Accuracy of computer-aided ultrasound as compared with magnetic resonance imaging in the evaluation of nonalcoholic fatty liver disease in obese and eutrophic adolescents

AbstractObjective:To compare the accuracy of computer-aided ultrasound (US) and magnetic resonance imaging (MRI) by means of hepatorenal gradient analysis in the evaluation of nonalcoholic fatty liver disease (NAFLD) in adolescents.Materials and Methods:This prospective, cross-sectional study evaluated 50 adolescents (aged 11–17 years), including 24 obese and 26 eutrophic individuals. All adolescents underwent computer-aided US, MRI, laboratory tests, and anthropometric evaluation. Sensitivity, specificity, positive and negative predictive values and accuracy were evaluated for both imaging methods, with subsequent generation of the receiver operating characteristic (ROC) curve and calculation of the area under the ROC curve to determine the most appropriate cutoff point for the hepatorenal gradient in order to predict the degree of steatosis, utilizing MRI results as the gold-standard.Results:The obese group included 29.2% girls and 70.8% boys, and the eutrophic group, 69.2% girls and 30.8% boys. The prevalence of NAFLD corresponded to 19.2% for the eutrophic group and 83% for the obese group. The ROC curve generated for the hepatorenal gradient with a cutoff point of 13 presented 100% sensitivity and 100% specificity. As the same cutoff point was considered for the eutrophic group, false-positive results were observed in 9.5% of cases (90.5% specificity) and false-negative results in 0% (100% sensitivity).Conclusion:Computer-aided US with hepatorenal gradient calculation is a simple and noninvasive technique for semiquantitative evaluation of hepatic echogenicity and could be useful in the follow-up of adolescents with NAFLD, population screening for this disease as well as for clinical studies.

An improved silver staining procedure for schizodeme analysis in polyacrylamide gradient gels

A simple protocol is described for the silver staining of polyacrylamide gradient gels used for the separation of restriction fragments of kinetoplast DNA [schizodeme analysis of trypanosomatids (Morel et al., 1980)]. The method overcomes the problems of non-uniform staining and strong background color which are frequently encountered when conventional protocols for silver staining of linear gels. The method described has proven to be of general applicability for DNA, RNA and protein separations in gradient gels.

Analysis of genetic diversity of Trypanosoma cruzi: an application of riboprinting and gradient gel electrophoresis methods

Analysis of restriction fragment length polymorphism (RFLP) profiles derived from digestion of polymerase chain reaction (PCR) products of the ribosomal 18S from Trypanosoma cruzi yields a typical `riboprint' profile that can vary intraspecifically. A selection of 21 stocks of T. cruzi and three outgroup taxa: T. rangeli, T. conorhini and Leishmania braziliensis were analysed by riboprinting to assess divergence within and between taxa. T. rangeli, T. conorhini and L. braziliensis could be easily differentiated from each other and from T. cruzi. Phenetic analysis of PCR-RFLP profiles indicated that, with one or two exceptions, stocks of T. cruzi could be broadly partitioned into two groups that formally corresponded to T. cruzi I and T. cruzi II respectively. To test if ribosomal 18S sequences were homogeneous within each taxon, gradient gel electrophoresis methods were employed utilising either chemical or temperature gradients. Upon interpretation of the melting profiles of riboprints and a section of the 18S independently amplified by PCR, there would appear to be at least two divergent 18S types present within T. cruzi. Heterogeneity within copies of the ribosomal 18S within a single genome has therefore been demonstrated and interestingly, this dimorphic arrangement was also present in the outgroup taxa. Presumably the ancestral duplicative event that led to the divergent 18S types preceded that of speciation within this group. These divergent 18S paralogues may have, or had, different functional pressures or rates of molecular evolution. Whether or not these divergent types are equally transcriptionally active throughout the life cycle, remain to be assessed.

