. Non-selective blocking of DGK with R59022 and R59949 led to acceleration of differentiation, reducing the time necessary to observe upregulation of CD11b, GPI-80 and generation of ROS by 50%. Likewise, the cell cycle was disrupted when DGK isoforms were inhibited. These results provide evidence that DGK levels are dynamically regulated throughout differentiation and that expression of DGKs play an important regulatory function during the differentiation of neutrophils. © 2004 Wiley-Liss, Inc.

Research paper thumbnail of Protective Effects of Resveratrol on Hepatotoxicity Induced by Isoniazid and Rifampicin via SIRT1 Modulation

Journal of Natural Products, 2014

Acute liver injury was induced in male BALB/c mice by coadministering isoniazid and rifampicin. I... more Acute liver injury was induced in male BALB/c mice by coadministering isoniazid and rifampicin. In this work, the effects of resveratrol (1) were investigated in the hepatotoxicity caused by isoniazid−rifampicin in mice. Compound 1 was administered 30 min prior to isoniazid−rifampicin. Serum biochemical tests, liver histopathological examination, oxidative stress, myeloperoxidase activity, cytokine production (TNF-α, IL-12p70, and IL-10), and mRNA expression of SIRT1−7 and PPAR-γ/ PGC1-α were evaluated. The administration of 1 significantly decreased aspartate transaminase and alanine aminotransferase levels, myeloperoxidase activity, and cytokine levels. Furthermore, 1 reverted the decrease of catalase and glutathione activities and ameliorated the histopathological alterations associated with antituberculosis drugs. Modulation of SIRT1 and PPAR-γ/ PGC1-α expression is likely involved in the protective effects of 1. The results presented herein show that 1 was able to largely prevent the hepatotoxicity induced by isoniazid and rifampicin in mice, mainly by modulating SIRT1 mRNA expression.

Research paper thumbnail of Porphyromonas gingivalis infection accelerates the progression of atherosclerosis in a heterozygous apolipoprotein E-deficient murine model

Circulation, Jan 19, 2002

Background-Current epidemiological data suggest that dental infections affecting tooth-supporting... more Background-Current epidemiological data suggest that dental infections affecting tooth-supporting tissues (periodontitis) can disseminate into the systemic circulation and thereby contribute to atherosclerosis progression. To test this hypothesis, we investigated the effect of repeated systemic inoculations with Porphyromonas gingivalis (Pg), a putative periodontal pathogen, on the progression of atherosclerosis in heterozygous apolipoprotein E-deficient (ApoE ϩ/Ϫ ) mice. Methods and Results-Ten-week-old, male ApoE ϩ/Ϫ mice fed either a high-fat diet or regular chow were inoculated intravenously with live Pg (10 7 CFU) or vehicle once per week for 10, 14, or 24 consecutive weeks. Histomorphometry of plaque cross-sectional area in the proximal aortas, en face measurement of plaque area over the aortic trees, Pg 16S ribosomal DNA amplification with polymerase chain reaction, ELISA for systemic proinflammatory mediators, and immunolocalization of macrophages in the proximal aorta were performed. Atherosclerotic lesions of the proximal aortas and aortic trees were more advanced in Pg-challenged animals than in vehicle control animals and occurred earlier (at 10 weeks) when no lesions were apparent in control animals. At 24 weeks after inoculation, proximal aortic lesion size quantified by histomorphometry was 9-fold greater in chow-fed mice inoculated with Pg than in noninoculated mice (PϽ0.001) and was 2-fold greater in Pg-inoculated versus noninoculated high-fat diet-fed mice (PϽ0.001); all atherosclerotic lesions were macrophage-rich. Pg ribosomal DNA was found in the aortas, livers, and hearts 24 weeks after inoculation.

Research paper thumbnail of Priming of neutrophil oxidative burst in diabetes requires preassembly of the NADPH oxidase

Journal of Leukocyte Biology - J LEUKOCYTE BIOL, 2008

Research paper thumbnail of Analysis of select members of the E26 (ETS) transcription factors family in colorectal cancer

Virchows Archiv, 2011

The E-twenty-six (ETS) family of transcription factors is known to act as positive or negative re... more The E-twenty-six (ETS) family of transcription factors is known to act as positive or negative regulators of the expression of genes that are involved in diverse biological processes, including those that control cellular proliferation, differentiation, hematopoiesis, apoptosis, metastasis, tissue remodeling, and angiogenesis. Identification of target gene promoters of normal and oncogenic transcription factors provides new insights into the regulation of genes that are involved in the control of normal cell growth and differentiation. The aim of the present investigation was to analyze the differential expression of 11 ETS (ELF-3, ESE3, ETS1, ETV3, ETV4, ETV6, NERF, PDEF, PU1, Spi-B, and Spi-C) as potential markers for prognostic of colorectal cancer. A series of paired tissue biopsies consisting of a tumor and a non-affected control sample were harvested from 28 individuals suffering from diagnosed colorectal lesions. Total RNA was isolated from the samples, and after reverse transcription, differential expression of the select ETS was carried out through real-time polymerase chain reaction. Tumor staging as determined by histopathology was carried out to correlate the degree of tumor invasiveness with the expression of the ETS genes. The results demonstrated a different quantitative profile of expression in tumors and normal tissues. ETV4 was significantly upregulated with further increase in the event of lymph node involvement. PDEF and Spi-B presented downregulation, which was more significant when lymph node involvement was present. These findings were supported by immunohistochemistry of tumoral tissues. The results suggest that select ETS may serve as potential markers of colorectal cancer invasiveness and metastasis.

