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Biomedical subjects

M R Leonardo

Publications and source records attributed to M R Leonardo.

At least 37 records · Page 2Linked to original sources
PubMed1997

Calcium hydroxide root canal sealers--histopathologic evaluation of apical and periapical repair after endodontic treatment.

Eighty root canals of the premolars of 4 dogs, with vital pulp, were instrumented and filled during the same session with the Sealapex, CRCS, Sealer 26, and Apexit sealers. The animals were sacrificed 180 days after root canal filling and their maxillae and mandibles were removed and fixed in 10% formalin. After routine histologic processing, the sections were stained with hematoxylin-eosin and Mallory trichrome. Histopathologic analysis showed that Sealapex was the sealer that best permitted the deposition of mineralized tissue at the apical level and was the only sealer that provided complete sealing (37.5% of cases). With the use of Sealapex, no inflammatory infiltrate occurred and there was no reabsorption of mineralized tissues. In contrast, partial sealing and a moderate inflammatory infiltrate occurred with the use of CRCS. When Apexit and Sealer 26 were used the absence of sealing was frequent and active reabsorption of mineralized tissues occurred in most cases. The inflammatory infiltrate predominating with the use of Apexit was of the severe type, whereas with the use of Sealer 26 the inflammatory infiltrate was mild or absent.

Animals↗
PubMed1997

Determination of the p-monochlorophenol residue in the calcium hydroxide + P-monochlorophenol combination used as an intracanal dressing in pulpless teeth of dogs with induced chronic periapical lesion.

To determine the presence of p-monochlorophenol in the calcium hydroxide (Calen) + p-monochlorophenol combination after its use as intracanal dressing, periapical lesions were induced in 60 root canals of upper and lower premolars of four dogs. After biomechanical preparation, the root canals received the intracanal medication, which was removed from the apical third after 2, 4, 7, and 14 days for chemical analysis by spectrophotometry. The results showed a p-monochlorophenol loss of approximately 50.0% in the dressing after 48 h, with no further significant loss after longer periods of times. p-Monochlorophenol was still present in the medication after 14 days.

Analysis of Variance↗
PubMed1997

Calcium hydroxide root canal sealers: evaluation of pH, calcium ion concentration and conductivity.

The calcium hydroxide ionization of four root canal sealers (Sealapex, CRCS, Sealer 26, and Apexit) was studied by measuring conductivity and pH and by conducting atomic absorption spectrophotometry. Samples 6 mm in diameter and 15 mm long were prepared from these sealers. After setting and 48 h storage in a desiccator, five samples of each material were placed in 50 mL distilled water and analysed after 0,1,2,4,6 and 24 h and 5, 15 and 30 days. The results showed that Sealapex was the root canal sealer showing the highest pH, ionic calcium and total calcium values (P < 0.05) throughout the experimental period, followed by CRCS, Apexit and Sealer 26.

Bismuth↗
PubMed1996

Re-examination of the role of the periplasmic domain of EnvZ in sensing of osmolarity signals in Escherichia coli.

In Escherichia coli, EnvZ senses changes in the osmotic conditions of the growth environment and controls the phosphorylated state of the regulatory protein, OmpR. OmpR-phosphate regulates the expression of the porin genes, ompF and ompC. To investigate the role of the periplasmic domain of EnvZ in sensing of osmolarity signals, portions of this domain were deleted. Cells containing the EnvZ mutant proteins were able to regulate normally the production of OmpF and OmpC in response to changes in osmolarity. The periplasmic domain of EnvZ was also replaced with the non-homologous periplasmic domain of the histidine kinase PhoR of Bacillus subtilis. Osmoregulation of OmpF and OmpC production in cells containing the PhoR-EnvZ hybrid protein was indistinguishable from that in cells containing wild-type EnvZ. Identical results were obtained with an envZ-pta/ack strain, which could not synthesize acetyl phosphate. Thus, acetyl phosphate was not involved in the regulation of ompF and ompC observed in this study. These results indicate that the periplasmic domain of EnvZ is not essential for sensing of osmolarity signals.

