Isolation of micropropagated strawberry endophytic bacteria and assessment of their potential for plant growth promotion

Twenty endophytic bacteria were isolated from the meristematic tissues of three varieties of strawberry cultivated in vitro, and further identified, by FAME profile, into the genera Bacillus and Sphingopyxis. The strains were also characterized according to indole acetic acid production, phosphate solubilization and potential for plant growth promotion. Results showed that 15 strains produced high levels of IAA and all 20 showed potential for solubilizing inorganic phosphate. Plant growth promotion evaluated under greenhouse conditions revealed the ability of the strains to enhance the root number, length and dry weight and also the leaf number, petiole length and dry weight of the aerial portion. Seven Bacillus spp. strains promoted root development and one strain of Sphingopyxis sp. promoted the development of plant shoots. The plant growth promotion showed to be correlated to IAA production and phosphate solubilization. The data also suggested that bacterial effects could potentially be harnessed to promote plant growth during seedling acclimatization in strawberry.     

Optimization and validation of an alternative method to evaluate total reactive antioxidant potential

The total reactive antioxidant potential (TRAP) is one of the methods most employed to estimate the antioxidant capacity of samples in vitro. This method is based on the quenching of luminol-enhanced chemiluminescence derived from the thermolysis of 2,2′-azo-bis(2-amidinopropane)dihydrochloride (AAPH) as the free radical source. However, this method can present limitations when the sample does not present a lag phase. In addition, there are no studies regarding TRAP assay validation. In this context, the aim of this work was to optimize and validate this method and to propose another evaluation method using the area under the curve (AUC). The main condition established was the need for the stabilization of the system, at 7000 s, before the addition of the antioxidant to be tested. Both evaluation methods were validated using Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) as a calibrator in the range of 50 to 250 nM, and all parameters showed satisfactory results: specificity, linearity (r > 0.99), precision (intra and interassay relative standard deviations <5%), robustness, and the limits of detection and quantitation (low and similar for both methods). The main advantage of the use of AUC is to evaluate the antioxidant potential of samples that do not present lag phase.     

Kinematics and kinetics of unanticipated misstep conditions: Femoral fracture implications in the elderly

Most hip fractures are thought to occur after falling during everyday activities. We speculated that hip fractures might also occur because of excessive loading of the hip joint during an unexpected misstep consequently leading to a fall. The aims of this study were to explore the kinematics and kinetics of the lower extremity joints during missteps as compared with regular stepping, as well as to compare the magnitude of forces acting upon the hip joint with the threshold forces expected to fracture the hip. Fourteen healthy adults performed two forward steps on a 17.8 cm high platform under the following four conditions: forward with and without vision, as well as a misstep (the box for the final step unexpectedly removed without participant awareness), and regular stepping down with eyes open. The results revealed no differences between stepping forward with and without vision. When compared with both stepping forward and regular stepping down, the misstep revealed altered joint positions accompanied by increased forces and moments acting upon the hip joint. For example, the peak vertical proximal thigh segment force was 3.05±0.55 BW vs. 1.23±0.14 BW and 0.91±0.09 BW (p<.001; misstep vs. regular stepping down and stepping forward, respectively), while the proximal thigh segment moment in frontal plane was 1.39±0.70 Nm/kg vs. 0.18±0.32 Nm/kg of adduction and 0.16±0.19 Nm/kg of abduction (p<.001). When compared with the literature data, the forces during misstep were within the range of those forces that could result in hip fractures in the elderly. Therefore, it may be possible for the elderly to experience hip/proximal femur fractures during missteps prior to falling.     

IL2-330G polymorphic allele is associated with decreased risk of Helicobacter pylori infection in adulthood

We evaluated whether polymorphisms in genes coding molecules linked to the innate and adaptive immune response are associated with susceptibility to Helicobacter pylori infection. IL1B-511C → T, IL1B-31 T → C, IL1RN allele 2, IL2-330 T → G, TNFA-307 G → A, TLR2Arg677Trp, TLR2Arg753Gln, TLR4Asp299Gly, and TLR5^392STOP polymorphisms were determined in 541 blood donors. IL2-330 T → G allele carriers had a decreased H. pylori infection risk (OR = 0.63, 95% CI = 0.43–0.93) after adjustment for demographic and environmental factors. Hence, we investigated whether the polymorphism is functional by evaluating IL-2 serum concentration in 150 blood donors and 100 children. IL-2 pro-inflammatory and anti-inflammatory properties were indirectly investigated by determining serum IFN-γ and IL-10/TGF-β levels. The polymorphism was associated with increased mean IL-2 levels in H. pylori-positive adults (2.65 pg/mL vs. 7.78 pg/mL) and children (4.19 pg/mL vs. 8.03 pg/mL). Increased IL-2 was associated with pro-inflammatory activity in adults (IFN-γ = 18.61 pg/mL vs. 25.71 pg/mL), and with anti-inflammatory activity in children (IL-10 = 6.99 vs. 14.17 pg/mL, TGF-β = 45.88 vs. 93.44 pg/mL) (p < 10⁻³ for all). In conclusion, in the context of H. pylori infection, IL2-330 T → G polymorphism is functional and is associated with decreased risk of infection in adults.     

