Structural complexes of human adenine phosphoribosyltransferase reveal novel features of the APRT catalytic mechanism

Adenine phosphoribosyltransferase (APRT) is an important enzyme component of the purine recycling pathway. Parasitic protozoa of the order Kinetoplastida are unable to synthesize purines de novo and use the salvage pathway for the synthesis of purine bases rendering this biosynthetic pathway an attractive target for antiparasitic drug design. The recombinant human adenine phosphoribosyltransferase (hAPRT) structure was resolved in the presence of AMP in the active site to 1.76 angstrom resolution and with the substrates PRPP and adenine simultaneously bound to the catalytic site to 1.83 angstrom resolution. An additional structure was solved containing one subunit of the dimer in the apo-form to 2.10 angstrom resolution. Comparisons of these three hAPRT structures with other `type I` PRTases revealed several important features of this class of enzymes. Our data indicate that the flexible loop structure adopts an open conformation before and after binding of both substrates adenine and PRPR Comparative analyses presented here provide structural evidence to propose the role of Glu 104 as the residue that abstracts the proton of adenine N9 atom before its nucleophilic attack on the PRPP anomeric carbon. This work leads to new insights to the understanding of the APRT catalytic mechanism.

SOME REMARKS ON THE YOGASUTRA

In this article are discussed some problems raised by T. S. Rukmani regarding the Yogasutra. According to Rukmani, yoga is not a coherent logical system. After stating the general goals of the yoga system, interpretations of some passages of the Yogasutra are formulated, and the defense is given that one must at least acknowledge the necessity of more exegetical investigation before judging the logical incoherence of the text. Finally, the problematic doctrine of living liberation in Yogasutra is exposed.

DESCRIPTION OF ADULTS AND NYMPH, AND REDESCRIPTION OF THE LARVA, OF ORNITHODOROS MARINKELLEI (ACARI: ARGASIDAE), WITH DATA ON ITS PHYLOGENETIC POSITION

The argasid tick Ornithodoros marinkellei Kohls, Clifford, and Jones, 1969 was described 4 decades ago based on larval specimens collected from bats (Pteronotus spp.) in Colombia and Panama. Thereafter, larval O. marinkellei parasitizing bats were reported from Venezuela, Guyana, and Brazil. Herein, we describe the adults and nymph, and redescribe the larva of O. marinkellei based on specimens recently collected in the western Brazilian Amazon region. In contrast to all other known adult argasids, the idiosoma of both males and females of O. marinkellei is covered with sclerotized plaques. The idiosoma of the nymph of O. marinkellei is entirely micromamillated, and differs from the adults by the absence of plaques. The larva of O. marinkellei is morphologically similar to the larvae of the 2 other species belonging to the subgenus Subparmatus, i.e., Ornithodoros viguerasi Cooley and Kohls, 1941 and Ornithodoros mormoops Kohls, Clifford, and Jones, 1969. Because of the long and narrow dorsal plate, the larva of O. marinkellei is readily distinguished from O. viguerasi and O. mormoops. Comparison of our larvae from Brazil with O. marinkellei paratype specimens from Colombia confirmed their taxonomic identification. However, a few morphological differences, particularly in the size of the gnathosoma, were observed. Further studies are necessary to clarify whether O. marinkellei is a complex of different species, or a single species represented by morphologically polymorphic, and geographically distinct populations. Partial mitochondrial 16S rDNA gene sequences were generated for O. marinkellei specimens from Brazil, and compared with available homologous sequences in GenBank. Phylogenetic analyses revealed O. marinkellei to be distinct from the remaining argasid species available in GenBank, including other bat-associated tick species that are found in sympatry with O. marinkellei in the Neotropical region.