Research paper thumbnail of Real time PCR quantification of viable Mycobacteriumtuberculosis from sputum samples treated with propidium monoazide

Tuberculosis, 2014

s u m m a r y Diagnostic methods of TB, nowadays, are prone to delay in diagnosis, increased fals... more s u m m a r y Diagnostic methods of TB, nowadays, are prone to delay in diagnosis, increased false negative results and are not sensitive to many forms of paucibacillary disease. The aims of this study were to implement a quantitative nucleic acid-based diagnostic test for paucibacillary tuberculosis, enabling the identification and quantification of viable Mycobacterium tuberculosis bacilli by quantitative Real-Time PCR (qRT-PCR). The intergenic region of the single-copy inhA-mabA gene was chosen as the target region for design of primers and probes conjugated with fluorophores. The construction of synthetic DNA flanking the target region served as standards for absolute quantification of nucleic acids. Using the intercaling dye, propidium monoazide, we were able to discriminate between viable and dead cells of M. tuberculosis. The diagnosis method showed a broad sensitivity (96.1%) when only compared to samples of smear-positive sputum and ROC analyses shows that our approach performed well and yielded a specificity of 84.6% and a sensitivity of 84.6% when compared to M. tuberculosis colony-forming units counting.

Research paper thumbnail of Increased Tissue Factor Expression and Poor Nephroblastoma Prognosis

The Journal of Urology, 2009

NWTS ϭ National WT Study SIOP ϭ Societé Internationale d'Urologie Pédiatrique TF ϭ tissue factor ... more NWTS ϭ National WT Study SIOP ϭ Societé Internationale d'Urologie Pédiatrique TF ϭ tissue factor WT ϭ Wilms tumor Purpose: There is potential interaction between malignant cell growth and the coagulation pathway. Recent studies suggest that tissue factor, a primary initiator of the extrinsic coagulation pathway, is expressed in various solid tumors in association with increased angiogenesis. To our knowledge we report for the first time the detection of tissue factor expression by immunohistochemistry in Wilms tumors and its correlation with clinical outcomes. Material and Methods: Tissue factor expression detected by immunohistochemistry was assessed in 41 formalin fixed, paraffin embedded Wilms tumor cases treated at university hospitals. We correlated findings with tumor recurrence and cancer specific survival. Results: Positive immunohistochemistry detection of tissue factor was observed in 88.3% of the tumors analyzed. Tissue factor on immunohistochemistry was associated with tumor recurrence and survival (p ϭ 0.01 and 0.02, respectively). Increased immunohistochemical detection of tissue factor was the most important risk factor for recurrence and mortality in our population on bivariate and multivariate analysis. Conclusions: Tissue factor is a promising research subject as a prognostic factor for Wilms tumor. More studies are needed to clarify the mechanisms by which tissue factor affects cancer progression and outcome, and its potential role as a therapeutic target.

Research paper thumbnail of Use of Bovine-Derived Anorganic Bone Associated With Guided Tissue Regeneration in Intrabony Defects. Six-Month Evaluation at Re-Entry

Journal of Periodontology, 1999

Background: Different filling materials have been associated with guided tissue regeneration (GTR... more Background: Different filling materials have been associated with guided tissue regeneration (GTR) in order to improve its regenerative potential and predictability. Anorganic bovine bone (ABB) has demonstrated biocompatibility and osteoconductive properties; however, there are limited data regarding its performance in the treatment of intrabony defects. This investigation aimed to evaluate the clinical outcome of the association of anorganic bovine bone with cellulose membranes in intrabony defects after 6 months.

Research paper thumbnail of Manifestations of Insulin-Dependent Diabetes Mellitus in the Periodontium of Young Brazilian Patients. A 10-Year Follow-Up Study

Journal of Periodontology, 1997

Eight female and 3 male patients from a group of 30 patients studied 10 years earlier and who had... more Eight female and 3 male patients from a group of 30 patients studied 10 years earlier and who had received no periodontal treatment during this period, in order to evaluate the progression of untreated periodontal disease in patients with insulin-dependent diabetes mellitus, were evaluated in terms of plaque accumulation, gingival inflammation, probing depth, and alveolar bone loss. The total number of dental surfaces that presented clinically detectable plaque deposits increased significantly (29% to 43%; P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.01; chi 2 = 46.36). Site-specific comparisons for plaque index between studies showed a significant variation (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.01) in the upper arch only for palatal surfaces and in the lower arch for the buccal and lingual surfaces. The total dental surfaces with inflamed surrounding gingiva increased from 11% to 33% in this study (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.01; chi 2 = 175.78). Site-specific comparison for gingival index showed a significant variation for all upper surfaces, while such difference for the lower arch was significant only for the buccal and lingual surfaces. The arithmetic means for the probing depth for the upper buccal, upper palatal, lower buccal, and lower lingual surfaces increased significantly (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.01). The arithmetic means of alveolar bone loss also increased significantly for the upper posterior and lower regions (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.01) and for the upper anterior and lower anterior regions (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05). The correlation between age and probing depth was significant only for the upper palatal region (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.01). The correlation between age and bone loss was significant only for the upper posterior region (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05). The results of this follow-up study suggest that despite little variation in plaque accumulation, gingival inflammation, probing depth, and bone loss increased after a 10-year interval in patients who had received no periodontal treatment during this period.

