Quantitative analysis of culturable bacteria, levels of endotoxins, inflammatory mediators and substance P in teeth with symptomatic irreversible pulpitis and in teeth with vital normal pulp tissues.

AIM: To comparatively analyse the levels of culturable bacteria, endotoxins (LPS), tumour necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1ß) and substance P in teeth with symptomatic irreversible pulpitis (SIP) and vital normal pulp (VNP) tissues. METHODOLOGY: Thirty-two patients were included (20 teeth with SIP and 12 teeth with VNP tissues) in this cross-sectional study. Samples were collected from the full length of the root canals (microbial analysis) and periapical tissues (2 mm beyond the apex for immunological analysis), using sterile absorbent paper points. The levels of culturable bacteria (culture method), endotoxins (LAL Pyrogent 5000), TNF-α, IL-1ß and substance P (ELISA) were assessed. The Mann-Whitney test was used for comparisons between the levels of CFU/mL, LPS, TNF-α, IL-1ß and substance P in the SIP and VNP groups. The statistical analysis was performed with the significance level set at 5%. RESULTS: Culturable bacteria were recovered from all teeth with SIP. On the other hand, no positive cultures were observed in the VNP tissues group (p > .05). The levels of LPS were approximately four times higher in teeth with SIP than in teeth with VNP tissues (p < .05). Higher levels of TNF-α and substance P were detected in teeth with SIP (p < .05). On the other hand, no difference in the levels of IL-1ß was detected between the two groups (p > .05). CONCLUSION: Teeth with symptomatic irreversible pulpitis present higher levels of culturable bacteria, endotoxins, TNF-α and substance P than those with vital normal pulp tissues. On the other hand, the levels of IL-1ß were similar in teeth from both groups suggesting reduced implications of this inflammatory mediator in the early stages of infection.

Investigation of microbial profile, levels of endotoxin and lipoteichoic acid in teeth with symptomatic irreversible pulpitis: a clinical study.

AIM: To investigate the microbial profile, and levels of endotoxin (LPS) and lipoteichoic acid (LTA), in infected dentine (ID) and root canals (RC) at different phases of root canal treatment in teeth with symptomatic irreversible pulpitis. METHODOLOGY: Ten volunteers were included, and samples were collected from infected dentine (ID) and the root canal lumen (RC) using sterile excavators and paper points, respectively. RC samples were taken before (S1) and after (S2) chemo-mechanical canal preparation (CMP), and after intracanal medication (ICM; S3). Checkerboard DNA-DNA hybridization was used for microbial analysis. The levels of LPS and LTA were evaluated using the limulus amebocyte lysate assay and ELISA, respectively. Shapiro-Wilk's test was used to verify data normality. Friedman's test was used to evaluate statistical differences using checkerboard DNA-DNA hybridization in the ID and RC at the different phases of the RC treatment. Post hoc Dunn's multiple comparison test was used to verify significant differences recorded at the different time-points. The levels of LPS and LTA were analysed statistically by using repeated measures anova and Tukey's post hoc test to evaluate differences in both sites. The significance level was set at 5% (P < 0.05). RESULTS: A total of 40 DNA probes were used for microbial investigation of ID and RC samples using checkerboard DNA-DNA hybridization. The levels and complexity of bacteria were similar in the ID and initial RC samples. The levels of LPS and LTA in ID were significantly higher than the initial RC samples (S1; P < 0.05). Canal preparation was effective in significantly decreasing the levels of bacteria, LPS and LTA (P < 0.05). ICM did not provide additional reduction in the levels of bacteria and LPS (P > 0.05). However, a significant reduction in the levels of LTA was observed after ICM (P < 0.05). CONCLUSION: The microbial profile of infected dentine and root canals of teeth with irreversible pulpitis was complex, harbouring different species including Gram-positive and Gram-negative, cocci and bacilli, and facultative and strict anaerobes. Root canal preparation was effective in reducing the levels of bacteria, LPS and LTA from the root canals of teeth with pulpitis.

Lateral and axial cutting efficiency of instruments manufactured with conventional nickel-titanium and novel gold metallurgy.

