Histology of the regenerate and docking site in bone transport

Bone transport is based on the principle of distraction osteogenesis described by Ilizarov and is a consecrated method for the treatment of segmental bone defects. One of its most problematic and, paradoxically, least studied aspects is the consolidation of the docking site. We studied histologically the ossification of the docking site and regenerate to determine any difference between them. Nine adult sheep were submitted to correction of a 1-cm tibial diaphyseal defect using a system of plate-fixed bone transport, with latency period of 1 week and 0.2 mm distraction of the transported segment four times a day. The sheep were divided into three groups of three animals each, according to the observation period of 3, 6 or 12 weeks between the fixation of the transported fragment and the euthanasia. The docking site and the regenerate were studied histologically on sections stained with Masson trichrome. The main mode of docking site ossification was the endochondral one and although intramembranous ossification was also observed simultaneously, it was limited to rare and small foci. In contrast, intramembranous ossification played the major role in the regenerate, with bone formation evolving from the base segment to the target segment. The experimental bone transport model proposed in the present study permits us to conclude that there is a clear difference between the ossification of the docking site and of the regenerate.

Salt iodination and excessive iodine intake among schoolchildren

Salt iodination and excessive iodine intake among schoolchildren. The objective of the present study was to evaluate the urinary excretion of iodine and relate it to the amount present in salt for human consumption. The study involved 145 children from two schools: a rural one and an urban one. We performed anthropometric measurements and collected a urine sample and a kitchen salt sample from each child. In the rural school, 3.38% of children had iodine deficiency. However, most of the values of urinary iodine were above 300 mu g/L (62.03%) and 59.49% of the kitchen salt samples contained 20 to 60 mg iodine per kilo of salt. In the urban school, 3.03% of the children had urinary iodine excretion of less than 100 mu g/L and 90.91% of the children had urinary iodine values exceeding 300 mu g/L. Of this total, 84.85% of the kitchen salt samples contained 20 to 60 mg iodine per kilo of salt. Iodine deficiency is controlled in this population, with the current reality showing a high prevalence of excess urinary iodine.

Polysaccharide fraction of Agaricus brasiliensis avoids tumor-induced IL-10 production and changes the microenvironment of subcutaneous Ehrlich adenocarcinoma

Subcutaneous Ehrlich tumor-bearing mice were treated with in situ inoculation of a P-glucan-rich extract of Agaricus brasiliensis (ATF), which reduced tumor growth. Histopathological analysis showed that the tumor masses of control mice (Ehr) presented giant tumor cells and many mitotic figures whereas the tumor tissue obtained from ATF-treated animals (Ehr-ATF) presented a lower frequency of both mitotic and giant cells, associated with a higher frequency of apoptotic cells than Ehr. Analysis of the lymphoproliferative activity of spleen cells showed that the treatment had a suppressive rather than a stimulatory effect. Spleen cells of the Ehr group produced higher in vitro levels of IL-10 than normal controls and this occurrence was partially avoided by treatment with ATF. Analysis of cytokine production by tumor-infiltrating cells (ELISpot) showed that ATF induced a higher number of IFN-gamma-producing cells at 7 and 14 days as well as reduction of IL-10-secreting cells at the latter time. Confocal microscopy analysis showed higher intensity of labeling of CD4+ and Mac-3+ cells in ATF-treated mice. Analysis of in situ expression of angiogenic growth factors showed a slight decrease of FGF-2 mRNA in Ehr-ATF animals (7th day) but not of VEGF-A or TGF-beta expression. This fraction could not directly lyse either lymphocytes or tumor cells and we speculate that antitumor effect of ATF could be due to induction of a selective migration of immunocompetent cells from the spleen to the tumor site and to the switch of cytokine production. (C) 2009 Elsevier Inc. All rights reserved.

