An Alternative Method of Assessing d/DHH Secondary-School Students' L2 Language Development.

Little information is available on d/Deaf and hard of hearing (d/DHH) learners' L2 development. Their limited auditory access may discourage them from taking standardized tests, highlighting the need for alternative ways of assessing their L2 development and proficiency. Therefore, this study suggests adopting processability theory, which demonstrates a universal order of L2 development. Interviews with d/DHH learners and their teachers were conducted to explore their current difficulties in regard to understanding their L2 development. Also, we conducted brief speaking tasks to suggest alternatives to testing the L2 development of learners who are d/DHH in comparison to typical literacy learners. The result showed d/DHH students' L2 developmental patterns are similar to those of typical hearing peers, suggesting that d/DHH students and hearing learners share difficulties in similar areas when learning English. Teachers highlighted the lack of appropriate English tests to determine the d/DHH students' L2 development.

Enhancement of preimplantation mouse embryo development with optimized in vitro culture dish via stabilization of medium osmolarity.

OBJECTIVE: We evaluated the efficacy of the newly developed optimized in vitro culture (OIVC) dish for cultivating preimplantation mouse embryos. This dish minimizes the need for mineral oil and incorporates microwells, providing a stable culture environment and enabling independent monitoring of individual embryos. METHODS: Mouse pronuclear (PN) zygotes and two-cell-stage embryos were collected at 18 and 46 hours after human chorionic gonadotropin injection, respectively. These were cultured for 120 hours using potassium simplex optimized medium (KSOM) to reach the blastocyst stage. The embryos were randomly allocated into three groups, each cultured in one of three dishes: a 60-mm culture dish, a microdrop dish, and an OIVC dish that we developed. RESULTS: The OIVC dish effectively maintained the osmolarity of the KSOM culture medium over a 5-day period using only 2 mL of mineral oil. This contrasts with the significant osmolarity increase observed in the 60-mm culture dish. Additionally, the OIVC dish exhibited higher blastulation rates from two-cell embryos (100%) relative to the other dish types. Moreover, blastocysts derived from both PN zygotes and two-cell embryos in the OIVC dish group demonstrated significantly elevated mean cell numbers. CONCLUSION: Use of the OIVC dish markedly increased the number of cells in blastocysts derived from the in vitro culture of preimplantation mouse embryos. The capacity of this dish to maintain medium osmolarity with minimal mineral oil usage represents a breakthrough that may advance embryo culture techniques for various mammals, including human in vitro fertilization and embryo transfer programs.

Mechanical effects of canes on standing posture: beyond perceptual information.

BACKGROUND: Numerous studies showed that postural balance improves through light touch on a stable surface highlighting the importance of haptic information, seemingly downplaying the mechanical contributions of the support. The present study examined the mechanical effects of canes for assisting balance in healthy individuals challenged by standing on a beam. METHODS: Sixteen participants supported themselves with two canes, one in each hand, and applied minimal, preferred, or maximum force onto the canes. They positioned the canes in the frontal plane or in a tripod configuration. Statistical analysis used a linear mixed model to evaluate the effects on the center of pressure and the center of mass. RESULTS: The canes significantly reduced the variability of the center of pressure and the center of mass to the same level as when standing on the ground. Increasing the exerted force beyond the preferred level yielded no further benefits, although in the preferred force condition, participants exploited the altered mechanics by resting their arms on the canes. The tripod configuration allowed for larger variability of the center of pressure in the task-irrelevant anterior-posterior dimension. High forces had a destabilizing effect on the canes: the displacement of the hand on the cane handle increased with the force. CONCLUSIONS: Given this static instability, these results show that using canes can provide not only mechanical benefits but also challenges. From a control perspective, effort can be reduced by resting the arms on the canes and by channeling noise in the task-irrelevant dimensions. However, larger forces exerted onto the canes can also have destabilizing effects and the instability of the canes needs to be counteracted, possibly by arm and shoulder stiffness. Insights into the variety of mechanical effects is important for the design of canes and the instructions of how to use them.

