Evaluating Patellar Kinematics Through Magnetic Resonance Imaging During Open- and Closed-Kinetic-Chain Exercises

Purpose: To evaluate patellar kinematics of volunteers Without knee pain at rest and during isometric contraction in open- and closed-kinetic-chain exercises. Methods: Twenty individuals took part in this study. All were submitted to magnetic resonance imaging (MRI) during rest and voluntary isometric contraction (VIC) in the open anti closed kinetic chain at 15 degrees, 30 degrees, and 45 degrees of knee flexion. Through MRI and using medical e-film software, the following measurements were evaluated: sulcus angle, patellar-tilt angle, and bisect offset. The mixed-effects linear model was used for comparison between knee positions, between rest and isometric contractions, and between (he exercises. Results: Data analysis revealed that the sulcus angle decreased as knee flexion increased and revealed increases with isometric contractions in both the open and closed kinetic chain for all knee-flexion angles. The patellar-tilt angle decreased with isometric contractions in both the open and closed kinetic chain for every knee position. However, in the closed kinetic chain, patellar tilt increased significantly with the knee flexed at 15 degrees. The bisect offset increased with the knee flexed at 15 degrees during isometric contractions and decreased as knee flexion increased during both exercises. Conclusion: VIC in the last degrees of knee extension may compromise patellar dynamics. On the other hand, it is possible to favor patellar stability by performing muscle contractions with the knee flexed at 30 degrees and 45 degrees in either the open or closed kinetic chain.

A simplified minimal residual disease polymerase chain reaction method at early treatment points can stratify children with acute lymphoblastic leukemia into good and poor outcome groups

Background Minimal residual disease is an important independent prognostic factor in childhood acute lymphoblastic leukemia. The classical detection methods such as multiparameter flow cytometry and real-time quantitative polymerase chain reaction analysis are expensive, time-consuming and complex, and require considerable technical expertise. Design and Methods We analyzed 229 consecutive children with acute lymphoblastic leukemia treated according to the GBTLI-99 protocol at three different Brazilian centers. Minimal residual disease was analyzed in bone marrow samples at diagnosis and on days 14 and 28 by conventional homo/heteroduplex polymerase chain reaction using a simplified approach with consensus primers for IG and TCR gene rearrangements. Results At least one marker was detected by polymerase chain reaction in 96.4%, of the patients. By combining the minimal residual disease results obtained on days 14 and 28, three different prognostic groups were identified: minimal residual disease negative on days 14 and 28, positive on day 14/negative on day 28, and positive on both. Five-year event-free survival rates were 85%, 75.6%,, and 27.8%, respectively (p<0.0001). The same pattern of stratification held true for the group of intensively treated children. When analyzed in other subgroups of patients such as those at standard and high risk at diagnosis, those with positive B-derived CD10, patients positive for the TEL/AML1 transcript, and patients in morphological remission on a day 28 marrow, the event-free survival rate was found to be significantly lower in patients with positive minimal residual disease on day 28. Multivariate analysis demonstrated that the detection of minimal residual disease on day 28 is the most significant prognostic factor. Conclusions This simplified strategy for detection of minimal residual disease was feasible, reproducible, cheaper and simpler when compared with other methods, and allowed powerful discrimination between children with acute lymphoblastic leukemia with a good and poor outcome.

RNA interference-mediated knockdown of CD49e (alpha 5 integrin chain) in human thymic epithelial cells modulates the expression of multiple genes and decreases thymocyte adhesion