Testing the performance of beta diversity measures based on incidence data: the robustness to undersampling

Aim Researchers measuring beta diversity have rarely concerned themselves with the problems of how complete the species lists of studied communities are, and of how the varying degrees of completeness can actually change estimates of beta diversity. No comprehensive assessment has been made regarding the behaviour of most beta diversity indices when applied to incomplete samples, a situation which is more common than usually recognized. Our objective was to assess the behaviour and robustness of a number of beta diversity measures for incidence data from undersampled communities. Location Mainland Portugal and the Azorean archipelago (North Atlantic). Methods Data from intensive sampling of spiders in mainland Portugal and arthropods in Azores were collected. We examined the properties of 15 beta diversity measures developed for incidence data. We simulated varying degrees of completeness, whereas computing beta diversity for selected pairs of samples. The robustness of these beta diversity accumulation curves was assessed for the purpose of finding the best measures for undersampled communities. Results The Harrison et al.β_‐2 and the Williams β_‐3 are particularly robust to undersampling. These measures are also insensitive to differences of alpha diversity (species richness) between communities, and therefore to nestedness. Colwell & Coddington β_cc and the related Jaccard β_j and Gaston et al.β_g performed best of the measures sensitive to alpha diversity. They performed poorly, however, when compared communities exhibited very low values of beta diversity. In such cases, the Routledge β_r performed the best. Main conclusions No index was found to perform without bias in all circumstances. Overall, β_‐2, β_‐3 and β_cc (or related measures β_j and β_g) are recommended as they seem to be the most robust to undersampling.     

An examination of the flexion-relaxation phenomenon in the cervical spine in lumbo-pelvic sitting

The flexion-relaxation phenomenon (FRP) is well documented at end-range lumbar spine flexion in both standing and sitting however, the FRP has been insufficiently investigated in cervico-thoracic musculature. The aim of this study was to determine whether the FRP occurs during forward flexion of the neck, in lumbo-pelvic sitting, amongst a pain-free population. Surface electromyography (EMG) was used to measure muscle activation in 20 (10 men, 10 women) asymptomatic subjects in selected cervico-thoracic muscles during four, 5-s phases (upright posture, forward flexion, full flexion and return to upright) while subjects were positioned in lumbo-pelvic sitting. Spinal kinematics were simultaneously measured using an electromagnetic motion tracking device. No FRP was observed in upper trapezius or thoracic erector spinae (T4). When using visual methods to determine the presence/absence of the FRP, five subjects were believed to show evidence of the FRP in the cervical erector spinae. However, when using various non-visual criteria to determine the existence of the FRP, substantial variations (0–13 subjects) were evident. We recommend that criteria based upon relatively large differences in muscle activation should be considered when defining the FRP. These findings are of significance for future investigations examining specific cervical pain disorders.     

Analysis of the subproteomes of proteinases and heparin‐binding toxins of eight Bothrops venoms

Viperid snakes show the most complex snake‐venom proteomes and offer an intriguing challenge in terms of understanding the nature of their components and the pathological outcomes of envenomation characterized by local and systemic effects. In this work, the venom complexity of eight Bothrops species was analyzed by 2‐DE, and their subproteomes of proteinases were explored by 2‐D immunostaining and 2‐D gelatin zymography, demonstrating the diversity of their profiles. Heparin, a highly sulfated glycosaminoglycan released from mast cells, is involved in anti‐coagulant and anti‐inflammatory processes. Here, we explored the hypothesis that heparin released upon envenomation could interact with toxins and interfere with venom pathogenesis. We first identified the Bothrops venom subproteome of toxins that bind with high‐affinity for heparin as composed of mainly serine proteinases and C‐type lectins. Next, we explored the Bothrops jararaca toxins that bind to heparin under physiological conditions and identified a relationship between the subproteomes of proteinases, and that of heparin‐binding toxins. Only the non‐bound fraction, composed mainly of metalloproteinases, showed lethal and hemorrhagic activities, whereas the heparin‐bound fraction contained mainly serine proteinases associated with coagulant and fibrinogenolytic activities. These data suggest that heparin binding to B. jararaca venom components in vivo has a minor protective effect to venom toxicity.