The identification of Anopheles (Nyssorhynchus) rondoni (Diptera: Culicidae) in Mato Grosso, Brazil: an analysis of key character variability

A morphological study was made of a population of Anopheles (Nyssorhynchus) rondoni (Neiva and Pinto) from northern Mato Grosso, Brazil. This population usually lacked the primary key character of a dark basal band on hindtarsomere 3, i.e., hindtarsomere 3 was all white as in most other members of the subgenus. It was determined that this species can be recognized instead by the presence of a dark spot on the thorax made up of a large dark prescutellar space that is contiguous with a concolorous central area on the scutellum. A secondary character of a dark area on the costa created by the fusion of the humeral dark, presector dark and sector dark proximal spots is also usually reliable. Regression analyses comparing the lengths and ratios of the dark bands on hindtarsomeres 2 to those on 3 describe a straight line relationship. This suggests that the "atypical" population is at one end of a character gradient. We propose that in the subgenus Nyssorhynchus individuals that have a long basal band on hindtarsomere 2 are more likely to also have a basal band on hindtarsomere 3. The pupal stage of this species has not been previously described. Reared-associated specimens from this study show that the pupa can be easily differentiated from all other Nyssorhynchus by the relatively stout, usually 2 or 3 branched (1-5), setae 1 and 5 on segments IV-VII.

Spatial analysis of the distribution of leprosy in the State of Ceará, Northeast Brazil

The aim of this study was to describe spatial patterns of the distribution of leprosy and to investigate spatial clustering of incidence rates in the state of Ceará, Northeast Brazil. The average incidence rate of leprosy for the period of 1991 to 1999 was calculated for each municipality of Ceará. Maps were used to describe the spatial distribution of the disease, and spatial statistics were applied to explore large- and small-scale variations of incidence rates. Three regions were identified in which the incidence of leprosy was particularly high. A spatial gradient in the incidence rates was identified, with a tendency of high rates to be concentrated on the North-South axis in the middle region of the state. Moran's I statistic indicated that a significant spatial autocorrelation also existed. The spatial distribution of leprosy in Ceará is heterogeneous. The reasons for spatial clustering of disease rates are not known, but might be related to an heterogeneous distribution of other factors such as crowding, social inequality, and environmental characteristics which by themselves determine the transmission of Mycobacterium leprae.

The sample processing time interval as an influential factor in flow cytometry analysis of lymphocyte subsets

The objective of this paper is to propose a protocol to analyze blood samples in yellow fever 17DD vaccinated which developed serious adverse events. We investigated whether or not the time between sample collection and sample processing could interfere in lymphocyte subset percentage, for it is often impossible to analyze blood samples immediately after collection due to transport delay from collection places to the flow cytometry facility. CD4+CD38+ T, CD8+CD38+ T, CD3+ T, CD19+ B lymphocyte subsets were analyzed by flow cytometry in nine healthy volunteers immediately after blood collection and after intervals of 24 and 48 h. The whole blood lysis method and gradient sedimentation by Histopaque were applied to isolate peripheral blood mononuclear cells for flow cytometry analyses. With the lysis method, there was no significant change in lymphocyte subset percentage between the two time intervals (24 and 48 h). In contrast, when blood samples were processed by Histopaque gradient sedimentation, time intervals for sample processing influenced the percentage in T lymphocyte subsets but not in B cells. From the results obtained, we could conclude that the whole blood lysis method is more appropriate than gradient sedimentation by Histopaque for immunophenotyping of blood samples collected after serious adverse events, due to less variation in the lymphocyte subset levels with respect to the time factor.

Soil infiltration based on bp neural network and grey relational analysis

Soil infiltration is a key link of the natural water cycle process. Studies on soil permeability are conducive for water resources assessment and estimation, runoff regulation and management, soil erosion modeling, nonpoint and point source pollution of farmland, among other aspects. The unequal influence of rainfall duration, rainfall intensity, antecedent soil moisture, vegetation cover, vegetation type, and slope gradient on soil cumulative infiltration was studied under simulated rainfall and different underlying surfaces. We established a six factor-model of soil cumulative infiltration by the improved back propagation (BP)-based artificial neural network algorithm with a momentum term and self-adjusting learning rate. Compared to the multiple nonlinear regression method, the stability and accuracy of the improved BP algorithm was better. Based on the improved BP model, the sensitive index of these six factors on soil cumulative infiltration was investigated. Secondly, the grey relational analysis method was used to individually study grey correlations among these six factors and soil cumulative infiltration. The results of the two methods were very similar. Rainfall duration was the most influential factor, followed by vegetation cover, vegetation type, rainfall intensity and antecedent soil moisture. The effect of slope gradient on soil cumulative infiltration was not significant.

