Anterior Pelvic Ring Fracture Pattern Predicts Subsequent Displacement in Lateral Compression Sacral Fractures.

OBJECTIVE: To determine if anterior pelvic fracture pattern in lateral compression (LC) sacral fractures correlates with subsequent displacement on examination under anesthesia (EUA) or follow-up in both nonoperative and operative cases. DESIGN: Retrospective cohort study. SETTING: Level 1 trauma center. PATIENTS: Two hundred twenty-seven skeletally mature patients with traumatic LC (OTA/AO 61B1.1, 61B2.1-2, and 61B3.1-2) pelvic ring injuries treated nonoperatively, with EUA, or with pelvic fixation were included. INTERVENTION: The study intervention included retrospective review of patients' charts and radiographs. MAIN OUTCOME MEASUREMENT: Displacement on EUA or follow-up radiographs (both operative and nonoperative) correlated with anterior pelvic ring fracture pattern. RESULTS: Independent of sacral fracture pattern (complete or incomplete), risk of subsequent displacement on EUA or at follow-up after both nonoperative and operative treatments correlated strongly with ipsilateral superior and inferior pubic rami fractures that were either comminuted (95.6%, P < 0.001) or oblique (100%, P < 0.001). Patients with transverse or lack of inferior pubic ramus fracture did not displace (0%, P < 0.001). Out of 21 LC injuries treated with posterior-only fixation, displacement at follow-up occurred in all 11 patients (100%) with comminuted and/or oblique superior and inferior pubic rami fractures. Nakatani zone I and II rami fractures correlated most with risk of subsequent displacement. CONCLUSIONS: Unstable anterior fracture patterns are characterized as comminuted and/or oblique fractures of ipsilateral superior and inferior pubic rami. EUA should be strongly considered in these patients to disclose occult instability, for both complete and incomplete sacral fracture patterns. Additionally, these unstable anterior fracture patterns are poor candidates for posterior-only fixation and supplemental anterior fixation should be considered. Irrespective of sacral fracture pattern (complete or incomplete), nonoperative management is successful in patients with transverse or lack of inferior pubic ramus fractures. LEVEL OF EVIDENCE: Prognostic Level III. See Instructions for Authors for a complete description of levels of evidence.

Preliminary Report on the Train the Brain Project, Part I: Sensorimotor Neural Correlates of Anterior Cruciate Ligament Injury Risk Biomechanics.

CONTEXT: Anterior cruciate ligament injury commonly occurs via noncontact motor coordination errors that result in excessive multiplanar loading during athletic movements. Preventing motor coordination errors requires neural sensorimotor integration activity to support knee-joint neuromuscular control, but the underlying neural mechanisms driving injury-risk motor control are not well understood. OBJECTIVE: To evaluate brain activity differences for knee sensorimotor control between athletes with high or low injury-risk mechanics. DESIGN: Case-control study. SETTING: Research laboratory. PATIENTS OR OTHER PARTICIPANTS: Of 38 female high school soccer players screened, 10 were selected for analysis based on magnetic resonance imaging compliance, injury-risk classification via 3-dimensional biomechanics during a drop vertical jump, and matching criteria to complete neuroimaging during knee motor tasks. MAIN OUTCOME MEASURE(S): Peak knee-abduction moment during landing was used for group allocation into the high (≥21.74 newton meters [Nm], n = 9) or low (≤10.6 Nm, n = 11) injury-risk classification (n = 11 uncategorized, n = 7 who were not compliant with magnetic resonance imaging). Ten participants (5 high risk, 5 low risk) with adequate data were matched and compared across 2 neuroimaging paradigms: unilateral knee-joint control and unilateral multijoint leg press against resistance. RESULTS: Athletes with high injury-risk biomechanics had less neural activity in 1 sensory-motor cluster for isolated knee-joint control (precuneus, peak Z score = 4.14, P ≤ .01, 788 voxels) and greater brain activity for the multijoint leg press in 2 cognitive-motor clusters: the frontal cortex (peak Z score = 4.71, P < .01, 1602 voxels) and posterior cingulate gyrus (peak Z score = 4.43, P < .01, 725 voxels) relative to the low injury-risk group. CONCLUSIONS: The high injury-risk group's lower relative engagement of neural sensory resources controlling the knee joint may elevate demand on cognitive motor resources to control loaded multijoint action. The neural activity profile in the high injury-risk group may manifest as a breakdown in neuromuscular coordination, resulting in elevated knee-abduction moments during landing.

