Immunolocalization of matrix metalloproteinases-2 and-9 during apical periodontitis development

Objective: The objective of this study was to determine the expression of matrix metalloproteinase-2 (MMP-2) and -9 (MMP-9) during apical periodontitis development. Methods: Using an experimental design of induced periapical lesions in rats and immunohistochemistry assay as investigative tool, the MMP-2 and MMP-9 expression and distribution were evaluated at 3, 7,14, 21, 30,60 and 90 days after coronary access and pulp exposure of the first left mandibular molar to the oral environment. Two blind observers scored the immunoreactivity. A semi-quantitative analysis was performed. Results: Except at day 3, MMP-2 and MMP-9 immunostaining was observed in all experimental periods. The MMP-2 (p = 0.004) and MMP-9 (p = 0.005) immunostaining was higher in the period between 7 and 21 days. They were mainly observed in cells surrounding the apical foramen and adjacent periapical areas. Cells into the hypercementosis areas were strongly stained while both osteoblasts and osteoclasts; presented discrete staining along of this study. No staining was observed on epithelial walls. At 30, 60 and 90 days, the subjacent connective tissue presented intense MMP-2 and MMP-9 immunostaining in mononuclear cells (suggestive of fibroblasts, macrophages, infiltrating neutrophils and lymphocytes). Conclusion: The results observed in this study suggest that MMP-2 and MMP-9 play a critical role in the development of inflammatory periapical lesions, probably involved in the extracellular matrix (ECM) degradation during the initial phase of the lesion development. (C) 2009 Elsevier Ltd. All rights reserved.

Eruption Cysts in the Neonate

Disturbances of the dental development may result in anomalies, which may be apparent as soon as the child is born. Eruption cysts are rarely observed in neonates considering that at this stage of the child`s life teeth eruption is uncommon. Thus, the aim of this report is to describe a case of eruption cysts in a neonate. A male neonate was brought to the emergency service with the chief complaint of an elevated area on the anterior region of the inferior alveolar ridge. The lesion was clinically characterized as a compressive and floating swelling. Through a radiographic exam two mandibular primary incisors could be seen superficially located. Due to the patient`s age and the initial diagnosis of eruption cysts the conduct adopted was clinical surveillance. Forty-five days after the first visit the lesions had significantly decreased in size, and completely disappeared after 4 months. at that age, both mandibular central incisors were already in the oral cavity exhibiting small hypoplastic areas in the incisal edges. The clinical and radiographic follow-up of eruption cysts in neonates appears to be an adequate conduct without differing from that recommended for older children.

Apical limit of root canal filling and its relationship with success on endodontic treatment of a mandibular molar: 11-year follow-up

Objective. This article discusses the relationship between apical limit of root canal filling and success on endodontic treatment of a mandibular molar. Study design. A mandibular right first molar with vital pulp was endodontically treated, and 3 years later periapical lesions on mesial and distal roots were detected. The canals were retreated and obturated to the same levels as in the previous treatment. Results. An 8-year radiographic follow-up showed repair of the periapical lesions on both roots. Conclusions. Results suggest that the apical limit of obturation seems to have no influence in the repair of periapical tissues in mandibular molars. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2011; 112: e48-e50)

The anatomy of two-rooted mandibular canines determined using micro-computed tomography

