Molecular determinants of improved cathepsin B inhibition by new cystatins obtained by DNA shuffling

Background: Cystatins are inhibitors of cysteine proteases. The majority are only weak inhibitors of human cathepsin B, which has been associated with cancer, Alzheimer's disease and arthritis. Results: Starting from the sequences of oryzacystatin-1 and canecystatin-1, a shuffling library was designed and a hybrid clone obtained, which presented higher inhibitory activity towards cathepsin B. This clone presented two unanticipated point mutations as well as an N-terminal deletion. Reversing each point mutation independently or both simultaneously abolishes the inhibitory activity towards cathepsin B. Homology modeling together with experimental studies of the reverse mutants revealed the likely molecular determinants of the improved inhibitory activity to be related to decreased protein stability. Conclusion: A combination of experimental approaches including gene shuffling, enzyme assays and reverse mutation allied to molecular modeling has shed light upon the unexpected inhibitory properties of certain cystatin mutants against Cathepsin B. We conclude that mutations disrupting the hydrophobic core of phytocystatins increase the flexibility of the N-terminus, leading to an increase in inhibitory activity. Such mutations need not affect the inhibitory site directly but may be observed distant from it and manifest their effects via an uncoupling of its three components as a result of increased protein flexibility.

Sequential injection analysis implementing multiple standard additions for As speciation by liquid chromatography and atomic fluorescence spectrometry (SIA-HPLC-AFS)

An analytical procedure for multiple standard additions of arsenic species using sequential injection analysis (SIA) is proposed for their quantification in seafood extracts. SIA presented flexibility for generating multiple specie standards at the ng mL(-1) concentration level by adding different volumes of As(III), As(V), monomethylarsonic (MMA) and dimethylarsinic (DMA) to the sample. The mixed sample plus standard solutions were delivered from SIA to fill the HPLC injection loop. Subsequently, As species were separated by HPLC and analyzed by atomic fluorescence spectrometry (AFS). The proposed system comprised two independently controlled modules, with the HPLC loop acting as the intermediary device. The analytical frequency was enhanced by combining the actions of both modules. While the added sample was flowing through the chromatographic column towards the detection system, the SIA program started performing the standard additions to another sample. The proposed method was applied to spoiled seafood extracts. Detection limits based on 3 sigma for As(III), As(V), MMA and DMA were 0.023, 0.39, 0.45 and 1.0 ng mL(-1), respectively. (C) 2011 Elsevier B.V. All rights reserved.

Quantitative assessment of postural alignment in young adults based on photographs of anterior, posterior, and lateral views

Objective: Postural assessment through photography is a simple method that allows the acquisition of quantitative values to define the alignment of body segments. The purpose of this study was to quantitatively assess the postural alignment of several body segments in standing through anterior, posterior, and lateral views. Methods: In this cross-sectional study, 122 subjects were initially evaluated. Seven subjects were excluded from the study after cluster analysis. The final sample had 115 subjects, 75% women with a mean age of 26 + 7 years. Photographs were taken from anterior, posterior, and lateral views after placement of markers on specific anatomical points. Photographs were analyzed using free Postural Analysis Software/Software of Postural Analysis (PAS/SAPO). Quantitative values for postural analysis variables were ascertained for head, upper and lower limbs, and trunk, along with the frequency of inclinations to the left and to the right. Results: Regarding the head, 88% of the sample presented some inclination, 67% of which was to the right. There was a predominance of right inclination of the shoulder and pelvis in 68% and 43% of study subjects, respectively. Lower limbs presented mean alignment of 178 in the anterior view, and the trunk showed predominant right inclination in 66% of participants. Conclusion: Small asymmetries were observed in anterior and posterior views. This study suggests that there is no symmetry in postural alignment and that small asymmetries represent the normative standard for posture in standing. (J Manipulative Physiol Ther 2011;34:371-380)

Is acute static stretching able to reduce the time to exhaustion at power output corresponding to maximal oxygen uptake?