Pixel MTF response effect on non-null interferometry.

The pixel modulation transfer function response degrades the contrast of non-null interferometric surface figure measurements. We experimentally quantify this effect for spatial frequencies ranging from 0 to 363 lp/mm (≈3.33 times the Nyquist limit). Our results show a low SNR spatial frequency band that behaves like a low-pass filter for sub-Nyquist interferometry and a stop-band filter for multiple-wavelength phase-shifting interferometry. We also introduce a multiple-mode, multiple-wavelength interferometry approach to measure optical surfaces with slope departure angles mapping to spatial frequencies in this low SNR band. The extended measurement range of this approach is achieved without using a sparse-array detector.

Temporal evolution of the biological response to laser-induced refractive index change (LIRIC) in rabbit corneas.

Laser-induced refractive index change (LIRIC) is a new, non-incisional, non-ablative, femtosecond photo-modification technique being developed for vision correction in humans. Prior, exvivo studies showed intra-tissue refractive index change to induce minimal cell death, restricted to the laser focal zone in the corneal stroma, and with no observable damage to the epithelium or endothelium. Here, we used live rabbits to ascertain longer-term consequences of LIRIC in vivo. Specifically, we assessed cell death, fibrosis, corneal nerve distribution, endothelial cell density, and corneal structure for up to 3 months after LIRIC. A +2.5 D gradient-index LIRIC Fresnel lens was inscribed inside 20 applanated corneas of Dutch Belted rabbits, over a circular region of the mid-stroma measuring 4.5 mm in diameter. Twelve additional rabbit eyes were used as applanation-only controls to differentiate the effects of laser treatment and suction applanation on biological and structural parameters. In vivo optical measurements were performed pre-operatively, then immediately, 2, 4, and 12 weeks after the procedure, to measure endothelial cell density and changes in corneal structure. Groups of four rabbits were sacrificed at 4 hours, 2, 4, and 12 weeks after LIRIC for histological determinations; the TUNEL assay was used to evaluate cell death, H&E staining was used to assess inflammatory infiltration, and immunostaining for α-smooth muscle actin (α-SMA) and ßIII tubulin (Tuj-1) was performed to assess myofibroblast differentiation and corneal nerve distribution, respectively. Consistent with prior ex vivo data, only minimal cell death was observed in the laser focal zone, with TUNEL-positive cells restricted to the stromal region of refractive index change 4 h after LIRIC. No TUNEL-positive cells were evident anywhere in the cornea 2, 4, or 12 weeks after LIRIC. Applanation-only corneas were completely TUNEL-negative. Neither LIRIC-treated nor applanation-only eyes exhibited α-SMA-positive staining or altered corneal nerve distributions at any of the time points examined. In vivo confocal imaging revealed normal endothelial cell densities in all eyes (whether LIRIC-treated or applanation-only) at all time points. Optical coherence tomography showed suction applanation to cause a temporary decrease in central corneal thickness, which returned to normal within 4 h. Corneas into which LIRIC Fresnel lenses were written while applanated did not undergo major structural or shape changes beyond the temporary thinning already described for suction applanation. The present findings suggest that LIRIC patterns, which generated a clinically-relevant refractive correction in the mid-stromal region of live rabbit corneas, induced little-to-no disruption to corneal structure and biology for 3 months after the procedure. This affirms the relative safety of LIRIC and predicts that compared to traditional laser vision correction surgeries, common post-operative complications such as dry eye, haze, or patient discomfort may be entirely avoided.

