Multi-Stage Modeling of the Kinetics of Activation of CaMKII

Ca 2+ /calmodulin-dependent protein kinase 2 (CaMKII) plays an important role in induction of long-term potentiation and formation of memory. It is abundant in dendritic spines, and is activated when Ca 2+ flows into the postsynaptic cytosol through open NMDA-type glutamate receptors. Its function is fine-tuned through interaction with other proteins as well as through subunit interactions and regulatory autophosphorylation. We have undertaken a multi-stage project to study the critical kinetics of activation of CaMKII in the spine by combining modeling and experimental studies. We are using computational modeling and simulations on various platforms, coupled with biochemical experiments in vitro, and eventually in vivo, to understand CaMKII regulation. The project includes the following steps: 1. Determining the parameters governing activation of a monomeric subunit. The CaMKII holoenzyme is a large dodecamer of similar, homologous subunits held together by interactions between the association domains located at the carboxyl end of each subunit. Individual, monomeric subunits can be expressed recombinantly by removing the association domain. Computer simulations of activation of monomeric CaMKII by Ca 2+ /calmodulin at both saturating and non-saturating concentrations in a test tube have helped to identify the binding parameters that are most crucial for modeling of regulation of CaMKII and thus have indicated the most useful biochemical assays to measure those parameters (Pepke et al., 2010). We are using these measurements to fine-tune our model of activation of individual catalytic subunits. 2. Building a model of the holoenzyme. Because a CaMKII holoenzyme contains 12 similar subunits, each of which can exist in several states, the holoenzyme can have a large number of state combinations. Thus, modeling the entire holoenzyme requires a computational framework that avoids the ensuing combinatorial complexity. The stochastic simulator MCell provides an elegant, rule-based way of modeling state changes in the CaMKII holoenzyme. 3. Modeling cooperativity that arises from the dodecameric structure of CaMKII. Autophosphorylation at threonine-286, which activates CaMKII subunits, is an inter-subunit event. Thus, it is greatly facilitated by the close proximity of subunits in the holoenzyme. In addition, the subunits within the holoenzyme are arranged as dimers which appears to result in cooperativity in the binding of Ca 2+ /CaM to individual subunits of the dimer (Chao et al., 2010). An accurate model of activation of subunits in the holoenzyme and their autophosphorylation will allow us to explore the effects of cooperativity on CaMKII activation on various time scales. 4. Modeling CaMKII within the context of a postsynaptic spine CaMKII interacts with a variety of other proteins, both in the postsynaptic density (PSD), close to major sources of Ca 2+ influx, and in other parts of the spine. In the fourth stage of this project we plan to implement kinetic models of activation of CaMKII in the context of an MCell model of Ca 2+ influx into a spine upon activation of NMDA-type glutamate receptors (Keller et al., 2008; Keller et al., 2011, submitted). We will explore the effects of different localization and numbers of CaMKII holoenzymes in the spine on CaMKII activation. References: Pepke, S., Kinzer-Ursem, T., Mihalas, S., and Kennedy, M.B. (2010). A dynamic model of interactions of Ca 2+ , calmodulin, and catalytic subunits of Ca 2+ /calmodulin-dependent protein kinase II. PLoS Comput Biol 6, e1000675. Chao, L.H., Pellicena, P., Deindl, S., Barclay, L.A., Schulman, H., and Kuriyan, J. (2010). Intersubunit capture of regulatory segments is a component of cooperative CaMKII activation. Nat Struct Mol Biol 17, 264-272. Keller, D.X., Franks, K.M., Bartol, T.M., Jr., and Sejnowski, T.J. (2008). Calmodulin activation by calcium transients in the postsynaptic density of dendritic spines. PLoS ONE 3, e2045. Keller, D.X., Bartol, T.M., Kinney, J.P, Kennedy, M.B., Bajaj, C., Harris, K.M., and Sejnowski, T.J. Regulation of synaptic calcium transients in reconstructed dendritic spines of hippocampal CA1 pyramidal neurons, submitted

The Ontogeny of Muscle Structure and Locomotory Function in the Long-Finned Squid Doryteuthis Pealeii