Samogin Lopes, FA, Menegon, EM, Franchini, E, Tricoli, V, and de M. Bertuzzi, RC. Is acute static stretching able to reduce the time to exhaustion at power output corresponding to maximal oxygen uptake? J Strength Cond Res 24(6): 1650-1656, 2010-This study analyzed the effect of an acute static stretching bout on the time to exhaustion (T(lim)) at power output corresponding to (V) over dotO(2)max. Eleven physically active male subjects (age 22.3 +/- 2.8 years, (V) over dotO(2)max 2.7 +/- 0.5 L . min(-1)) completed an incremental cycle ergometer test, 2 muscle strength tests, and 2 maximal tests to exhaustion at power output corresponding to (V) over dotO(2)max with and without a previous static stretching bout. The T(lim) was not significantly affected by the static stretching (164 +/- 28 vs. 150 +/- 26 seconds with and without stretching, respectively, p = 0.09), but the time to reach (V) over dotO(2)max (118 +/- 22 vs. 102 +/- 25 seconds), blood-lactate accumulation immediately after exercise (10.7 +/- 2.9 vs. 8.0 +/- 1.7 mmol . L(-1)), and oxygen deficit (2.4 +/- 0.9 vs. 2.1 +/- 0.7 L) were significantly reduced (p <= 0.02). Thus, an acute static stretching bout did not reduce T(lim) at power output corresponding to (V) over dotO(2)max possibly by accelerating aerobic metabolism activation at the beginning of exercise. These results suggest that coaches and practitioners involved with aerobic dependent activities may use static stretching as part of their warm-up routines without fear of diminishing high-intensity aerobic exercise performance.

Testing motor fitness in karate

Background and Study Aim: Evaluation of sport skills test can be very useful tool for coach practice. The aim of the present paper was: (a) to evaluate the reliability and accuracy of the Specific Physical Fitness Tests (SPFT) (b) to review the results of karate athletes who represent different weight categories, and who are at different stages of schooling; (c) to establish grading criteria of physical fitness preparation. Material/Methods: The reseach was conducted among 219 Kyokushin karate players, whose profiles were presented as (chi) over bar +/- SD and their main characteristics were the following: age 26.8 +/- 4.67 (19-39) years, body mass 75.2 +/- 8.35 (50-97) kg and body height 176.4 +/- 5.67 (160-196) cm. The value of the BMI amounted to 24.1 +/- 2.17 (17.9-29.4) kg/m(2). All the subjects of the research had training experience of 10.5 +/- 3.71 (4-20) years and their degree of proficiency ranged from 4(th) kyu to 3(rd) dan. The physical fitness trials proposed by Story (1989) included: hip turning speed, speed punches, flexibility, rapid kicks, agility, and evasion actions. It was supplemented by a test of local strength endurance, composing a battery of the SPFT, which was implemented by first of the authors between 1991 and 2006. Results: SPFT is characterized by high reliability and it can be used to diagnose the physical fitness preparation and monitor the individual results of training. It discriminates accurately competitors with different sports level and it is characterized by very high accuracy, it is correlated with the test results of motor general physical fitness abilities and coordination abilities as well as it is connected with the somatic build of the athlete. The performance classification table was developed on the basis of our research. Discussion: Results obtained in SPFT were shortly discussed. Conclusions: The collected results of our research allowed us to come to, the conclusion: The table can be applied not only to assess karate fighters, but also adepts in taekwondo, kick-boxing, ju-jitsu, hapkido or other mixed martial arts.

Transient effects of stretching exercises on gait parameters of elderly women

This study aimed to analyse the effects of a single stretching exercise session on a number of gait parameters ill elderly participants in all attempt to determine whether these exercises can influence the risk of fall. Fifteen healthy women living in the community Volunteered to participate in the study. A kinematic gait analysis was performed immediately before and after a session of static stretching exercises applied oil hip flexor/extensor muscles. Results showed a significant influence of stretching exercises on a number of gait parameters, which have previously been proposed as fall predictors. Participants showed increased gait velocity, greater step length and reduced double Support time during stance after performing stretching exercises, suggesting improved stability and mobility. Changes around the pelvis (increased anterior-posterior tilt and rotation range of motion) resulting from the stretching exercises were suggested to influence the gait parameters (velocity, step length and double support time). Therefore, stretching exercises were shown to be a promising strategy to facilitate changes in gait parameters related to the risk of fall. Some other gait variables related to the risk of fall remained Unaltered (e.g., toe clearance). The stable pattern of segmental angular velocities was proposed to explain the stability of these unchanged gait variables. The results indicate that stretching exercises, performed oil a regular (daily) basis, result in gait adaptations which can be considered as indicative of reduced fall risk. Other Studies to determine whether regular stretching routines are an effective strategy to reduce the risk of fall are required. (C) 2008 Elsevier Ltd. All rights reserved.