Background: The thymus is a central lymphoid organ, in which bone marrow-derived T cell precursors undergo a complex process of maturation. Developing thymocytes interact with thymic microenvironment in a defined spatial order. A component of thymic microenvironment, the thymic epithelial cells, is crucial for the maturation of T-lymphocytes through cell-cell contact, cell matrix interactions and secretory of cytokines/chemokines. There is evidence that extracellular matrix molecules play a fundamental role in guiding differentiating thymocytes in both cortical and medullary regions of the thymic lobules. The interaction between the integrin alpha 5 beta 1 (CD49e/CD29; VLA-5) and fibronectin is relevant for thymocyte adhesion and migration within the thymic tissue. Our previous results have shown that adhesion of thymocytes to cultured TEC line is enhanced in the presence of fibronectin, and can be blocked with anti-VLA-5 antibody. Results: Herein, we studied the role of CD49e expressed by the human thymic epithelium. For this purpose we knocked down the CD49e by means of RNA interference. This procedure resulted in the modulation of more than 100 genes, some of them coding for other proteins also involved in adhesion of thymocytes; others related to signaling pathways triggered after integrin activation, or even involved in the control of F-actin stress fiber formation. Functionally, we demonstrated that disruption of VLA-5 in human TEC by CD49e-siRNA-induced gene knockdown decreased the ability of TEC to promote thymocyte adhesion. Such a decrease comprised all CD4/CD8-defined thymocyte subsets. Conclusion: Conceptually, our findings unravel the complexity of gene regulation, as regards key genes involved in the heterocellular cell adhesion between developing thymocytes and the major component of the thymic microenvironment, an interaction that is a mandatory event for proper intrathymic T cell differentiation.

Leucine supplementation improves adiponectin and total cholesterol concentrations despite the lack of changes in adiposity or glucose homeostasis in rats previously exposed to a high-fat diet

Background: Studies suggest that leucine supplementation (LS) has a therapeutic potential to prevent obesity and to promote glucose homeostasis. Furthermore, regular physical exercise is a widely accepted strategy for body weight maintenance and also for the prevention of obesity. The aim of this study was to determine the effect of chronic LS alone or combined with endurance training (ET) as potential approaches for reversing the insulin resistance and obesity induced by a high-fat diet (HFD) in rats. Methods: Forty-seven rats were randomly divided into two groups. Animals were fed a control diet-low fat (n = 10) or HFD (n = 37). After 15 weeks on HFD, all rats received the control diet-low fat and were randomly divided according to treatment: reference (REF), LS, ET, and LS+ET (n = 7-8 rats per group). After 6 weeks of treatment, the animals were sacrificed and body composition, fat cell volume, and serum concentrations of total cholesterol, HDL-cholesterol, triacylglycerol, glucose, adiponectin, leptin and tumor necrosis factor-alpha (TNF-alpha) were analyzed. Results: At the end of the sixth week of treatment, there was no significant difference in body weight between the REF, LS, ET and LS+ET groups. However, ET increased lean body mass in rats (P = 0.019). In addition, ET was more effective than LS in reducing adiposity (P = 0.019), serum insulin (P = 0.022) and TNF-alpha (P = 0.044). Conversely, LS increased serum adiponectin (P = 0.021) levels and reduced serum total cholesterol concentration (P = 0.042). Conclusions: The results showed that LS had no beneficial effects on insulin sensitivity or adiposity in previously obese rats. On the other hand, LS was effective in increasing adiponectin levels and in reducing total cholesterol concentration.

Chain motifs: The tails and handles of complex networks

A great part of the interest in complex networks has been motivated by the presence of structured, frequently nonuniform, connectivity. Because diverse connectivity patterns tend to result in distinct network dynamics, and also because they provide the means to identify and classify several types of complex network, it becomes important to obtain meaningful measurements of the local network topology. In addition to traditional features such as the node degree, clustering coefficient, and shortest path, motifs have been introduced in the literature in order to provide complementary descriptions of the network connectivity. The current work proposes a different type of motif, namely, chains of nodes, that is, sequences of connected nodes with degree 2. These chains have been subdivided into cords, tails, rings, and handles, depending on the type of their extremities (e.g., open or connected). A theoretical analysis of the density of such motifs in random and scale-free networks is described, and an algorithm for identifying these motifs in general networks is presented. The potential of considering chains for network characterization has been illustrated with respect to five categories of real-world networks including 16 cases. Several interesting findings were obtained, including the fact that several chains were observed in real-world networks, especially the world wide web, books, and the power grid. The possibility of chains resulting from incompletely sampled networks is also investigated.