P>Aim To investigate the internal and external anatomy of extracted human mandibular canines with two roots and two distinct canals using micro-computed tomography (mu CT). Methodology Fourteen two-rooted human mandibular canines were scanned using a high-resolution mu CT system (SkyScan 1174v2; SkyScan N.V., Kontich, Belgium). The images were processed to evaluate the size of the roots, the furcation regions, the presence of accessory canals, the mean distances between several anatomical landmarks, the position of the apical foramina, the direction of root curvatures, the cross-sectional appearances (SMI index), the volume and surface areas of the root canals. Results Root bifurcation was located in both apical (44%, n = 6) and middle (58%, n = 8) thirds of the root. The size of the buccal and lingual roots was similar in 29% of the sample. From a buccal view, no curvature towards the lingual or buccal direction occurred in either roots. From a proximal view, no straight lingual root occurred. In both views, S-shaped roots were found in 21% of the specimens. Location of the apical foramen varied considerably, tending to the mesio-buccal aspect of both roots. Lateral and furcation canals were observed mostly in the cervical third in 29% and 65% of the sample, respectively. The structure model index (SMI) index ranged from 1.87 to 3.86, with a mean value of 2.93 +/- 0.46. Mean volume and area of the root canals were 11.52 +/- 3.44 mm3 and 71.16 +/- 11.83 mm2, respectively. Conclusions The evaluation of two-rooted mandibular canines revealed that bifurcations occurred in the apical and middle third. S-shaped roots were found in 21% of the specimens. Mean volume, surface area and SMI index of the root canals were 11.52 mm3, 71.16 mm2 and 2.93, respectively.

Further studies on an intermediate host murine model showing that a primary Echinococcus granulosus infection is protective against subsequent oncospheral challenge

We describe the use of a murine model to evaluate resistance against subsequent challenge following a primary infection with oncospheres of Echinococcus granulosus. Mice (Kunming strain) were infected with hatched oncospheres of Echinococcus granulosus; 21 days later a second challenge was given by a different route of infection. A primary infection by intraperitoneal (i.p.) injection stimulated 100 and 90.5% protection in terms of reduced cyst numbers against a secondary infection given subcutaneously (s.c.) or intravenously (i.v.) respectively. A primary infection given s.c. followed by i.p. or i.v. challenge resulted in 84.0 and 100% protection, respectively. Intravenous infection followed by i.p. or s.c. challenge resulted in 98.5 and 69.4% protection, respectively. With the i.v. route of infection, almost all resultant cysts were present in the lungs. The data show that a primary infection with oncospheres can induce total or a high degree of protection against a subsequent challenge and confirms that natural (concomitant) immunity can be stimulated in the intermediate host as the result of a primary infection. This may explain the decline in hydatid infection in sheep older than 2 years in hyper-endemic areas such as those found in Xingjiang, China. These older sheep may have been earlier infected and have subsequently self-cured, with the primary infection stimulating an immune response that protects the intermediate host animals from further infection. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved.

Human trypsinogen in colorectal cancer

Trypsinogen (TRY), the precursor to the serine protease trypsin, is found in the pancreas and mediates digestive proteolysis in the small intestine. Differential display of cDNAs expressed by human colorectal tumor tissues compared with adjacent normal colonic mucosa identified an isoform of TRY (TRY2) up-regulated in colorectal cancers. Northern blot analysis of RNA isolated from a series of 28 malignant colon tumors and corresponding normal mucosa showed that TRY transcripts were up-regulated 2- to 33-fold in 29% of tumors. Further, TRY mRNA was expressed in 6 colorectal cancer cell lines, with highest levels detected in the metastatic tumor lines SW620 and HT29. Immunostaining for TRY protein expression showed intense immunoreactivity in the supranuclear cytoplasm of colon tumors in 16% of tissue specimens. To evaluate the relative contributions of 2 isoforms of TRY, TRY1 and TRY2, to total TRY mRNA expression, a semiquantitative multiplex RT-PCR assay was developed. TRY2 mRNA was detected in all 6 colorectal tumor cell lines, whereas TRY1 mRNA was expressed only in the metastatic tumor lines, showing that the high levels of TRY expression in the metastatic tumor lines are likely due to up-regulation of TRY1. Evaluation of TRY1 and TRY2 mRNA expression by multiplex RT-PCR in a series of 20 colon tumor tissues representative of the range of tumor progression showed that TRY2 mRNA was expressed much more commonly than TRY1 mRNA in normal mucosa (26% vs. 6%) as well as in primary tumor tissues (65% vs. 15%). These data demonstrate that TRY2 is the dominant TRY in colon tissue and suggest that up-regulation of TRY1 expression in colon tumors may be associated with a metastatic phenotype. (C) 2001 Wiley-Liss, Inc.