A compact high-precision periodic-error-free heterodyne interferometer.

We present the design, bench-top setup, and experimental results of a compact heterodyne interferometer that achieves picometer-level displacement sensitivities in air over frequencies above 100 MHz. The optical configuration with spatially separated beams prevents frequency and polarization mixing, and therefore eliminates periodic errors. The interferometer is designed to maximize common-mode optical laser beam paths to obtain high rejection of environmental disturbances, such as temperature fluctuations and acoustics. The results of our experiments demonstrate the short- and long-term stabilities of the system during stationary and dynamic measurements. In addition, we provide measurements that compare our interferometer prototype with a commercial system, verifying our higher sensitivity of 3 pm, higher thermal stability by a factor of two, and periodic-error-free performance.

Advances in optical metrology and instrumentation: introduction.

Optical measurement and characterization are two of the pillars of metrology. The ability to measure precisely with high dynamic range and accuracy betters our understanding of nature and the universe. In this feature issue, we present a collection of articles that delves into the fundamental techniques used to advance the field.

Prospective longitudinal investigation shows correlation of event-related potential to mild traumatic brain injury in adolescents.

STUDY DESIGN: Prospective longitudinal cohort study. BACKGROUND: Adolescent athletes may be more susceptible to the long-term effects of mild traumatic brain injury (mTBI). A diagnostic and prognostic neuromarker may optimize management and return-to-activity decision-making in athletes who experience mTBI. OBJECTIVE: Measure an event-related potential (ERP) component captured with electroencephalography (EEG), called processing negativity (PN), at baseline and post-injury in adolescents who suffered mTBI and determine their longitudinal response relative to healthy controls. METHODS: Thirty adolescents had EEG recorded during an auditory oddball task at a pre-mTBI baseline session and subsequent post-mTBI sessions. Longitudinal EEG data from patients and healthy controls (n= 77) were obtained from up to four sessions in total and processed using Brain Network Analysis algorithms. RESULTS: The average PN amplitude in healthy controls significantly decreased over sessions 2 and 3; however, it remained steady in the mTBI group's 2nd (post-mTBI) session and decreased only in sessions 3 and 4. Pre- to post-mTBI amplitude changes correlated with the time interval between sessions. CONCLUSION: These results demonstrate that PN amplitude changes may be associated with mTBI exposure and subsequent recovery in adolescent athletes. Further study of PN may lead to it becoming a neuromarker for mTBI prognosis and return-to-activity decision-making in adolescents.

Real-time biofeedback integrated into neuromuscular training reduces high-risk knee biomechanics and increases functional brain connectivity: A preliminary longitudinal investigation.

Prospective evidence indicates that functional biomechanics and brain connectivity may predispose an athlete to an anterior cruciate ligament injury, revealing novel neural linkages for targeted neuromuscular training interventions. The purpose of this study was to determine the efficacy of a real-time biofeedback system for altering knee biomechanics and brain functional connectivity. Seventeen healthy, young, physically active female athletes completed 6 weeks of augmented neuromuscular training (aNMT) utilizing real-time, interactive visual biofeedback and 13 served as untrained controls. A drop vertical jump and resting state functional magnetic resonance imaging were separately completed at pre- and posttest time points to assess sensorimotor adaptation. The aNMT group had a significant reduction in peak knee abduction moment (pKAM) compared to controls (p = .03, d = 0.71). The aNMT group also exhibited a significant increase in functional connectivity between the right supplementary motor area and the left thalamus (p = .0473 after false discovery rate correction). Greater percent change in pKAM was also related to increased connectivity between the right cerebellum and right thalamus for the aNMT group (p = .0292 after false discovery rate correction, r2  = .62). No significant changes were observed for the controls (ps > .05). Our data provide preliminary evidence of potential neural mechanisms for aNMT-induced motor adaptations that reduce injury risk. Future research is warranted to understand the role of neuromuscular training alone and how each component of aNMT influences biomechanics and functional connectivity.

