Genome-wide and structural analyses of pseudokinases encoded in the genome of Arabidopsis thaliana provide functional insights

Protein Kinase-Like Non-Kinases (PKLNKs), commonly known as “pseudokinases”, are homologous to eukaryotic Ser/Thr/Tyr protein kinases (PKs) but lack the crucial aspartate residue in the catalytic loop, indispensable for phosphotransferase activity. Therefore, they are predicted to be “catalytically inactive” enzyme homologs. Analysis of protein-kinase like sequences from Arabidopsis thaliana led to the identification of more than 120 pseudokinases lacking catalytic aspartate, majority of which are closely related to the plant-specific receptor-like kinase family. These pseudokinases engage in different biological processes, enabled by their diverse domain architectures and specific subcellular localizations. Structural comparison of pseudokinases with active and inactive conformations of canonical PKs, belonging to both plant and animal origin, revealed unique structural differences. The currently available crystal structures of pseudokinases show that the loop topologically equivalent to activation segment of PKs adopts a distinct-folded conformation, packing against the pseudoenzyme core, in contrast to the extended and inhibitory geometries observed for active and inactive states, respectively, of catalytic PKs. Salt-bridge between ATP-binding Lys and DFG-Asp as well as hydrophobic interactions between the conserved nonpolar residue C-terminal to the equivalent DFG motif and nonpolar residues in C-helix mediate such a conformation in pseudokinases. This results in enhanced solvent accessibility of the pseudocatalytic loop in pseudokinases that can possibly serve as an interacting surface while associating with other proteins. Specifically, our analysis identified several residues that may be involved in pseudokinase regulation and hints at the repurposing of pseudocatalytic residues to achieve mechanistic control over noncatalytic functions of pseudoenzymes.     

Growth-band counts from elephantfish Callorhinchus milii fin spines do not correspond with independently estimated ages

Fin spines from elephantfish Callorhinchus milii were sectioned and viewed with transmitted white light under a compound microscope. The sections displayed growth bands but their interpretation and significance were unclear. Three different methods were used for counting growth bands. The results were compared with reference growth curves based on length-at-age estimates for six juvenile year classes derived from length-frequency distributions, and tagging data that showed longevity is at least 20 years. None of the three ageing methods showed good correspondence with the reference curves and all methods departed markedly from the reference curves at ages above 2 years old. Therefore, growth bands present in C. milii spines are not useful for ageing, at least with the three methods tested here. Spine bands may not represent age marks, but instead may be layers of material deposited irregularly to strengthen the spine.     

Spinal stiffness in asymptomatic subjects

The objective of the study was to measure postero-anterior stiffness of thoracolumbar spine from normal asymptomatic subjects at T₄, T₉, and L₂ levels using a standardized device and protocol. Sixteen volunteer subjects (eight males and eight females) meeting the inclusion and exclusion criteria were recruited for the study. Their T₄, T₉, and L₂ spinous processes were identified and marked. These spinous processes were cyclically loaded with 22.5 N, 45 N, 90 N, and 135 N fixed forces at 0.1 Hz in postero-anterior direction by a computer controlled and operated Therapeutic Spinal Mobilizer (TSM) for five cycles to the level of subject’s acceptance of the load magnitude. The magnitude of the force and displacement experienced at the spinal level were recorded using a load cell and linear variable differential transducer. The stiffness was obtained from the slope of the load/deformation curve. The stiffness values were subjected to analysis of variance to determine the effect of independent variables. The stiffness at different levels was significantly different (p < 0.0001) and it was significantly affected by the testing load (p < 0.0001). The age, sex, height and weight were not significantly associated with the stiffness, neither were the cycles at the same load. The postero-anterior stiffness of the thoracolumbar spine is different at different spinal levels and varies with testing loads.     

Premaxillary-maxillary suture fusion and anterior nasal tubercle morphology in the chimpanzee

The anterior nasal spine (ANS) is considered by some to be a unique characteristic of modern humans; however, it has been reported to occur in chimpanzees. Among human populations, ANS size correlates with facial prognathism and timing of premaxillary-maxillary sutural fusion, which may have implications for models of midfacial growth. To investigate these questions, skulls from 55 chimpanzees (Pan troglodytes) (29 male, 26 female), ranging in age from infants to adults, were examined. None of the 55 specimens examined had a true ANS, but a number of specimens exhibited raised anterior nasal prominences or tubercles (ANT) on the nasal surface of the premaxilla. Female specimens had significantly (P less than .05) more ANTs and more extensive fusion of both the palatal and facial components of the suture than did the male specimens. Results from the present study document the presence of an ANT in a sample of chimpanzee crania and are consonant with associations predicted by a septal-traction model of midfacial growth.     