Aerobic profile of climbers during maximal arm test

This study compared measurements of upper body aerobic fitness in elite (EC; n = 7) and intermediate rock climbers (IC; n = 7), and a control group (C; n = 7). Subjects underwent an upper limb incremental test on hand cycle ergometer, with increments of 23 W.min(-1), until exhaustion. Ventilation (VE) data were smoothed to 10 s averages and plotted against time for the visual determination of the first (VT1) and second (VT2) ventilatory thresholds. Peak power output was not different among groups [EC = 130.9 (+/- 11.8) W; IC = 122.1 (+/- 28.4) W; C = 115.4 (+/- 15.1) W], but time to exhaustion was significantly higher in EC than IC and C. VO(2PEAK) was significantly higher in EC [36.8 (+/- 5.7) mL.kg(-1).min(-1)] and IC [35.5 (+/- 5.2) mL.kg(-1).min(-1)] than C [28.8 (+/- 5.0) mL.kg(-1).min(-1)], but there was no difference between EC and IC. VT1 was significantly higher in EC than C [EC = 69.0 (+/- 9.4) W; IC = 62.4 (+/- 13.0) W; C = 52.1 (+/- 11.8) W], but no significant difference was observed in VT2 [EC = 103.5 (+/- 18.8) W; IC = 92.0 (+/- 22.0) W; C = 85.6 (+/- 19.7) W]. These results show that elite indoor rock climbers elicit higher aerobic fitness profile than control subjects when measured with an upper body test.

Physiological, performance, and nutritional profile of the Brazilian olympic wushu (kung-fu) team

Artioli, GG, Gualano, B, Franchini, E, Batista, RN, Polacow, VO, and Lancha, AH Jr. Physiological, performance, and nutritional profile of the Brazilian Olympic Wushu (kung-fu) team. J Strength Cond Res 23(1): 20-25, 2009-The purpose of the present study was to determine physiological, nutritional, and performance profiles of elite Olympic Wushu (kung-fu) athletes. Ten men and four women elite athletes took part in the study. They completed the following tests: body composition, nutritional assessment, upper-body Wingate Test, vertical jump, lumbar isometric strength, and flexibility. Blood lactate was determined at rest and after the Wingate Test. Blood lactate was also determined during a training session (combat and Taolu training). We found low body fat (men: 9.5 +/- 6.3%; women: 18.0 +/- 4.8%), high flexibility (sit-and-reach-men: 45.5 +/- 6.1 cm; women: 44.0 +/- 6.3 cm), high leg power (vertical jump-men: 37.7 +/- 8.4 cm; women: 32.3 +/- 1.1 cm), high lumbar isometric strength (men: 159 6 13 cm; women: 94 6 6 cm), moderate arm mean and peak power (Wingate Test-men: 4.1 +/- 0.4 and 5.8 +/- 0.5 W.kg(-1), respectively; women: 2.5 +/- 0.3 and 3.4 +/- 0.3 W.kg(-1), respectively), and elevated blood lactate after the Wingate Test (men: 10.8 +/- 2.0 mmol.L(-1); women: 10.2 +/- 2.0 mmol.L(-1)) and during training (combat: 12.0 +/- 1.8 mmol.L(-1); Taolu: 7.7 +/- 3.3 mmol.L(-1)). Men athletes consume a high-fat, low-carbohydrate diet, whereas women consume a moderate, high-carbohydrate diet. Energy consumption was markedly variable. In conclusion, Olympic Wushu seems to be a highly anaerobic-dependent combat sport. Low body fat, high flexibility, leg anaerobic power, isometric strength, and moderately high arm anaerobic power seem to be important for successful competitive performance.

Utility of Quantitative Ultrasound of the Calcaneus in Diagnosing Osteoporosis in Spinal Cord Injury Patients

Bittar CK, Cliquet A Jr, dos Santos Floter M: Utility of quantitative ultrasound of the calcaneus in diagnosing osteoporosis in spinal cord injury patients. Am J Phys Med Rehabil 2011;90:477-481. Objective: The aim of this study was to assess the utility of quantitative ultrasound of the calcaneus in diagnosing osteoporosis in spinal cord injury patients in a Brazilian Teaching Hospital. Design: This is a diagnostic test criterion standard comparison study. Between January 2008 and October 2009, the bone density of 15 spinal cord injury patients was assessed for analysis before beginning rehabilitation using muscle stimulation. The bone density was assessed using bone densitometry examination (DEXA) and ultrasound examination of the calcaneus (QUS). The measurements acquired using QUS and DEXA were compared between patients with spinal cord injury and a control group of ten healthy individuals. Results: The T-score values for femoral neck using DEXA (P < 0.0022) and those using QUS of the calcaneus (P < 0.0005) differed significantly between the groups, and the means in the normal subjects were higher than those in spinal cord injury patients who would receive electrical stimulation. In spinal cord injury patients, the significant differences were found between the QUS T-score for calcaneus and the DEXA scores for the lumbar spine and femoral neck. Conclusions: Because of the low level of mechanical stress on the calcaneus, the results of the QUS could not be correlated with the DEXA results for diagnosing osteoporosis. Therefore, QUS seems to be not a good choice for diagnosis and follow-up.