AIM: To isolate the effect of metallurgy in lateral and axial cutting efficacy against plastic and bovine dentine substrates by comparing two rotary systems with identical design but manufactured with either conventional nickel-titanium or heat-treated gold alloy. METHODOLOGY: A total of 258 ProTaper Universal (PTU) and ProTaper Gold (PTG) Shaping instruments were used. Bending behaviour was assessed to determine the appropriate displacement associated with a 2 N force in lateral cutting. Ten instruments of each type were used in lateral action for 60 s against bovine dentine or plastic substrates four consecutive times producing four notches in each specimen. Ten further instruments of each type were used in on axial action in four standardized simulated root canals fabricated from 4-mm thick plastic or dentine discs. Both tests were performed at 300 rpm in a computer-controlled testing platform. Notch area and torsional load were compared with Student's t-tests. Repeated measures ANOVA was used to compare cutting efficiency across the four different time-points. Pearson correlation coefficients between substrates were also determined. RESULTS: For lateral action, all three PTG instruments cut significantly more effectively (P < 0.05) than PTU on the plastic substrate. S1 and S2 PTG cut significantly more after 120 and 180 s (P < 0.05) on bovine dentine substrate. For axial action, S1 and S2 PTG were significantly more efficient in cutting at 180 s on plastic and 120 s on bovine dentine (P < 0.05). CONCLUSION: Instruments made from heat-treated nickel-titanium gold alloy had equal or greater cutting efficiency when compared to those made from conventional nickel-titanium.

Ion chambers compliance results of Brazilian radiation therapy facilities.

The Brazilian Nuclear Energy Commission (cnen) has been making a constant effort to keep up to date with international standards and national needs to strengthen the status of radiological protection of the country. The guidelines related to radiation therapy facilities have been revised in the last five years in order to take into consideration the most relevant aspects of the growing technology as well as to mitigate the accidents or incidents observed in practice. Hence, clinical dosimeters have gained special importance in this matter. In the present work, we discuss the effectiveness of regulation and inspections to the enforcement of instrument calibration accuracy for the improvement of patient dosimetry and quality control. As a result, we observed that the number of calibrated instruments, mainly well chambers, is increasing each year. The same behavior is observed for instruments employed in technologically advanced radiation treatments such as intensity modulated radiotherapy, volumetric therapy and stereotatic radiosurgery. We ascribe this behavior to the new regulation.

Interleukin-10 and tumour necrosis factor-alpha serum levels in chronic Chagas disease patients.

In Chagas disease, chronically infected individuals may be asymptomatic or may present cardiac or digestive complications, and it is well known that the human immune response is related to different clinical manifestations. Different patterns of cytokine levels have been previously described in different clinical forms of this disease, but contradictory results are reported. Our aim was to evaluate the serum levels of interleukin-10 and tumour necrosis factor-alpha in patients with asymptomatic and cardiac Chagas disease. The serum interleukin-10 levels in patients with cardiomyopathy were higher than those in asymptomatic patients, mainly in those without heart enlargement. Although no significant difference was observed in serum tumour necrosis factor-alpha levels among the patients, we found that cardiac patients also present high levels of this cytokine, largely those with heart dilatation. Therefore, these cytokines play an important role in chronic Chagas disease cardiomyopathy. Follow-up investigations of these and other cytokines in patients with chronic Chagas disease need to be conducted to improve the understanding of the immunopathology of this disease.

Cardiac lesions in 30 dogs naturally infected with Leishmania infantum chagasi.

The hearts of 30 dogs naturally infected with Leishmania infantum chagasi were evaluated histologically and immunohistochemically. Myocardial lesions were detected in all dogs, including lymphoplasmacytic myocarditis (27/30), myonecrosis (24/30), increased interstitial collagen (22/30), lepromatous-type granulomatous myocarditis (7/30), fibrinoid vascular change (3/30), and vasculitis (1/30). The parasite was detected in the hearts of 20 of 30 dogs. The number of parasitized cells correlated with the intensity of the inflammation and with the number of granulomas. The results indicate that cardiac lesions are prevalent in dogs with naturally occurring leishmaniasis even in the absence of clinical signs of cardiac disease.