Short report: Molecular genetic characterization of an unusually severe case of hydatid disease in Alaska caused by the cervid strain of Echinococcus granulosusi

Distinct Echinococcus granulosus life cycle patterns have been described in North America: domestic and sylvatic. Gene sequences of the sylvatic E. granulosus indicate that it represents a separate variant. Case-based data have suggested that the course of sylvatic disease is less severe than that of domestic disease. which led to the recommendation to treat cystic echinococcosis patients in the Arctic by careful medical management rather than by aggressive surgery. We recently reported the first two documented E. granalosus human cases in Alaska with accompanying severe sequelae. Here we describe the results of molecular genetic analysis of the cyst material of one of the subjects that supported identification of the parasite as the sylvatic (cervid) strain and not the domestic (common sheep strain), which was initially thought to be implicated in these unusually severe Alaskan cases.

Diagnosis and management of unusual dental abscesses in children

Although the majority of dental abscesses in children originate from dental caries or trauma, a few are associated with unusual conditions which challenge diagnosis and management. Recent research findings have shed light on these unusual entities and greatly improved understanding of their clinical implications. These conditions include developmental abnormalities such as dens invaginatus in which there is an invagination of dental tissues into the pulp chamber and dens evaginatus in which a tubercle containing pulp is found on the external surface of a tooth crown. In addition, inherited conditions which show abnormal dentine such as dentine dysplasia, dentinogenesis imperfecta, and osteogenesis imperfecta predispose the dentition to abscess formation. Furthermore, 'spontaneous' dental abscesses are frequently encountered in familial hypophosphataemia, also known as vitamin D-resistant rickets, in which there is hypomineralization of dentine and enlargement of the pulp. In addition to developmental conditions, there are also acquired conditions which may cause unusual dental abscesses,. These include pre-eruptive intracoronal resorption which was previously known as 'pre-eruptive caries' or the 'fluoride bomb'. In addition, some undiagnosed infections associated with developing teeth are now thought to be the mandibular infected buccal cysts which originate from infection of the developing dental follicles. In the present paper, these relatively unknown entities Which cause unusual abscesses in children are reviewed with the aim of updating the general practitioner in their diagnosis and management.

Mast cells and oral inflammation

Mast cells are mobile granule-containing secretory cells that are distributed preferentially about the microvascular endothelium in oral mucosa and dental pulp. The enzyme profile of mast cells in oral tissues resembles that of skin, with most mast cells expressing the serine proteases tryptase and chymase. Mast cells in oral tissues contain the pro-inflammatory cytokine tumour necrosis factor-alpha in their granules, and release of this promotes leukocyte infiltration during evolving inflammation in several conditions, including lichen planus, gingivitis, pulpitis, and periapical inflammation, through induction of endothelial-leukocyte adhesion molecules. Mast cell synthesis and release of other mediators exerts potent immunoregulatory effects on other cell types, while several T-lymphocyte-derived cytokines influence mast cell migration and mediator release. Mast cell proteases may contribute to alterations in basement membranes in inflammation in the oral cavity, such as the disruptions that allow cytotoxic lymphocytes to enter the epithelium in oral lichen planus. A close relationship exists among mast cells, neural elements, and laminin, and this explains the preferential distribution of mast cells in tissues. Mast cells are responsive to neuropeptides and, through their interaction with neural elements, form a neural immune network with Langerhans cells in mucosal tissues. This facilitates mast cell degranulation in response to a range of immunological and non-immunological stimuli. Because mast cells play a pivotal role in inflammation, therapies that target mast cell functions could have value in the treatment of chronic inflammatory disorders in the oral cavity.