Research paper thumbnail of Management of Soft Tissue Ridge Deformities With Acellular Dermal Matrix. Clinical Approach and Outcome After 6 Months of Treatment

Journal of Periodontology, 2001

Soft tissue ridge defects often hamper ideally shaped artificial crowns and are basically treated... more Soft tissue ridge defects often hamper ideally shaped artificial crowns and are basically treated using autogenous soft tissue grafts or alloplastic materials. These approaches present disadvantages such as the necessity of creating additional surgical fields to harvest the graft and the requirement of primary closure, which may reduce ridge height. This investigation evaluated the use of acellular dermal matrix (ADM) in the treatment of soft tissue ridge defects. Eight patients, non-smokers with non-contributory medical history, provided 18 sites corresponding to missing teeth in the anterior maxillary arch. The ideal horizontal gain (desired gain) was waxed up in study casts, which served as templates for construction of modified acrylic stents with orthodontic wires. These stents served as references for ideal horizontal gain and also as fixed reference points for further evaluation. The distance from the orthodontic wire to the buccal plate of the defect also represented its baseline horizontal component. Vertical variations were evaluated with another stent and, in this case, no desired gain was considered. After raising partial-thickness flaps, the ADM material was rehydrated and folded to fill the defect and reproduce the desired gain. Flaps were sutured with no tension, and part of the material was intentionally left exposed to avoid pressure on the incision line and prevent height loss. Patients used local and systemic antimicrobials, and the sutures were removed at 7 days. Evaluations were carried out at 30 days, and 3 and 6 months, and all sites healed uneventfully. Neither infection nor significant pain was reported by the patients, and the material was covered by tissue at about 21 days. Mean horizontal gain of 1.72 +/- 0.59 mm (58.5%) at 6 months and mean shrinkage of 1.22 +/- 0.46 mm (41.4%) were observed. There was a mean improvement in vertical gain of only 0.61 +/- 0. 77 mm, although 66. 7% of the treated sites showed a 1 to 2 mm gain. Clinically, the total gain in the subjects was very effective and matched the receptor tissues nicely. ADM may be a suitable material for the treatment of soft tissue ridge deformities due to its biocompatibility, color matching, and horizontal gain. Additional controlled, comparative trials are necessary to establish its advantages and potential compared to autogenous soft tissue techniques.

Research paper thumbnail of Guided Tissue Regeneration Associated With Bovine-Derived Anorganic Bone in Mandibular Class II Furcation Defects. 6-Month Results at Re-Entry

Journal of Periodontology, 2000

The use of graft materials with guided tissue regeneration (GTR) in Class II furcation defects is... more The use of graft materials with guided tissue regeneration (GTR) in Class II furcation defects is aimed at improving the outcome of the regenerative technique. In this regard, however, there are a limited number of studies discussing the results obtained when GTR and graft materials are used in the treatment of Class II furcation defects. Furthermore, most studies employ either allogeneic or autogenous materials. The present trial sought to determine whether the use of a bovine-derived anorganic bone (ABB) in conjunction with GTR influenced the outcome of mandibular Class II furcation treatment. This study included 14 patients who provided 15 pairs of similar periodontal defects. Each defect was randomly assigned to treatment with either a cellulose membrane in combination with bovine-derived anorganic bone (GTR+ABB) or membrane alone (GTR). Following basic therapy, baseline measurements were recorded including probing depth (PD), clinical attachment level (CAL), and gingival margin position (GMP). Hard tissue measurements were performed during surgery to determine alveolar crestal height (CEJ-AC), and vertical (VDD) and horizontal defect depth (HDD). Membranes remained in position for at least 4 weeks. After 6 months, all sites were re-entered and soft and hard tissue measurements were recorded. Both surgical procedures resulted in statistically significant probing depth reduction and gain in clinical attachment levels, with no significant difference between groups. Gingival recession was more pronounced in the GTR+ABB group (0.87 +/- 0.83 mm), but not statistically different from the GTR group (0.46 +/- 1.19 mm). Vertical defect resolution was significant in both groups (GTR: 1.60 +/- 1.50 mm; GTR+ABB: 1.80 +/- 2.11 mm), without differences between groups. Only horizontal furcation resolution (GTR: 2.47 +/- 0.99 mm; GTR+ABB: 3.27 +/- 1.39 mm) was significantly different between groups (P &amp;amp;lt;0.05). The use of ABB with GTR techniques improved horizontal defect resolution in mandibular Class II furcation defects, but did not yield superior results regarding soft tissue changes when compared to sites treated with GTR alone. Evaluation of a larger sample could indicate differences and advantages between the evaluated approaches and confirm the real necessity of associating filling materials with GTR.

Research paper thumbnail of parte I) Amado

Research paper thumbnail of Apical Periodontium Response to Enamel Matrix Derivative as an Intracanal Medication in Rat Immature Teeth with Pulp Necrosis: Radiographic and Histologic Findings

Journal of Endodontics, 2012

Introduction: The aim of this study was to evaluate the enamel matrix derivative (EMD) biomateria... more Introduction: The aim of this study was to evaluate the enamel matrix derivative (EMD) biomaterial in nonvital immature teeth. Methods: To arrest root development, pulpectomies were performed in the lower first molars of 36 4-week-old rats; the cavities were left exposed to the oral environment for 3 weeks. Then, chemical disinfection was performed, and triple antibiotic paste (TAP) or EMD was applied in the root canals. A control group did not receive any treatment. Radiographic and histological data were evaluated after 3 and 6 weeks. Results: At 3 weeks, TAP promoted a milder inflammatory response and increased root lengths compared with the control group. At 6 weeks, root development and reduced periapical lesions could be observed in both test groups, mainly because of the deposition of a cementum-like tissue. EMD promoted narrower canals compared with TAP (P < .05). Conclusions: EMD deserves attention as a potential tool in the treatment of nonvital immature teeth. The ingrowth of cementum-like tissues into canal spaces favored dental wall thickness and may contribute to tooth resistance and support. (J Endod 2012;38:449-453)

Research paper thumbnail of Response to Intracanal Medication in Immature Teeth with Pulp Necrosis: An Experimental Model in Rat Molars

