Molecular modeling and structural analysis of nAChR variants uncovers the mechanism of resistance to snake toxins

Nicotinic acetylcholine receptors (nAChRs) are neuromuscular proteins responsible for muscle contraction upon binding with chemical stimulant acetylcholine (ACh). The α-neurotoxins of snake mimic the structure of ACh and attacks nAChRs, which block the flow of ACh and leads to numbness and paralysis. The toxin-binding site of alpha subunit in the nAChRs is highly conserved throughout chordate lineages with few exceptions in resistance organisms. In this study, we have analyzed the sequence and structures of toxin-binding/resistant nAChRs and their interaction stability with toxins through molecular docking and molecular dynamics simulation (MDS). We have reported the potential glycosylation residues within the toxin-binding cleft adding sugar moieties through N-linked glycosylation in resistant organisms. Residue variations at key positions alter the secondary structure of binding cleft, which might interfere with toxin binding and it could be one of the possible explanations for the resistance to snake venoms. Analysis of nAChR-α-neurotoxin complexes has confirmed the key interacting residues. In addition, drastic variation in the binding stability of Mongoose nAChR-α-Bungarotoxin (α-BTX) and human nAChR-α-BTX complexes were found at specific phase of MDS. Our findings suggest that specific mutations in the binding site of toxin are potentially preventing the formation of stable complex of receptor-toxin, which might lead to mechanism of resistance. This in silico study on the binding cleft of nAChR and the findings of interacting residues will assist in designing potential inhibitors as therapeutic targets.     

Effects of application resistive and flex resistance to breath in training sportsmen

The research on studying influence of a rate of muscular training in conditions of additional aerodynamic and flex resistance to breath on a level of physical working capacity, aerobic productivity and a functional condition of system of breath is carried out. It is shown, that regular muscular training on a background of use of the dosed out breath with increased aerodynamic and flex resistance to respiratory streams and respiratory movements, first of all, provides essential growth of functionalities of the respiratory muscles expressed in authentic increase of power and speed-power parameters, shortening of time of impellent reaction of respiratory muscles. Improvement of a functional condition of respiratory system, and, in particular, respiratory muscles, causes significant growth of physical working capacity and physical readiness of young sportsmen.     

细菌在油水界面粘附的理论、特性及应用

细菌在油水界面粘附主要应用于烷烃污染的环境修复、石油开采与加工、食品加工等方面,其研究已经引起各界的广泛关注。本文基于XDLVO理论阐述细菌在油水界面的粘附机理;总结细菌表面特性、油相及水相性质对细菌粘附的影响;介绍细菌粘附作用在微生物驱油、生物乳化与破乳以及烷烃降解方面的应用,并对今后的研究方向提出展望。     

Estrogen receptor beta and truncated variants enhance the expression of transfected MMP-1 promoter constructs in response to specific mechanical loading

Background

Joint diseases such as osteoarthritis (OA) predominantly afflict post-menopausal women, suggesting a pertinent role for female hormones. Estrogen receptor beta (ER-β) has been detected in connective tissues of the knee joint suggesting that these tissues are responsive to the hormone estrogen. Matrix metalloproteinase-1 (MMP-1) activity contributes to cartilage degradation, a key factor leading to OA development in synovial joints. Two polymorphic forms of MMP-1 exist due to a deletion/insertion of the guanine residue in the promoter, and the 2G allelic variant of MMP-1 exhibits more activity than the 1G allele. Previous studies have demonstrated that the polymorphic forms of the human MMP-1 are influenced by the modulating effects of estrogen receptor isoforms. In addition to hormonal influences, physiological factors such as altered mechanical loading are also contributory features of OA. In the present study, the combined influence of biomechanical and hormonal variables on the activity of MMP-1 isoforms was evaluated. We hypothesized that the combined effects of ER-β and sheer stress will differentially activate the two allelic forms of MMP-1 in a hormone-independent manner.

Methods

HIG-82 synoviocytes were transiently transfected with 1G or 2G alleles (±) ER-β and subjected to either shear or equibiaxial stress. Next, 1G/2G promoter activity was measured to determine the combined influence of physiological stimuli. Truncated ER-β constructs were used to determine the importance of different domains of ER-β on 1G/2G activation.

Results

The 2G allele exhibited a constitutively higher activity than the 1G allele, which was further increased when the transfected cells were subject to shear stress, but not equibiaxial stress. Moreover, the combination of ER-β and shear stress further increased the activity levels of the 1G/2G allelic variants. Additionally, select AF-2 truncated ER-β variants led to increased activity levels for the 2G allele, indicating the AF-1 domain was likely involved in the response to mechanical stimulation.