Relationship between attack and skipping in taekwondo contests

Santos, VGF, Franchini, E, and Lima-Silva, AE. Relationship between attack and skipping in Taekwondo contests. J Strength Cond Res 25(6): 1743-1751, 2011-The purpose of this study was to determine the relationship between attack time (AT) and skipping time (ST) during the 2007 Taekwondo World Championship and 2008 Beijing Olympic Games. A total of 22 matches (65 rounds, 13 semifinals, and 8 finals) from the World Championship and 23 matches (63 rounds, 22 rounds with 16 athletes each and 1 quarterfinal round) from the Olympic Games, both in the male category, were assessed using time-motion analysis. The AT was considered as the total time during which the athlete attacked or tried to attack, whereas ST was the total time without attempting to attack. The ratio of AT to ST was similar to 1:7 based on the data pooled from the 2 competitions. The AT/ST ratio was significantly lower for the World Championship than for the Olympic Games (p <= 0.05). In the Olympic Games, no consistent differences across weight divisions were found. However, during the World Championship, the heavier weight divisions (>78 kg) exhibited a lower average AT, lower summed AT, lower attack numbers (ANs) and higher average ST than lighter weight divisions (<58 kg, p <= 0.05). For both competitions, the ST was lower, and the ANs and AT/ST ratio were higher in round 3 than in round 1 or 2. In conclusion, the results of this study suggest that matches in the Olympic Games were less cadenced than in the World Championship, but that in both competitions, the intensity of the match increased in round 3. Practically, these data suggest that coaches need to structure Taekwondo training sessions in a manner that allows the work/pause ratio to mirror the physical demand imposed during competitions.

Effect of hemicellulose and lignin on enzymatic hydrolysis of cellulose from brewer`s spent grain

Enzymatic hydrolysis of brewer`s spent grain in three different forms: original (untreated), pretreated by dilute acid (cellulignin), and pretreated by a sequence of dilute acid and dilute alkali (cellulose pulp), was studied to verify the effect of hemicellulose and lignin on cellulose conversion into glucose. The hydrolysis was carried out using a commercial cellulase concentrate (Celluclast 1.5 L) in an enzyme/substrate ratio of 45 FPU/g, 2% (w/v) substrate concentration, 45 degrees C for 96 h. According to the results, the cellulose hydrolysis was affected by the presence of hemicellulose and/or lignin in the sample. The cellulose conversion ratio (defined as glucose yield + cellobiose yield) from cellulignin was 3.5-times higher than that from untreated sample, whereas from cellulose pulp such value was 4-times higher, correspondent to 91.8% (glucose yield of 85.6%). This best result was probably due to the strong modification in the material structure caused by the hemicellulose and lignin removal from the sample. As a consequence, the cellulose fibers were separated being more susceptible to the enzymatic attack. It was concluded that the lower the hemicellulose and lignin contents in the sample, the higher the efficiency of cellulose hydrolysis. (C) 2007 Elsevier Inc. All rights reserved.

Influence of the slice position on chloride migration tests for concrete in marine conditions

Chloride migration tests are used to measure the concrete capacity to inhibit chloride attack. Many researchers carry out this test in a slice of concrete extracted from the central part of cylindrical specimens, discarding about 75% of the concrete used to mold the specimens. This fact generated the question: would it be possible to extract more slices from a same specimen without losing the confidence in the results? The main purpose of this work is to answer this question. Moreover, another aim of this study was to show the difference of chloride penetration between finished faces and the formwork surfaces of concrete beams and slabs. The results indicated that it is possible to use more slices of a single specimen for a chloride migration test. Moreover, it was demonstrated that there is a significant difference of chloride penetration between the finished surface and the formwork surface of the specimens. (C) 2008 Elsevier Ltd. All rights reserved.

Wood CO(2) efflux and foliar respiration for Eucalyptus in Hawaii and Brazil

We measured CO(2) efflux from wood for Eucalyptus in Hawaii for 7 years and compared these measurements with those on three-and four-and-a-half-year-old Eucalyptus in Brazil. In Hawaii, CO(2) efflux from wood per unit biomass declined similar to 10x from age two to age five, twice as much as the decline in tree growth. The CO(2) efflux from wood in Brazil was 8-10x lower than that for comparable Hawaii trees with similar growth rates. Growth and maintenance respiration coefficients calculated from Hawaii wood CO(2) efflux declined with tree age and size (the growth coefficient declined from 0.4 mol C efflux mol C(-1) wood growth at age one to 0.1 mol C efflux mol C(-1) wood growth at age six; the maintenance coefficient from 0.006 to 0.001 mu mol C (mol C biomass)(-1) s(-1) at 20 degrees C over the same time period). These results suggest interference with CO(2) efflux through bark that decouples CO(2) efflux from respiration. We also compared the biomass fractions and wood CO(2) efflux for the aboveground woody parts for 3- and 7-year-old trees in Hawaii to estimate how focusing measurements near the ground might bias the stand-level estimates of wood CO(2) efflux. Three-year-old Eucalyptus in Hawaii had a higher proportion of branches < 0.5 cm in diameter and a lower proportion of stem biomass than did 7-year-old trees. Biomass-specific CO(2) efflux measured at 1.4 m extrapolated to the tree could bias tree level estimates by similar to 50%, assuming no refixation from bark photosynthesis. However, the bias did not differ for the two tree sizes. Foliar respiration was identical per unit nitrogen for comparable treatments in Brazil and Hawaii (4.2 mu mol C mol N(-1) s(-1) at 20 degrees C).