Journal of Endodontics, 2011

Introduction: The present study aimed at developing an experimental model in rat molars for evalu... more Introduction: The present study aimed at developing an experimental model in rat molars for evaluating treatment strategies in necrotic immature teeth. Methods: To define the periods to be adopted in the experimental procedures and to confirm induction of periapical lesions and interruption of root embryogenesis, the left lower first molars of 4-weeks-old Wistar rats underwent pulpectomy and were left open to the oral environment. Comparisons with the right lower first molars (vital teeth) were performed in animals with ages of 7, 10, 13, and 16 weeks. In another group of animals the teeth were left open for 3 weeks, and then interventions for disinfection including the use of an antibiotic paste were carried out. Root formation was then assessed after 3 and 6 weeks on the basis of radiographic and histologic evaluation. Results: Vital teeth showed increase of root length and hard tissue thickness throughout the experimental periods. On the other hand, induction of necrosis arrested root formation. Teeth subjected to disinfection with sodium hypochlorite associated with the triple antibiotic paste showed significant reduction of periapical lesions, gain in root length, and increased wall thickness compared with the control (P < .05). Conclusions: The root canal disinfection protocol used was able to reduce periapical lesion size and improve root development. The experimental model presented should contribute to studies that aim at improving therapeutic strategies for necrotic immature teeth by using a rat model. (J Endod 2011;37:1069-1073

Research paper thumbnail of Moesin-induced signaling in response to lipopolysaccharide in macrophages: Moesin-mediated signaling in macrophages

Journal of Periodontal Research, 2010

Zawawi KH, Kantarci A, Schulze-Späte U, Fujita T, Batista EL Jr, Amar S, Van Dyke TE. Moesin-indu... more Zawawi KH, Kantarci A, Schulze-Späte U, Fujita T, Batista EL Jr, Amar S, Van Dyke TE. Moesin-induced signaling in response to lipopolysaccharide in macrophages. J Periodont Res 2010; 45: 589–601.©2010 John Wiley & Sons A/SBackground and Objective: Many physiological and pathophysiological conditions are attributable in part to cytoskeletal regulation of cellular responses to signals. Moesin (membrane-organizing extension spike protein), an ERM (ezrin, radixin and moesin) family member, is involved in lipopolysaccharide (LPS)-mediated events in mononuclear phagocytes; however, its role in signaling is not fully understood. The aim of this study was to investigate the LPS-induced moesin signaling pathways in macrophages.Material and Methods: Macrophages were stimulated with 500 ng/mL LPS in macrophage serum-free medium. For blocking experiments, cells were pre-incubated with anti-moesin antibody. Moesin total protein and phosphorylation were studied with western blotting. Moesin mRNA was assessed using quantitative real-time PCR. To explore binding of moesin to LPS, native polyacrylamide gel electrophoresis (PAGE) gel shift assay was performed. Moesin immunoprecipitation with CD14, MD-2 and Toll-like receptor 4 (TLR4) and co-immunoprecipitation of MyD88–interleukin-1 receptor-associated kinase (IRAK) and IRAK–tumor necrosis factor receptor-activated factor 6 (TRAF6) were analyzed. Phosphorylation of IRAK and activities of MAPK, nuclear factor κB (NF-κB) and IκBα were studied. Tumor necrosis factor α, interleukin-1β and interferon β were measured by ELISA.Results: Moesin was identified as part of a protein cluster that facilitates LPS recognition and results in the expression of proinflammatory cytokines. Lipopolysaccharide stimulates moesin expression and phosphorylation by binding directly to the moesin carboxyl-terminus. Moesin is temporally associated with TLR4 and MD-2 after LPS stimulation, while CD14 is continuously bound to moesin. Lipopolysaccharide-induced signaling is transferred downstream to p38, p44/42 MAPK and NF-κB activation. Blockage of moesin function interrupts the LPS response through an inhibition of MyD88, IRAK and TRAF6, negatively affecting subsequent activation of the MAP kinases (p38 and ERK), NF-κB activation and translocation to the nucleus.Conclusion: These results suggest an important role for moesin in the innate immune response and TLR4-mediated pattern recognition in periodontal disease.

Research paper thumbnail of The role of CXCR2 chemokine receptors in the oral squamous cell carcinoma

Investigational New Drugs, 2000

This study evaluated the relevance of CXCR2 chemokine receptors in oral squamous cell carcinoma, ... more This study evaluated the relevance of CXCR2 chemokine receptors in oral squamous cell carcinoma, by means of in vitro and in vivo approaches. The in vitro incubation of the selective and non-peptide CXCR2 receptor antagonist N-(2-hydroxy-4-nitrophenyl)-N9-(2-bromophenyl) Urea (SB225002; 25 to 800 nM) produced a time- and concentration-dependent inhibition of SCC158 (rat) and HN30 (human) cell lines viability. Conversely, this antagonist did not significantly affect the viability of the immortalized keratinocyte lineage, HaCaT. Additionally, the incubation of human IL-8 and rat CINC-1 CXCR2 agonists produced a concentration-related increase on HN30 and SCC158 proliferation. The submucosal injection of SCC158 cells (5 × 106 cells) into the tongue of Fischer 344 rats induced tumor development, which displayed typical clinical features. Immunohistochemical analysis of rat tongue biopsies revealed a marked increase of CXCR2 receptor immunoreactivity, which was accompanied by augumented expression of VEGF and caspase-3. Our data suggests an important role for CXCR2 receptors in oral squamous cell carcinoma.

Research paper thumbnail of Response to Intracanal Medication in Immature Teeth with Pulp Necrosis: An Experimental Model in Rat Molars

Journal of Endodontics, 2011

The present study aimed at developing an experimental model in rat molars for evaluating treatmen... more The present study aimed at developing an experimental model in rat molars for evaluating treatment strategies in necrotic immature teeth.To define the periods to be adopted in the experimental procedures and to confirm induction of periapical lesions and interruption of root embryogenesis, the left lower first molars of 4-weeks-old Wistar rats underwent pulpectomy and were left open to the oral environment. Comparisons with the right lower first molars (vital teeth) were performed in animals with ages of 7, 10, 13, and 16 weeks. In another group of animals the teeth were left open for 3 weeks, and then interventions for disinfection including the use of an antibiotic paste were carried out. Root formation was then assessed after 3 and 6 weeks on the basis of radiographic and histologic evaluation.Vital teeth showed increase of root length and hard tissue thickness throughout the experimental periods. On the other hand, induction of necrosis arrested root formation. Teeth subjected to disinfection with sodium hypochlorite associated with the triple antibiotic paste showed significant reduction of periapical lesions, gain in root length, and increased wall thickness compared with the control (P < .05).The root canal disinfection protocol used was able to reduce periapical lesion size and improve root development. The experimental model presented should contribute to studies that aim at improving therapeutic strategies for necrotic immature teeth by using a rat model.