Alterations in knee sensorimotor brain functional connectivity contributes to ACL injury in male high-school football players: a prospective neuroimaging analysis.

OBJECTIVE: This study's purpose was to utilize a prospective dataset to examine differences in functional brain connectivity in male high school athletes who suffered an anterior cruciate ligament (ACL) injury relative to their non-injured peers. METHODS: Sixty-two male high school football players were evaluated using functional magnetic resonance imaging prior to their competitive season to evaluate resting-state functional brain connectivity. Three athletes later experienced an ACL injury and were matched to 12 teammates who did not go on to sustain an ACL injury (controls) based on school, age, height, weight, and year in school. Twenty-five knee-motor regions of interest (ROIs) were created to identify differences in connectivity between the two groups. Between-subject F and t tests were used to identify significant ROI differences using a false discovery rate correction for multiple comparisons. RESULTS: There was significantly less connectivity between the left secondary somatosensory cortex and the left supplementary motor area (p = 0.025), right pre-motor cortex (p = 0.026), right supplementary motor area (p = 0.026), left primary somatosensory cortex (superior division; p = 0.026), left primary somatosensory cortex (inferior division; p = 0.026), and left primary motor cortex (p = 0.048) for the ACL-injured compared to the control subjects. No other ROI-to-ROI comparisons were significantly different between the groups (all p > 0.05). CONCLUSION: Our preliminary data indicate a potential sensorimotor disruption for male football players who go on to experience an ACL injury. Future studies with larger sample sizes and complementary measures of neuromuscular control are needed to support these findings.

A Novel Approach to Evaluate Brain Activation for Lower Extremity Motor Control.

BACKGROUND AND PURPOSE: The purpose of this study was to assess the consistency of a novel MR safe lower extremity motor control neuroimaging paradigm to elicit reliable sensorimotor region brain activity. METHODS: Participants completed multiple sets of unilateral leg presses combining ankle, knee, and hip extension and flexion movements against resistance at a pace of 1.2 Hz while lying supine in a 3T MRI scanner. Regions of Interest (ROI) consisted of regions primarily involved in lower extremity motor control (right and left primary motor cortex, primary somatosensory cortex, premotor cortex, secondary somatosensory cortex, basal ganglia, and the cerebellum). RESULTS: The group analysis based on mixed effects paired samples t-test revealed no differences for brain activity between sessions (P > .05). Intraclass correlation coefficients in the sensorimotor regions were good to excellent for average percent signal change (.621 to .918) and Z-score (.697 to .883), with the exception of the left secondary somatosensory cortex percent signal change (.165). CONCLUSIONS: These results indicate that a loaded lower extremity force production and attenuation task that simulates the range of motion of squatting, stepping, and landing from a jump is reliable for longitudinal neuroimaging applications and support the use of this paradigm in further studies examining therapeutic interventions and changes in dynamic lower extremity motor function.

Does brain functional connectivity contribute to musculoskeletal injury? A preliminary prospective analysis of a neural biomarker of ACL injury risk.

OBJECTIVES: We aimed to present a unique prospective neurological dataset for participants who experienced an ACL injury. DESIGN: Prospective longitudinal case-control. METHODS: High school female soccer athletes were evaluated using functional magnetic resonance imaging to capture resting-state brain connectivity prior to their competitive season. Two of these athletes later experienced an ACL injury (ACLI). We matched these ACLI participants with eight teammates who did not go on to sustain an ACL injury (uninjured controls, Con) based on age, grade, sex, height, and weight to examine differences in preseason connectivity. Knee-motor regions of interest (ROIs) were created based on previously published data from which five specific areas were selected as seeds for analysis. Independent-samples t-tests with a false discovery rate correction for multiple comparisons determined differences in connectivity between the ACLI and Con. RESULTS: There was significantly greater connectivity between the left primary sensory cortex (a brain region responsible for proprioception) and the right posterior lobe of the cerebellum (a brain region responsible for balance and coordination) for the Con relative to ACLI, t (8)=4.53, p=0.03 (false discovery rate corrected). CONCLUSIONS: These preliminary data indicate that those who do not later sustain an ACL injury exhibit a stronger functional connection between a cortical sensory-motor region and a cerebellar region responsible for balance and coordination. These findings may help to guide development of brain-driven biofeedback training that optimizes and promotes adaptive neuroplasticity to reduce motor coordination errors and injury risk.

