Postoperative Sensitivity of Composite Replacement of Amalgam Restoration: A Randomized Clinical Trial.

PURPOSE: This randomized clinical trial aimed to evaluate the postoperative sensitivity of different resin composite/adhesive materials, placed either by an incremental or bulk-fill technique in posterior resin composite replacement of amalgam restorations. METHODS AND MATERIALS: A total of 47 patients with good overall health and at least four class I or class II amalgam restorations to be replaced participated in this study. The characteristics of 188 cavities were registered and randomly restored using incremental (Z350XT, 3M) or bulk-fill (Filtek Bulk Fill, 3M; Sonic Fill, Kerr; and Opus Bulk Fill, FGM) technique. The postoperative sensitivity was assessed using a Visual Analog Scale (0-100) after 24 hours, 7 days, and 30 days. Pain scores were temporally analyzed using Friedman test followed by Dunn post hoc test (α=0.05). The frequency of tests was calculated according to the frequency and percentage of the McNemar test. RESULTS: The restorative technique and the bulk-fill system used did not affect the postoperative sensitivity, except for Filtek Bulk Fill group, which presented less postoperative sensitivity after 24 hours (p=0.037). Regardless of the restorative material, the postoperative sensitivity decreased after 24 hours, and no differences were found after 7 and 30 days. CONCLUSIONS: After 1 week, the filling technique and the bulk-fill system have no influence on the postoperative sensitivity.

Phenotype comparison among individuals with developmental delay/intellectual disability with or without genomic imbalances.

BACKGROUND: The chromosomal microarray analysis (CMA) is recommended as a first-tier test for individuals with developmental delay (DD)/intellectual disability (ID) and/or multiple congenital anomalies. However, owing to high costs, this technique is not widely performed for diagnostic purposes in several countries. The aim of this study was to identify clinical features that could favour the hypothesis of genomic imbalances (GIs) in individuals with DD/ID. METHODS: The sample consisted of 63 individuals, and all of them underwent a detailed evaluation by a clinical geneticist and were investigated by the CMA. They were divided into two groups. Group A composed of 20 individuals with pathogenic copy number variants (CNVs); and group B composed of 43 individuals with normal CMA results or variants of uncertain clinical significance (VUS). RESULTS: Pathogenic GIs were found in 20 cases (32%), including 11 individuals with an abnormal karyotype, VUS was found in five individuals (8%) and the results were normal in 38 individuals (60%). Major anomalies were found in 15/20 (75%) individuals in group A against 35/43 (81%) in group B. Dysmorphisms (≥5) were found in 17/20 (85%) in group A and 41/43 (95%) in group B. The most frequent major anomalies detected in group A were congenital heart disease, epilepsy and renal malformation; and in group B, they were malformations of central nervous system, congenital heart disease, microcephaly, epilepsy and hearing impairment. There was no significant statistical difference among the frequencies in groups A and B. CONCLUSIONS: Evidences point that every individual with DD/ID, with no specific clinical suspicion, should have screening for GIs as a first-tier test, regardless of the presence or absence of additional major anomalies or dysmorphisms. Future studies with a similar design would be helpful, especially in countries where the access to new technologies is still limited.

Cell toxicity by ricin and elucidation of mechanism of Ricin inactivation.

Castor cake is a by-product of the extraction of oil from from seeds of castor plants (Ricinus communis). This by-product contains high levels of proteins, but a toxic protein, ricin, limits its use as an animal feed. Ricin can be efficiently inactivated by treatment with calcium oxide (CaO), which can be evaluated by a cytotoxicity assay using LLC-MK2 cells. The mechanism by which the CaO treatment inactivates ricin, however, is unclear. We report the structural changes responsible for ricin inactivation. Purified ricin was treated with 0.6% CaO and then analyzed by mass spectrometry. This treatment degraded the ricin at preferential sites. The aqueous CaO solution had a pH >12, which preferentially cleaved asparagine residues, followed by glutamine, serine and glycine residues. The alkaline pH affected the tertiary structure of the ricin, cleaving its polypeptide chains and thereby eliminating its cytotoxic activity.