Histological alterations in liver and testis of Astyanax aff. bimaculatus caused by acute exposition to zinc

This study investigated the effect of acute exposition to zinc (Zn) on histology of the liver and testes of yellow tail lambari (Astyanax aff. bimaculatus). The exposure consisted of six concentrations of Zn (0, 3, 5, 10, 15, and 20 mg/L) for 96 hours of exposure. Fragments of liver and testis were routinely processed and embedded in plastic resin based on glycol methacrylate. Fragments of bones, muscles, liver and testis were dehydrated and digested to quantify the absorption levels of Zn in the tissue. Acute exposure to concentrations above 10mg/L has produced structural changes in the liver and gonads. The changes found in the liver were vascular congestion; decrease of cellular volume; displacement of the hepatocyte nucleus; necrosis; disarrangement of cordon structure; leukocyte infiltrate and vacuolization. The changes found in the gonads were ruptured cyst, delayed development of germ cells, pyknotic nucleus, cell cluster, displacement of cyst wall and vacuolization. The histological changes observed were compatible with the increasing concentration of zinc in environment, compromising liver and reproductive functions, because there was an increase in relative frequency of hepatocytes and reduced sperm production

Citotoxicidade do ácido peracético: avaliação metabólica, estrutural e de morte em fibroblastos L929

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Molecular epidemiology of Aspergillus collected from cystic fibrosis patients

Background - Aspergillus respiratory infection is a common complication in cystic fibrosis (CF) and is associated with loss of pulmonary function and allergic disease. Methods - Fifty-three Aspergillus isolates recovered from CF patients were identified to species by Internal Transcribed Spacer Region (ITS), β-tubulin, and calmodulin sequencing. Results - Three species complexes (Terrei, Nigri, and Fumigati) were found. Identification to species level gave a single Aspergillus terreus sensu stricto, one Aspergillus niger sensu stricto and 51 Aspergillus fumigatus sensu stricto isolates. No cryptic species were found. Conclusions - To our knowledge, this is the first prospective study of Aspergillus species in CF using molecular methods. The paucity of non-A. fumigatus and of cryptic species of A. fumigatus suggests a special association of A. fumigatus sensu stricto with CF airways, indicating it likely displays unique characteristics making it suitable for chronic residence in that milieu. These findings could refine an epidemiologic and therapeutic approach geared to this pathogen.

Immunodiagnostic confirmation of hydatid disease in patients with a presumptive diagnosis of infection

Information obtained from the routine application of hydatid immunodiagnostic techniques in different clinical situations over a seven-year period is presented. The Immunoelectrophoresis test was used until it was replaced by the simpler, more sensitive and equally specific arc 5 double diffusion (DD5) test. Examination of sera from 1,888 patients with signs and/or symptoms compatible with hydatid disease revealed that the presurgical confirmation of Echinococcus granulosus infection is only obtained by detection of anti-antigen 5 antibodies. The latter were not found in 1,539 presumptive hydatidosis patients whose definitive diagnoses corresponded to other disease conditions. However, false positive latex agglutination test results were obtained in two cases. In all patients whose preoperative serum showed three or more uncharacteristic bands in the absence of anti-antigen 5 antibodies, hydatid cysts were found sur gically. DD5 testing of a fluid sample collected by puncture established its hydatid etiology. Post-operative monitoring of hydatidosis patients demonstrated that persistence of DD5-positivity two years after surgery established the presence of other cysts. Further evidence was obtained in patients with hydatid cysts in intrathoracic, abdominal or other locations associating cyst membrane integrity, antigen release and immunodiagnostic test positivity.

Sedimentation in parasitological coproscopy

A sedimentation technique is described, in which a fecal suspension is placed on top of an aqueous sucrose solution of specific gravity 1.015 g/cm³. Using 100 by 15 mm test tubes, duplicate gravity sedimentation experiments were made using homogenized fecal suspensions (single-columns) and fecal suspensions placed on top of clear columns (double-columns). Egg- and cyst-counts, and turbidity determinations were made in the sediments obtained after definite time intervals. Most Ascaris lumbricoides, Trichuris trichiura and Ancylostomidae eggs sedimented within 20 minutes in single - and between 30 and 60 minutes in double-columns. Giardia duodenalis cysts required longer periods to sediment in double - than in single-columns; after 180 minutes (the maximum period of observation), double-column sediments produced 60.0% of the counts of single-columns. Double-column sediments were consistently less turbid than single-column ones