Isabel Mello

Hepatitis B virus infection in Haemodialysis Centres from Santa Catarina State, Southern Brazil. Predictive risk factors for infection and molecular epidemiology

BackgroundPatients under haemodialysis are considered at high risk to acquire hepatitis B virus (HBV) infection. Since few data are reported from Brazil, our aim was to assess the frequency and risk factors for HBV infection in haemodialysis patients from 22 Dialysis Centres from Santa Catarina State, south of Brazil.MethodsThis study includes 813 patients, 149 haemodialysis workers and 772 healthy controls matched by sex and age. Serum samples were assayed for HBV markers and viraemia was detected by nested PCR. HBV was genotyped by partial S gene sequencing. Univariate and multivariate statistical analyses with stepwise logistic regression analysis were carried out to analyse the relationship between HBV infection and the characteristics of patients and their Dialysis Units.ResultsFrequency of HBV infection was 10.0%, 2.7% and 2.7% among patients, haemodialysis workers and controls, respectively. Amidst patients, the most frequent HBV genotypes were A (30.6%), D (57.1%) and F (12.2%). Univariate analysis showed association between HBV infection and total time in haemodialysis, type of dialysis equipment, hygiene and sterilization of equipment, number of times reusing the dialysis lines and filters, number of patients per care-worker and current HCV infection. The logistic regression model showed that total time in haemodialysis, number of times of reusing the dialysis lines and filters, and number of patients per worker were significantly related to HBV infection.ConclusionsFrequency of HBV infection among haemodialysis patients at Santa Catarina state is very high. The most frequent HBV genotypes were A, D and F. The risk for a patient to become HBV positive increase 1.47 times each month of haemodialysis; 1.96 times if the dialysis unit reuses the lines and filters ≥ 10 times compared with haemodialysis units which reuse < 10 times; 3.42 times if the number of patients per worker is more than five. Sequence similarity among the HBV S gene from isolates of different patients pointed out to nosocomial transmission.

Effectiveness of Photodynamic Therapy Against Enterococcus faecalis, With and Without the Use of an Intracanal Optical Fiber: An In Vitro Study

BACKGROUND AND OBJECTIVE Photodynamic therapy (PDT) is a new technique introduced in endodontics that combines the action of a photosensitizer (dye) and a low intensity light source. Currently, there are no PDT studies evaluating the microbial disinfection of root canals in order to compare the effects of light delivery systems in the photosensitizer activation. The aim of this study was to evaluate the PDT effectiveness in reducing Enterococcus faecalis, with and without the aid of an intracanal optical fiber. METHODS Extracted single-rooted teeth were selected, instrumented, inoculated with E. faecalis and divided into six groups: one control group (untreated), one conventionally-treated group (1% NaOCl irrigation) and four PDT-treated groups. Irradiation (diode laser) was performed with (OF) or without an intracanal optical fiber (NOF) using two different irradiation times: 1 min and 30 sec (IT(90)) or 3 min (IT(180)). Samples were collected before and after testing procedures and CFU/mL was determined. RESULTS The greatest reduction of E. faecalis (99.99%) was achieved with irrigation with 1% NaOCl. PDT also significantly reduced E. faecalis in the following decreasing order: OF/IT(180), NOF/IT(180), OF/IT(90) and NOF/IT(90), with no significant statistical difference among the groups. CONCLUSIONS These results suggest that PDT was effective against E. faecalis, regardless of the use of an intracanal optical fiber.

Molecular models of NS3 protease variants of the Hepatitis C virus

BackgroundHepatitis C virus (HCV) currently infects approximately three percent of the world population. In view of the lack of vaccines against HCV, there is an urgent need for an efficient treatment of the disease by an effective antiviral drug. Rational drug design has not been the primary way for discovering major therapeutics. Nevertheless, there are reports of success in the development of inhibitor using a structure-based approach. One of the possible targets for drug development against HCV is the NS3 protease variants. Based on the three-dimensional structure of these variants we expect to identify new NS3 protease inhibitors. In order to speed up the modeling process all NS3 protease variant models were generated in a Beowulf cluster. The potential of the structural bioinformatics for development of new antiviral drugs is discussed.ResultsThe atomic coordinates of crystallographic structure 1CU1 and 1DY9 were used as starting model for modeling of the NS3 protease variant structures. The NS3 protease variant structures are composed of six subdomains, which occur in sequence along the polypeptide chain. The protease domain exhibits the dual beta-barrel fold that is common among members of the chymotrypsin serine protease family. The helicase domain contains two structurally related beta-alpha-beta subdomains and a third subdomain of seven helices and three short beta strands. The latter domain is usually referred to as the helicase alpha-helical subdomain. The rmsd value of bond lengths and bond angles, the average G-factor and Verify 3D values are presented for NS3 protease variant structures.ConclusionsThis project increases the certainty that homology modeling is an useful tool in structural biology and that it can be very valuable in annotating genome sequence information and contributing to structural and functional genomics from virus. The structural models will be used to guide future efforts in the structure-based drug design of a new generation of NS3 protease variants inhibitors. All models in the database are publicly accessible via our interactive website, providing us with large amount of structural models for use in protein-ligand docking analysis.