Sagittal spinal alignment in paraplegics: a new paradigm for the rehabilitation under neuromuscular electrical stimulation

Study design: Radiographic analysis of sagittal spinal alignment of paraplegics in a standing position under surface neuromuscular electrical stimulation (NMES). Objectives: Describing the radiographic parameters of the sagittal spinal alignment of paraplegics going through a rehabilitation program with NMES. Setting: The University Hospital`s Ambulatory (UNICAMP), Campinas, Sao Paulo, Brazil. Methods: Panoramic X-ray images in profile were taken for 10 paraplegics. All patients participated in the rehabilitation program and were able to perform gait through NMES of the femoral quadriceps muscles. The radiographic parameters used for the analysis were the same as those described in the literature for healthy people. The results were didactically organized into three groups: anatomical shape of the spine, morphology and kinetics of the pelvis and spinopelvic alignment. Results: The physiological curvature of the spine in paraplegics showed average values similar to those described in the literature for healthy patients. The inversion of the pelvic tilt and the increase in the sacral slope were defined by the anterior backward rotation of the pelvis. The existing theoretical mathematical formulas that define lumbar lordosis, pelvic incidence and pelvic tilt showed normal values, despite the anterior intense sagittal imbalance. Conclusions: The adaptive posture of the spine in paraplegics standing through the stimulation of the femoral quadriceps does not allow for a neutral sagittal alignment. This novel radiographic detailed description of the various segments of the spine can be of assistance toward the understanding of the global postural control for such subjects. Spinal Cord (2010) 48, 251-256; doi: 10.1038/sc.2009.123; published online 29 September 2009

Effect of Fill Time on the Performance of Pilot-scale ASBR and AnSBBR Applied to Sanitary Wastewater Treatment

Many lab-scale studies have been carried out regarding the effect of feed strategy on the performance of anaerobic sequencing batch reactors (ASBR); however, more detailed pilot-scale studies should be performed to assess the real applicability of this type of operation. Therefore, the objective of this work was to assess the effect of feed strategy or fill time in a 1-m(3) mechanically stirred pilot-scale sequencing batch reactor, treating 0.65 m(3) sanitary wastewater in 8-h cycles at ambient temperature. Two reactor configurations were used: one containing granular biomass (denominated ASBR) and the other immobilized biomass on polyurethane foam as inert support (denominated anaerobic sequencing batch biofilm reactor (AnSBBR)). The reactors were operated under five distinct feed strategies, namely: typical batch and fed-batch for 25%, 50%, 75%, and 100% of the cycle length. Stirring frequency in the ASBR was 40 rpm with two flat-blade turbine impellers and 80 rpm in the AnSBBR with two helix impellers. The results showed that both the ASBR and AnSBBR when operated under typical batch, fed-batch for 50% and 75% of the cycle length, presented improved organic matter removal efficiencies, without significant differences in performance, thus showing important operational flexibility. In addition, the reactors presented operation stability under all conditions.

Effect of specific feed volume on the performance of an anaerobic sequencing batch biofilm reactor (AnSBBR) with circulation treating different wastewaters under different organic loads

The objective of this research was to study the behavior of two anaerobic sequencing batch reactors, containing immobilized biomass (AnSBBR), as a function of the ratio of the volume of treated medium in each cycle to the total volume of reaction medium. The reactors, in which mixing was accomplished by recirculation of the liquid phase, were maintained at 30 +/- 1 degrees C and treated different wastewaters in 8-h cycles. The operational conditions imposed had the objective to investigate whether maintenance of a residual volume in the reactor would affect, at the end of each cycle, process efficiency and stability, as well as to verify the intensity of the effect for different types of wastewaters and organic loading rates. The first reactor, with work volume of 2.5 L, treated reconstituted cheese whey at an organic loading rate of 12 g COD.L(-1).d(-1) and presented similar effluent quality for the four conditions under which it was operated: renewal of 100, 70, 50 and 25 % of its work volume at each cycle. Despite the fact that reduction in the renewed volume did not significantly affect effluent quality, in quantitative terms, this reduction resulted in an increase in the amount of organic matter removed by the first reactor. The second reactor, with work volume of 1.8 L, treated synthetic wastewater at organic loading rates of 3 and 5 g COD.L(-1).d(-1) and operated under two conditions for each loading: renewal of 100 and 50 % of its work volume. At the organic loading rate of 3 g COD.L(-1).d(-1), the results showed that both effluent quality and amount of organic matter removed by the second reactor were independent of the treated volume per cycle. At the organic loading rate of 5 g COD.L(-1).d(-1), although the reduction in the renewed volume did not affect the amount of organic matter removed by the reactor, effluent quality improved during reactor operation with total discharge of its volume. In general, results showed process stability under all conditions, evidencing reactor flexibility and the potential to apply this technology in the treatment of different types of wastewater.