Conclusions

These results suggest that the 1G/2G alleles of MMP-1 are influenced by specific mechanical stimuli like shear stress, as well as the ER-β receptor. These findings contribute to the potential allelic involvement in connective tissue diseases such as OA in females compared to males.

     

The application of training to failure in periodized multiple-set resistance exercise programs

Few studies and reports in the body of literature have directly addressed the issue of whether resistance exercise sets should be performed to failure. Research has clearly demonstrated the superiority of performing multiple sets vs. single sets for increases in maximal strength. However, there is little direct evidence to decide conclusively whether or not multiple sets should be performed to failure. Therefore, the purpose of this research note was to discuss what is currently known concerning the application of training to failure and to stimulate further research on this topic. Although not essential for increases in muscular characteristics such as strength and hypertrophy, training to failure might allow advanced lifters to break through training plateaus when incorporated periodically into short-term microcycles. Because muscular hypertrophy is a key contributor to long-term increases in maximal strength, advanced lifters should consider training to failure occasionally. The potential mechanisms by which training to failure might provide an advantage are through greater activation of motor units and secretion of growth-promoting hormones. However, training to failure is not an effective stimulus without lifting at a sufficient intensity (percentage of 1 repetition maximum). Furthermore, training to failure should not be performed repeatedly over long periods, due to the high potential for overtraining and overuse injuries. Therefore, the training status and the goals of the lifter should guide the decision-making process on this issue.     

Cellular accommodation and the response of bone to mechanical loading

Several mathematical rules by which bone adapts to mechanical loading have been proposed. Previous work focused mainly on negative feedback models, e.g., bone adapts to increased loading after a minimum strain effective (MES) threshold has been reached. The MES algorithm has numerous caveats, so we propose a different model, according to which bone adapts to changes in its mechanical environment based on the principle of cellular accommodation. With the new algorithm we presume that strain history is integrated into cellular memory so that the reference state for adaptation is constantly changing. To test this algorithm, an experiment was performed in which the ulnae of Sprague-Dawley rats were loaded in axial compression. The animals received loading for 15 weeks with progressively decreasing loads, increasing loads, or a constant load. The results showed the largest increases in geometry in the decreasing load group, followed by the constant load group. Bone formation rates (BFRs) were significantly greater in the decreasing load group during the first 2 weeks of the study as compared to all other groups (P<0.05). After the first few weeks of mechanical loading, the BFR in the loaded ulnae returned to the values of the nonloaded ulnae. These experimental results closely fit the predicted results of the cellular accommodation algorithm. After the initial weeks of loading, bone stopped responding so the degree of adaptation was proportional to the initial peak load magnitude.     

In vivo human tendon mechanical properties: effect of resistance training in old age

Recent advances in ultrasound scanning have made it possible to obtain the mechanical properties of human tendons in vivo. Application of the in vivo method in elderly individuals showed that their patellar tendons stiffened in response to a 14-week resistance training program by approximately 65% both structurally and materially. The rate of muscle torque development increased by approximately 27%, indicating faster contractile force transmission to the skeleton. The present findings suggest that strength training in old age can, at least partly, reverse the deteriorating effect of ageing on tendon properties and function.     

Chronic resistance training in women potentiates growth hormone in vivo bioactivity: characterization of molecular mass variants

This investigation determined the influence of acute and chronic resistance exercise on responses of growth hormone (GH) molecular variants in women. Seventy-four healthy young women (23 +/- 3 yr, 167 +/- 7 cm, 63.8 +/- 9.3 kg, 26.3 +/- 4.0% body fat) performed an acute bout of resistance exercise (6 sets of 10 repetition maximum squat). Blood samples were obtained pre- and postexercise. Resulting plasma was fractionated by molecular mass (fraction A, >60 kDa; fraction B, 30-60 kDa; and fraction C, <30 kDa) using chromatography. Fractionated and unfractionated (UF) plasma was then assayed for GH using three different detection systems (monoclonal immunoassay, polyclonal immunoassay, and rat tibial line in vivo bioassay). Subjects were then matched and randomly placed into one of four resistance exercise training groups or a control group for 24 wk. All experimental procedures were repeated on completion of the 24-wk resistance training programs. After acute exercise, immunoassays showed consistent increases in UF GH samples and fractions B and C; increases in fraction A using immunoassay were seen only in the monoclonal assay. No consistent changes in bioactive GH were found following acute exercise. Conversely, chronic exercise induced no consistent changes in immunoassayable GH of various molecular masses, whereas, in general, bioassayable GH increased. In summary, although acute exercise increased only immunoactive GH, chronic physical training increased the biological activity of circulating GH molecular variants. Increased bioactive GH was observed across all fractions and training regimens, suggesting that chronic resistance exercise increased a spectrum of GH molecules that may be necessary for the multitude of somatogenic and metabolic actions of GH.     