Streptococcus pneumoniae infection regulates expression of neurotrophic factors in the olfactory bulb and cultured olfactory ensheathing cells.

Streptococcus pneumoniae is the causative agent of numerous diseases including severe invasive infections such as bacteremia and meningitis. It has been previously shown that strains of S. pneumoniae that are unable to survive in the bloodstream may colonize the CNS. However, information on cellular components and pathways involved in the neurotropism of these strains is still scarce. The olfactory system is a specialized tissue in which olfactory receptor neurons (ORNs) are interfacing with the external environment through several microvilli. Olfactory ensheathing cells (OECs) which also form the glial limiting membrane at the surface of the olfactory bulb (OB) are the only cells that ensheathe the ORNs axons. Since previous data from our group showed that OECs may harbor S. pneumoniae, we decided to test whether infection of the OB or OEC cultures modulates the expression levels of neurotrophic factor's mRNA and its putative effects on the activation and viability of microglia. We observed that neurotrophin-3 (NT-3) and glial cell-line-derived neurotrophic factor (GDNF) expression was significantly higher in the OB from uninfected mice than in infected mice. A similar result was observed when we infected OEC cultures. Brain-derived neurotrophic factor (BNDF) expression was significantly lower in the OB from infected mice than in uninfected mice. In contrast, in vitro infection of OECs resulted in a significant increase of BDNF mRNA expression. An upregulation of high-mobility group box 1 (HMGB1) expression was observed in both OB and OEC cultures infected with S. pneumoniae. Moreover, we found that conditioned medium from infected OEC cultures induced the expression of the pro-apoptotic protein cleaved-caspase-3 and an apparently continuous nuclear factor-kappa B (NF-κB) p65 activation in the N13 microglia. Altogether, our data suggest the possible existence of an OEC-pathogen molecular interface, through which the OECs could interfere on the activation and viability of microglia, favoring the access of non-hematogenous S. pneumoniae strains to the CNS in the absence of bacteremia.

Plasma DYRK1A as a novel risk factor for Alzheimer's disease.

To determine whether apparent involvement of DYRK1A in Alzheimer's disease (AD) pathology makes it a candidate plasma biomarker for diagnosis, we developed a method to quantify plasma DYRK1A by immunoblot in transgenic mouse models having different gene dosages of Dyrk1a, and, consequently, different relative protein expression. Then, we measured plasma DYRK1A levels in 26 patients with biologically confirmed AD and 25 controls (negative amyloid imaging available on 13). DYRK1A was detected in transgenic mouse brain and plasma samples, and relative levels of DYRK1A correlated with the gene copy number. In plasma from AD patients, DYRK1A levels were significantly lower compared with controls (P<0.0001). Results were similar when we compared AD patients with the subgroup of controls confirmed by negative amyloid imaging. In a subgroup of patients with early AD (CDR=0.5), lower DYRK1A expression was confirmed. In contrast, no difference was found in levels of DYRK1B, the closest relative of DYRK1A, between AD patients and controls. Further, AD patients exhibited a positive correlation between plasma DYRK1A levels and cerebrospinal fluid tau and phosphorylated-tau proteins, but no correlation with amyloid-ß42 levels and Pittsburgh compound B cortical binding. DYRK1A levels detected in lymphoblastoid cell lines from AD patients were also lower when compared with cells from age-matched controls. These findings suggest that reduced DYRK1A expression might be a novel plasma risk factor for AD.

Neural correlates of the mini-SEA (Social cognition and Emotional Assessment) in behavioral variant frontotemporal dementia.