Occupational injuries in children and adolescents in emergency services of Aracaju, Brazil.

BACKGROUND: The incidence of occupational injuries among children and adolescents is not well known in Brazil. This study aims to identify occurrences and characteristics of occupational injuries in children and adolescents at the two major public emergency hospitals in Aracaju, Brazil. METHODS: In a cross sectional study, all children and adolescents (5-17 years) attending the services because of accidents of external cause in October-November 2006 and May-June 2007 were evaluated. The patients and their responsibles were interviewed to obtain information about the injury, family and patient characteristics and relation to work in the last 7 days. Data were analysed in number and percentages using descriptive statistics and chi-squared and Fisher's exact tests. RESULTS: There were 917 patients with injuries of external cause in the period studied. Seventy-one (8%) had worked in the last 7 days and in 40 (4%) the injury was related to work, with 2 (0.21%) deaths. Occupational injuries occurred more frequently in boys (95%); in the 14- to 17-year-old age group (77%); in agricultural (46%) and commercial activities (18%); to help with family budget (54%); without any safety training (73.5%) and not using personal protective equipment (94%). Main mechanisms of injuries were transportation incidents (42.5%) and contact with tools and equipment (27.5%). Injuries were mostly fractures (37.5%) and lacerations (22.5%). Upper extremities were injured in 49% of cases and the lower extremities in 26.5%. Comparison between the two groups (accident related or not related to labour) shows that there were significant differences for gender, age group, the main mechanism of injury and the part of the body injured, but not for the nature of injury. CONCLUSION: Working children and adolescents have been injured during their occupational activities and work-related injuries represent 4% of injuries of external causes in the emergency departments of Aracaju with 5% lethality.

Leukocyte entry into the CNS of Leishmania chagasi naturally infected dogs.

In dogs, there is an association of chronic visceral leishmaniasis with neurological symptoms, and very few publications have investigated whether these neurological manifestations correlate with specific alterations in brain. A total of 42 mixed-breed adult dogs were selected from the Veterinary Hospital of UNESP-Araçatuba and the Control Zoonosis Center in Araçatuba, São Paulo State, Brazil, which is an endemic area for visceral leishmaniasis. Animals presenting positive ELISA and/or positive parasitological diagnosis of Leishmania were enrolled in the group of infected dogs (n=32). Animals with negative ELISA results and parasitological tests for Leishmania, including a negative immunofluorescence test for toxoplasmosis and neosporosis, were included as the control group (n=10). Brain samples were collected, stored in 10% buffered formalin and subjected to routine histological procedures, following by staining with haematoxylin-eosin (HE) and immunohistochemical examination for T and B lymphocytes and phagocytic cells. Cerebrospinal fluid was collected to determine the anti-Leishmania antibody titers. Histological examination of HE stains demonstrated intense inflammatory infiltrate, primarily in the choroid plexus, which was composed of mononuclear cells with no detectable parasites. Immunohistochemistry revealed that CD3(+) T lymphocytes were the major components of the inflammatory infiltrate at the choroid plexus and in the brain. Infected dogs had more CD3(+) T cells than uninfected animals (P=0.0002). Cerebrospinal fluid from infected dogs contained high titers of anti-Leishmania antibodies in comparison with control animals (P<0.0001), which suggests a compromise of the blood-cerebrospinal fluid barrier. Leukocyte entry into the brain suggests the participation of these cells in the pathogenesis of neurological disorders during the advanced stages of leishmaniasis and confirms that the choroid plexus is an important structure for T cell influx.

Effect of cryopreservation on sperm DNA fragmentation and apoptosis rates in the testicular tissue of domestic cats.