Leptin and leptin receptor genetic variants associate with habitual physical activity and the arm body composition response to resistance training

Purpose

We investigated the influence of Leptin (LEP) and leptin receptor (LEPR) SNPs on habitual physical activity (PA) and body composition response to a unilateral, upper body resistance training (RT) program.

Methods

European-derived American volunteers (men = 111, women = 131, 23.4 ± 5.4 yr, 24.4 ± 4.6 kg·m^− 2) were genotyped for LEP 19 G>A (rs2167270), and LEPR 326 A>G (rs1137100), 668 A>G (rs1137101), 3057 G>A (rs1805096), and 1968 G>C (rs8179183). They completed the Paffenbarger PA Questionnaire. Arm muscle and subcutaneous fat volumes were measured before and after 12 wk of supervised RT with MRI. Multivariate and repeated measures ANCOVA tested differences among phenotypes by genotype and gender with age and body mass index as covariates.

Results

Adults with the LEP 19 GG genotype reported more kcal/wk in vigorous intensity PA (1273.3 ± 176.8, p = 0.017) and sports/recreation (1922.8 ± 226.0, p < 0.04) than A allele carriers (718.0 ± 147.2, 1328.6 ± 188.2, respectively). Those with the LEP 19 GG genotype spent more h/wk in light intensity PA (39.7 ± 1.6) than A allele carriers (35.0 ± 1.4, p = 0.03). In response to RT, adults with the LEPR 668 G allele gained greater arm muscle volume (67,687.05 ± 3186.7 vs. 52,321.87 ± 5125.05 mm³, p = 0.01) and subcutaneous fat volume (10,599.89 ± 3683.57 vs. − 5224.73 ± 5923.98 mm³, p = 0.02) than adults with the LEPR 668 AA genotype, respectively.

Conclusion

LEP19 G>A and LEPR 668 A>G associated with habitual PA and the body composition response to RT. These LEP and LEPR SNPs are located in coding exons likely influencing LEP and LEPR function. Further investigation is needed to confirm our findings and establish mechanisms for LEP and LEPR genotype and PA and body composition associations we observed.     

Lateral specificity in resistance training: the effect of bilateral and unilateral training

 Maximal voluntary strength of simultaneous bilateral exertion has been shown to be small compared to the sum of the unilateral exertions. Three experiments were conducted to determine the effects of bilateral and unilateral resistance training on this bilateral deficit and to compare these in hands, arms, and legs. In each experiment, the subjects were divided into three groups: unilateral training group, bilateral training group, and control group. The subjects of the training group performed maximal isometric handgrip training in experiment I, and maximal isokinetic arm and leg extension training in experiments II and III. In each experiment, the subjects of the training group continued one of these resistance training exercises three times a week, for 6 weeks. The increase in handgrip strength of the bilateral training group produced in the bilateral condition [5.1 (SEM 2.4)%, after 3 weeks, 6.4 (SEM 2.3) %, after 6 weeks] was significantly greater compared with the control group [−1.1 (SEM 1.0) %, after 3 weeks, −1.5 (SEM 1.1) %, after 6 weeks. The increase in leg extension power of the bilateral training group produced in the bilateral condition [16.1 (SEM 9.6) %, after 3 weeks, 24.1 (SEM 7.4) %, after 6 weeks] was significantly greater compared with the unilateral training group [−5.0 (SEM 3.4) %, after 3 weeks, −3.4 (SEM 4.2) %, after 6 weeks] and the control group [−4.3 (SEM 2.5) %, after 3 weeks, 1.5 (SEM 5.5) %, after 6 weeks]. The increase in handgrip strength of the unilateral training group produced in the unilateral condition [7.3 (SEM 1.7) %, after 3 weeks] was significantly greater compared with the control group [−0.9 (SEM 1.8) %, after 3 weeks]. The increase in arm extension power of the unilateral training group produced in the unilateral condition [7.2 (SEM 1.8) %, after 6 weeks] was significantly greater compared with the bilateral training group [−3.0 (SEM 2.3) %, after 6 weeks] and the control group [−2.1 (SEM 2.6) %, after 6 weeks]. Bilateral indexes (BI) were shifted in a positive direction by bilateral training and tended to shift in a negative direction by unilateral training. With regard to the magnitude of change in BI, there were no significant differences among handgrip, arm extension, and leg extension training. It is suggested that there is lateral specificity in resistance training and that there is no difference among body parts in the modification of bilateral deficit by lateral training. Accepted: 27 August 1996     