Genome-wide CRISPR-Cas9 Interrogation of Splicing Networks Reveals a Mechanism for Recognition of Autism-Misregulated Neuronal Microexons.

Alternative splicing is crucial for diverse cellular, developmental, and pathological processes. However, the full networks of factors that control individual splicing events are not known. Here, we describe a CRISPR-based strategy for the genome-wide elucidation of pathways that control splicing and apply it to microexons with important functions in nervous system development and that are commonly misregulated in autism. Approximately 200 genes associated with functionally diverse regulatory layers and enriched in genetic links to autism control neuronal microexons. Remarkably, the widely expressed RNA binding proteins Srsf11 and Rnps1 directly, preferentially, and frequently co-activate these microexons. These factors form critical interactions with the neuronal splicing regulator Srrm4 and a bi-partite intronic splicing enhancer element to promote spliceosome formation. Our study thus presents a versatile system for the identification of entire splicing regulatory pathways and further reveals a common mechanism for the definition of neuronal microexons that is disrupted in autism.

Anomalous vibration suppression in a weak-value-emulated heterodyne roll interferometer.

We experimentally validate the vibration suppression capabilities of a weak-value-like protocol. The phase-sensitive heterodyne technique exhibits advantageous characteristics of a weak measurement including anomalous amplification in sensitivity and technical noise suppression. It does not, however, leverage the entanglement between the system and meter to amplify the signal of interest, as is typical in a weak measurement. In this formalism, we demonstrate an amplification in sensitivity to the roll angle of over 700 times. High precision roll experiments anchor numerical simulations to show that the interferometer outperforms standard interferometry by a factor of 500 in terms of peak-to-peak noise amplitude. During the measurement of a rolling stage, technical noise - primarily in the form of vibrations - is substantially attenuated. This is the first demonstration of vibration suppression capabilities that are inherent to the light from a metrology instrument instead of achieved via mechanical damping. The emulation presented in this work also identifies an avenue to achieve anomalous amplification outside of the standard weak measurement protocol.

Brain-Behavior Mechanisms for the Transfer of Neuromuscular Training Adaptions to Simulated Sport: Initial Findings From the Train the Brain Project.

CONTEXT: A limiting factor for reducing anterior cruciate ligament injury risk is ensuring that the movement adaptions made during the prevention program transfer to sport-specific activity. Virtual reality provides a mechanism to assess transferability, and neuroimaging provides a means to assay the neural processes allowing for such skill transfer. OBJECTIVE: To determine the neural mechanisms for injury risk-reducing biomechanics transfer to sport after anterior cruciate ligament injury prevention training. DESIGN: Cohort study. SETTING: Research laboratory. PARTICIPANTS: Four healthy high school soccer athletes. INTERVENTIONS: Participants completed augmented neuromuscular training utilizing real-time visual feedback. An unloaded knee extension task and a loaded leg press task were completed with neuroimaging before and after training. A virtual reality soccer-specific landing task was also competed following training to assess transfer of movement mechanics. MAIN OUTCOME MEASURES: Landing mechanics during the virtual reality soccer task and blood oxygen level-dependent signal change during neuroimaging. RESULTS: Increased motor planning, sensory and visual region activity during unloaded knee extension and decreased motor cortex activity during loaded leg press were highly correlated with improvements in landing mechanics (decreased hip adduction and knee rotation). CONCLUSION: Changes in brain activity may underlie adaptation and transfer of injury risk-reducing movement mechanics to sport activity. Clinicians may be able to target these specific brain processes with adjunctive therapy to facilitate intervention improvements transferring to sport.