Influence of curing rate on softening in ethanol, degree of conversion, and wear of resin composite

Purpose: To investigate the effect of curing rate on softening in ethanol, degree of conversion, and wear of resin composites. Methods: With a given energy density and for each of two different light-curing units (QTH or LED), the curing rate was reduced by modulating the curing mode. Thus, the irradiation of resin composite specimens (Filtek Z250, Tetric Ceram, Esthet-X) was performed in a continuous curing mode and in a pulse-delay curing mode. Wallace hardness was used to determine the softening of resin composite after storage in ethanol. Degree of conversion was determined by infrared spectroscopy (FTIR). Wear was assessed by a three-body test. Data were submitted to Levene`s test, one and three-way ANOVA, and Tukey HSD test (alpha= 0.05). Results: Immersion in ethanol, curing mode, and material all had significant effects on Wallace hardness. After ethanol storage, resin composites exposed to the pulse-delay curing mode were softer than resin composites exposed to continuous cure (P< 0.0001). Tetric Ceram was the softest material followed by Esthet-X and Filtek Z250 (P< 0.001). Only the restorative material had a significant effect on degree of conversion (P< 0.001): Esthet-X had the lowest degree of conversion followed by Filtek Z250 and Tetric Ceram. Curing mode (P= 0.007) and material (P< 0.001) had significant effect on wear. Higher wear resulted from the pulse-delay curing mode when compared to continuous curing, and Filtek Z250 showed the lowest wear followed by Esthet-X and Tetric Ceram. (Am J Dent 2011;24:115-118).

Induced seismicity in the Castarthao reservoir, NE Brazil - Preliminary results

The Castanhao reservoir was built in the state of Ceara, a dry region in Northeastern Brazil, to regulate the flow of the Jaguaribe River, for irrigation, and for power generation. It is an earth-filled dam, 60 m high, with a water capacity of 4.5 x 10(9) m(3). The seismicity in the area has been monitored since 1998, with a few interruptions, using one analog or one digital station and, during a few periods, a three-station network. The first earthquakes likely to be induced events were detected in 2003, when the water level was about 20 in high. In early 2004 a very heavy rainfall season quickly filled the reservoir. Shortly after, an increase in the seismic activity occurred and many micro-earthquakes were recorded. We suggest that this activity resulted from an increase in pore pressure due to undrained response. Therefore, we may classify this cluster of microearthquakes as ""initial seismicity."" We deployed a network with four analog stations in the area, following this activity, to determine the epicentral zone. At least three epicentral areas under the reservoir were detected. The spatio-temporal analysis of the available data revealed that the seismicity occurs in clusters and that these were activated at different periods. We identified four sets of faults (N-S-, E-W-, NW-SE-, and NE-SW-oriented), some of which moved in shallow crustal levels and as recently as the Quaternary (1.8 Ma). Under the present-day stress regime, the last two sets moved as strike-slip structures. We suggest a possible correlation between dormant faults and the observed induced seismicity. (c) 2008 Elsevier B.V. All rights reserved.

Differential proteome analysis of mature and germinated embryos of Araucaria angustifolia

Araucaria angustifolia is an endangered Brazilian native conifer tree. The aim of the present work was to identify differentially expressed proteins between mature and germinated embryos of A. angustifolia, using one and two dimensional gel electrophoresis approaches followed by protein identification by tandem mass spectrometry. The identities of 32 differentially expressed protein spots from two dimensional gel maps were successfully determined, including proteins and enzymes involved in storage mobilization such as the vicilin-like storage protein and proteases. A label free approach, based on spectral counts, resulted in detection of 10 and 14 mature and germinated enriched proteins, respectively. Identified proteins were mainly related to energetic metabolism pathways, translational processes. oxidative stress regulation and cellular signaling. The integrated use of both strategies permitted a comprehensive protein expression overview of changes in germinated embryos in relation to matures, providing insights into the this process in a recalcitrant seed species. Applications of the data generated on the monitoring and control of in vitro somatic embryos were discussed. Published by Elsevier Ltd.