Ant species distribution along a topographic gradient in a "terra-firme" forest reserve in Central Amazonia

In Amazonia, topographical variations in soil and forest structure within "terra-firme" ecosystems are important factors correlated with terrestrial invertebrates' distribution. The objective of this work was to assess the effects of soil clay content and slope on ant species distribution over a 25 km² grid covering the natural topographic continuum. Using three complementary sampling methods (sardine baits, pitfall traps and litter samples extracted in Winkler sacks), 300 subsamples of each method were taken in 30 plots distributed over a wet tropical forest in the Ducke Reserve (Manaus, AM, Brazil). An amount of 26,814 individuals from 11 subfamilies, 54 genera, 85 species and 152 morphospecies was recorded (Pheidole represented 37% of all morphospecies). The genus Eurhopalothrix was registered for the first time for the reserve. Species number was not correlated with slope or clay content, except for the species sampled from litter. However, the Principal Coordinate Analysis indicated that the main pattern of species composition from pitfall and litter samples was related to clay content. Almost half of the species were found only in valleys or only on plateaus, which suggests that most of them are habitat specialists. In Central Amazonia, soil texture is usually correlated with vegetation structure and moisture content, creating different microhabitats, which probably account for the observed differences in ant community structure.

Diversity of springtails (Hexapoda) according to a altitudinal gradient

The objective of this work was to elevate gradient effect on diversity of Collembola, in a temperate forest on the northeast slope of Iztaccíhuatl Volcano, Mexico. Four expeditions were organized from November 2003 to August 2004, at four altitudes (2,753, 3,015, 3,250 and 3,687 m a.s.l.). In each site, air temperature, CO2 concentration, humidity, and terrain inclination were measured. The influence of abiotic factors on faunal composition was evaluated, at the four collecting sites, with canonical correspondence analyses (CCA). A total of 24,028 specimens were obtained, representing 12 families, 44 genera and 76 species. Mesaphorura phlorae, Proisotoma ca. tenella and Parisotoma ca. notabilis were the most abundant species. The highest diversity and evenness were recorded at 3,250 m (H' = 2.85; J' = 0.73). Canonical analyses axes 1 and 2 of the CCA explained 67.4% of the variance in species composition, with CO2 and altitude best explaining axis 1, while slope and humidity were better correlated to axis 2. The results showed that CO2 is an important factor to explain Collembola species assemblage, together with slope and humidity.

COMPARISON BETWEEN WOOD DRYING DEFECT SCORES: SPECIMEN TESTING X ANALYSIS OF KILN-DRIED BOARDS

It is important to develop drying technologies for Eucalyptus grandis lumber, which is one of the most planted species of this genus in Brazil and plays an important role as raw material for the wood industry. The general aim of this work was to assess the conventional kiln drying of juvenile wood of three clones of Eucalyptus grandis. The specific aims were to compare the behavior between: i) drying defects indicated by tests with wood specimens and conventional kiln-dried boards; and ii) physical properties and the drying quality. Five 11-year-old trees of each clone were felled, and only flatsawn boards of the first log were used. Basic density and total shrinkage were determined, and the drying test with wood specimens at 100 °C was carried out. Kiln drying of boards was performed, and initial and final moisture content, moisture gradient in thickness, drying stresses and drying defects were assessed. The defect scoring method was used to verify the behavior between the defects detected by specimen testing and the defects detected in kiln-dried boards. As main results, the drying schedule was too severe for the wood, resulting in a high level of boards with defects. The behavior between the defects in the drying test with specimens and the defects of kiln-dried boards was different, there was no correspondence, according to the defect scoring method.

A contribution to the phytogeography of Brazilian campos: an analysis based on Poaceae

Twenty areas from eight Brazilian states were compared according to a list of 224 species of Poaceae. In order to determinate affinity patterns between the areas, a binary matrix was submitted to cluster and ordination analysis. The patterns found were then faced to climate and geographic position. The scores corresponding to the areas obtained from the cluster analysis showed a strong correlation to temperature. The scores corresponding to the species suggest a gradient that associates distribution patterns to the photosynthetic pathway (C3 or C4). The current results suggest that the traditional classification of the Southern American grasslands might require some modification in order to be broadly applicable in the Brazilian context.

Floristic composition and similarity analysis of an Atlantic rain forest fragment in Cananéia, São Paulo State, Brazil

The aim of this study is to investigate the floristic composition of an Atlantic rain forest fragment located in Cananéia, São Paulo, Brazil, and to contribute to the knowledge on Atlantic forest through the comparative analysis of this and other floristic surveys both on the southern and southeastern Brazil, in different soil and relief types. We surveyed 215 species in 132 genera and 51 families. Classification and ordination analysis were applied to a binary matrix in order to analyze the similarity among 24 surveys, including the present one, of Atlantic forest from the south and southeast coast of Brazil. Higher floristic similarity was observed among this area and the ones where there was marine influence and more rugged relief. The surveys in areas with greater marine influence (sandy soil) were separated from those in other conditions, possibly indicating a species replacement gradient from the steep slopes towards the lowland and were probably related to different edaphic conditions. A latitudinal gradient was found among the surveys apparently confirming a continuous species replacement along the Atlantic forest, related to a restricted distribution of the species. This suggests that it is essential to preserve areas from the whole Atlantic coast. Atlantic forest distribution is quite complex and its composition cannot be adequately represented by small localized areas.