Although Frontotemporal Dementia (FTD) is the second most common form of dementia after Alzheimer's disease, its diagnosis remains particularly challenging today. This is particularly true for the behavioral variant (bvFTD), the most common phenotype of FTD, which is characterised by dramatic changes in personal and social conduct. Novel clinical cognitive tests have been recently proposed to diagnose and assess these patients. Among them, the mini-SEA (Social cognition & Emotional Assessment) has shown promising results. This quick clinical tool evaluates emotion recognition and theory of mind deficits, both recognized as hallmark features of bvFTD. In this study, we investigated the neural correlates of the mini-SEA in twenty bvFTD patients, using single photon emission computed tomography (SPECT) and focusing on the mPFC. Results showed that detection of faux pas during a theory of mind evaluation was related to rostral mPFC perfusion (BA 10) while recognition of emotion involved more dorsal regions within the mPFC (BA 9). As significant and early dysfunction of the mPFC has been extensively described in bvFTD, this study supports the use of the mini-SEA in evaluation and diagnosis purposes in bvFTD.

[Markers of prodromal Alzheimer's disease]. / Les marqueurs de la maladie d'Alzheimer prodromale.

The diagnosis of Alzheimer's disease has long been considered a diagnosis of probability, as the definitive diagnosis can only be established by histopathological examination. However, the development of in-vivo biomarkers, considered a reflection of physiopathological processes, has changed our view of the disease. New criteria have recently been proposed that integrate such biomarkers as found in the cerebrospinal fluid (CSF) using new diagnostic tools such as magnetic resonance imaging (MRI), brain scintigraphy, FDG-positron emission tomography (PET) and PET amyloid ligand uptake studies. The value of these new criteria for the diagnosis of prodromal Alzheimer's disease and the prospect of disease-modifying drugs are also discussed.

[Semantic dementia associated with amyotrophic lateral sclerosis]. / Démence sémantique associée à une sclérose latérale amyotrophique.

The association semantic dementia-amyotrophic lateral sclerosis has been rarely reported in the literature. We report a case study of a patient with a severe prosopagnosia in relation with semantic dementia associated, 12 months later, with a motor neuron disease.

Dithiocarbamates residues in Brazilian food and the potential risk for consumers.

Dithiocarbamates are a non-systemic group of pesticides widely used to protect crops from fungal diseases. The current methodology used by monitoring laboratories to determine dithiocarbamates in food involves the analysis of CS(2) generated after hydrolysis of the compound present in the sample. This method does not identify the origin of the CS(2) detected, which may or may not be related to the presence of pesticides leading to a potential overestimation of the dietary dithiocarbamate intake. In this paper, 520 food samples (papaya, banana, apple, strawberry, orange, potato, tomato, rice and dry beans) collected in the local market of the Federal District, Brazil, were analyzed for dithiocarbamate content. Detectable levels (> or =10.10 mg/kg CS(2)) were found in 60.8% of the samples, with the highest levels (up to 3.8 mg/kg) found in strawberry, papaya and banana. No residues were found in rice (polished) and only one dry bean sample had detectable levels of the fungicides. Detectable residues were found in the pulp of banana, papaya (including the seeds) and orange (50-62% of the analyzed samples). An exposure assessment, based on dithiocarbamate levels detected in the food crops analyzed in this study, confirms that the intake of dithiocarbamates through food consumption in the country does not represent a health risk to consumers, i.e., the estimated daily intake is less than the acceptable daily intake. Furthermore, the implementation of more selective methodologies to individually analyze these compounds in food monitoring programs in Brazil is not necessary.

Thermal characteristics of recombinant green fluorescent protein (GFPuv) extracted from Escherichia coli.

AIMS: The thermal stability of isolated and extracted recombinant green fluorescent protein (GFPuv) was evaluated by analysing the loss of fluorescence intensity. METHODS AND RESULTS: GFPuv was expressed by Escherichia coli, extracted by the three-phase partitioning method and purified by elution through an hydrophobic interaction column. The collected fractions were further diluted in Tris-HCl-EDTA (pH 8.0) and subjected to continuous heating at set temperatures (45-95 degrees C). From a standard curve relating fluorescence intensity to GFPuv concentration, the loss of fluorescence intensity was converted to denatured GFPuv concentration (microg ml-1). To determine the extent of the thermal stability of GFPuv, decimal reduction times (D-values), z-value and energy of activation (Ea) were calculated. CONCLUSIONS: For temperatures between 45 and 70 degrees C, extracted native GFPuv activity decreased from 11 to 75% relative to initial native protein concentration above 70 degrees C, the average decrease in GFPuv fluorescence was between 72 to 83%. SIGNIFICANCE AND IMPACT OF THE STUDY: The thermal stability of GFPuv provides the basis for its potential utility as a fluorescent biological indicator to assess the efficacy of the treatment of liquids and materials exposed to steam.