Alleles↗
PubMed1996

Anaerobic microorganisms in root canals of human teeth with chronic apical periodontitis detected by indirect immunofluorescence.

Aiming to assess the presence of selected anaerobic microorganisms in root canals of human teeth with chronic apical periodontitis. 25 central and lateral upper incisors presenting with radiographic evidence of chronic apical periodontitis were studied. The pulp chamber was opened under aseptic conditions and samples of the root canal content were collected with sterile absorbent paper points, which were placed and dispersed in test tubes containing reduced transport medium RTT. Aliquots were dried on glass slides and stained by indirect immunofluorescence for detection of Actinomyces viscosus, Fusobacterium nucleatum, Porphyromonas gingivalis and Prevotella intermedia. The results showed a positive indirect immunofluorescence reaction in 24 of the 25 samples. Fourteen were positive for the specie Actinomyces viscosus, 12 for Prevotella intermedia, 10 for Fusobacterium nucleatum and 4 for Porphyromonas gingivalis. A semiquantitative assay was easily implemented for assessment of degree of infection by the organisms in individual cases.

Actinomyces viscosus↗
PubMed1996

Role of NAD in regulating the adhE gene of Escherichia coli.

The fermentative alcohol dehydrogenase of Escherichia coli is encoded by the adhE gene, which is induced under anaerobic conditions but repressed in air. Previous work suggested that induction of adhE might depend on NADH levels. We therefore directly measured the NAD+ and NADH levels for cultures growing aerobically and anaerobically on a series of carbon sources whose metabolism generates different relative amounts of NADH. Expression of adhE was monitored both by assay of alcohol dehydrogenase activity and by expression of phi(adhE'-lacZ) gene fusions. The expression of the adhE gene correlated with the ratio of NADH to NAD+. The role of NADH in eliciting adhE induction was supported by a variety of treatments known to change the ratio of NADH to NAD+ or alter the total NAD+-plus-NADH pool. Blocking the electron transport chain, either by mutation or by chemical inhibitors, resulted in the artificial induction of the adhE gene under aerobic conditions. Conversely, limiting NAD synthesis, by introducing mutational blocks into the biosynthetic pathway for nicotinic acid, decreased the expression of adhE under anaerobic conditions. This, in turn, was reversed by supplementation with exogenous NAD or nicotinic acid. In merodiploid strains carrying deletion or insertion mutations abolishing the synthesis of AdhE protein, an adhE-lacZ fusion was expressed at nearly 10-fold the level observed in an adhE+ background. Introduction of mutant adhE alleles producing high levels of inactive AdhE protein gave results equivalent to those seen in absence of the AdhE protein. This finding implies that it is the buildup of NADH due to lack of enzyme activity, rather than the absence of the AdhE protein per se, which causes increased induction of the phi(adhE'-lacZ) fusion. Moreover, mutations giving elevated levels of active AdhE protein decreased the induction of the phi(adhE'-lacZ) fusion. This finding suggests that the enzymatic activity of the AdhE protein modulates the level of NADH under anaerobic conditions, thus indirectly regulating its own expression.

Aerobiosis↗
PubMed1995

Histopathological observations of periapical repair in teeth with radiolucent areas submitted to two different methods of root canal treatment.

Dogs' teeth with induced chronic periapical periodontitis were treated endodontically by two different methods, and the results were compared. A total of 40 root canals from the upper and lower premolars of two dogs were prepared chemomechanically. In method 1, a high-concentration (5.25%) hypochlorite solution was used during the instrumentation of the root canal, and an antibacterial dressing was applied between sessions, followed by filling of the root canal. In method 2, a low-concentration (0.5%) sodium hypochlorite solution was used as an adjunct to mechanical debridement, and the root canal was filled during the same session. The histopathological results showed that method 1 led to better periapical repair than method 2.