In vivo qualitative analysis of the biocompatibility of different cyanoacrylate-based adhesives

Cyanocrylates have been widely used in the medical and dental fields for several years. In Dentistry, cyanoacrylates have been used for suturing, pulp capping, as retrofilling material in endodontic surgeries, and as cervical plug for pulpless teeth bleaching. The biocompatibility of these adhesives has been the topic of many researches and subcutaneous implantation is an effective methodology for these studies. The present study evaluated the biocompatibility of three different cyanoacrylate-based adhesives. Thirty-six Wistar rats were used, divided into four groups of 9 animals each: A (control)--distilled water, B--cyanoacrylate ester (Super Bonder), C - n-butyl-cyanoacrylate (Histoacryl) and D--alpha-cyanoacrylate (Three Bond). The materials were dispensed in sponges of polyvinyl chloride, the animals were incised and the sponges were inserted in the subcutaneous tissue and sutured. Each group was sub-divided according to the time of sacrifice of the animals: 7, 21 and 45 days. Subjective analysis of the histologic material showed that all groups presented some degree of irritability, but the inflammatory reaction decreased with the experimental time in all groups. Group D showed an inflammatory reaction which was closer to that of the control group and was considered to have good biocompatibility. Groups B and C were similar and presented more aggressive inflammatory reactions when compared to the control group. Based on the results, it was concluded that alpha-cyanoacrylate (Three Bond) was the most biocompatible adhesive because it caused the lowest levels of inflammation.

Influence of different volumes of EDTA for final rinse on smear layer removal

OBJECTIVE The aim of this study was to analyze the influence of different volumes of 17% EDTA for final rinse on smear layer removal on the different areas of the root canal. STUDY DESIGN Forty single-rooted teeth were instrumented using rotary instruments. The teeth were divided into 3 test groups according to the EDTA volume for final rinse (5 mL, 10 mL, 15 mL) and 1 control group (10 mL of 1% sodium hypochlorite). The roots were axially split into halves, and the smear layer removal from the canals was determined under scanning electron microscope. The data were analyzed using Kruskal Wallis and Dunn tests. RESULTS The 3 experimental groups showed no statistical difference (P > .05); however, when the test groups were compared to the control group statistically significant differences were found (P < .01). The root canal wall surfaces of teeth in the control group showed the presence of heavy smear layer through the entire length of the root canals. The other groups showed mainly smear layer-free surfaces or a small amount of debris. When coronal, middle and apical thirds were compared, there was no statistically significant difference (P > .05). CONCLUSIONS Based on the results, it appears that a final rinse with 5 mL of EDTA per canal provides good smear layer removal, with root canal walls free of debris and mostly open dentinal tubules in all areas.

Acute hepatitis C virus infection assessment among chronic hemodialysis patients in the Southwest Parana State, Brazil

BackgroundChronic hemodialysis patients are at higher risk for acquiring hepatitis C virus (HCV). The prevalence varies among different countries and hemodialysis centers. Although guidelines for a comprehensive infection control program exist, the nosocomial transmission still accounts for the new cases of infection. The aim of this study was analyze the follow up of newly acquired acute hepatitis C cases, during the period from January 2002 to May 2005, in the Hemodialysis Center, located in the Southwest region of Parana State, Brazil and to analyze the effectiveness of the measures to restrain the appearance of new cases of acute hepatitis C.MethodsPatients were analyzed monthly with anti-HCV tests and ALT measurements. Patients with ALT elevations were monitored for possible acute hepatitis C.ResultsDuring this period, 32 new cases were identified with acute hepatitis C virus infection. Blood screening showed variable ALT levels preceding the anti-HCV seroconversion. HCV RNA viremia by PCR analysis was intermittently and even negative in some cases. Ten out of 32 patients received 1 mcg/kg dose of pegylated interferon alfa-2b treatment for 24 weeks. All dialysis personnel were re-trained to strictly follow the regulations and recommendations regarding infection control, proper methods to clean and disinfect equipment were reviewed and HCV-positive patients were isolated.ConclusionLaboratory tests results showed variable ALT preceding anti-HCV seroconversion and intermittent viremia. The applied recommendations contributed importantly to restrain the appearance of new cases of acute hepatitis C in this center and the last case was diagnosed in May 2004.