Power-law creep behavior of a semiflexible chain

Rheological properties of adherent cells are essential for their physiological functions, and microrheological measurements on living cells have shown that their viscoelastic responses follow a weak power law over a wide range of time scales. This power law is also influenced by mechanical prestress borne by the cytoskeleton, suggesting that cytoskeletal prestress determines the cell's viscoelasticity, but the biophysical origins of this behavior are largely unknown. We have recently developed a stochastic two-dimensional model of an elastically joined chain that links the power-law rheology to the prestress. Here we use a similar approach to study the creep response of a prestressed three-dimensional elastically jointed chain as a viscoelastic model of semiflexible polymers that comprise the prestressed cytoskeletal lattice. Using a Monte Carlo based algorithm, we show that numerical simulations of the chain's creep behavior closely correspond to the behavior observed experimentally in living cells. The power-law creep behavior results from a finite-speed propagation of free energy from the chain's end points toward the center of the chain in response to an externally applied stretching force. The property that links the power law to the prestress is the chain's stiffening with increasing prestress, which originates from entropic and enthalpic contributions. These results indicate that the essential features of cellular rheology can be explained by the viscoelastic behaviors of individual semiflexible polymers of the cytoskeleton.

Unusual magneto-optical phenomenon reveals low energy spin dispersion in the spin-1 anisotropic Heisenberg antiferromagnetic chain system NiCl(2)-4SC(NH(2))(2)

Electron paramagnetic resonance measurements of NiCl(2)-4SC(NH(2))(2) reveal the low-energy spin dispersion, including a magnetic-field interval in which the two-magnon continuum is within k(B)T of the ground state, allowing a continuum of excitations over a range of k states, rather than only the k=0 single-magnon excitations. This produces a novel Y shape in the frequency-field EPR spectrum measured at T >= 1.5 K. Since the interchain coupling J(perpendicular to)< k(B)T, this shape can be reproduced by a single S=1 antiferromagnetic Heisenberg chain with a strong easy-plane single-ion anisotropy. Importantly, the combination of experiment and modeling we report herein demonstrates a powerful approach to probing spin dispersion in a wide range of interacting magnetic systems without the stringent sample requirements and complications associated with inelastic scattering experiments.

Environmental stresses inhibit splicing in the aquatic fungus Blastocladiella emersonii

Background: Exposure of cells to environmental stress conditions can lead to the interruption of several intracellular processes, in particular those performed by macromolecular complexes such as the spliceosome. Results: During nucleotide sequencing of cDNA libraries constructed using RNA isolated from B. emersonii cells submitted to heat shock and cadmium stress, a large number of ESTs with retained introns was observed. Among the 6,350 ESTs obtained through sequencing of stress cDNA libraries, 181 ESTs presented putative introns (2.9%), while sequencing of cDNA libraries from unstressed B. emersonii cells revealed only 0.2% of ESTs containing introns. These data indicate an enrichment of ESTs with introns in B. emersonii stress cDNA libraries. Among the 85 genes corresponding to the ESTs that retained introns, 19 showed more than one intron and three showed three introns, with intron length ranging from 55 to 333 nucleotides. Canonical splicing junctions were observed in most of these introns, junction sequences being very similar to those found in introns from genes previously characterized in B. emersonii, suggesting that inhibition of splicing during stress is apparently a random process. Confirming our observations, analyses of gpx3 and hsp70 mRNAs by Northern blot and S1 protection assays revealed a strong inhibition of intron splicing in cells submitted to cadmium stress. Conclusion: In conclusion, data indicate that environmental stresses, particularly cadmium treatment, inhibit intron processing in B. emersonii, revealing a new adaptive response to cellular exposure to this heavy metal.