Histopathological analysis of rat mesentery as a method for evaluating neutrophil migration: differential effects of dexamethasone and pertussis toxin

In the present study, histopathological analysis of rat mesentery was used to quantify the effect of two anti-inflammatory agents, dexamethasone (Dex) and pertussis toxin (Ptx), on leukocyte migration. The intravenous injection of Dex (1 mg/kg) and Ptx (1,200 ng) 1 h prior to the intraperitoneal injection of the inflammatory stimuli lipopolysaccharide (LPS) or formyl-methionyl-leucyl-phenylalanine (fMLP) significantly reduced the neutrophil diapedesis (LPS: Ptx = 0.86 ± 0.19 and Dex = 0.35 ± 0.13 vs saline (S) = 2.85 ± 0.59; fMLP: Ptx = 0.43 ± 0.09 and Dex 0.01 ± 0.01 vs S = 1.08 ± 0.15 neutrophil diapedesis/field) and infiltration (LPS: Ptx = 6.29 ± 1.4 and Dex = 3.06 ± 0.76 vs S = 15.94 ± 3.97; fMLP: Ptx = 3.85 ± 0.56 and Dex = 0.40 ± 0.16 vs S = 7.15 ± 1.17 neutrophils/field) induced by the two agonists in the rat mesentery. The inhibitory effect of Dex and Ptx was clearly visible in the fields nearest the venule (up to 200 µm), demonstrating that these anti-inflammatory agents act preferentially in the transmigration of neutrophils from the vascular lumen into the interstitial space, but not in cell movement in response to a haptotactic gradient. The mesentery of rats pretreated with Dex showed a decreased number of neutrophils within the venules (LPS: Dex = 1.50 ± 0.38 vs S = 4.20 ± 1.01; fMLP: Dex = 0.25 ± 0.11 vs S = 2.20 ± 0.34 neutrophils in the lumen/field), suggesting that this inhibitor may be acting at a step that precedes neutrophil arrival in the inflamed tissue. In contrast to that observed with Dex treatment, the number of neutrophils found in mesenteric venules was significantly elevated in animals pretreated with Ptx (LPS: Ptx = 9.85 ± 2.25 vs S = 4.20 ± 1.01; fMLP: Ptx = 4.66 ± 1.24 vs S = 2.20 ± 0.34 neutrophils in the lumen/field). This discrepancy shows that Ptx and Dex act via different mechanisms and suggests that Ptx prevents locomotion of neutrophils from the vascular lumen to the interstitial space. In conclusion, the method described here is useful for quantifying the inflammatory and anti-inflammatory effect of different substances. The advantage of this histopathological approach is that it provides additional information about the steps involved in leucocyte migration.

Characterization of spoilage bacteria in pork sausage by PCR-DGGE analysis

To investigate microbial diversity and identify spoilage bacteria in fresh pork sausages during storage, twelve industrial pork sausages of different trademarks were stored at 4 ºC for 0, 14, 28 and 42 days, 80% relative humidity and packaged in sterile plastic bags. Microbiological analysis was performed. The pH and water activity (a w) were measured. The culture-independent method performed was the Polymerase Chain Reaction - Denaturing Gradient Gel Electrophoresis (PCR-DGGE). The culture-dependent method showed that the populations of mesophilic bacteria and Lactic Acid Bacteria (LAB) increased linearly over storage time. At the end of the storage time, the average population of microorganisms was detected, in general, at the level of 5 log cfu g-1. A significant (P < 0.005) increase was observed in pH and a w values at the end of the storage time. The PCR-DGGE allowed a rapid identification of dominant communities present in sausages. PCR-DGGE discriminated 15 species and seven genera of bacteria that frequently constitute the microbiota in sausage products. The most frequent spoilage bacteria identified in the sausages were Lactobacillus sakei and Brochothrix thermosphacta. The identification of dominant communities present in fresh pork sausages can help in the choice of the most effective preservation method for extending the product shelf-life.