The rib cage reduces intervertebral disc pressures in cadaveric thoracic spines by sharing loading under applied dynamic moments

The effects of the rib cage on thoracic spine loading are not well studied, but the rib cage may provide stability or share loads with the spine. Intervertebral disc pressure provides insight into spinal loading, but such measurements are lacking in the thoracic spine. Thus, our objective was to examine thoracic intradiscal pressures under applied pure moments, and to determine the effect of the rib cage on these pressures. Human cadaveric thoracic spine specimens were positioned upright in a testing machine, and Dynamic pure moments (0 to ±5 N·m) with a compressive follower load of 400 N were applied in axial rotation, flexion - extension, and lateral bending. Disc pressures were measured at T4-T5 and T8-T9 using needle-mounted pressure transducers, first with the rib cage intact, and again after the rib cage was removed. Changes in pressure vs. moment slopes with rib cage removal were examined. Pressure generally increased with applied moments, and pressure-moment slope increased with rib cage removal at T4-T5 for axial rotation, extension, and lateral bending, and at T8-T9 for axial rotation. The results suggest the intact rib cage carried about 62% and 56% of axial rotation moments about T4-T5 and T8-T9, respectively, as well as 42% of extension moment and 36–43% of lateral bending moment about T4-T5 only. The rib cage likely plays a larger role in supporting moments than compressive loads, and may also play a larger role in the upper thorax than the lower thorax.     

Influence of follower load application on moment-rotation parameters and intradiscal pressure in the cervical spine

The objective of this study was to implement a follower load (FL) device within a robotic (universal force-moment sensor) testing system and utilize the system to explore the effect of FL on multi-segment cervical spine moment-rotation parameters and intradiscal pressure (IDP) at C45 and C56. Twelve fresh-frozen human cervical specimens (C3-C7) were biomechanically tested in a robotic testing system to a pure moment target of 2.0 Nm for flexion and extension (FE) with no compression and with 100 N of FL. Application of FL was accomplished by loading the specimens with bilateral cables passing through cable guides inserted into the vertebral bodies and attached to load controlled linear actuators. FL significantly increased neutral zone (NZ) stiffness and NZ width but resulted in no change in the range of motion (ROM) or elastic zone stiffness. C45 and C56 IDP measured in the neutral position were significantly increased with application of FL. The change in IDP with increasing flexion rotation was not significantly affected by the application of FL, whereas the change in IDP with increasing extension rotation was significantly reduced by the application of FL. Application of FL did not appear to affect the specimen’s quantity of motion (ROM) but did affect the quality (the shape of the curve). Regarding IDP, the effects of adding FL compression approximates the effect of the patient going from supine to a seated position (FL compression increased the IDP in the neutral position). The change in IDP with increasing flexion rotation was not affected by the application of FL, but the change in IDP with increasing extension rotation was, however, significantly reduced by the application of FL.     

Biomechanical analysis of lumbar interbody fusion cages with various lordotic angles: a finite element study

Inappropriate lordotic angle of lumbar fusion cage could be associated with cage damage or subsidence. The biomechanical influence of cage lordotic angle on lumbar spine has not been fully investigated. Four surgical finite element models were constructed by inserting cages with various lordotic angles at L3-L4 disc space. The four motion modes were simulated. The range of motion (ROM) decreased with increased lordotic angle of cage in flexion, extension, and rotation, whereas it was not substantially changed in bending. The maximum stress in cage decreased with increased lordotic angle of cage in all motion modes. The maximum stress in endplate at surgical level increased with increased lordotic angle of cage in flexion and rotation, whereas it was not substantially changed in extension and bending. The facet joint force (FJF) was much smaller than that for the intact conditions in extension, bending, and rotation, while it was not substantially changed in flexion. In conclusion, the ROM, stresses in the cage and endplate at surgical level are sensitive to the lordotic angle of cage. The increased cage lordotic angle may provide better stability and reduce the risk of cage damage, whereas it may increase the risk of subsidence in flexion and rotation.     

珠江下游颗粒沟链藻Aulacoseira granulata (Ehrenberg) Simonsen及形态相近种赫组基沟链藻Aulacoseira herzogii (Lemmerman) Simonsen的形态学研究

采用扫描电镜(SEM)技术,对采自珠江下游的沟链藻属2个物种颗粒沟链藻Aulacoseira granulata(Ehrenberg)Simonsen和赫组基沟链藻A.herzogii(Lemmerman)Simonsen的形态特征进行了观察和描述,并与其他相近种的形态异同点进行了比较分析。     

Four different head shapes in Daphnia hyalina (Leydig) induced by the presence of larvae of Chaoborus flavicans (Meigen)

In many species of Daphnia spines, neck teeth, and enlarged or reshaped helmets are well-known as defences against invertebrate predators. Until now, Daphnia hyalina (Leydig, 1860), a common species in many European lakes, has appeared to be an exception to this rule.Here, we provide evidence that the larvae of Chaoborus flavicans (Meigen) also can induce morphological changes in D. hyalina. Specimens react morphologically to the presence of larvae in three ways: (a) by changes in the shape of head shield, (b) by the formation of spine(s) on the head, and (c) by the elongation of the tail spine. The frequency and intensity of these morphological changes are correlated positively with midge larvae densities. The most pronounced reactions occurred in young D. hyalina.The predator kairomone also induced changes in Daphnia body size.