The objectives of the present study were to evaluate the damage caused by cryopreservation on sperm DNA and estimate the percentage of cell apoptosis in tissue after thawing. Testicles of cats were sectioned into of 0.3 cm3 and 0.5 cm3 fragments and evaluated for DNA damage using acridine orange and semi-quantitatively through histo-morphological and immunohistochemical methods (caspase-3). Other fragments were placed in cryotubes with diluent containing either 3% glycerol or 3% propanediol, and were cryopreserved. Evaluation using acridine orange indicated there was a difference with use of propanediol and glycerol on DNA damage in 0.5 cm3fragments, with the latter being more effective than the former for cryopreservation. Results from histomorphological evaluations indicated there was a greater cell integrity among germ cells that were not cryopreserved, based on criteria assessed (detachment of cells from basal membrane, retraction of seminiferous tubule epithelium, visibility of the spermatogonia nucleoli and nuclear spermatogonia condensation), for both sizes of fragments. The values for these variables decreased after cryopreservation, with there being no differences as a result of size of fragment stored or between cryoprotectants used (P > 0.05). The staining for caspase-3 differed for the cytoplasm, nuclei and germ cells. Assessment of these staining patterns indicated the fresh fragments had an amount of cell damage and there was a similar amount of damage detected in cryopreserved fragments. This finding indicated that there was considerable efficacy in preserving the tissue fragments with use of the freezing protocols that were evaluated in this study.

Assessing anthropogenic pressures on estuarine fish nurseries along the Portuguese coast: a multi-metric index and conceptual approach.

Estuaries are among the most productive ecosystems and simultaneously among the most threatened by conflicting human activities which damage their ecological functions, namely their nursery role for many fish species. A thorough assessment of the anthropogenic pressures in Portuguese estuarine systems (Douro, Ria de Aveiro, Mondego, Tejo, Sado, Mira, Ria Formosa and Guadiana) was made applying an aggregating multi-metric index, which quantitatively evaluates influences from key components: dams, population and industry, port activities and resource exploitation. Estuaries were ranked from most (Tejo) to least pressured (Mira), and the most influential types of pressure identified. In most estuaries overall pressure was generated by a dominant group of pressure components, with several systems being afflicted by similar problematic sources. An evaluation of the influence of anthropogenic pressures on the most important sparidae, soleidae, pleuronectidae, moronidae and clupeidae species that use these estuaries as nurseries was also performed. To consolidate information and promote management an ecological conceptual model was built to identify potential problems for the nursery function played by these estuaries, identifying pressure agents, ecological impacts and endpoints for the anthropogenic sources quantified in the assessment. This will be important baseline information to safeguard these vital areas, articulating information and forecasting the potential efficacy of future management options.

Host-Microbiome Cross-talk in Oral Mucositis.

Oral mucositis (OM) is among the most common, painful, and debilitating toxicities of cancer regimen-related treatment, resulting in the formation of ulcers, which are susceptible to increased colonization of microorganisms. Novel discoveries in OM have focused on understanding the host-microbial interactions, because current pathways have shown that major virulence factors from microorganisms have the potential to contribute to the development of OM and may even prolong the existence of already established ulcerations, affecting tissue healing. Additional comprehensive and disciplined clinical investigation is needed to carefully characterize the relationship between the clinical trajectory of OM, the local levels of inflammatory changes (both clinical and molecular), and the ebb and flow of the oral microbiota. Answering such questions will increase our knowledge of the mechanisms engaged by the oral immune system in response to mucositis, facilitating their translation into novel therapeutic approaches. In doing so, directed clinical strategies can be developed that specifically target those times and tissues that are most susceptible to intervention.

Impact of Model for End-stage Liver Disease Score in Post-Liver Transplantation Prognosis.

Although the Model for End-Stage Liver Disease (MELD) score has been used to evaluate mortality of patients on the waiting list for liver transplantation, it has not established if it correlates with post-liver transplantation prognosis. The aim of this study is to assess if there is a relationship between pretransplantation MELD and post-transplantation results, especially regarding 30-day post-LT survival rates. We analyzed data from 172 patients who underwent deceased-donor liver transplantation between February 2013 and May 2015 in a single center. The sample was divided in two groups according to their MELD scores: group 1, with MELD <25 (n = 134), and group 2, with MELD ≥25 (n = 38). The groups were compared with regard to 1-, 3-, and 12-month postoperative survival; total length of stay and in intensive care unit; and perioperative blood transfusion. The global 30-day post-liver transplantation survival rate was 87.2% (89.55% for group 1 and 78.99% for group 2 (P = .090). The evaluation of 3- and 12-month survival showed that in group 2 the mortality was higher (P = .01). Our data suggests a positive relationship between the higher MELD group and longer length of stay in intensive care unit (P = .0345), but not for total time of hospital admission (P = .524). Perioperative blood product transfusion was needed in 35.82% of patients in group 1 and 71.05% in group 2; this correlated with a higher 30- day postoperative mortality (P < .001).