Research paper thumbnail of Increased Tissue Factor Expression and Poor Nephroblastoma Prognosis

Journal of Urology, 2009

There is potential interaction between malignant cell growth and the coagulation pathway. Recent ... more There is potential interaction between malignant cell growth and the coagulation pathway. Recent studies suggest that tissue factor, a primary initiator of the extrinsic coagulation pathway, is expressed in various solid tumors in association with increased angiogenesis. To our knowledge we report for the first time the detection of tissue factor expression by immunohistochemistry in Wilms tumors and its correlation with clinical outcomes.Tissue factor expression detected by immunohistochemistry was assessed in 41 formalin fixed, paraffin embedded Wilms tumor cases treated at university hospitals. We correlated findings with tumor recurrence and cancer specific survival.Positive immunohistochemistry detection of tissue factor was observed in 88.3% of the tumors analyzed. Tissue factor on immunohistochemistry was associated with tumor recurrence and survival (p = 0.01 and 0.02, respectively). Increased immunohistochemical detection of tissue factor was the most important risk factor for recurrence and mortality in our population on bivariate and multivariate analysis.Tissue factor is a promising research subject as a prognostic factor for Wilms tumor. More studies are needed to clarify the mechanisms by which tissue factor affects cancer progression and outcome, and its potential role as a therapeutic target.

Research paper thumbnail of Sequential Chemotactic and Phagocytic Activation of Human Polymorphonuclear Neutrophils

Infection and Immunity, 2007

Human polymorphonuclear neutrophils (PMN) chemotax to a foreign entity. When the chemoattractants... more Human polymorphonuclear neutrophils (PMN) chemotax to a foreign entity. When the chemoattractants' origins are reached, specific receptors bind to the invader's surface, initiating phagocytosis, phagosome formation, and fusion with granule membranes, generating the bactericidal oxidative burst, and releasing lytic enzymes, specific peptides, and proteins. We explored the initial signaling involved in these functions by observing naïve, unprimed PMN in suspension using fluorescent indicators of cytoplasmic signals (⌬[Ca 2؉ ] i and ⌬pH i ) and of bactericidal entities (oxidative species and elastase) exposed to N-formyl-methionyl-leucylphenylalanine (fMLP) and/or multivalent immune complexes (IC). fMLP and IC each initiate a rapid transient rise in [Ca 2؉ ] i , mostly from intracellular stores, simultaneously with a drop in pH i ; these are followed by a drop in [Ca 2؉ ] i and a rise in pH i , with the latter being due to a Na ؉ /H ؉ antiport. The impact of a second stimulation depends on the order in which stimuli are applied, on their dose, and on their nature. Provided that [Ca 2؉ ] i is restored, 10 ؊7 M fMLP, previously shown to elicit maximal ⌬[Ca 2؉ ] i but no bactericidal functions, did not prevent the cells' responses with ⌬[Ca 2؉ ] i to a subsequent high dose of fMLP or IC; conversely, cells first exposed to 120 g/ml IC, previously shown to elicit maximal ⌬[Ca 2؉ ] i and bactericidal functions, exhibited no subsequent ⌬[Ca 2؉ ] i or ⌬pH i to either stimulus. While exposure to 10 ؊7 M fMLP, which saturates the PMN high-affinity receptor, did not elicit bactericidal release from these naïve unprimed PMN in suspension, 10 ؊5 M fMLP did, presumably via the low-affinity receptor, using a different Ca 2؉ source. on February 9, 2015 by guest http://iai.asm.org/ Downloaded from FIG. 4. Sequential stimulation of PMN by saturating doses (120 g/ml) of IC followed 5 min later by 10 Ϫ7 M fMLP. (a) Control; (b) 5 mM EGTA added 15 s before IC injection; (c) 5 mM EGTA added before fMLP injection; (d) release of elastase (shown as F 460 ) and of oxidative products (shown as F 530 ). Each figure is representative of five independent experiments. VOL. 75, 2007 SEQUENTIAL CHEMOTACTIC AND IC ACTION ON PMN 3995 on February 9, 2015 by guest

Research paper thumbnail of Identification of mRNAs for the various diacylglycerol kinase isoforms in neutrophils from patients with localized aggressive periodontitis

Journal of Periodontal Research, 2003

Background: Diacylglycerol kinase (DGK) metabolizes diacylglycerol (DAG), an endogenous activato... more Background: Diacylglycerol kinase (DGK) metabolizes diacylglycerol (DAG), an endogenous activator of protein kinase C, to phosphatidic acid. We have previously reported increased DAG in neutrophils from patients with localized aggressive periodontitis (LAP) associated with reduced DGK activity. This reduction could be related to a mutation, post-translational modification, differential expression, or lack of expression of a particular isoform(s).Objective: The aim of this study was to identify the mRNAs for DGK isoforms in normal and LAP neutrophils.Methods: The α-, γ-, and δ-isoforms of DGK were identified by polymerase chain reaction (PCR) using specific oligonucleotide primers for each isoform. The PCR products were isolated and sequenced for comparison to published sequences to confirm the validity of the PCR reaction. Total RNA was isolated from LAP and normal neutrophils, and northern blotting and semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was used to examine the level of mRNA for each isoform.Results: No major differences were found in the isoform pattern between resting normal and LAP neutrophils. However, the levels of mRNA for the α- and γ-isoforms of DGK were increased in normal neutrophils while slightly decreased in LAP cells upon stimulation with N-formyl-methionyl-leucyl-phenylalanine (fMLP).Conclusion: These data suggest that alterations in the mRNAs for the various isoforms of DGK during cell stimulation and the involvement of DGK that is expressed in multiple forms are subject to a variety of regulatory/control mechanisms and these mechanisms may explain the role of the ‘primed’ neutrophil phenotype associated with LAP.