Bioinsecticidal activity of Talisia esculenta reserve protein on growth and serine digestive enzymes during larval development of Anticarsia gemmatalis

Plants synthesize a variety of molecules to defend themselves against an attack by insects. Talisin is a reserve protein from Talisia esculenta seeds, the first to be characterized from the family Sapindaceae. In this study, the insecticidal activity of Talisin was tested by incorporating the reserve protein into an artificial diet fed to the velvetbean caterpillar Anticarsia gemmatalis, the major pest of soybean crops in Brazil. At 1.5% (w/w) of the dietary protein, Talisin affected larval growth, pupal weight, development and mortality, adult fertility and longevity, and produced malformations in pupae and adult insects. Talisin inhibited the trypsin-like activity of larval midgut homogenates. The trypsin activity in Talisin-fed larvae was sensitive to Talisin, indicating that no novel protease-resistant to Talisin was induced in Talisin-fed larvae. Affinity chromatography showed that Talisin bound to midgut proteinases of the insect A. gemmatalis, but was resistant to enzymatic digestion by these larval proteinases. The transformation of genes coding for this reserve protein could be useful for developing insect resistant crops. (C) 2010 Elsevier Inc. All rights reserved.

Predation and reproductive output of the ladybird beetle Stethorus tridens preying on tomato red spider mite Tetranychus evansi

Predatory behaviour and reproductive output of the ladybird beetle Stethorus tridens Gordon as function of the tomato red spider mite (TRSM), Tetranychus evansi Baker & Pritchard, densities was investigated in the laboratory. Adult female of S. tridens were isolated in cylindrical plastic arenas, containing a leaf disc of Solanum americanum Mill. with 5, 20, 40, 60, 80 or 100 T. evansi nymphs. The number of prey consumed and eggs laid were evaluated daily for ten consecutive days, starting at the oviposition. Oviposition of S. tridens was positively correlated with prey consumption and lower threshold prey consumption for S. tridens laying eggs was 16.3 mites per day. The instantaneous rate of attack (ca. discovery area) and the handling time were 0.0062 h(-1) and 0.83 h, and 0.00254 h(-1) and 0.78 h, respectively, for predators at the 1st- and 10th-oviposition day. The predator exhibited a type II functional response at 1st- and 10th-oviposition day with a maximum consumption per predator of 33 T. evansi nymphs per day at the highest prey density. The ladybird beetle S. tridens is often collected associated with red spider mite colonies on solanaceous wild plants and the results suggest the potential of this ladybird beetle to control T. evansi in tomatoes crops.

Regulatory effects of an inhibitor from Plathymenia foliolosa seeds on the larval development of Anagasta kuehniella (Lepidoptera)

The Mediterranean flour moth, Anagasta kuehniella, is one of the most important insect pests of grains, reported worldwide, feeding on stored grains and products of rice, rye, corn and wheat. Plants synthesize a variety of molecules, including trypsin inhibitors, to defend themselves against attack by insects. In this study, a trypsin inhibitor (PFTI) was purified from Plathymenia foliolosa (Benth.) seeds and was tested for insect growth regulatory effect. The survival and mass of A. kuehniella larvae feeding on control seeds were about 82.7% and 5 ring, respectively, whereas survival on seeds containing 0.7% PFTI was about 56%, while a 66.1% reduction in the average mass of the larvae was observed. The results from dietary utilization experiments with A. kuehniella larvae showed a reduction in efficiency of conversion of ingested food and digested food, and an increase in approximate digestibility and metabolic cost. The level of trypsin was significantly decreased in larval midgut and increased in the feces of larvae reared on a diet containing 0.7% PFTI. Results indicate that PFTI possesses a toxic effect against A. kuehniella larvae. (C) 2008 Elsevier Inc. All rights reserved.

