FAP206 is a Microtubule-Docking Adapter for Ciliary Radial Spoke 2 and Dynein c

Radial spokes are conserved macromolecular complexes that are essential for ciliary motility. A triplet of three radial spokes, RS1, RS2, and RS3, repeats every 96 nm along the doublet microtubules. Each spoke has a distinct base that docks to the doublet and is linked to different inner dynein arms. Little is known about the assembly and functions of individual radial spokes. A knockout of the conserved ciliary protein FAP206 in the ciliate Tetrahymena resulted in slow cell motility. Cryo–electron tomography showed that in the absence of FAP206, the 96-nm repeats lacked RS2 and dynein c. Occasionally, RS2 assembled but lacked both the front prong of its microtubule base and dynein c, whose tail is attached to the front prong. Overexpressed GFP-FAP206 decorated nonciliary microtubules in vivo. Thus FAP206 is likely part of the front prong and docks RS2 and dynein c to the microtubule

Lack of Antinociceptive Cross-Tolerance With Co-Administration of Morphine and Fentanyl Into the Periaqueductal Gray of Male Sprague-Dawley Rats

Tolerance to the antinociceptive effect of mu-opioid receptor (MOPr) agonists, such as morphine and fentanyl, greatly limits their effectiveness for long-term use to treat pain. Clinical studies have shown that combination therapy and opioid rotation can be used to enhance opioid-induced antinociception once tolerance has developed. The mechanism and brain regions involved in these processes are unknown. The purpose of this study was to evaluate the contribution of the ventrolateral periaqueductal gray (vlPAG) to antinociceptive tolerance and cross-tolerance between administration and co- administration of morphine and fentanyl. Tolerance was induced by pretreating rats with morphine or fentanyl or low-dose combination of morphine and fentanyl into the vlPAG followed by assessment of cross-tolerance to the other opioid. In addition, tolerance to the combined treatment was assessed. Cross-tolerance did not develop between repeated vlPAG microinjections of morphine and fentanyl. Likewise, there was no evidence of cross-tolerance from morphine or fentanyl to co-administration of morphine and fentanyl. Co-administration did not cause cross-tolerance to fentanyl. Cross- tolerance was only evident to morphine or morphine and fentanyl combined in rats pretreated with co-administration of low-doses of morphine and fentanyl. In conclusion, cross-tolerance does not develop between morphine and fentanyl within the vlPAG. This finding is consistent with the functionally selective signaling that has been reported for antinociception and tolerance following morphine and fentanyl binding to the MOPr. This research supports the notion that combination therapy and opioid rotation may be useful clinical practices to reduce opioid tolerance and other side effects. Perspective: This preclinical study shows that there is a reduction in cross tolerance between morphine and fentanyl within the periaqueductal gray which is key brain region in opioid antinociception and tolerance

De novo variants in the RNU4-2 snRNA cause a frequent neurodevelopmental syndrome.

Around 60% of individuals with neurodevelopmental disorders (NDD) remain undiagnosed after comprehensive genetic testing, primarily of protein-coding genes1. Large genome-sequenced cohorts are improving our ability to discover new diagnoses in the non-coding genome. Here we identify the non-coding RNA RNU4-2 as a syndromic NDD gene. RNU4-2 encodes the U4 small nuclear RNA (snRNA), which is a critical component of the U4/U6.U5 tri-snRNP complex of the major spliceosome2. We identify an 18 base pair region of RNU4-2 mapping to two structural elements in the U4/U6 snRNA duplex (the T-loop and stem III) that is severely depleted of variation in the general population, but in which we identify heterozygous variants in 115 individuals with NDD. Most individuals (77.4%) have the same highly recurrent single base insertion (n.64_65insT). In 54 individuals in whom it could be determined, the de novo variants were all on the maternal allele. We demonstrate that RNU4-2 is highly expressed in the developing human brain, in contrast to RNU4-1 and other U4 homologues. Using RNA sequencing, we show how 5 splice-site use is systematically disrupted in individuals with RNU4-2 variants, consistent with the known role of this region during spliceosome activation. Finally, we estimate that variants in this 18 base pair region explain 0.4% of individuals with NDD. This work underscores the importance of non-coding genes in rare disorders and will provide a diagnosis to thousands of individuals with NDD worldwide

The response of relativistic outflowing gas to the inner accretion disk of a black hole