Difference in kinematics and kinetics between high- and low-velocity resistance loading equated by volume: implications for hypertrophy training

Although it is generally accepted that a high load is necessary for muscle hypertrophy, it is possible that a low load with a high velocity results in greater kinematics and kinetics than does a high load with a slow velocity. The purpose of this study was to determine if 2 training loads (35 and 70% 1 repetition maximum [1RM]) equated by volume, differed in terms of their session kinematic and kinetic characteristics. Twelve subjects were recruited in this acute randomized within-subject crossover design study. Two bouts of a half-squat exercise were performed 1 week apart, one with high load-low velocity (HLLV = 3 sets of 12 reps at 70% 1RM) and the other with low-load high-velocity (LLHV = 6 sets of 12 reps at 35% 1RM). Time under tension (TUT), average force, peak force (PF), average power (AP), peak power (PP), work (TW), and total impulse (TI) were calculated and compared between loads for the eccentric and concentric phases. For average eccentric and concentric single repetition values, significantly (p < 0.05) greater (～15-22%) PP outputs were associated with the LLHV loading, whereas significantly greater (～7-61%) values were associated with the HLLV condition for most other variables of interest. However, in terms of total session kinematics and kinetics, the LLHV protocol resulted in significantly greater (～16-61%) eccentric and concentric TUT, PF, AP, PP, and TW. The only variable that was significantly greater for the HLLV protocol than for the LLHV protocol was TI (～20-24%). From these results, it seems that the LLHV protocol may offer an equal if not better training stimulus for muscular adaptation than the HLLV protocol, because of the greater time under tension, power, force, and work output when the total volume of the exercise is equated.     

Optimization of paper bridge loading for 2-DE analysis in the basic pH region: application to the mitochondrial subproteome

Separation of basic proteins with 2-DE presents technical challenges involving protein precipitation, load limitations, and streaking. Cardiac mitochondria are enriched in basic proteins and difficult to resolve by 2-DE. We investigated two methods, cup and paper bridge, for sample loading of this subproteome into the basic range (pH 6-11) gels. Paper bridge loading consistently produced improved resolution of both analytical and preparative protein loads. A unique benefit of this technique is that proteins retained in the paper bridge after loading basic gels can be reloaded onto lower pH gradients (pH 4-7), allowing valued samples to be analyzed on multiple pH ranges.     

A mechanical analysis of woodpecker drumming and its application to shock-absorbing systems

A woodpecker is known to drum the hard woody surface of a tree at a rate of 18 to 22 times per second with a deceleration of 1200 g, yet with no sign of blackout or brain damage. As a model in nature, a woodpecker is studied to find clues to develop a shock-absorbing system for micromachined devices. Its advanced shock-absorbing mechanism, which cannot be explained merely by allometric scaling, is analyzed in terms of endoskeletal structures. In this analysis, the head structures (beak, hyoid, spongy bone, and skull bone with cerebrospinal fluid) of the golden-fronted woodpecker, Melanerpes aurifrons, are explored with x-ray computed tomography images, and their shock-absorbing mechanism is analyzed with a mechanical vibration model and an empirical method. Based on these analyses, a new shock-absorbing system is designed to protect commercial micromachined devices from unwanted high-g and high-frequency mechanical excitations. The new shock-absorbing system consists of close-packed microglasses within two metal enclosures and a viscoelastic layer fastened by steel bolts, which are biologically inspired from a spongy bone contained within a skull bone encompassed with the hyoid of a woodpecker. In the experimental characterizations using a 60 mm smoothbore air-gun, this bio-inspired shock-absorbing system shows a failure rate of 0.7% for the commercial micromachined devices at 60?000 g, whereas a conventional hard-resin method yields a failure rate of 26.4%, thus verifying remarkable improvement in the g-force tolerance of the commercial micromachined devices.     