Determination of dithiocarbamate fungicide residues in food by a spectrophotometric method using a vertical disulfide reaction system.

Dithiocarbamates are a class of fungicides extensively used in many crops worldwide. The current residue definition of dithiocarbamates in food for compliance with maximum residue limits, at national and international levels, is total residues arising from the use of any or each dithiocarbamate fungicide, determined as CS(2). The analytical method most frequently used to analyze dithiocarbamate residues in food for monitoring purposes was proposed more than 30 years ago. In this method, total dithiocarbamates are decomposed to CS(2), which is purified and reacted with a cupric reagent. The yellow complex formed is quantified by spectrophotometry. In this paper, a new reaction system for the purification and complexation of CS(2) is proposed. The new system is less fragile than the traditional design, is easier to assemble, and allows for a higher sample throughput, in addition to being of low cost. Recovery of added mancozeb, thiram, or ziram (0.15-8.0 mg/kg) in rice, beans, apple, banana, orange, papaya, tomato, cucumber, and potato ranged from 82 to 120%, with relative standard deviations from 0 to 10% (n = 3 or 5). Analysis of apple, tomato, and papaya samples with field-incurred dithiocarbamate residues showed comparable results using both the traditional and the new reaction systems.

Influence of hypoxia on nitric oxide synthase activity and gene expression in children with congenital heart disease: a novel pathophysiological adaptive mechanism.

BACKGROUND: Chronic hypoxia has been shown to modulate nitric oxide (NO) responses in different cell models, but the relationship between hypoxia and NO synthase (NOS) regulation in humans was not studied. We studied the relationship between endothelial and inducible NOS (eNOS and iNOS) activities and expression and chronic hypoxia in children with cyanotic and acyanotic congenital heart defects. METHODS AND RESULTS: Right atrial tissue was excised from 18 patients during cardiac surgery. eNOS and iNOS activities were measured by conversion of L-[H(3)]arginine to L-[H(3)]citrulline. Gene expression of eNOS and iNOS was quantified by competitive reverse transcription-polymerase chain reaction. The eNOS activity and expression were significantly reduced in cyanotic hearts compared with acyanotic hearts: 0.38+/-0.14 versus 1.06+/-0.11 pmol. mg(-1). min(-1) (P<0.0001) and 0.54+/-0.08 versus 0.80+/-0.10 relative optical density (ROD) of cDNA (P<0.0001), respectively. In contrast, iNOS activity and expression were significantly higher in cyanotic than in acyanotic children: 7.04+/-1.20 versus 4.17+/-1.10 pmol. mg(-1). min(-1) (P<0.0001) and 2.55+/-0.11 versus 1.91+/-0.18 ROD of cDNA (P<0.0001), respectively. CONCLUSIONS: Hypoxia downregulates eNOS activity and gene expression in cardiac tissue from patients with cyanotic congenital heart defects. By contrast, iNOS activity and expression are increased in cyanotic children and may represent an alternative mechanism to counteract the effects of hypoxia in the cardiovascular system. Therefore, a novel adaptive mechanism during hypoxia is suggested.

[Assessment of the chronic risk for ingestion of pesticide residues in the Brazilian diet]. / Avaliação de risco crônico da ingestão de resíduos de pesticidas na dieta brasileira.