Cytotype-specific ISSR profiles and karyotypes in the Neotropical genus Eigenmannia (Teleostei: Gymnotiformes)

The genus Eigenmannia (Teleostei: Gymnotiformes), a widely distributed fish genus from the Neotropical region, presents very complex morphological patterns and many taxonomic problems. It is suggested that this genus harbors a species complex that is hard to differentiate using only morphological characteristics. As a result, many species of Eigenmannia may be currently gathered under a common name. With the objective of providing new tools for species characterization in this group, an analysis of the polymorphism of DNA inter-simple sequence repeats (ISSR), obtained by single primer amplification reaction (SPAR), combined with karyotype identification, was carried out in specimens sampled from populations of the Upper Parana, So Francisco and Amazon river basins (Brazil). Specific ISSR patterns generated by primers (AAGC)(4) and (GGAC)(4) were found to characterize the ten cytotypes analyzed, even though the cytotypes 2n = 38 and 2n = 38 XX:XY, from the Upper Parana basin, share some ISSR amplification patterns. The geographical distribution of all Eigenmannia specimens sampled was inferred, showing the cytotype 2n = 31/2n = 32 as the most frequent and largely distributed in the Upper Parana basin. The cytotype 2n = 34 was reported for the first time in the genus Eigenmania, restricted to the So Francisco basin. Polymorphic ISSR patterns were also detected for each cytotype. Considering our results and the data reported previously in the literature, it is suggested that many of the forms of Eigenmannia herein analyzed might be regarded as different species. This work reinforces the importance of employing diverse approaches, such as molecular and cytogenetic characterization, to address taxonomic and evolutionary issues.

A new species of Moenkhausia from the rio Amazonas and rio Orinoco basins (Characiformes: Characidae)

A new species of Moenkhausia is described from the rio Amazonas and rio Orinoco basins. The new species can be distinguished from congeners mainly by the combination of a conspicuous, relatively small and circular humeral spot, a black spot on the upper caudal-fin lobe, lower caudal-fin lobe without spot or a faint one, and middle caudal-fin rays hyaline or with dark tips. Mature males have a unique combination of two large-sized bony hooks on the anal-fin rays and tiny spines on the distal portion of all fins, which distinguishes the new species from any other species of Characidae.

The Snake Venom Peptide Bj-PRO-7a Is a M1 Muscarinic Acetylcholine Receptor Agonist

Proline-rich peptides from Bothrops jararaca venom (Bj-PRO) were characterized based on the capability to inhibit the somatic angiotensin-converting enzyme. The pharmacological action of these peptides resulted in the development of Captopril, one of the best examples of a target-driven drug discovery for treatment of hypertension. However, biochemical and biological properties of Bj-PROs were not completely elucidated yet, and many recent studies have suggested that their activity relies on angiotensin-converting enzyme-independent mechanisms. Here, we show that Bj-PRO-7a (and even for pharmacological applications. (C) 2010 International Society for Advancement of Cytometry

Identification and lineage genotyping of South American trypanosomes using fluorescent fragment length barcoding

Trypanosoma cruzi and Trypanosoma rangeli are human-infective blood parasites, largely restricted to Central and South America. They also infect a wide range of wild and domestic mammals and are transmitted by a numerous species of triatomine bugs. There are significant overlaps in the host and geographical ranges of both species. The two species consist of a number of distinct phylogenetic lineages. A range of PCR-based techniques have been developed to differentiate between these species and to assign their isolates into lineages. However, the existence of at least six and five lineages within T. cruzi and T. rangeli, respectively, makes identification of the full range of isolates difficult and time consuming. Here we have applied fluorescent fragment length barcoding (FFLB) to the problem of identifying and genotyping T. cruzi, T. rangeli and other South American trypanosomes. This technique discriminates species on the basis of length polymorphism of regions of the rDNA locus. FFLB was able to differentiate many trypanosome species known from South American mammals: T. cruzi cruzi. T. cruzi marinkellei, T. dionisii-like, T. evansi, T. lewisi, T. rangeli, T. theileri and T. vivax. Furthermore, all five T. rangeli lineages and many T. cruzi lineages could be identified, except the hybrid lineages TcV and TcVI that could not be distinguished from lineages III and II respectively. This method also allowed identification of mixed infections of T. cruzi and T. rangeli lineages in naturally infected triatomine bugs. The ability of FFLB to genotype multiple lineages of T. cruzi and T. rangeli together with other trypanosome species, using the same primer sets is an advantage over other currently available techniques. Overall, these results demonstrate that FFLB is a useful method for species diagnosis, genotyping and understanding the epidemiology of American trypanosomes. (C) 2010 Elsevier B.V. All rights reserved.