Erratic migration of Ascaridia hermaphrodita (Nematoda: Ascaridida) in a blue-fronted Amazon parrot (Amazona aestiva) from illegal wildlife trade: case report / [Migração errática de Ascaridia hermaphrodita (Nematoda: Ascaridida) em papagaio-verdadeiro (Amazona aestiva) oriundo do comércio ilegal: relato de caso]

Ascaridiasis by Ascaridia hermaphrodita is a common parasitic disease in blue-fronted Amazon parrots (Amazona aestiva) and the contamination occurs by ingestion of eggs. Illegal trade of wild birds can increase the frequency of visceral erratic migration by this parasite. Hence, this report describes the erratic migration of A. hermaphrodita and associated lesions in a blue-fronted Amazon parrot. This is a retrospective study, which originated from several apprehension of illegal birds in the state of Paraíba, Brazil. Necropsy revealed intense intestinal ascarid infestation. Hepatic lesions were also observed associated with the parasite migration. Stereoscopy allowed the identification of the morphology of the parasites. Summarizing, this condition in wild birds is not fully understood and may be associated with illegal trade.(AU)

Ascaridíase por Ascaridia hermaphrodita é uma doença parasitária comum em papagaios-verdadeiros. Sua contaminação ocorre pela ingestão de ovos. O tráfico de aves silvestres pode facilitar a migração errática visceral desse parasita. Este relato descreve a migração errática de A. hermaphrodita e lesões associadas em papagaio-verdadeiro. Trata-se de um relato retrospectivo, analisado a partir de uma série de apreensões de aves no estado da Paraíba, Brasil. A necropsia revelou intensa infestação ascarídea intestinal. Também foram observadas lesões hepáticas em uma ave, associadas à migração do parasita. A estereoscopia permitiu distinguir a morfologia dos parasitas. Resumidamente, essa condição em aves silvestres é pouco conhecida e pode ser considerada, inclusive, associada à comercialização ilegal.(AU)

Experimental infection of chickens by a flagellated motile strain of Salmonella enterica serovar Gallinarum biovar Gallinarum.

Salmonella enterica subsp. enterica serovar Gallinarum biovar Gallinarum (SG) causes fowl typhoid (FT), a septicaemic disease which can result in high mortality in poultry flocks. The absence of flagella in SG is thought to favour systemic invasion, since bacterial recognition via Toll-like receptor (TLR)-5 does not take place during the early stages of FT. In the present study, chicks susceptible to FT were inoculated with a wild type SG (SG) or its flagellated motile derivative (SG Fla(+)). In experiment 1, mortality and clinical signs were assessed, whereas in experiment 2, gross pathology, histopathology, systemic invasion and immune responses were evaluated. SG Fla(+) infection resulted in later development of clinical signs, lower mortality, lower bacterial numbers in the liver and spleen, and less severe pathological changes compared to SG. The CD8(+) T lymphocyte population was higher in the livers of chicks infected with SG at 4 days post-inoculation (dpi). Chicks infected with SG had increased expression of interleukin (IL)-6 mRNA in the caecal tonsil at 1 dpi and increased expression of IL-18 mRNA in the spleen at 4 dpi. In contrast, the CD4(+) T lymphocyte population was higher at 6 dpi in the livers of birds infected with SG Fla(+). Therefore, flagella appeared to modulate the chicken immune response towards a CD4(+) T profile, resulting in more efficient bacterial clearance from systemic sites and milder infection.

Identification of a novel R21X mutation in the liver-type arginase gene (ARG1) in four Portuguese patients with argininemia.