Determination of parathion and carbaryl pesticides in water and food samples using a self assembled monolayer/acetylcholinesterase electrochemical biosensor

An acetylcholinesterase (AchE) based amperometric biosensor was developed by immobilisation of the enzyme onto a self assembled modified gold electrode. Cyclic voltammetric experiments performed with the SAM-AchE biosensor in phosphate buffer solutions ( pH = 7.2) containing acetylthiocholine confirmed the formation of thiocholine and its electrochemical oxidation at E-p = 0.28 V vs Ag/AgCl. An indirect methodology involving the inhibition effect of parathion and carbaryl on the enzymatic reaction was developed and employed to measure both pesticides in spiked natural water and food samples without pre-treatment or pre-concentration steps. Values higher than 91-98.0% in recovery experiments indicated the feasibility of the proposed electroanalytical methodology to quantify both pesticides in water or food samples. HPLC measurements were also performed for comparison and confirmed the values measured amperometrically.

Flow injection analysis of paracetamol using a biomimetic sensor as a sensitive and selective amperometric detector

This work describes the coupling of a biomimetic sensor to a flow injection system for the sensitive determination of paracetamol. The sensor was prepared as previously described in the literature (M. D. P. T. Sotomayor, A. Sigoli, M. R. V. Lanza, A. A. Tanaka and L. T. Kubota, J. Braz. Chem. Soc., 2008, 19, 734) by modifying a glassy carbon electrode surface with a Nafion (R) membrane doped with iron tetrapyridinoporphyrazine (FeTPyPz), a biomimetic catalyst of the P450 enzyme. The performance of the sensor for paracetamol detection was investigated and optimized in a flow injection system (FIA) using a wall jet electrochemical cell. Under optimized conditions a wide linear response range (1.0 x 10(-5) to 5.0 x 10(-2) mol L(-1)) was obtained, with a sensitivity of 2579 (+/- 129) mu A L mu mol(-1). The detection and quantification limits of the sensor for paracetamol in the FIA system were 1.0 and 3.5 mu mol L(-1), respectively. The analytical frequency was 51 samples h(-1), and over a period of five days (320 determinations) the biosensor maintained practically the same response. The system was successfully applied to paracetamol quantification in seven pharmaceutical formulations and in water samples from six rivers in Sao Paulo State, Brazil.

Meiobenthic community underneath the carcass of a stingray: a snapshot after natural death

The impact of large food falls and carrion on meiobenthic communities remains little understood. The objective of the present study was to investigate whether the carcass of a stingray, encountered fortuitously in an Australian estuary, affects the underlying meiobenthic community, in particular nematode assemblages. The integrity of the skeleton and the low redox values observed under the carcass suggest that the cadaver had been slowly and chiefly decomposed by microbes. The abundance and number of meiofaunal taxa, as well as nematode abundance and nematode-species richness, were significantly lower under the carcass when compared to samples outside the carcass. Nonetheless, a few nematode species, typical of hypoxic/anoxic sediments, were more abundant under the carcass. Interestingly, all these species were absent or rare in samples near, but not under, the carcass, suggesting that they may take advantage of the reduced environment created by the carcass and the consequent lack of competition to prosper. As observed for other marine environments, carcasses in estuaries create a microhabitat that supports a characteristic meiobenthic fauna, distinct from those inhabiting the surrounding sediments, but similar to those of reduced habitats.

INAA with gamma-gamma coincidence for selenium determination in food

Selenium detection limits of INAA are normally above its concentration in most biological materials. Gamma-gamma coincidence methodology can be used to improve the detection limits and uncertainties in the determination of selenium. Here, some edible parts of plants were measured using a HPGe detector equipped with a NaI(Tl) active shielding, producing spectra both in normal and coincidence modes. The results presented the reduction of the detection limits of selenium by a factor of 2 to 3 times and improvement in the uncertainty of up to 2 times.