Greenhouse evaluation of the potential of sorghum, pearl millet and crotalaria in the management of Meloidogyne javanica

Crop rotation can play a valuable role in managing plant parasitic nematodes, depending on the availability of profitable non-host or poor host crops. Alternatively, non-host cover crops or green manures can be used in succession to summer cash Crops for this Purpose. The aim of the current study was to evaluate, under greenhouse conditions, the host status of commercial hybrids and cultivars of grain and silage sorghum (Sorghum bicolor) for Meloidogyne javanica, and to assess the effect of sorghum on nematode population in comparison with pearl millet (poor host for M. javanica), showy crotalaria and sunn hemp (both non-hosts). Based on two experiments, it was stated that, as a rule, grain sorghum is a poor host for M. javanica, but silage sorghum is a good host. Silage sorghum `BRS 601` was an exception. In other experiments, grain sorghum, pearl millet (Pennisetum glaucum `BN 2`), showy crotalaria (Crotolaria spectabilis `Comum`) and sunn hemp (C. juncea `IAC-KR-1`) reduced M. javanica population level, while silage sorghum increased the nematode density.

Effect of water renewal on dominance hierarchy of juvenile Nile tilapia

Nile tilapia social position (Oreochromis niloticus) can be mediated by multiple channels, including chemical communication. Absence of chemical cues in the environment prevents hierarchical settlement among pairs, and enhances time spent in confrontations. The aim of this study was to test the effect of continuously renewed water flow on the establishment of hierarchical dominance in Nile tilapia juveniles. In this condition, a high frequency of attacks and disruption on hierarchical stability were expected because chemical cues for hierarchy maintenance could be washed out. After 3 days in isolation, the fish were paired by standard size but not by sex, and submitted to two conditions: continuously renewed water flow (RENEWED, n = 7) and non-renewed water flow (NONRENEWED n = 8). The paired fish were placed in an aquarium (40 cm x 30 cm x 40 cm) for 3 h; four 10-min sessions were video-recorded: the first, immediately after the fish were paired and the others 1, 2, and 3 h after pairing. Hierarchy was identified by a dominance index (DI = given attacks/received + given attacks) For each fish. The hierarchical stability was achieved by analyzing the difference between dominant DI and subordinate DI (DI-D). Hierarchy was established in both groups after second session because the DI was significantly higher for one fish of the pair. The frequency of attacks of the dominant fish in RENEWED and NONRENEWED conditions was similar in all observation sessions. The attack frequency by subordinate fish was also similar during the first three sessions (2-h pairing). However, the frequency of attacks by subordinate fish in the RENEWED condition was higher than in the NONRENEWED situation at the fourth observation session (means +/- S.E.: RENEWED = 2.83 +/- 0.94 x 10 min(-1) and NONRENEWED = 0.25 +/- 0.16 x 10 min(-1); Mann-Whitney, p = 0.04). At this point, a significant reduction of the DI-D was observed (means +/- S.E.: RENEWED = 0.70 +/- 0.11 and NONRENEWED = 1,00 +/- 0.002; Mann-Whitney, p = 0.04). The changes in DI-D were related to more frequent attacks by the subordinated fish in renewed water flow. According to our results, the unsteady agonistic interaction under renewed water flow leads to social instability. Thus, continuous water renewing can wash out relevant chemical substances and therefore disturb the dominance recognition by subordinate fish. (C) 2007 Elsevier B.V. All rights reserved.

Acute and Chronic Effects of Epicardial Radiofrequency Applications Delivered on Epicardial Coronary Arteries