The brightness of an active galactic nucleus is set by the gas falling onto it from the galaxy, and the gas infall rate is regulated by the brightness of the active galactic nucleus; this feedback loop is the process by which supermassive black holes in the centres of galaxies may moderate the growth of their hosts. Gas outflows (in the form of disk winds) release huge quantities of energy into the interstellar medium, potentially clearing the surrounding gas. The most extreme (in terms of speed and energy) of these-the ultrafast outflows-are the subset of X-ray-detected outflows with velocities higher than 10,000 kilometres per second, believed to originate in relativistic (that is, near the speed of light) disk winds a few hundred gravitational radii from the black hole. The absorption features produced by these outflows are variable, but no clear link has been found between the behaviour of the X-ray continuum and the velocity or optical depth of the outflows, owing to the long timescales of quasar variability. Here we report the observation of multiple absorption lines from an extreme ultrafast gas flow in the X-ray spectrum of the active galactic nucleus IRAS 13224-3809, at 0.236 ± 0.006 times the speed of light (71,000 kilometres per second), where the absorption is strongly anti-correlated with the emission of X-rays from the inner regions of the accretion disk. If the gas flow is identified as a genuine outflow then it is in the fastest five per cent of such winds, and its variability is hundreds of times faster than in other variable winds, allowing us to observe in hours what would take months in a quasar. We find X-ray spectral signatures of the wind simultaneously in both low- and high-energy detectors, suggesting a single ionized outflow, linking the low- and high-energy absorption lines. That this disk wind is responding to the emission from the inner accretion disk demonstrates a connection between accretion processes occurring on very different scales: the X-ray emission from within a few gravitational radii of the black hole ionizing the disk wind hundreds of gravitational radii further away as the X-ray flux rises.M.L.P., C.P., A.C.F. and A.L. acknowledge support from the European Research Council through Advanced Grant on Feedback 340492. W.N.A. and G.M. acknowledge support from the European Union Seventh Framework Programme (FP7/2013-2017) under grant agreement number 312789, StrongGravity. D.J.K.B. acknowledges support from the Science and Technology Facilities Council. This work is based on observations with XMM-Newton, an ESA science mission with instruments and contributions directly funded by ESA Member States and NASA. D.R.W. is supported by NASA through Einstein Postdoctoral Fellowship grant number PF6-170160, awarded by the Chandra X-ray Center, operated by the Smithsonian Astrophysical Observatory for NASA under contract NAS8-03060. This work made use of data from the NuSTAR mission, a project led by the California Institute of Technology, managed by the Jet Propulsion Laboratory, and funded by NASA. This research has made use of the NuSTAR Data Analysis Software (NuSTARDAS) jointly developed by the ASI Science Data Center and the California Institute of Technology

Averting biodiversity collapse in tropical forest protected areas

The rapid disruption of tropical forests probably imperils global biodiversity more than any other contemporary phenomenon¹⁻³. With deforestation advancing quickly, protected areas are increasingly becoming final refuges for threatened species and natural ecosystem processes. However, many protected areas in the tropics are themselves vulnerable to human encroachment and other environmental stresses⁴⁻⁹. As pressures mount, it is vital to know whether existing reserves can sustain their biodiversity. A critical constraint in addressing this question has been that data describing a broad array of biodiversity groups have been unavailable for a sufficiently large and representative sample of reserves. Here we present a uniquely comprehensive data set on changes over the past 20 to 30 years in 31 functional groups of species and 21 potential drivers of environmental change, for 60 protected areas stratified across the world’s major tropical regions. Our analysis reveals great variation in reserve ‘health’: about half of all reserves have been effective or performed passably, but the rest are experiencing an erosion of biodiversity that is often alarmingly widespread taxonomically and functionally. Habitat disruption, hunting and forest-product exploitation were the strongest predictors of declining reserve health. Crucially, environmental changes immediately outside reserves seemed nearly as important as those inside in determining their ecological fate, with changes inside reserves strongly mirroring those occurring around them. These findings suggest that tropical protected areas are often intimately linked ecologically to their surrounding habitats, and that a failure to stem broad-scale loss and degradation of such habitats could sharply increase the likelihood of serious biodiversity declines.Keywords: Ecology, Environmental scienc

Developing a Gait Enhancing Mobile Shoe to Alter Over-Ground Walking Coordination

This paper presents a Gait Enhancing Mobile Shoe (GEMS) that mimics the desirable kinematics of a split-belt treadmill except that it does so over ground. Split-belt treadmills, with two separate treads running at different speeds, have been found useful in the rehabilitation of persons with asymmetric walking patterns. Although in preliminary testing, beneficial after-effects have been recorded, various drawbacks include the stationary nature of the split-belt treadmill and the inability to keep a person on the split-belt treadmill for an extended period of time. For this reason, the after-effects for long-term gait training are still unknown. The mobile ability of the GEMS outlined in this paper enables it to be worn in different environments such as in one\u27s own house and also enables it to be worn for a longer period of time since the GEMS is completely passive. Healthy subject testing has demonstrated that wearing this shoe for twenty minutes can alter the wearer\u27s gait and will generate after-effects in a similar manner as a split-belt treadmill does