Argininemia is a rare autossomal recessive disorder caused by deficiency in the cytosolic liver-type arginase enzyme (L-arginine urea-hydrolase; E.C. 3.5.3.1). In order to investigate the molecular basis for argininemia in four unrelated Portuguese patients (two from northern Portugal and two from Madeira Island) we performed a DNA sequence analysis of all the exons and exon/intron boundaries of the liver-type arginase gene (ARG1). All patients were found to be homozygous for a newly identified C ->T transition in codon 21 (exon 2) substituting arginine for a premature stop codon (R21X: CGA to TGA) and generating a NlaIII restriction site. Restriction digestion following PCR amplification of ARG1 exon 2 confirmed the presence of the mutation.

Delayed evoked potentials in children exposed to methylmercury from seafood.

Methylmercury poisoning may cause constriction of visual fields and deafness, especially if exposure occurs prenatally. However, the risks associated with exposure from contaminated seafood is unclear. We examined 149 children attending first grade in a Madeiran fishing community. As maternal dietary habits were relatively unchanged, current maternal hair concentrations were used as indicator of the child's prenatal exposure to methylmercury (geometric average, 9.64 microg/g [48.2 nmol/g]). After adjustment for age and sex, the mean (+/-SD) latency of peak III of the brainstem auditory evoked potentials at 40 Hz was increased by 0.128+/-0.047 ms when maternal hair-mercury concentrations exceeded 10 microg/g (50 nmol/g) (p for association, 0.002), and the increase of the N145 pattern-reversal visual evoked potential latency at 15 minutes of arc was 3.16+/-1.57 ms (p for association, 0.002). No such relationships were seen with the child's own hair-mercury concentration, and other clinical examinations revealed no mercury-associated deficits. Neurophysiological evidence of adverse effects on brain function are relatively independent of confounders, and should be considered in the risk assessment of this seafood pollutant.

Retrospective analysis of 15 cases of nasolabial cyst.

The nasolabial cyst is classified as a nonodontogenic cyst and is the only nonodontogenic cyst to occur in soft tissue. The aim of this retrospective analysis was to gather demographic, clinical, radiographic, and histopathologic data on the nasolabial cysts described in the files of the Oral Pathology Laboratory of Minas Gerais University School of Dentistry. Fifteen cases were included in the analysis. The mean age of the patients at the time of diagnosis was 42 years, and there was a female predilection. All the cysts were asymptomatic. Pseudostratified columnar (respiratory) epithelium, alone or in combination with other types of epithelium, was present in all cysts. Recurrence was not observed after surgical excision.

[Ovarian pregnancy. Presentation of a case and review of the literature]. / Embarazo ovárico. Presentación de un caso y revisión de la literatura.

Ovarian pregnancy is a rare phenomenon with an incidence of 1 case in 7000 deliveries. We present a case implanted in the corpus luteum and discuss the possible etiology and the classification. We also review the diagnostic problems that it presents, and the advances made with transvaginal ultrasonography and hCG determinations in urine. The surgical management is presented, with special interest in conservative procedures.

Modeling fish biological responses to contaminants and natural variability in estuaries.

Understanding the factors that influence biological responses to contaminants has long been a major goal in marine environmental research. Seven estuarine sites along the Portuguese coast were sampled over a year, and different biological responses of Pomatoschistus microps and Atherina presbyter were determined: superoxide dismutase, catalase, ethoxyresorufin O-deethylase, glutathione S-transferase, metallothioneins, lipid peroxidation, RNA:DNA ratio and condition factor K. Generalized linear models (GLM) were developed for each biological variable per species in relation to sediment chemical characterization (metals and polycyclic aromatic hydrocarbons concentration) and environmental conditions (month, site, water temperature, salinity, depth and mud percentage in the sediment). GLM varied in explanatory power and in the set of predictor variables included in the models. Environmental factors were frequently selected as predictor variables. Individual metals concentration and sediment quality guidelines (integrating all metals) were the major contaminants explaining biological variability. Accordingly, models for metallothioneins and lipid peroxidation had highest explanatory power. Species-specific responses and dataset size were the basis of observed differences between GLM for the two species.