Animals↗
PubMed1994

Radiographic and microbiologic evaluation of posttreatment apical and periapical repair of root canals of dogs' teeth with experimentally induced chronic lesion.

The objective of the present study was to evaluate radiographically and bacteriologically apical and periapical repair in dogs' teeth with induced chronic periapical lesions with the use of two different operative techniques (techniques 1 and 2). The study was conducted on 40 root canals of upper and lower premolars from two dogs aged approximately 12 months. Periapical lesions were induced by leaving the root canals exposed to the oral environment for 5 days and then sealing them with zinc oxide-eugenol for 45 days. After this period, radiographic examination revealed the occurrence of a radiolucent lesion and endodontic treatment was started. The two techniques did not differ in terms of chemomechanical preparation, final filling, or type of cement, but differed in terms of irrigating solution and the presence of an antibacterial dressing. Thus 4% to 6% hypochlorite and hydrogen peroxide (10 volumes) were used in technique 1 during chemomechanical preparation and an antibacterial dressing based on calcium hydroxide was applied between sessions, whereas Dakin's fluid (0.5% sodium hypochlorite solution) and a final filling with no antibacterial dressing were used in technique 2. After chemomechanical preparation, the root canals were filled with gutta-percha cones and Sealapex (Sealapex-Sybron, Kerr, Sao Paulo, Brazil), and the animals were killed 270 days after the final filling. Blocks were cut into 6-microns sections and stained by the Brown and Brenn method.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Histological evaluation of therapy using a calcium hydroxide dressing for teeth with incompletely formed apices and periapical lesions.

Periapical tissue response was evaluated histologically in teeth of 6-month-old dogs with incomplete root formation and periapical lesions in order to compare the effects of two calcium hydroxide-containing pastes prepared with different vehicles on induced root apex formation and repair of the periapical region. A group of teeth with no root canal dressing was used as control. After induction of the periapical lesion, the root canals were instrumented and filled with one of the two pastes (Calen and Calasept), which were renewed monthly for 90 days. At the end of the follow-up period (3 months), the animals were killed. Block sections from the actual areas were dissected free and prepared for histopathological analysis. Both calcium hydroxide pastes helped induce apical sealing and repair of the region, but the Calen paste yielded better results than Calasept. The induced mineralized tissue was predominantly of the cementoid type after the use of both pastes, but the inflammatory infiltrate was significantly less intense with the use of the Calen paste. In the control group, the absence of a root canal dressing impaired the process of periapical repair and yielded no apical root closure in any of the cases analyzed.

Animals↗
PubMed1993

Effect of intracanal dressings on repair and apical bridging of teeth with incomplete root formation.

Periapical repair and apical bridging were studied in dog's teeth with incomplete root formation and induced chronic periapical lesions treated with different dressings. A total of 75 root canals from the upper and lower premolars of 4 dogs approximately 6 months of age were chemo-mechanically prepared and filled with the following dressings: antibacterial dressing consisting of a calcium hydroxide+camphorated p-monochlorophenol paste applied for 7 days and followed by monthly renewed calcium hydroxide paste as temporary dressing at 30, 60 and 90 days (Group A); antibacterial dressing consisting of camphorated p-monochlorophenol alone for 7 days, followed by temporary dressing with calcium hydroxide paste renewed at 30, 60 and 90 days (Group B). A control group (Group C) received no dressings. Ninety days after the last calcium hydroxide paste (Groups A, B) and after the last irrigation/aspiration (Group C), the animals were killed, the maxillae and mandibles were removed, and the material submitted to routine histological processing and examination. Both root canal dressings, were of fundamental importance for apical repair and bridging. The apical bridging was predominantly complete in Group A, incomplete in Group B, and absent in Group C. The calcium hydroxide+camphorated p-monochlorophenol combination gave better results than camphorated p-monochlorophenol alone.