Anaerobic sequencing batch biofilm reactor applied to automobile industry wastewater treatment: Volumetric loading rate and feed strategy effects

This paper presents a technological viability study of wastewater treatment in an automobile industry by an anaerobic sequencing batch biofilm reactor containing immobilized biomass (AnSBBR) with a draft tube. The reactor was operated in 8-h cycles, with agitation of 400 rpm, at 30 degrees C and treating 2.0 L wastewater per cycle. Initially the efficiency and stability of the reactor were studied when supplied with nutrients and alkalinity. Removal efficiency of 88% was obtained at volumetric loading rate (VLR) of 3.09 mg COD/L day. When VLR was increased to 6.19 mg COD/L day the system presented stable operation with reduction in efficiency of 71%. In a second stage the AnSBBR was operated treating wastewater in natura, i.e., without nutrients supplementation, only with alkalinity, thereby changing feed strategy. The first strategy consisted in feeding 2.0 L batch wise (10 min), the second in feeding 1.0 L of influent batch wise (10 min) and an additional 1.0 L fed-batch wise (4 h), both dewatering 2.0 L of the effluent in 10 min. The third one maintained 1.0 L of treated effluent in the reactor, without discharging, and 1.0 L of influent was fed fed-batch wise (4 h) with dewatering 1.0 L of the effluent in 10 min. For all implemented strategies (VLR of 1.40, 2.57 and 2.61 mg COD/L day) the system presented stability and removal efficiency of approximately 80%. These results show that the AnSBBR presents operational flexibility, as the influent can be fed according to industry availability. In industrial processes this is a considerable advantage, as the influent may be prone to variations. Moreover, for all the investigated conditions the kinetic parameters were obtained from fitting a first-order model to the profiles of organic matter, total volatile acids and methane concentrations. Analysis of the kinetic parameters showed that the best strategy is feeding 1.0 L of influent batchwise (10 min) and 1.0 L fed-batch wise (4 h) in 8-h cycle. (c) 2007 Elsevier B.V. All rights reserved.

Bond strength and transversal deformation aging on cement-polymer adhesive mortar

Polymer-modified mortar is widely used to set ceramic tiles used as external finishing for high rise buildings in countries such as Brazil, Israel, Singapore and Portugal, mainly because it shows better bond strength and flexibility as compared to the traditional ones. Despite this, the results in the literature already published concerning the long-term performance of those composite mortars are is not conclusive. This paper, based on a laboratory program, compared the performance over time of four commercial polymer-modified adhesive mortars exposed to a typical Brazilian outdoor aging environment and to an indoor environment in terms of mortar flexibility and the bond strength to porcelain tiles. The results show that under laboratory condition, the mortars are more flexible and have higher bond strength than under external condition, and that there is an important correlation between the transversal deformability and the bond strength. (C) 2008 Elsevier Ltd. All rights reserved.

The influence of moisture on the deformability of cement-polymer adhesive mortar

Adhesive mortars are widely used to set porcelain stoneware tiles on buildings because their bond strength and flexibility properties increase the cladding serviceability. However, their long-term performance is not well understood, mainly the degradation of the polymeric matrix. The influence of moisture content on the flexibility of six adhesive mortars is investigated, based on standard EN 12002. Four of them have defined formulations and the other two are commercial and are widely used to set porcelain stoneware tiles on building facades in Brazil. The results show that moisture content above 6% is sufficient to reduce 50% of the mortar deformability, but that the drying process allows it to recover to a value similar to that prior to saturation; a logarithmic function best fits the correlation between moisture content and flexibility; water immersion increases matrix rigidity. It is suggested that standards should consider flexibility tests on both dried and wet samples as a requirement for polymer-modified mortars. (C) 2010 Elsevier Ltd. All rights reserved.