Capillariaisis (Trichurida, Trichinellidae, Capillaria hepatica) in the Brazilian Amazon: low pathogenicity, low infectivity and a novel mode of transmission

Background: Human capillariasis caused by Capillaria hepatica (syn. Calodium hepaticum) is a rare disease with no more than 40 cases registered around the world. Classically, the disease has severe symptoms that mimic acute hepatitis. Natural reservoirs of C. hepatica are urban rodents (Mus musculus and Rattus novergicus) that harbor their eggs in the liver. After examining the feces of 6 riverine inhabitants (Rio Preto area, 8 degrees 03`S and 62 degrees 53`W to 8 degrees 14`S and 62 degrees 52`W) of the State of Rondonia, Brazil, and identifying C. hepatica eggs in their feces, the authors decided to investigate the real dimension of these findings by looking for two positive signals. Methods: Between June 1(st) and 15(th), 2008, 246 out of 304 individuals were clinically examined. Blood samples were collected, kept under -20 degrees C, and test by the indirect immunofluorescence technique. Results: The first positive signal was the presence of specific antibodies at 1: 150 dilution, which indicates that the person is likely to have been exposed to eggs, most likely non-infective eggs, passing through the food chain or via contaminated food (total prevalence of 34.1%). A second more specific signal was the presence of antibodies at higher titers, thus indicating true infection. Conclusions: The authors concluded that only two subjects were really infected (prevalence of 0.81%); the rest was false-positives that were sensitized after consuming non-embryonated eggs. The present study is the first one carried out in a native Amazonian population and indicates the presence of antibodies against C. hepatica in this population. The results further suggest that the transmission of the parasite occurs by the ingestion of embryonated eggs from human feces and/or carcasses of wild animals. The authors propose a novel mode of transmission, describing the disease as a low pathogenic one, and showing low infectivity.

Degenerate Two-Photon Absorption in All-Trans Retinal: Nonlinear Spectrum and Theoretical Calculations

In this work we investigate the degenerate two-photon absorption spectrum of all-trans retinal ill ethanol employing the Z-scan technique with femtosecond pulses, The two-photon absorption (2PA) spectrum presents a monotonous increase as the excitation wavelength approaches the one-photon absorption band and it peak at 790 nm. We attribute the 2PA hand to the mixing of states (1)B(u)+-like and vertical bar S(1)>, which are strongly allowed by one- and two-photon, respectively. We modeled the 2PA spectrum by using the sum-over-states approach and obtained spectroscopic parameters of the electronic transitions to vertical bar S >, vertical bar S(2)> (""(1)Bu(+)""), vertical bar S(3)>, and vertical bar S(4)> singlet-excited states. The results were compared with theoretical predictions of one- and two-photon transition calculations using the response Functions formalism within the density functional theory framework with the aid of the CAM-B3LYP functional.

The development of blowouts and foredunes in the Ilha Comprida barrier (Southeastern Brazil): the influence of Late Holocene climate changes on coastal sedimentation

Middle to Late Holocene barriers are conspicuous landforms in southeastern and southern Brazilian regions. The barriers in the coastal zones of northern Santa Catarina, Parana and Sao Paulo states (27 degrees 19`-24 degrees 00`S) are formed mainly by beach ridge alignments and many barriers present foredune and blowout alignments in their seaward portion. The development of these eolian landforms appears to record a regional shift in coastal dynamics and barrier building. In this context, the Ilha Comprida barrier stands out for its well-developed and well-preserved foredunes and blowouts. Based on the presence or not and type of eolian landforms, the Ilha Comprida barrier can be divided seaward into inner, middle and outer units. The inner unit is formed entirely by beach ridges. The middle unit comprises a narrow belt of blowouts (up to 15 m high) aligned alongshore. Blowout lobes pointing NNW are indicative of their generation by southern winds. The outer unit is represented by low (<= 1 m high) active or stabilized foredunes and a small transgressive dunefield (similar to 1 km(2)). Twenty-seven luminescence ages (SAR protocol) obtained for the beach ridges, foredunes, and blowouts of these three units allow definition of a precise chronology of these landforms and calculation of rates of coastal progradation. The inner unit presents ages greater than 1004 +/- 88 years. The blowouts of the middle unit show ages from 575 +/- 47 to 172 +/- 18 years. The ages of the outer unit are less than 108 +/- 10 years. Rates of coastal progradation for the inner and outer units are 0.71-0.82 m/year and 0.86-2.23 m/year, respectively. The main phase of blowout development correlates well with the Little Ice Age (LIA) climatic event. These results indicate that southern winds in subtropical Brazil became increasingly more intense and/or frequent during the LIA. These conditions persist to the present and are responsible for the development of the eolian landforms in the outer unit. Thus, barrier geomorphology can record global climatic events. The sensitivity of barrier systems in subtropical Brazil to Late Holocene climate changes was favored by the relative sea level stillstand during this time. Luminescence dating makes it possible to analyze barrier geomorphology during Late Holocene climate changes operating on timescales of a hundred to thousand years. These results improve our knowledge of barrier building and will help in the evaluation of the impact of future climate changes on coastal settings. (C) 2008 Elsevier Ltd. All rights reserved.