OBJECTIVE: To conduct a chronic dietary risk assessment of the pesticides registered in Brazil up until 1999. METHODS: The Theoretical Maximum Daily Intake (TMDI) for each pesticide was calculated using the Brazilian maximum residue limits and food consumption data from IBGE, the Brazilian Statistical Institute. The risk characterization was done comparing the TMDI with the acceptable daily intakes (ADI) from other countries and from the Codex Alimentarius. RESULTS: The TMDI was higher than the ADI (%ADI>100) at least in one Brazilian metropolitan region for 23 pesticides. Sixteen compounds are organophosphate insecticides, with methyl parathion having the TMDI exceeding the most toxicological parameter (%ADI N=9,300). Rice, beans, citrus and tomato were the commodities which most contributed to the ingestion. From the compounds under higher risk, only 6 were registered according to the Law 98.816/90, which concerns the use of pesticides in the country. CONCLUSIONS: The compounds identified in the study as presenting a potential health concern to the Brazilian consumers, and the commodities which most contributed to the ingestion, should be prioritized by the government in pesticide residue monitoring programs and in the re-registration process. In addition, residue data in food as consumed, processing factors and appropriate consumption data should be generated to allow further studies.

Evaluation of pH and calcium ion release of three root canal sealers.

The pH and calcium ion release of three root canal sealers--Sealapex, Sealer 26, and Apexit--were assessed at 24 and 48 hr, and at 7 and 30 days after spatulation. After manipulation, the sealers were placed in 1-cm-long tubes measuring 4 mm in diameter and immersed in a glass flask containing 10 ml of deionized water, which was hermetically sealed and stored at 37 degrees C. The tubes were removed at the scheduled times and the water in which they were immersed was tested for pH with a pHmeter and for released calcium by atomic absorption spectrophotometry. Starting 48 hr after immersion, Selapex produced an alklaine pH and released significantly higher calcium amounts compared with the other two sealers, with even more pronounced results after 30 days. On the basis of the results obtained, we conclude that Sealapex presented the highest calcium and hydroxyl release, especially after longer time intervals, whereas Sealer 26 showed highest release during the initial periods (i.e. during its setting time). Apexit presented the least satisfactory results.

The flexible endaural tympanoplasty: pathology-guided, pathogenesis-oriented surgery for the middle ear.

The flexible approach to tympanoplasty has been found to be adaptable to various forms of pathologic conditions found in the temporal bone, including inflammation and infection, congenital stenosis, benign and malignant tumors, and traumatic injuries. This approach finds its best indication among all pathologic conditions in the temporal bone, in the surgical treatment of otitis media, and its sequelae. A procedure conceived to treat this dynamic process must be adaptable to new circumstances and new findings and be ready to manage unexpected situations. The flexible tympanoplasty is a step-wise approach designed to explore the contents of the middle ear methodically and in the process disclose, confirm, and often treat disease.

Incidence of leptospiral abortion in Brazilian dairy cattle.

We examined dams' paired serum samples and foetal kidneys (histopathologically and attempting leptospiral cultures) from 120 Brazilian Holstein abortions from 10 herds near Sao Paulo, Brazil, in 1996. Leptospiras was isolated from 15 foetuses. The Leptospira interrogans serovar hardjo was obtained from four, pomona from three and wolffi from eight foetuses. Ten of these 15 foetuses showed a four-fold rise in titres between the day of abortion and the second samples taken 15 days later. Fifty-seven other foetuses had histologic evidence of leptospires; 27 of their dams (47%) had four-fold rises in titres. In total, 72 of 120 aborted foetuses had evidence of leptospiral infection.

[Perinatal deaths in the city of Fortaleza: how many deaths could be prevented?]. / Mortes perinatais no município de Fortaleza, Ceará: o quanto é possível evitar?