Research paper thumbnail of Differentiation of HL60 cells to granulocytes involves regulation of select diacylglycerol kinases (DGKs)

Journal of Cellular Biochemistry, 2005

AbstractDiacylglycerol Kinases (DGKs) are a family of enzymes that regulate the levels of differe... more AbstractDiacylglycerol Kinases (DGKs) are a family of enzymes that regulate the levels of different pools of diacylglycerol (DAG), affecting DAG-mediated signal transduction. Since DAG is known to play several important regulatory roles in granulocyte physiology, we investigated the expression pattern of DGK isoforms throughout differentiation of HL-60 cells to granulocytes. HL-60 cells were incubated with 1.25% dimethyl-sulfoxide (DMSO) to initiate differentiation and total RNA isolated at different time points. DGK expression was assessed through Northern blot, end-point PCR, and real-time PCR. The non-selective inhibitors R59022 and R59949 were used to block DGK at different time points throughout differentiation. CD11b and GPI-80, reactive oxygen species (ROS) generation, changes in the cell cycle, and apoptosis were used as markers of differentiation. Of the nine isoforms of DGK evaluated (α, δ, ε, γ, ζ, β, θ, ι, η), only five (α, δ, ε, γ, and ζ) were expressed in HL-60 cells. DGKα was virtually absent in non-differentiated cells, but was markedly upregulated throughout differentiation. The other isoforms (δ, ε, γ, and ζ) were expressed in undifferentiated HL-60 cells but were substantially decreased throughout differentiation. Non-selective blocking of DGK with R59022 and R59949 led to acceleration of differentiation, reducing the time necessary to observe upregulation of CD11b, GPI-80 and generation of ROS by 50%. Likewise, the cell cycle was disrupted when DGK isoforms were inhibited. These results provide evidence that DGK levels are dynamically regulated throughout differentiation and that expression of DGKs play an important regulatory function during the differentiation of neutrophils. © 2004 Wiley-Liss, Inc.

Research paper thumbnail of Protective Effects of Resveratrol on Hepatotoxicity Induced by Isoniazid and Rifampicin via SIRT1 Modulation

Journal of Natural Products, 2014

Acute liver injury was induced in male BALB/c mice by coadministering isoniazid and rifampicin. I... more Acute liver injury was induced in male BALB/c mice by coadministering isoniazid and rifampicin. In this work, the effects of resveratrol (1) were investigated in the hepatotoxicity caused by isoniazid−rifampicin in mice. Compound 1 was administered 30 min prior to isoniazid−rifampicin. Serum biochemical tests, liver histopathological examination, oxidative stress, myeloperoxidase activity, cytokine production (TNF-α, IL-12p70, and IL-10), and mRNA expression of SIRT1−7 and PPAR-γ/ PGC1-α were evaluated. The administration of 1 significantly decreased aspartate transaminase and alanine aminotransferase levels, myeloperoxidase activity, and cytokine levels. Furthermore, 1 reverted the decrease of catalase and glutathione activities and ameliorated the histopathological alterations associated with antituberculosis drugs. Modulation of SIRT1 and PPAR-γ/ PGC1-α expression is likely involved in the protective effects of 1. The results presented herein show that 1 was able to largely prevent the hepatotoxicity induced by isoniazid and rifampicin in mice, mainly by modulating SIRT1 mRNA expression.

Research paper thumbnail of Porphyromonas gingivalis infection accelerates the progression of atherosclerosis in a heterozygous apolipoprotein E-deficient murine model

Circulation, Jan 19, 2002

Background-Current epidemiological data suggest that dental infections affecting tooth-supporting... more Background-Current epidemiological data suggest that dental infections affecting tooth-supporting tissues (periodontitis) can disseminate into the systemic circulation and thereby contribute to atherosclerosis progression. To test this hypothesis, we investigated the effect of repeated systemic inoculations with Porphyromonas gingivalis (Pg), a putative periodontal pathogen, on the progression of atherosclerosis in heterozygous apolipoprotein E-deficient (ApoE ϩ/Ϫ ) mice. Methods and Results-Ten-week-old, male ApoE ϩ/Ϫ mice fed either a high-fat diet or regular chow were inoculated intravenously with live Pg (10 7 CFU) or vehicle once per week for 10, 14, or 24 consecutive weeks. Histomorphometry of plaque cross-sectional area in the proximal aortas, en face measurement of plaque area over the aortic trees, Pg 16S ribosomal DNA amplification with polymerase chain reaction, ELISA for systemic proinflammatory mediators, and immunolocalization of macrophages in the proximal aorta were performed. Atherosclerotic lesions of the proximal aortas and aortic trees were more advanced in Pg-challenged animals than in vehicle control animals and occurred earlier (at 10 weeks) when no lesions were apparent in control animals. At 24 weeks after inoculation, proximal aortic lesion size quantified by histomorphometry was 9-fold greater in chow-fed mice inoculated with Pg than in noninoculated mice (PϽ0.001) and was 2-fold greater in Pg-inoculated versus noninoculated high-fat diet-fed mice (PϽ0.001); all atherosclerotic lesions were macrophage-rich. Pg ribosomal DNA was found in the aortas, livers, and hearts 24 weeks after inoculation.