Background-Epicardial coronary injury is by far the most feared complication of epicardial ablation. Little information is available regarding the chronic effects of delivering radiofrequency in the vicinity of large coronary vessels, and the long-term impact of this approach for mapping and ablation on epicardial vessel integrity is poorly understood. Therefore, the aim of this study was to characterize the acute and chronic histopathologic changes produced by in vivo epicardial pulses of radiofrequency ablation on coronary artery of porcine hearts. Methods and Results-Seven pigs underwent a left thoracotomy. The catheter was sutured adjacent to the left anterior descending artery and left circumflex artery, and 20 pulses of radiofrequency energy were applied. Radiofrequency lesions located no more than 1 mm of the vessel were used for this analysis. Three animals were euthanized 20 days (acute phase) after the procedure and 4 animals after 70 days (chronic phase). The following parameters were obtained in each vessel analyzed: (1) internal and external perimeter; (2) vessel wall thickness; (3) tunica media thickness, and (4) tunica intima thickness. The presence of adipose tissue around the coronary arteries, the distance between the artery and the epicardium, and the anatomic relationship of the artery with the coronary vein was also documented for each section. Sixteen of 20 (80%) sections analyzed, showed intimal thickening with a mean of 0.18 +/- 0.14 mm compared with 0.13 +/- 0.16 mm in the acute phase (P=0.331). The mean tunica media thickness was 0.25 +/- 0.10 mm in the chronic phase animals compared with 0.18 +/- 0.03 mm in the acute phase animals (P=0.021). A clear protective effect of pericardial fat and coronary veins was also present. A positive correlation between depth of radiofrequency lesion and the degree of vessel injury expressed as intimal and media thickening (P=0.001) was present. A negative correlation was identified (r = -0.83; P=0.002) between intimal thickening and distance between epicardium and coronary artery. Conclusions-In this porcine model of in vivo epicardial radiofrequency ablation in proximity to coronary arteries leads to acute and chronic histopathologic changes characterized by tunica intima and media thickening, with replacement of smooth muscle cells with extracellular matrix, but no significant stenosis was observed up to 70 days after the ablation. The absence of acute coronary occlusion or injury does not preclude subsequent significant arterial damage, which frequently occurs when epicardial radiofrequency applications are delivered in close vicinity to the vessels. (Circ Arrhythm Electrophysiol. 2011;4:526-531.)

Effects of Erosive, Cariogenic or Combined Erosive/Cariogenic Challenges on Human Enamel An in situ/ex vivo Study

Individuals with cariogenic diet can also consume erosive beverages. Thus, it seems necessary to investigate a possible caries/erosion interaction. To test in situ/ex vivo a combination of these challenges, 11 subjects wore intraoral appliances containing four enamel blocks randomly assigned. In the first 2-week phase, the appliances were immersed in a cola drink 3 times/day. Two blocks were free of plaque (erosion only: EO) and two blocks were covered with plaque (erosion + plaque: EP). In the second 2-week phase, four new blocks were all covered with plaque and subjected to a sucrose solution 8 times/day. Among the four new blocks, two were also subjected to the cola drink 3 times/day (erosion + caries: EC) while the other two were not (caries only: CO). Thus, in EO, the specimens were fixed at the intraoral appliance level. In EP, EC and CO they were fixed 1.0 mm under the appliance level and covered with plastic meshes for dental plaque accumulation. Changes in wear and hardness were measured. Data were tested using ANOVA and Tukey`s test (p < 0.05). Mean values of wear (mu m) and change in hardness (kp/mm(2)) were: EO 4.82/310; EP 0.14/48; EC 0.34/245; CO 0.42/309. With respect to surface softening, EP and EC differed significantly from each other and from EO and CO, which did not differ significantly. EO presented significantly higher wear than the other groups. The data suggest that the presence of dental plaque can decrease the acid attack of an erosive drink and the association of erosive and cariogenic challenges showed less enamel alterations when compared to erosive or cariogenic challenges only. Copyright (C) 2008 S. Karger AG, Basel

Effects of let-7 microRNA on Cell Growth and Differentiation of Papillary Thyroid Cancer

Papillary thyroid carcinoma (PTC) is the most common endocrine malignancy and RET/PTC rearrangements represent key genetic events frequently associated to this cancer, enhancing proliferation and dedifferentiation by activation of the RET/PTC-RAS-BRAF-mitogen-activated protein kinase (MAPK) pathway. Recently, let-7 microRNA was found to reduce RAS levels in lung cancer, acting as a tumor suppressor gene. Here, we report that RET/PTC3 oncogenic activation in PCCL3 rat thyroid cells markedly reduces let-7f expression. Moreover, stable transfection of let-7 microRNA in TPC-1 cells, which harbor RET/PTC1 rearrangement, inhibits MAPK activation. As a result, let-7f was capable of reducing TPC-1 cell growth, and this might be explained, at least in part, by decreased messenger RNA (mRNA) expression of cell cycle stimulators such as MYC and CCND1 (cyclin D1) and increased P21 cell cycle inhibitor mRNA. In addition, let-7 enhanced transcriptional expression of molecular markers of thyroid differentiation such as TITF1 and TG. Thus, reduced expression of let-7f might be an essential molecular event in RET/PTC malignant transformation. Moreover, let-7f effects on thyroid growth and differentiation might attenuate neoplastic process of RET/PTC papillary thyroid oncogenesis through impairment of MAPK signaling pathway activation. This is the first functional demonstration of an association of let-7 with thyroid cancer cell growth and differentiation.