Mild Jugular Compression Collar Ameliorated Changes in Brain Activation of Working Memory after One Soccer Season in Female High School Athletes.

Recent neuroimaging studies have suggested that repetitive subconcussive head impacts, even after only one sport season, may lead to pre- to post-season structural and functional alterations in male high school football athletes. However, data on female athletes are limited. In the current investigation, we aimed to (1) assess the longitudinal pre- to post-season changes in functional MRI (fMRI) of working memory and working memory performance, (2) quantify the association between the pre- to post-season change in fMRI of working memory and the exposure to head impact and working memory performance, and (3) assess whether wearing a neck collar designed to reduce intracranial slosh via mild compression of the jugular veins can ameliorate the changes in fMRI brain activation observed in the female high school athletes who did not wear collars after a full soccer season. A total of 48 female high school soccer athletes (age range: 14.00-17.97 years) were included in the study. These athletes were assigned to the non-collar group (n = 21) or to the collar group (n = 27). All athletes undewent MRI at both pre-season and post-season. In each session, a fMRI verbal N-Back task was used to engage working memory. A significant pre- to post-season increase in fMRI blood oxygen level dependent (BOLD) signal was demonstrated when performing the N-back working memory task in the non-collar group but not in the collar group, despite the comparable exposure to head impacts during the season between the two groups. The collar group demonstrated significantly smaller pre- to post-season change in fMRI BOLD signal than the non-collar group, suggesting a potential protective effect from the collar device. Significant correlations were also found between the pre- to post-season increase in fMRI brain activation and the decrease in task accuracy in the non-collar group, indicating an association between the compensatory mechanism in underlying neurophysiology and the alteration in the behavioral outcomes.

Two-dimensional straightness measurement based on optical knife-edge sensing.

Straightness error is a parasitic translation along a perpendicular direction to the primary displacement axis of a linear stage. The parasitic translations could be coupled into other primary displacement directions of a multi-axis platform. Hence, its measurement and compensation are critical in precision multi-axis metrology, calibration, and manufacturing. This paper presents a two-dimensional (2D) straightness measurement configuration based on 2D optical knife-edge sensing, which is simple, light-weight, compact, and easy to align. It applies a 2D optical knife-edge to manipulate the diffraction pattern sensed by a quadrant photodetector, whose output voltages could derive 2D straightness errors after a calibration process. This paper analyzes the physical model of the configuration and performs simulations and experiments to study the system sensitivity, measurement nonlinearity, and error sources. The results demonstrate that the proposed configuration has higher sensitivity and insensitive to beam's vibration, compared with the conventional configurations without using the knife-edge, and could achieve ±0.25 µm within a ±40 µm measurement range along a 40 mm primary axial motion.

Contrasting cellular damage after Blue-IRIS and Femto-LASIK in cat cornea.