Effects of the training dataset characteristics on the performance of nine species distribution models: application to Diabrotica virgifera virgifera

Many distribution models developed to predict the presence/absence of invasive alien species need to be fitted to a training dataset before practical use. The training dataset is characterized by the number of recorded presences/absences and by their geographical locations. The aim of this paper is to study the effect of the training dataset characteristics on model performance and to compare the relative importance of three factors influencing model predictive capability; size of training dataset, stage of the biological invasion, and choice of input variables. Nine models were assessed for their ability to predict the distribution of the western corn rootworm, Diabrotica virgifera virgifera, a major pest of corn in North America that has recently invaded Europe. Twenty-six training datasets of various sizes (from 10 to 428 presence records) corresponding to two different stages of invasion (1955 and 1980) and three sets of input bioclimatic variables (19 variables, six variables selected using information on insect biology, and three linear combinations of 19 variables derived from Principal Component Analysis) were considered. The models were fitted to each training dataset in turn and their performance was assessed using independent data from North America and Europe. The models were ranked according to the area under the Receiver Operating Characteristic curve and the likelihood ratio. Model performance was highly sensitive to the geographical area used for calibration; most of the models performed poorly when fitted to a restricted area corresponding to an early stage of the invasion. Our results also showed that Principal Component Analysis was useful in reducing the number of model input variables for the models that performed poorly with 19 input variables. DOMAIN, Environmental Distance, MAXENT, and Envelope Score were the most accurate models but all the models tested in this study led to a substantial rate of mis-classification.     

Effects of low-load resistance training with vascular occlusion on the mechanical properties of muscle and tendon

The present study aimed to investigate the effects of low-load resistance training with vascular occlusion on the specific tension and tendon properties by comparing with those of high-load training. Nine participants completed 12 weeks (3 days/week) of a unilateral isotonic training program on knee extensors. One leg was trained using low load (20% of 1 RM) with vascular occlusion (LLO) and other leg using high load (80% of 1 RM) without vascular occlusion (HL). Before and after training, maximal isometric knee extension torque (MVC) and muscle volume were measured. Specific tension of vastus lateralis muscle (VL) was calculated from MVC, muscle volume, and muscle architecture measurements. Stiffness of tendon-aponeurosis complex in VL was measured using ultrasonography during isometric knee extension. Both protocols significantly increased MVC and muscle volume of quadriceps femoris muscle. Specific tension of VL increased significantly 5.5% for HL, but not for LLO. The LLO protocol did not alter the stiffness of tendon-aponeurosis complex in knee extensors, while the HL protocol increased it significantly. The present study demonstrated that the specific tension and tendon properties were found to remain following low-load resistance training with vascular occlusion, whereas they increased significantly after high-load training.     

On the conditional equivalence of chemical loading and mechanical loading on articular cartilage

Osmotic pressure loading of articular cartilage has been customarily invoked to be equivalent to mechanical loading. In the literature, this equivalence is defined by the amount of water squeezed from the tissue, i.e. if the amount of water content lost by these two modes of loading are the same, it has been generally regarded that the two loadings are equivalent. This assumption has never been proven. Using the water content lost concept, in this paper, we derived the exact conditions under which an osmotic pressure loading of cartilage can be considered to be equivalent to a mechanical loading. However, the mechanical loading condition satisfying this equivalancy criterion, i.e. an isotropic loading delivered via a porous–permeable rigid platen uniformily applied all around the specimen, is not practically achievable. Moreover, even if this were achieved experimentally, the interstitial fluid pressure caused by the two loading conditions are not the same. This result has important ramifications for interpretation of experimental data from mechanical stimulations of cartilage explant studies.     

Methods for detecting associations with rare variants for common diseases: application to analysis of sequence data

Although whole-genome association studies using tagSNPs are a powerful approach for detecting common variants, they are underpowered for detecting associations with rare variants. Recent studies have demonstrated that common diseases can be due to functional variants with a wide spectrum of allele frequencies, ranging from rare to common. An effective way to identify rare variants is through direct sequencing. The development of cost-effective sequencing technologies enables association studies to use sequence data from candidate genes and, in the future, from the entire genome. Although methods used for analysis of common variants are applicable to sequence data, their performance might not be optimal. In this study, it is shown that the collapsing method, which involves collapsing genotypes across variants and applying a univariate test, is powerful for analyzing rare variants, whereas multivariate analysis is robust against inclusion of noncausal variants. Both methods are superior to analyzing each variant individually with univariate tests. In order to unify the advantages of both collapsing and multiple-marker tests, we developed the Combined Multivariate and Collapsing (CMC) method and demonstrated that the CMC method is both powerful and robust. The CMC method can be applied to either candidate-gene or whole-genome sequence data.