Effect of Chemical Substances in Removing Methylene Blue After Photodynamic Therapy in Root Canal Treatment

BACKGROUND AND OBJECTIVE The disadvantage of photodynamic therapy (PDT) is in the photosensitizing agents that may stain the tooth structure. There is no register of PDT studies evaluating protocols to minimize that concern. The present study evaluated the efficiency of chemical adjuncts in methylene blue dye (MB) removal after PDT. MATERIALS AND METHODS Forty single-rooted teeth, after root canal preparation, were filled with 0.01% MB for 5  min, and irradiated with diode laser 660  nm at 40  mW for 240  sec (total energy 9.6  J). The specimens were divided into four groups (n = 10), according to the chemical adjuncts used for dye removal: (a) 2.5% sodium hypochlorite (NaOCl); (b) 2.5% NaOCl + Endo-PTC cream; (c) 70% ethyl alcohol and (d) saline (control). The crowns were sectioned and fixed in a device. Photographs were taken before the PDT (T0), immediately after (T1) and upon dye removal (T2). The chromatic alterations were evaluated using Adobe Photoshop and K values were determined in four fixed points of each crown. RESULTS K values (dental staining) increased in all groups when comparing T0 and T1. The effectiveness of the tested adjuncts was, in decreasing order: G1 (-3.11) > G2 (-2.97) > G3 (-1.28) > G4 (-1.19), not observing significant statistical differences (p < 0.05) between G1 and G2 and also between G3 and G4. CONCLUSIONS Protocols to remove photosensitizing dyes should be applied after PDT in order to minimize dental stain. The protocols tested in this study by using 2.5% NaOCl, associated or not with Endo-PTC cream, were effective in avoiding tooth staining caused by MB during PDT.

Coronal and apical leakage analysis of two different root canal obturation systems

This study compared the coronal and apical leakage of AH Plus with gutta-percha to that of Epiphany with Resilon. Twenty-four single rooted teeth were instrumented and divided into 2 groups according to the solutions for smear layer removal and the obturation materials employed: Group A - 17% EDTA-T and AH Plus with gutta-percha; Group B - primer and Epiphany with Resilon. The Group B specimens were light-cured in the coronal area for 20 s. The external root surfaces were covered with a double layer of ethyl cyanoacrylate, except for the apical foramen and the cavity access. The teeth were immersed in 0.5% methylene blue for 48 h. The specimens were rinsed, dried and axially split for dye penetration measurement with the ImageLab 2.3 software. The t-test showed no significant differences for coronal leakage between the groups, but there were significant differences for apical leakage between the groups (P < 0.05). AH Plus with gutta-percha and Epiphany with Resilon provided the same coronal seal, whereas Epiphany with Resilon provided the best apical seal.

Influence of Er:YAG laser irradiation on apical sealing of four different sealers.

The sealing of the root canal system is of fundamental importance for successful endodontic treatment. To obtain an adequate apical seal, many factors must be considered such as the presence of smear layer and the sealer applied. After canal preparation, this layer must be removed because it prevents close contact between the dentinal walls and the sealing material. The goal of this study was to evaluate the sealing ability of four different sealers after smear layer removal with either 17% EDTA-T irrigation or Er:YAG laser irradiation of 46 teeth. The canals were sealed with four different sealers: Sealapex, Ketac Endo, AH Plus and N-Rickert. The method for smear layer removal did not influence apical sealing. AH Plus and N-Rickert allowed less dye leakage when compared to Sealapex and Ketac Endo.

Tooth Discoloration Induced by Endodontic Phenothiazine Dyes in Photodynamic Therapy

OBJECTIVE This study sought to assess if discoloration of tooth structures occurs after photodynamic therapy (PDT) and to determine the efficacy of a protocol to remove the photosensitizers. BACKGROUND DATA PDT has been used in root canal treatment to enhance cleaning and disinfection of the root canal system. PDT uses a low power laser in association with a dye as a photosensitizer. Photosensitizers can induce staining of the dental structures, resulting in an unaesthetic appearance. METHODS Forty teeth were randomly divided into four groups according to the photosensitizer used and pre-irradiation time: 0.01% methylene blue for 5 min (MB5); 0.01% methylene blue for 10 min (MB10); 0.01% toluidine blue for 5 min (TB5); and 0.01% toluidine blue for 10 min (TB10). Specimens were irradiated with a 660 nm diode laser with a 300 μm diameter optical fiber, at 40 mW power setting for 3 min. Immediately after, the photosensitizers were removed with Endo-PTC cream+2.5% sodium hypochlorite (NaOCl). The shade was measured by a Vita Easyshade spectrophotometer based on the CIELAB color system (L*a*b* values) at three different experimental times: before PDT (T0), immediately after PDT (T1), and after removal of the photosensitizer (T2). RESULTS The results showed a decrease in the averages of the L*a*b* coordinate values after PDT (T1) in all the groups, when compared with the number at T0, with a significant statistical difference in group MB10. After photosensitizer removal (T2), all the values of the coordinates increased with significant statistical differences (p<0.05) between T1 and T2 in L* and a*. CONCLUSIONS It can be concluded that both methylene blue and toluidine blue dyes cause tooth discoloration, and that Endo-PTC cream associated with 2.5% NaOCl effectively remove these dyes, regardless of the pre-irradiation time used for PDT.