Animals↗
PubMed1993

Anaerobic regulation of the adhE gene, encoding the fermentative alcohol dehydrogenase of Escherichia coli.

The regulation of the adhE gene, which encodes the trifunctional fermentative acetaldehyde-alcohol dehydrogenase of Escherichia coli, was investigated by the construction of gene fusions and by two-dimensional protein gel electrophoresis. Both operon and protein fusions of adhE to lacZ were induced 10- to 20-fold by anaerobic conditions, and both fusions were repressed by nitrate, demonstrating that regulation is at the level of transcription. Nitrate repression of phi (adhE-lacZ) expression, as well as of alcohol dehydrogenase enzyme activity, was partly relieved by a mutation in narL. Mutations in rpoN or fnr had no effect on the expression of adhE. Two-dimensional protein gels demonstrated that increases in the amount of adhE protein correlated with increases in enzyme activity, demonstrating that induction was due to synthesis of new protein, not to activation of preexisting protein. When oxidized sugar derivatives such as gluconate or glucuronate were used as carbon sources, the anaerobic expression of phi (adhE-lacZ) was greatly reduced, whereas when sugar alcohols such as sorbitol were used, the expression was increased compared with expression when glucose was the carbon source. This observation suggested that induction of phi (adhE-lacZ) might depend on the level of reduced NADH, which should be highest with sorbitol-grown cells and lowest with glucuronate-grown cells. When phi (adhE-lacZ) was present in a strain deleted for the adhE structural gene, anaerobic expression of phi (adhE-lacZ) was approximately 10-fold higher than in an adhE+ strain. Since the presence of alcohol dehydrogenase would serve to decrease NADH levels, this finding again implies that the adhE gene is regulated by the concentration of reduced NAD. Introduction of a pgi (phosphoglucose isomerase) mutation reduced the anaerobic induction of phi(adhE-lacZ) when the cells were grown on glucose, but had little effect on fructose-grown cells. Pyruvate did not overcome the pgi effect, but glycerol 3-phosphate did, which is again consistent with the possibility that adhE expression responds to the level of reduced NAD rather than to a glycolytic intermediate.

Alcohol Dehydrogenase↗
PubMed1991

Tissue reaction to endodontic materials: methods, criteria, assessment, and observations.

The purpose of this study was to develop methods and criteria for the testing of the biocompatibility of endodontic materials. One hundred twenty-one teeth from 12 baboons (Papio anubis) were used to test three sealers: AH26, Kerr pulp canal sealer, and Kloroperka N.O. Gutta-percha cones were used as solid core in all cases. Under disinfected conditions, access preparation was performed, and with working length approximately 1.0 mm short of the foramen, the instrumentation of the root canal was started, using sodium hypochlorite (1%) for irrigation. After the root canals were cleaned and shaped, they were obturated with gutta-percha cones and the sealer, and by lateral condensation technique. Histologic periapical reactions were evaluated at 1, 7, 30, 365, 730, and 1095 days. The methods and criteria used were adequate for ranking of the biocompatibility of the tested materials in the short and long periods. At short observation periods (1 to 7 days) AH26 caused severe reactions, and Kerr pulp canal sealer and Kloroperka N.O., moderate and mild reactions, respectively. At 2- and 3-year observation periods the ranking was AH26, mild; Kerr pulp canal sealer, moderate; and Kloropercha N.O., severe.

Animals↗
PubMed1990

Comparison of subcutaneous connective tissue responses among three different formulations of gutta-percha used in thermatic techniques.

A study of the subcutaneous connective tissue response of 24 white rats to three different formulations of gutta-percha was undertaken. The prepared specimens were examined under the light microscope after intervals of 7, 21, 60 and 120 days. The results showed identical tissue responses after the initial period of 7 days. However, after 120 days the gutta-percha supplied with the Ultrafil system presented mature granulation tissue with neither oedema nor vascular congestion, in contrast to the responses observed with the McSpadden and Obtura formulations.

Animals↗