Altered control strategy between leading and trailing leg increases knee adduction moment in the elderly while descending stairs

The aim of the study was to examine the external knee adduction moments in a group of older and younger adults while descending stairs and thus the possibility of an increased risk of knee osteoarthritis due to altered knee joint loading in the elderly. Twenty-seven older and 16 younger adults descended a purpose-built staircase. A motion capture system and a force plate were used to determine the subjects' 3D kinematics and ground reaction forces (GRF) during locomotion. Calculation of the leg kinematics and kinetics was done by means of a rigid, three-segment, 3D leg model. In the initial portion of the support phase, older adults showed a more medio-posterior GRF vector relative to the ankle joint, leading to lower ankle joint moments (P<0.05). At the knee, the older adults demonstrated a more medio-posterior directed GRF vector, increasing in knee flexion and adduction in the second part of the single support phase (P<0.05). Further, GRF magnitude was lower in the initial and higher in the mid-portions of the support phase for the elderly (P<0.05). The results show that older adults descend stairs by using the trailing leg before the initiation of the double support phase more compared to the younger ones. The consequence of this altered control strategy while stepping down is a more medially directed GRF vector increasing the magnitude of external knee adduction moment in the elderly. The observed changes between leading and trailing leg in the elderly may cause a redistribution of the mechanical load at the tibiofemoral joint, affecting the initiation and progression of knee osteoarthritis in the elderly.     

c-di-GMP Enhances Protective Innate Immunity in a Murine Model of Pertussis

Innate immunity represents the first line of defense against invading pathogens in the respiratory tract. Innate immune cells such as monocytes, macrophages, dendritic cells, NK cells, and granulocytes contain specific pathogen-recognition molecules which induce the production of cytokines and subsequently activate the adaptive immune response. c-di-GMP is a ubiquitous second messenger that stimulates innate immunity and regulates biofilm formation, motility and virulence in a diverse range of bacterial species with potent immunomodulatory properties. In the present study, c-di-GMP was used to enhance the innate immune response against pertussis, a respiratory infection mainly caused by Bordetella pertussis. Intranasal treatment with c-di-GMP resulted in the induction of robust innate immune responses to infection with B. pertussis characterized by enhanced recruitment of neutrophils, macrophages, natural killer cells and dendritic cells. The immune responses were associated with an earlier and more vigorous expression of Th1-type cytokines, as well as an increase in the induction of nitric oxide in the lungs of treated animals, resulting in significant reduction of bacterial numbers in the lungs of infected mice. These results demonstrate that c-di-GMP is a potent innate immune stimulatory molecule that can be used to enhance protection against bacterial respiratory infections. In addition, our data suggest that priming of the innate immune system by c-di-GMP could further skew the immune response towards a Th1 type phenotype during subsequent infection. Thus, our data suggest that c-di-GMP might be useful as an adjuvant for the next generation of acellular pertussis vaccine to mount a more protective Th1 phenotype immune response, and also in other systems where a Th1 type immune response is required.     

Crop Growth, Disease and Yield Components of Rusted Phaseolus Beans in Ethiopia

Crop growth and disease epidemics in sprayed and non-sprayed bean plots, artificially infected with rust (Uromyces appendiculatus) 3 weeks after emergence. were assessed weekly in two cultivars, at two locations for two seasons. Disease intensity was regulated by the application of a fungicide at 5 spray frequencies. Fungicide application influenced leaf area index (LAI) and reduced rust intensity. The fungicide had no significant effect on other diseases and dead leaf area. Fungicide application increased seed yield (SY) by increased numbers of pods per plant (PP). Rust severity was strongly correlated with pustule density but the overall relationships among rust assessment parameters depended on cultivar and location. Seed yield and pods per plant were highly correlated with LAI. The relationships between LAI and seeds per pod or seed weight depended on cultivar and location. Overall rust assessment parameters (rust severity and pustule density) showed close, negative relationships with seed yield. seed weight and pods per plant but not with seeds per pod. The relationships obtained in the partially resistant line 6-R-395 were less definite than those in the susceptible line Mexican 142. The yield parameters seed yield and pods per plant, showed strong positive relationships.     