FATTY ACID COMPOSITION OF ACID, BIOLOGICAL AND ENZYMATIC FISH SILAGE

The use of the fish silage as an ingredient in feed for aquatic organisms is an alternative to solve sanitary and environmental problems caused by the lack of an adequate destination for the residues generated by the fishing industry. It would also lower the costs with feed, and consequently the fish production costs, since the expenses with the feed account for approximately 60% of the total cost. The objective of this study was to evaluate the fatty acid composition of the acid silage (AS), biological silage (BS) and enzymatic silage (ES) produced from discardings of the culture and from processing residues of the Nile tilapia (Oreochromis niloticus). The values found for lipids (dry matter basis) were: 12.45; 12.25 and 12.17 g 100 g(-1) for BS, AS, and ES, respectively. The fatty acids present in the lipid fraction of the silages are predominantly unsaturated. Oleic acid was present in larger amounts (30.49, 28.60 and 30.60 g 100 g(-1) of lipids for BS, AS and ES, respectively). Among saturated fatty acids, palmitic and stearic acids were present in larger amounts. Only traces of eicosapentaenoic (EPA) and docosahexaenoic (DHA) fatty acids were found. The silages produced from discardings of the culture and processing residues of the Nile tilapia are not a good source of EPA and DHA for fish feeds.

Effect of Food Restriction and Intense Physical Training on Estrous Cyclicity and Plasma Leptin Concentrations in Rats

Intense physical training and dietary energy restriction have been associated with consequences such as nutritional amenorrhea. We investigated the effects of intense physical training, food restriction or the combination of both strategies on estrous cyclicity in female rats, and the relationship between leptin ad these effects. Twenty-seven female Wistar rats were distributed into four groups: SF: sedentary, fed ad libitum; SR: sedentary subjected to 50% food restriction (based on the food intake of their fed counterparts); TF: trained (physical training on a motor treadmill with a gradual increase in speed and time), fed ad libitum; TR; trained with 50% food restriction. We analysed estrous cyclicity, plasma leptin and estradiol as well as chemical composition of the carcass, body weight variation. and weight of ovaries and perirenal adipose tissue. Data demonstrate that physical training alone was not responsible for significant modifications in either carcass chemical composition or reproductive function. Food restriction reduced leptin levels in all animals and interrupted the estrous cyclicity in some animals, but only the combination of food restriction and physical training was capable of interrupting the estrous cyclicity in all animals. Leptin was not directly related to estrous cyclicity. From our findings, it may be concluded that there is an additive or synergistic effect of energy intake restriction and energy expenditure by intense physical training on estrous cyclicity. Leptin appears to be one among others factors related to estrous cycle, but it probably acts indirectly.

Effects of branched-chain amino acid (BCAA) supplementation on endurance exercise performance of pregnant rats

Aims. - The present study evaluated the effects of BCAA supplementation on exercise performance of pregnant rats. Methods. - In order to assess these effects, Wistar rats were divided into four groups: sedentary not-supplemented (SNS, n = 8); sedentary supplemented (SS, n = 8); trained not-supplemented (TNS, n = 8) and trained supplemented (TS, n = 8). All groups were submitted to the endurance test until exhaustion (ET) and post-effort lactate (PEL) determination before pregnancy (ET-B and PEL-B) and at the 19th day of pregnancy (ET-19 and PEL-19). Results. - The endurance training significantly increased the ET time to exhaustion (p<0.05). Regardless of BCAA supplementation, both endurance trained groups (TS and TNS) showed a longer time to exhaustion, assessed by ET, compared with the sedentary groups (SS and SNS) (p < 0.05). In the TNS, ET-19 time to exhaustion decreased when compared with the period before pregnancy. On the other hand, ET-19 time to exhaustion was not affected in the TS at the end of the pregnancy period. In addition, TS showed a marked PEL-19 reduction when compared with PEL-B. The data presented herein suggest that BCAA supplementation plays an ergogenic role in the maintenance of exercise performance during pregnancy in rats. (C) 2008 Elsevier Masson SAS. All rights reserved.