Research paper thumbnail of Priming of neutrophil oxidative burst in diabetes requires preassembly of the NADPH oxidase

Journal of Leukocyte Biology - J LEUKOCYTE BIOL, 2008

Research paper thumbnail of Analysis of select members of the E26 (ETS) transcription factors family in colorectal cancer

Virchows Archiv, 2011

The E-twenty-six (ETS) family of transcription factors is known to act as positive or negative re... more The E-twenty-six (ETS) family of transcription factors is known to act as positive or negative regulators of the expression of genes that are involved in diverse biological processes, including those that control cellular proliferation, differentiation, hematopoiesis, apoptosis, metastasis, tissue remodeling, and angiogenesis. Identification of target gene promoters of normal and oncogenic transcription factors provides new insights into the regulation of genes that are involved in the control of normal cell growth and differentiation. The aim of the present investigation was to analyze the differential expression of 11 ETS (ELF-3, ESE3, ETS1, ETV3, ETV4, ETV6, NERF, PDEF, PU1, Spi-B, and Spi-C) as potential markers for prognostic of colorectal cancer. A series of paired tissue biopsies consisting of a tumor and a non-affected control sample were harvested from 28 individuals suffering from diagnosed colorectal lesions. Total RNA was isolated from the samples, and after reverse transcription, differential expression of the select ETS was carried out through real-time polymerase chain reaction. Tumor staging as determined by histopathology was carried out to correlate the degree of tumor invasiveness with the expression of the ETS genes. The results demonstrated a different quantitative profile of expression in tumors and normal tissues. ETV4 was significantly upregulated with further increase in the event of lymph node involvement. PDEF and Spi-B presented downregulation, which was more significant when lymph node involvement was present. These findings were supported by immunohistochemistry of tumoral tissues. The results suggest that select ETS may serve as potential markers of colorectal cancer invasiveness and metastasis.

Research paper thumbnail of Real time PCR quantification of viable Mycobacteriumtuberculosis from sputum samples treated with propidium monoazide

Tuberculosis, 2014

s u m m a r y Diagnostic methods of TB, nowadays, are prone to delay in diagnosis, increased fals... more s u m m a r y Diagnostic methods of TB, nowadays, are prone to delay in diagnosis, increased false negative results and are not sensitive to many forms of paucibacillary disease. The aims of this study were to implement a quantitative nucleic acid-based diagnostic test for paucibacillary tuberculosis, enabling the identification and quantification of viable Mycobacterium tuberculosis bacilli by quantitative Real-Time PCR (qRT-PCR). The intergenic region of the single-copy inhA-mabA gene was chosen as the target region for design of primers and probes conjugated with fluorophores. The construction of synthetic DNA flanking the target region served as standards for absolute quantification of nucleic acids. Using the intercaling dye, propidium monoazide, we were able to discriminate between viable and dead cells of M. tuberculosis. The diagnosis method showed a broad sensitivity (96.1%) when only compared to samples of smear-positive sputum and ROC analyses shows that our approach performed well and yielded a specificity of 84.6% and a sensitivity of 84.6% when compared to M. tuberculosis colony-forming units counting.

Research paper thumbnail of Increased Tissue Factor Expression and Poor Nephroblastoma Prognosis

The Journal of Urology, 2009

NWTS ϭ National WT Study SIOP ϭ Societé Internationale d'Urologie Pédiatrique TF ϭ tissue factor ... more NWTS ϭ National WT Study SIOP ϭ Societé Internationale d'Urologie Pédiatrique TF ϭ tissue factor WT ϭ Wilms tumor Purpose: There is potential interaction between malignant cell growth and the coagulation pathway. Recent studies suggest that tissue factor, a primary initiator of the extrinsic coagulation pathway, is expressed in various solid tumors in association with increased angiogenesis. To our knowledge we report for the first time the detection of tissue factor expression by immunohistochemistry in Wilms tumors and its correlation with clinical outcomes. Material and Methods: Tissue factor expression detected by immunohistochemistry was assessed in 41 formalin fixed, paraffin embedded Wilms tumor cases treated at university hospitals. We correlated findings with tumor recurrence and cancer specific survival. Results: Positive immunohistochemistry detection of tissue factor was observed in 88.3% of the tumors analyzed. Tissue factor on immunohistochemistry was associated with tumor recurrence and survival (p ϭ 0.01 and 0.02, respectively). Increased immunohistochemical detection of tissue factor was the most important risk factor for recurrence and mortality in our population on bivariate and multivariate analysis. Conclusions: Tissue factor is a promising research subject as a prognostic factor for Wilms tumor. More studies are needed to clarify the mechanisms by which tissue factor affects cancer progression and outcome, and its potential role as a therapeutic target.

Research paper thumbnail of Use of Bovine-Derived Anorganic Bone Associated With Guided Tissue Regeneration in Intrabony Defects. Six-Month Evaluation at Re-Entry

Journal of Periodontology, 1999

Background: Different filling materials have been associated with guided tissue regeneration (GTR... more Background: Different filling materials have been associated with guided tissue regeneration (GTR) in order to improve its regenerative potential and predictability. Anorganic bovine bone (ABB) has demonstrated biocompatibility and osteoconductive properties; however, there are limited data regarding its performance in the treatment of intrabony defects. This investigation aimed to evaluate the clinical outcome of the association of anorganic bovine bone with cellulose membranes in intrabony defects after 6 months.