Motion Controlled Gait Enhancing Mobile Shoe for Rehabilitation

Walking on a split-belt treadmill, which has two belts that can be run at different speeds, has been shown to improve walking patterns post-stroke. However, these improvements are only temporarily retained once individuals transition to walking over ground. We hypothesize that longer-lasting effects would be observed if the training occurred during natural walking over ground, as opposed to on a treadmill. In order to study such long-term effects, we have developed a mobile and portable device which can simulate the same gait altering movements experienced on a split-belt treadmill. The new motion controlled gait enhancing mobile shoe improves upon the previous version\u27s drawbacks. This version of the GEMS has motion that is continuous, smooth, and regulated with on-board electronics. A vital component of this new design is the Archimedean spiral wheel shape that redirects the wearer\u27s downward force into a horizontal backward motion. The design is passive and does not utilize any motors. Its motion is regulated only by a small magnetic particle brake. Further experimentation is needed to evaluate the long-term after-effects

Split-Belt Treadmill Adaptation Shows Different Functional Networks for Fast and Slow Human Walking

New walking patterns can be learned over short time scales (i.e., adapted in minutes) using a split-belt treadmill that controls the speed of each leg independently. This leads to storage of a modified motor pattern that is expressed as an aftereffect in regular walking conditions and must be de-adapted to return to normal. Here we asked whether the nervous system adapts a general walking pattern that is used across many speeds or a specific pattern affecting only the two speeds experienced during split-belt training. In experiment 1, we tested three groups of healthy adult subjects walking at different split-belt speed combinations and then assessed aftereffects at a range of speeds. We found that aftereffects were largest at the slower speed that was used in split-belt training in all three groups, and it decayed gradually for all other speeds. Thus adaptation appeared to be more strongly linked to the slow walking speed. This result suggests a separation in the functional networks used for fast and slow walking. We tested this in experiment 2 by adapting walking to split belts and then determining how much fast regular walking washed out the slow aftereffect and vice versa. We found that 23–38% of the aftereffect remained regardless of which speed was washed out first. This demonstrates that there is only partial overlap in the functional networks coordinating different walking speeds. Taken together, our results suggest that there are some neural networks for controlling locomotion that are recruited specifically for fast versus slow walking in humans, similar to recent findings in other vertebrates

Gait Transitions in Human Infants: Coping with Extremes of Treadmill Speed

<div><p>Spinal pattern generators in quadrupedal animals can coordinate different forms of locomotion, like trotting or galloping, by altering coordination between the limbs (interlimb coordination). In the human system, infants have been used to study the subcortical control of gait, since the cerebral cortex and corticospinal tract are immature early in life. Like other animals, human infants can modify interlimb coordination to jump or step. Do human infants possess functional neuronal circuitry necessary to modify coordination <i>within</i> a limb (intralimb coordination) in order to generate distinct forms of alternating bipedal gait, such as walking and running? We monitored twenty-eight infants (7–12 months) stepping on a treadmill at speeds ranging between 0.06–2.36 m/s, and seventeen adults (22–47 years) walking or running at speeds spanning the walk-to-run transition. Six of the adults were tested with body weight support to mimic the conditions of infant stepping. We found that infants could accommodate a wide range of speeds by altering stride length and frequency, similar to adults. Moreover, as the treadmill speed increased, we observed periods of flight during which neither foot was in ground contact in infants and in adults. However, while adults modified other aspects of intralimb coordination and the mechanics of progression to transition to a running gait, infants did not make comparable changes. The lack of evidence for distinct walking and running patterns in infants suggests that the expression of different functional, alternating gait patterns in humans may require neuromuscular maturation and a period of learning post-independent walking.</p></div

The interdomain region of dengue NS5 protein interacts with NS3 and host proteins

Although dengue virus genome replication occurs in the cytoplasm of infected cells, it has been shown that the NS5 protein (RNA-dependent RNA polymerase) is hyperphosphorylated at a late stage in infection and localized to the cell nucleus. A 37 amino acid sequence of NS5 (residues 369-405) was shown to contain a functional nuclear localization signal (NLS) that interacted with the cellular nuclear transport factor, importin a/β heterodimer. Further studies using the yeast two-hybrid system revealed that the NS5 region (residues 320-368) immediately adjacent to the NLS contained an importin β-binding site that abuts or overlaps the binding site for the NS3 protein (protease-helicase). The importin β-binding site has also been shown to be a functional NLS (bNLS). Intriguingly, when both bNLS and NLS (residues 320-405) were present, the fused β -galactosidase protein did not accumulate in the nucleus. Here we provide a review of our studies on the NS5 interdomain region and compare it to other members of the Flavivirus genus in order to highlight the importance of this region as a possible target for developing broad-acting antiviral agent dengue and other mechanistically-related viruses