Understanding the extent to which changes in muscle form and function underlie ontogenetic changes in locomotory behaviors and performance is important in understanding the evolution of musculoskeletal systems and also the ecology of different life stages. We explored ontogenetic changes in the structure, myosin heavy chain (MHC) expression and contractile properties of the circular muscles that provide power for jet locomotion in the long-finned squid Doryteuthis pealeii. The circular muscle fibers of newly hatched paralarvae had different sizes, shapes, thick filament lengths, thin: thick filament ratio, myofilament organization and sarcoplasmic reticulum (SR) distribution than those of adults. Viewed in cross section, most circular muscle cells were roughly triangular or ovoid in shape with a core of mitochondria; however, numerous muscle cells with crescent or other unusual cross-sectional shapes and muscle cells with unequal distributions of mitochondria were present in the paralarvae. The frequency of these muscle cells relative to \u27normal\u27 circular muscle cells ranged from 1: 6 to 1: 10 among the 19 paralarvae we surveyed. The thick filaments of the two types of circular fibers, superficial mitochondria-rich (SMR) and central mitochondria-poor (CMP), differed slightly in length among paralarvae with thick filament lengths of 0.83+/-0.15 μm and 0.71+/-0.1μm for the SMR and CMP fibers, respectively (P 0.05; ANOVA). During ontogeny the thick filament lengths of both the CMP and SMR fibers increased significantly to 1.78+/-0.27 μm and 3.12+/-0.56 μm, respectively, in adults (P-1 (where L0 was the preparation length that generated the peak isometric stress), nearly twice that measured in other studies for the CMP fibers of adults. The mean peak isometric stress was 119+/-15mN mm-2 physiological cross section, nearly half that measured for the CMP fibers of adults. Reverse transcriptase-polymerase chain reaction analysis of paralarval and adult mantle samples revealed very similar expression patterns of the two known isoforms of squid MHC. The ontogenetic differences in the structure and physiology of the circular muscles may result in more rapid mantle movements during locomotion. This prediction is consistent with jet pulse durations observed in other studies, with shorter jet pulses providing hydrodynamic advantages for paralarvae

Choroidal thickness measured using swept-source optical coherence tomography is reduced in patients with type 2 diabetes

Objective To compare choroidal thickness between patients with type 2 diabetes (T2D) and healthy controls measured using swept-source optical coherence tomography (SS-OCT). Methods The sample comprised 157 eyes of 94 T2D patients, 48 eyes of which had diabetic macular edema (DME), and 71 normal eyes of 38 healthy patients. Subfoveal (SF) choroidal thickness, and choroidal thickness at 500-µm intervals up to 2500 µm nasal and temporal from the fovea were measured using the SS-OCT. Choroidal thicknesses were compared between groups using Student’s t-test. Additionally, Pearson correlations were calculated between diabetes duration, glycosylated hemoglobin (HbA1c) levels, and choroidal thickness. Results Mean diabetes duration was 16.6±9.5 years, while mean glycosylated hemoglobin was 7.7 ±1.3%. Overall, the choroid was significantly thinner in T2D patients. Individuals with DME had reduced choroidal thickness in all measurements, except at 2000 and 2500-µm nasal positions, compared to healthy controls. There was a moderate correlation between choroidal thickness and HbA1c levels in DME patients (SF: r = 0.342; p = 0.017). Diabetes duration did not correlate significantly with choroidal thickness. Conclusion SS-OCT measurements revealed that the choroid was significantly thinner in T2D patients, moderate non-proliferative diabetic retinopathy patients, and DME patients than in healthy individuals. Further studies are needed to clarify the effect of diabetes on this layer and the relationship between choroidal thickness and DME

Evidence of self-correcting spiral flows in swimming boxfishes

"The marine boxfishes have rigid keeled exteriors (carapaces) unlike most fishes, yet exhibit high stability, high maneuverability and relatively low drag given their large cross-sectional area. These characteristics lend themselves well to bioinspired design. Based on previous stereolithographic boxfish model experiments, it was determined that vortical flows develop around the carapace keels, producing self-correcting forces that facilitate swimming in smooth trajectories. To determine if similar self-correcting flows occur in live, actively swimming boxfishes, two species of boxfishes (Ostracion meleagris and Lactophrys triqueter) were induced to swim against currents in a water tunnel, while flows around the fishes were quantified using digital particle image velocimetry. Significant pitch events were rare and short lived in the fishes examined. When these events were observed, spiral flows around the keels qualitatively similar to those observed around models were always present, with greater vortex circulation occurring as pitch angles deviated from 0deg. Vortex circulation was higher in live fishes than models presumably because of pectoral fin interaction with the keel-induced flows. The ability of boxfishes to modify their underlying self-correcting system with powered fin control is important for achieving high levels of both stability and maneuverability. Although the challenges of performing stability and maneuverability research on fishes are significant, the results of this study together with future studies employing innovative new approaches promise to provide valuable inspiration for the designers of bioinspired aquatic vehicles."http://deepblue.lib.umich.edu/bitstream/2027.42/64212/1/bb8_1_014001.pd

Repeatability of choroidal thickness measurements assessed with swept-source optical coherence tomography in healthy and diabetic individuals

PURPOSE: To assess the intrasession repeatability of choroidal thickness measurements obtained using swept-source optical coherence tomography in Type 2 diabetic (T2D) patients and healthy controls. METHODS: This was a single-center, prospective, observational, cross-sectional study with consecutive inclusion of 33 healthy subjects and 43 T2D patients. Subjects underwent three consecutive swept-source optical coherence tomography scans in a single session. After automatic delineation of the choroid, subfoveal choroidal thickness, and thickness at 500-µm intervals up to 2, 500 µm nasal and temporal from the fovea were measured using the software caliper by the same operator. Intraclass correlation coefficients (ICCs), coefficients of variation, and test-retest variability were calculated. RESULTS: Mean subfoveal choroidal thickness in healthy subjects and in T2D patients was 229.97 ± 79.9 and 192.67 ± 74.3 µm, respectively (P = 0.013). All intrasession intraclass correlation coefficients were higher than 0.95 and 0.99, respectively. Coefficients of variations were less than 4.4% and 1.8%, respectively. Test-retest variability ranged from 0.76 µm to 11.12 µm and 0.64 µm to 6.29 µm, respectively. No significant differences were found in the intrasession repeatability of any choroidal measurement between healthy subjects and T2D patients. CONCLUSION: Swept-source optical coherence tomography provided excellent intrasession repeatability of choroidal thickness measurements in healthy subjects and T2D patients