OBJECTIVE: The main objective of this study was to explore the relationship between perinatal care and neonatal mortality as a component of the infant mortality. Indicators were selected to analyze some aspects of the quality of perinatal care attendance offered to pregnant women and their newborns. Specific attention was given to the avoidable perinatal deaths in the city of Fortaleza, Ceará. METHODS: Population-based, prospective study on incidence. The study was carried out collecting data of seventeen maternity units and two referral pediatric hospitals of the public health system in Fortaleza, the capital of the State of Ceará. All births, alive or dead, weighting 500 grams or more, along the whole year of 1995, were included in the study. The perinatal deaths' causes were studied according to the modified Wigglesworth classification (1989). RESULTS: Along the whole study period, 40,712 children were born and 1,337 perinatal deaths occurred. Among them 730 occurred during the fetal life and 607 during the first week of life. In spite of the low incidence of low-weight newborns (7.4%), the coefficients of fetal mortality were high (17.9 per thousand), as well as the perinatal mortality (32.8 per thousand) and neonatal mortality (15.1 per thousand). The number of avoidable perinatal deaths related to adequate perinatal care was estimated to be one third of the total deaths studied (458/1,323). CONCLUSIONS: The results indicate that the perinatal assistance in the city of Fortaleza is of low quality, with problems related to the health system organization and to the low quality of the care offered to women during pregnancy, at delivery and to the newborns in the delivery room as well as in neonatal wards.

[Salmonellas and fecal coliforms in drinking water for animals]. / Salmonelas e coliformes fecais em águas de bebida para animais.

Sixty small farms of Botucatu County, S. Paulo, Brazil, given over to cattle breeding were studied. These farms account for 15% of the total of 402 farms that exist in the county. The sample was drawn by simple probabilistic technique. There were found to be one hundred and thirteen drinking places located on the farms. Samples of water were taken from these drinking places and examined for bacteria of the genus Salmonella, for the determination of Most Probable Number (MNP) of fecal coliform bacteria as well as determination of the water's pH. Water temperature was measured before collection. Samples from 15 drinking places (13.3%) were positive for Salmonella. The drinking places belonged to 12 of the sixty farms studied (20%). The following serotypes were identified: S. dublin, S. newport, S. madelia, S. IV 43:g,z57:-, S. saphra, S. glostrup, S. IV ochsenzool; S. I9,12:i:- and two new serotypes S. IV 41:z52:- and S. IV 50:d:-. Of the 113 samples studied 14 (12.4%) presented MPN/100ml of fecal coliforms above 4,000. There was no relationship between MPN/100ml of fecal coliforms above 4,000 and positivity for Salmonella. Highest positivity both for Salmonella and MPN/100mL of fecal coliforms over 4,000 occurred at temperatures above 18 degrees C. As regards pH, in both situations the highest positivity occurred between 6.0 and 7.0.

[Surgical management of the aortic valve in patients older than 70 years of age]. / Manuseio cirúrgico da valva aórtica em pacientes acima de 70 anos.

PURPOSE: To assess the short and long-term benefits of patients who were submitted to isolated aortic valve replacement or valve replacement (VR) concomitant myocardial revascularization (MR); to evaluate the incidence of postoperative complications, hospital mortality and late mortality. METHODS: From January 1985, through December 1989, 20 consecutive patients underwent surgical intervention, 15 male (75%) and 5 female (25%), the mean age was 74.8% (ranging from 70 to 86 years old), and the aortic valve gradient ranged between 78 and 180 mmHg (mean = 97 mmHg). They presented preoperative diagnosis to have either isolated aortic stenosis (AS) or As and coronary artery disease (CAD). No patient was in NYHA functional class I; 3 patients (15%) were in class II, 14 (70%) in class III and 3 (15%) in class IV. RESULTS: The most frequent post-operative complications found were: extended intubation in 7 patients (35%), bleeding in 4 (20%), acute renal failure in 3 (15%) and ventricular arrhythmia in 3 (15%). Hospital mortality occurred in 2 patients (10%) who had been submitted to VR and concomitant MR. Late mortality occurred in 1 patient (5%). Through December 1989, 11 patients (64%) were in functional class I (NYHA), 3 (18%) in class II, 3 (18%) in class III and none in class IV. CONCLUSION: We concluded that the surgical treatment is indicate to elderly patients with isolated AS os with AS and concomitant CAD. There was a significant post-operative improvement of the functional class (NYHA) to the surviving patients.