Digestive physiology and characterization of digestive cathepsin L-like proteinase from the sugarcane weevil Sphenophorus levis

Sugarcane is an important crop that has recently become subject to attacks from the weevil Sphenophorus levis, which is not efficiently controlled with chemical insecticides. This demands the development of new control devices for which digestive physiology data are needed. In the present study, ion-exchange chromatography of S. levis whole midgut homogenates, together with enzyme assays with natural and synthetic substrates and specific inhibitors, demonstrated that a cysteine proteinase is a major proteinase, trypsin is a minor one and chymotrypsin is probably negligible. Amylase, maltase and the cysteine proteinase occur in the gut contents and decrease throughout the midgut; trypsin is constant in the entire midgut, whereas a membrane-bound aminopeptidase predominates in the posterior midgut. The cysteine proteinase was purified to homogeneity through ion-exchange chromatography. The purified enzyme had a mass of 37 kDa and was able to hydrolyze Z-Phe-Arg-MCA and Z-Leu-Arg-MCA with k(cat)/K(m) values of 20.0 +/- 1.1 mu M(-1) s(-1) and 30.0 +/- 0.5 mu M(-1) s(-1), respectively, but not Z-Arg-Arg-MCA. The combined results suggest that protein digestion starts in the anterior midgut under the action of a cathepsin L-like proteinase and ends on the surface of posterior midgut cells. All starch digestion takes place in anterior midgut. These data will be instrumental to developing S. levis-resistant sugarcane. (C) 2011 Elsevier Ltd. All rights reserved.

Development and Validation of a Mice liar Electrokinetic Chromatography Method for Quantitative Determination of Butenolides in Piper malacophyllum (C. Presl) C. DC.

Introduction - A large number of natural and synthetic compounds having butenolides as a core unit have been described and many of them display a wide range of biological activities. Butenolides from P. malacophyllum have presented potential antifungal activities but no specific, fast, and precise method has been developed for their determination. Objective - To develop a methodology based on micellar electrokinetic chromatography to determine butenolides in Piper species. Methodology - The extracts were analysed in an uncoated fused-silica capillaries and for the micellar system 20 mmol/L SDS, 20% (v/v) acetonitrile (ACN) and 10 mmol/L STB aqueous buffer at pH 9.2 were used. The method was validated for precision, linearity, limit of detection (LOD) and limit of quantitation (LOQ) and the standard deviations were determined from the standard errors estimated by the regression line. Results - A micellar electrokinetic chromatography (MEKC) method for determination of butenolides in extracts gave full resolution for 1 and 2. The analytical curve in the range 10.0-50.0 mu g/mL (r(2) = 0.999) provided LOD and LOQ for 1 and 2 of 2.1/6.3 and 1.1/3.5 mu g/mL, respectively. The RSD for migration times were 0.12 and 1.0% for peak area ratios with 100.0 +/- 1.4% of recovery. Conclusions - A novel high-performance MEKC method developed for the analysis of butenolides 1 and 2 in leaf extracts of P. malacophyllum allowed their quantitative determined within an analysis time shorter than 5 min and the results indicated CE to be a feasible analytical technique for the quantitative determination of butenolides in Piper extracts. Copyright (C) 2010 John Wiley & Sons, Ltd.