Use of SPR to Study the Interaction of G7-18NATE Peptide with the Grb7-SH2 Domain

Surface plasmon resonance (SPR) is a useful biosensor technique for the study of biomolecular interactions, with the potential for high-throughput screening of ligand interactions with drug targets. The key to its successful use, however, is in the appropriate design of the experiment, including the mode of immobilization to the biosensor chip. We report an investigation of the use of SPR for measuring the affinity of the G7-18NATE peptide ligand for its Grb7-SH2 domain target involved in the migratory and proliferative potential of cancer cells. Previous studies have shown that the cyclic non-phosphorylated peptide, G7-18NATE, inhibits Grb7 interactions with upstream binding partners and is able to inhibit both cell migration and proliferation of cancer cells. We report the synthesis of a biotinylated G7-18NATE covalently attached to a linker (G7-18NATE-ASASASK-Biotin) and compare its interaction with the Grb7-SH2 domain by SPR using three different immobilization strategies; immobilisation of the peptide via streptavidin, immobilization of glutathione S-transferase (GST)-Grb7-SH2 domain via anti-GST antibody, and immobilization of biotinylated Grb7-SH2 domain via streptavidin. This revealed that sensorgrams free from non-specific binding and displaying simple kinetics were most readily achieved by immobilising the protein rather than the peptide, in spite of the lower response associated with this method. K _D values of ~300 μM were determined for both strategies at pH 7.4. This compared with a K _D value of 4.4 μM at pH 6 demonstrating the importance of pH on this interaction. Overall, the immobilised protein systems are most suitable for future comparative screening efforts using SPR.     

Prevalence and Factors Associated with Road Traffic Crash among Taxi Drivers in Mekelle Town,Northern Ethiopia, 2014: A Cross Sectional Study

Objectives

The 2013 World Health Organization Status Report on Road Safety estimated that approximately 1.24 million deaths occur annually due to road traffic crashes with most of the burden falling on low- and middle-income countries. The objective of this research is to study the prevalence of road traffic crashes in Mekelle, Tigray, Northern Ethiopia and to identify risk factors with the ultimate goal of informing prevention activities and policies.

Methods

This study used a cross-sectional design to measure the prevalence and factors associated with road traffic crashes among 4-wheeled minibus (n = 130) and 3-wheeled Bajaj (n = 582) taxi drivers in Mekelle, Ethiopia. Bivariate and multivariate logistic regression were used to evaluate the association between risk factors and drivers’ involvement in a road traffic crash within the 3 years prior to the survey.

Findings

Among the 712 taxi drivers, 26.4% (n = 188) of them reported involvement in a road traffic crash within the past 3 years. Drivers who listened to mass media had decreased likelihood of road traffic crash involvement (AOR = 0.51, 0.33–0.78), while speedy driving (AOR = 4.57, 3.05–7.44), receipt of a prior traffic punishment (AOR = 4.57, 2.67–7.85), and driving a mechanically faulty taxi (AOR = 4.91, 2.81–8.61) were strongly associated with road traffic crash involvement. Receiving mobile phone calls while driving (AOR = 1.91, 1.24–2.92) and history of alcohol use (AOR = 1.51, 1.00–2.28) were also associated with higher odds of road traffic crash involvement.

Conclusion

The results of this study show that taxi drivers in Mekelle habitually place themselves at increased risk of road traffic crashes by violating traffic laws, especially related to speedy driving, mobile phone use, and taxi maintenance. This research can be used to support re-evaluation of the type, severity, and enforcement of traffic violation penalties.     

Benzopyrazine derivatives: A novel class of growth factor receptor bound protein 7 antagonists

Growth factor receptor bound protein 7 (Grb7) is an adapter protein that functions as a downstream effector of growth factor mediated signal transduction. Over-expression of Grb7 has been implicated in a variety of cancers such as breast, blood, pancreatic, esophageal, and gastric carcinomas. Inhibition of Grb7 has been shown to reduce the migratory and proliferative potential of these cancers, making it an attractive therapeutic target. Starting with a known peptide antagonist, the present work reports the application of a succession of computational ligand design tools comprising a ligand shape based similarity search, molecular docking and a 2D-similarity search to identify small molecular antagonists of the Grb7-SH2 domain from the NCI chemical database. Binding to the Grb7-SH2 domain was then experimentally tested using melting point shift assays and isothermal titration calorimetry. Overall, a total of 11 benzopyrazine based small molecular antagonists were identified with affinity for the Grb7-SH2 domain. Representative compounds tested using ITC were revealed to possess moderate binding affinity in the low micromolar range. Finally, the lead compound (NSC642056) was found to reduce the growth of a Grb7-expressing breast cancer cell line with an IC₅₀ of 86 ??M. It is expected that the identified antagonists will be useful additions to further explore the function of Grb7 and for the development of inhibitors with therapeutic potential.     