Research paper thumbnail of Manifestations of Insulin-Dependent Diabetes Mellitus in the Periodontium of Young Brazilian Patients. A 10-Year Follow-Up Study

Journal of Periodontology, 1997

Eight female and 3 male patients from a group of 30 patients studied 10 years earlier and who had... more Eight female and 3 male patients from a group of 30 patients studied 10 years earlier and who had received no periodontal treatment during this period, in order to evaluate the progression of untreated periodontal disease in patients with insulin-dependent diabetes mellitus, were evaluated in terms of plaque accumulation, gingival inflammation, probing depth, and alveolar bone loss. The total number of dental surfaces that presented clinically detectable plaque deposits increased significantly (29% to 43%; P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.01; chi 2 = 46.36). Site-specific comparisons for plaque index between studies showed a significant variation (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.01) in the upper arch only for palatal surfaces and in the lower arch for the buccal and lingual surfaces. The total dental surfaces with inflamed surrounding gingiva increased from 11% to 33% in this study (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.01; chi 2 = 175.78). Site-specific comparison for gingival index showed a significant variation for all upper surfaces, while such difference for the lower arch was significant only for the buccal and lingual surfaces. The arithmetic means for the probing depth for the upper buccal, upper palatal, lower buccal, and lower lingual surfaces increased significantly (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.01). The arithmetic means of alveolar bone loss also increased significantly for the upper posterior and lower regions (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.01) and for the upper anterior and lower anterior regions (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05). The correlation between age and probing depth was significant only for the upper palatal region (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.01). The correlation between age and bone loss was significant only for the upper posterior region (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05). The results of this follow-up study suggest that despite little variation in plaque accumulation, gingival inflammation, probing depth, and bone loss increased after a 10-year interval in patients who had received no periodontal treatment during this period.

Research paper thumbnail of Management of Soft Tissue Ridge Deformities With Acellular Dermal Matrix. Clinical Approach and Outcome After 6 Months of Treatment

Journal of Periodontology, 2001

Soft tissue ridge defects often hamper ideally shaped artificial crowns and are basically treated... more Soft tissue ridge defects often hamper ideally shaped artificial crowns and are basically treated using autogenous soft tissue grafts or alloplastic materials. These approaches present disadvantages such as the necessity of creating additional surgical fields to harvest the graft and the requirement of primary closure, which may reduce ridge height. This investigation evaluated the use of acellular dermal matrix (ADM) in the treatment of soft tissue ridge defects. Eight patients, non-smokers with non-contributory medical history, provided 18 sites corresponding to missing teeth in the anterior maxillary arch. The ideal horizontal gain (desired gain) was waxed up in study casts, which served as templates for construction of modified acrylic stents with orthodontic wires. These stents served as references for ideal horizontal gain and also as fixed reference points for further evaluation. The distance from the orthodontic wire to the buccal plate of the defect also represented its baseline horizontal component. Vertical variations were evaluated with another stent and, in this case, no desired gain was considered. After raising partial-thickness flaps, the ADM material was rehydrated and folded to fill the defect and reproduce the desired gain. Flaps were sutured with no tension, and part of the material was intentionally left exposed to avoid pressure on the incision line and prevent height loss. Patients used local and systemic antimicrobials, and the sutures were removed at 7 days. Evaluations were carried out at 30 days, and 3 and 6 months, and all sites healed uneventfully. Neither infection nor significant pain was reported by the patients, and the material was covered by tissue at about 21 days. Mean horizontal gain of 1.72 +/- 0.59 mm (58.5%) at 6 months and mean shrinkage of 1.22 +/- 0.46 mm (41.4%) were observed. There was a mean improvement in vertical gain of only 0.61 +/- 0. 77 mm, although 66. 7% of the treated sites showed a 1 to 2 mm gain. Clinically, the total gain in the subjects was very effective and matched the receptor tissues nicely. ADM may be a suitable material for the treatment of soft tissue ridge deformities due to its biocompatibility, color matching, and horizontal gain. Additional controlled, comparative trials are necessary to establish its advantages and potential compared to autogenous soft tissue techniques.

Research paper thumbnail of Guided Tissue Regeneration Associated With Bovine-Derived Anorganic Bone in Mandibular Class II Furcation Defects. 6-Month Results at Re-Entry

Journal of Periodontology, 2000

The use of graft materials with guided tissue regeneration (GTR) in Class II furcation defects is... more The use of graft materials with guided tissue regeneration (GTR) in Class II furcation defects is aimed at improving the outcome of the regenerative technique. In this regard, however, there are a limited number of studies discussing the results obtained when GTR and graft materials are used in the treatment of Class II furcation defects. Furthermore, most studies employ either allogeneic or autogenous materials. The present trial sought to determine whether the use of a bovine-derived anorganic bone (ABB) in conjunction with GTR influenced the outcome of mandibular Class II furcation treatment. This study included 14 patients who provided 15 pairs of similar periodontal defects. Each defect was randomly assigned to treatment with either a cellulose membrane in combination with bovine-derived anorganic bone (GTR+ABB) or membrane alone (GTR). Following basic therapy, baseline measurements were recorded including probing depth (PD), clinical attachment level (CAL), and gingival margin position (GMP). Hard tissue measurements were performed during surgery to determine alveolar crestal height (CEJ-AC), and vertical (VDD) and horizontal defect depth (HDD). Membranes remained in position for at least 4 weeks. After 6 months, all sites were re-entered and soft and hard tissue measurements were recorded. Both surgical procedures resulted in statistically significant probing depth reduction and gain in clinical attachment levels, with no significant difference between groups. Gingival recession was more pronounced in the GTR+ABB group (0.87 +/- 0.83 mm), but not statistically different from the GTR group (0.46 +/- 1.19 mm). Vertical defect resolution was significant in both groups (GTR: 1.60 +/- 1.50 mm; GTR+ABB: 1.80 +/- 2.11 mm), without differences between groups. Only horizontal furcation resolution (GTR: 2.47 +/- 0.99 mm; GTR+ABB: 3.27 +/- 1.39 mm) was significantly different between groups (P &amp;amp;lt;0.05). The use of ABB with GTR techniques improved horizontal defect resolution in mandibular Class II furcation defects, but did not yield superior results regarding soft tissue changes when compared to sites treated with GTR alone. Evaluation of a larger sample could indicate differences and advantages between the evaluated approaches and confirm the real necessity of associating filling materials with GTR.