Blue-intra-tissue refractive index shaping (Blue-IRIS) is a new approach to laser refractive correction of optical aberrations in the eye, which alters the refractive index of the cornea rather than changing its shape. Before it can be implemented in humans, it is critical to establish whether and to what extent, Blue-IRIS damages the cornea. Here, we contrasted the impact of -1.5 D cylinder refractive corrections inscribed using either Blue-IRIS or femtosecond laser in-situ keratomileusis (femto-LASIK) on corneal cell viability. Blue-IRIS was used to write a -1.5 D cylinder gradient index (GRIN) lens over a 2.5 mm by 2.5 mm area into the mid-stromal region of the cornea in six freshly-enucleated feline eyes. The same correction (-1.5 D cylinder) was inscribed into another four cat eyes using femto-LASIK. Six hours later, all corneas were processed for histology and stained for terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) and p-γ-H2AX to label damaged cells. In Blue-IRIS-treated corneas, no tissue was removed and TUNEL-stained cells were confined to the laser focal zone in the stroma. In femto-LASIK, photoablation removed 14 µm of anterior stroma, but in addition, TUNEL-positive cells clustered across the femto-flap, the epithelium at the flap edges and the stroma below the ablation zone. Keratocytes positive for p-γ-H2AX were seen adjacent to all Blue-IRIS focal zones, but were completely absent from femto-LASIK-treated corneas. Unlike femto-LASIK, Blue-IRIS attains refractive correction in the cornea without tissue removal and only causes minimal, localized keratocyte death within the laser focal zones. In addition, Blue-IRIS induced DNA modifications associated with phosphorylation of γ-H2AX in keratocytes adjacent to the laser focal zones. We posit that this p-γ-H2AX response is related to alterations in chromatin structure caused by localized changes in osmolarity, a possible mechanism for the induced refractive index changes.

Comparable change in stromal refractive index of cat and human corneas following blue-IRIS.

Blue intratissue refractive index shaping (blue-IRIS) is a method with potential to correct ocular refraction noninvasively in humans. To date, blue-IRIS has only ever been applied to cat corneas and hydrogels. To test the comparability of refractive index change achievable in cat and human tissues, we used blue-IRIS to write identical phase gratings in ex vivo feline and human corneas. Femtosecond pulses (400 nm) were focused ? 300 ?? ? m below the epithelial surface of excised cat and human corneas and scanned to write phase gratings with lines ? 1 ?? ? m wide, spaced 5 ?? ? m apart, using a scan speed of 5 ?? mm / s . Additional cat corneas were used to test writing at 3 and 7 ?? mm / s in order to document speed dependence of the refractive index change magnitude. The first-order diffraction efficiency was immediately measured and used to calculate the refractive index change attained. Our data show that blue-IRIS induces comparable refractive index changes in feline and human corneas, an essential requirement for further developing its use as a clinical vision correction technique.

LUMIER: A Discovery Tool for Mammalian Protein Interaction Networks.

Protein-protein interactions (PPIs) play an essential role in all biological processes. In vivo, PPIs occur dynamically and depend on extracellular cues. To discover novel protein-protein interactions in mammalian cells, we developed a high-throughput automated technology called LUMIER (LUminescence-based Mammalian IntERactome). In this approach, we co-express a Luciferase (LUC)-tagged fusion protein along with a Flag-tagged protein in an efficiently transfectable cell line such as HEK-293T cells. The interaction between the two proteins is determined by co-immunoprecipitation using an anti-Flag antibody, and the presence of the LUC-tagged interactor in the complex is subsequently detected via its luciferase activity. LUMIER can easily detect transmembrane protein partners, interactions that are signaling- or splice isoform-dependent, as well as those that may occur only in the presence of posttranslational modifications. Using various collections of Flag-tagged proteins, we have generated protein interaction networks for several TGF-ß family receptors, Wnt pathway members, and have systematically analyzed the effect of neural-specific alternative splicing on protein interaction networks. The results have provided important insights into the physiological and functional relevance of some of the novel interactions found. LUMIER is highly scalable and can be used for both low- and high-throughput strategies. LUMIER is thus a valuable tool for the identification and characterization of dynamically regulated PPIs in mammalian systems. Here, we describe a manual version of LUMIER in a 96-well format that can be easily implemented in any laboratory.

Simultaneous interferometric measurement of linear coefficient of thermal expansion and temperature-dependent refractive index coefficient of optical materials.

Characterizing the thermal properties of optical materials is necessary for understanding how to design an optical system for changing environmental conditions. A method is presented for simultaneously measuring both the linear coefficient of thermal expansion and the temperature-dependent refractive index coefficient of a sample interferometrically in air. Both the design and fabrication of the interferometer is presented as well as a discussion of the results of measuring both a steel and a CaF2 sample.