Mechanical properties of the triceps surae tendon and aponeurosis in relation to intensity of sport activity

The purpose of the present study was to investigate whether the mechanical properties (i.e. force strain relationship) of the triceps surae tendon and aponeurosis relate to the performed sport activity in an intensity-dependent manner. This was done by comparing sprinters with endurance runners and subjects not active in sports. Sixty-six young male subjects (26+/-5 yr; 183+/-6 cm; 77.6+/-6.7 kg) participated in the study. Ten of these subjects were adults not active in sports, 28 were endurance runners and 28 sprinters. All subjects performed isometric maximal voluntary plantar flexion contractions (MVC) on a dynamometer. The distal aponeuroses of the gastrocnemius medialis (GM) was visualised by ultrasound during the MVC. The results showed that only the sprinters had higher normalised stiffness (relationship between tendon force and tendon strain) of the triceps surae tendon and aponeurosis and maximal calculated tendon forces than the endurance runners and the subjects not active in sports. Furthermore, including the data of all 66 examined participants tendon stiffness correlated significantly (r=0.817, P<0.001) with the maximal tendon force achieved during the MVC. It has been concluded that the mechanical properties of the triceps surae tendon and aponeurosis do not show a graded response to the intensity of the performed sport activity but rather remain at control level in a wide range of applied strains and that strain amplitude and/or frequency should exceed a given threshold in order to trigger additional adaptation effects. The results further indicate that subjects with higher muscle strength possibly increase the margin of tolerated mechanical loading of the tendon due to the greater stiffness of their triceps surae tendon and aponeurosis.     

RWD Domain as an E2 (Ubc9)-Interaction Module

An RWD domain is a well conserved domain found through bioinformatic analysis of the human proteome sequence; however, its function has been unknown. Ubiquitin-like modifications require the catalysis of three enzymes generally known as E1, E2, and E3. We solved the crystal structure of the E2 for the small ubiquitin-like modifiers (SUMO) in complex with an RWD domain and confirmed the structure using solution NMR analysis. The binding surface of RWD on Ubc9 is located near the N terminus of Ubc9 that is known to be involved in noncovalent binding of the proteins in the conjugation machinery, including a domain of E1, SUMO, and an E3 ligase. NMR data indicate that the RWD domain does not bind to SUMO and E1. The interaction between RWD and Ubc9 has a K_d of 32 ± 4 μm. Consistent with the structure and binding affinity and in contrast to a previous report, the RWD domain and RWDD3 have minimal effects on global SUMOylation. The structural and biochemical information presented here forms the basis for further investigation of the functions of RWD-containing proteins.     

Symmetry and reproducibility of the components of dynamic stability in young adults at different walking velocities on the treadmill

In the literature, analysis of dynamic gait stability using the extrapolated center of mass concept is often an objective that assumes reproducible and symmetrical data. Here, we examined the validity of this assumption by analyzing subjects walking at different velocities. Eleven healthy young subjects walked on a treadmill at six different velocities (1.0-2.0m·s(-1)). Dynamic stability at touchdown of the left and right foot (10 gait trials for each body side) was investigated by using the margin of stability, determined as the difference between base of support and extrapolated center of mass. Dynamic stability parameters showed no significant differences (P>0.05) between gait trials, with a root mean square difference in margin of stability of less than 1.62cm. Correlation coefficients between trials were above 0.70 for all parameters, demonstrating that two gait trials are sufficient to obtain reproducible data. In more than 90% of the cases, the absolute symmetry index was below 8% with no relevant functional differences between body sides. We concluded that analyzing two gait trials for one body side is sufficient to determine representative characteristics of the components of dynamic stability in healthy young adults while walking on the treadmill at a wide range of velocities.     

Deficits in the way to achieve balance related to mechanisms of dynamic stability control in the elderly

The purpose of this study was to examine the postural corrections related to components of dynamic stability aimed to increase our understanding of successful postural control among the elderly population. This was done by comparing balance behaviour of older adults who were able to recover stability (stable) and others who failed to regain stability (unstable) with a single step after a forward fall. Thirty-eight old male adults (64+/-3yr, 176+/-6cm, 78.5+/-7.8kg) had to recover balance after a sudden induced forward fall. All participants performed maximal isometric ankle plantarflexion and knee extension contractions on a dynamometer. The elongation of the gastrocnemius medialis and the vastus lateralis tendon and aponeuroses during isometric contraction was examined by ultrasonography. There were no differences in leg-extensor muscle strength or tendon stiffness between the two groups showing that the muscle tendon capacities may not be the reason for the observed differences in dynamic stability control. The unstable participants created a higher horizontal ground reaction push-off force of the support limb in the second part ( approximately 260ms after release) of the phase until touchdown leading to an unstable body position at touchdown. The results indicate deficits in the way to achieve balance related to mechanisms responsible for dynamic stability control within the elderly population.