Comparison of visual and optical quality of monofocal versus multifocal intraocular lenses

Objective: To compare visual quality in patients implanted with Tecnis ® monofocal (ZCB00) and multifocal (ZMB00) intraocular lenses taking into account their optical quality measured in vitro with an eye model. Methods: In total, 122 patients participated in this study: 44 implanted with monofocal and 78 with multifocal intraocular lenses. Measurements of visual acuity and contrast sensitivity were performed. The optical quality of the intraocular lenses was evaluated in three image planes (distance, intermediate and near) using an eye model on a test bench. The metric considered was the area under the curve of the modulation transfer function. Results: Optical quality at the far focus of the monofocal intraocular lens (area under the curve of the modulation transfer function = 66.97) was considerably better than that with the multifocal lens (area under the curve of the modulation transfer function = 32.54). However, no significant differences were observed between groups at the distance-corrected visual acuity. Distance-corrected near vision was better in the multifocal (0.15 ± 0.20 logMAR) than that in the monofocal group (0.43 ± 0.21 logMAR, p < 0.001), which correlated with the better optical quality at near reached by the multifocal intraocular lens (area under the curve of the modulation transfer function = 29.11) in comparison with the monofocal intraocular lens (area under the curve of the modulation transfer function = 5.0). In intermediate vision, visual acuity was 0.28 ± 0.16 logMAR (multifocal) and 0.36 ± 0.14 logMAR (monofocal) with p = 0.014, also in good agreement with the values measured in the optical quality (area under the curve of the modulation transfer function = 10.69 (multifocal) and 8.86 (monofocal)). The contrast sensitivity was similar in almost all frequencies. Pelli–Robson was slightly better in the monofocal (1.73) than in the multifocal group (1.64; p = 0.023). Conclusion: Patients implanted with multifocal ZMB00 achieved a distance visual acuity similar to those implanted with monofocal ZCB00, but showed significantly better intermediate and near visual acuity. A correlation was found between intraocular lenses’ optical quality and patients’ visual acuity. Contrast sensitivity was very similar between the multifocal and monofocal groups

Influence of ocular biometric factors on the defocus curve in an enlarged depth-of-focus intraocular lens

Background: To assess the influence of biometric measurements on the defocus curve after the implantation of enlarged depth-of-focus (EDoF) intraocular lens (IOL). Methods: Patients who underwent cataract surgery with bilateral implantation of Tecnis Symfony IOL were enrolled. Preoperatively, axial length (AL), corneal keratometry (K), pupil size and corneal aberrations were measured. 1 month after surgery, distance, intermediate, and near visual acuities (VA) were recorded. At 3 months, monocular and binocular corrected contrast sensitivities under photopic and mesopic lighting conditions were measured with CSV-1000E test. At 6-months, the defocus curve between −5.00 to + 3.00 diopters (D) was assessed in steps of 0.50 D, and NEI-RQL-42 questionnaire was administered. Results: One hundred thirty one eyes of 66 patients were included. Binocular logMAR VA better than 0.1 for intermediate vision was obtained in 90% of patients, whereas only 17.7% obtained that result in near vision. The rate of satisfaction was high (96%) and most of them (85.5%) had no or little difficulties in near vision. The mean amplitude of the defocus curve was 2.35D ± 0.73D, and smaller AL, smaller pupils, younger age, and male sex were associated with wider range of clear vision. Conclusions: Tecnis Symfony IOL enables functional vision at all distances, but demographic variables and preoperative biometric measurements like AL and pupil size influence the postoperative amplitude of the defocus curve. These parameters could be used to predict the performance of EDoF IOLs

Comparison of visual and optical quality of monofocal versus multifocal intraocular lenses

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Avoiding obscure topics and generalising findings produces higher impact research

Much academic research is never cited and may be rarely read, indicating wasted effort from the authors, referees and publishers. One reason that an article could be ignored is that its topic is, or appears to be, too obscure to be of wide interest, even if excellent scholarship produced it. This paper reports a word frequency analysis of 874,411 English article titles from 18 different Scopus natural, formal, life and health sciences categories 2009-2015 to assess the likelihood that research on obscure (rarely researched) topics is less cited. In all categories examined, unusual words in article titles associate with below average citation impact research. Thus, researchers considering obscure topics may wish to reconsider, generalise their study, or to choose a title that reflects the wider lessons that can be drawn. Authors should also consider including multiple concepts and purposes within their titles in order to attract a wider audience