Gait Entrainment to Torque Pulses From a Hip Exoskeleton Robot.

Robot-aided locomotor rehabilitation has proven challenging. To facilitate progress, it is important to first understand the neuro-mechanical dynamics and control of unimpaired human locomotion. Our previous studies found that human gait entrained to periodic torque pulses at the ankle when the pulse period was close to preferred stride duration. Moreover, synchronized gait exhibited a constant phase relation with the pulses so that the robot provided mechanical assistance. To test the generality of mechanical gait entrainment, this study characterized unimpaired human subjects' responses to periodic torque pulses during overground walking. The intervention was applied by a hip exoskeleton robot, Samsung GEMS-H. Gait entrainment was assessed based on the time-course of the phase at which torque pulses occurred within each stride. Experiments were conducted for two consecutive days to evaluate whether the second day elicited more entrainment. Whether entrainment was affected by the difference between pulse period and preferred stride duration was also assessed. Results indicated that the intervention evoked gait entrainment that occurred more often when the period of perturbation was closer to subjects' preferred stride duration, but the difference between consecutive days was insignificant. Entrainment was accompanied by convergence of pulse phase to a similar value across all conditions, where the robot maximized mechanical assistance. Clear evidence of motor adaptation indicated the potential of the intervention for rehabilitation. This study quantified important aspects of the nonlinear neuro-mechanical dynamics underlying unimpaired human walking, which will inform the development of effective approaches to robot-aided locomotor rehabilitation, exploiting natural dynamics in a minimally-encumbering way.

Role of mTORC1 activity during early retinal development and lamination in human-induced pluripotent stem cell-derived retinal organoids.

Retinal organoids derived from human-induced pluripotent stem cells (hiPSC) are powerful tools for studying retinal development as they model spatial and temporal differentiation of retinal cell types. Vertebrate retinal development involves a delicate and coordinated process of retinal progenitor cell (RPC) differentiation, and the mammalian target of rapamycin complex 1 (mTORC1) has been reported to play a significant role in this complex process. Herein, using hiPSC-derived retinal organoids, we identify the time-dependent role of mTORC1 in retinal development, specifically in retinal ganglion cell (RGC) differentiation and the retinal lamination process, during the early stages of retinal organoid (RO) development. mTORC1 activity in ROs was the highest at 40 days of differentiation. MHY1485-induced hyperactivation of mTORC1 during this period resulted in a significant increase in the overall size of ROs compared to the untreated controls and rapamycin-treated Ros; there was also a marked increase in proliferative activity within the inner and outer layers of ROs. Moreover, the MHY1485-treated ROs showed a significant increase in the number of ectopic RGCs in the outer layers (indicating disruption of retinal laminar structure), with robust expression of HuC/D-binding proteins in the inner layers. These results demonstrate that mTORC1 plays a critical role in the development of hiPSC-derived ROs, especially during the early stages of differentiation.

Reliability and Validity of the Subjective Cognitive Complaints Questionnaire for Parkinson's Disease (SCCQ-PD).

BACKGROUND AND PURPOSE: Subjective cognitive complaints (SCCs) are gaining attention as a self-perceived symptom for cognitive impairment in patients with Parkinson's disease (PD), but there are few suitable tools for assessing SCCs in PD. This study aimed to develop and validate a questionnaire for assessing SCCs in PD, called the Subjective Cognitive Complaints Questionnaire for Parkinson's Disease (SCCQ-PD). METHODS: The SCCQ-PD consists of 12 yes/no questions on subjective cognitive function, and the questionnaire was completed by patients with PD (score-P) and their caregivers (score-C). The cognitive function of patients was examined using comprehensive neuropsychological tests. RESULTS: This study included 73 patients (38 cognitively normal, 25 with mild cognitive impairment [MCI], and 10 demented) and their caregivers. Score-P and score-C had excellent reliability (Kuder-Richardson formula 20 coefficients of 0.893 and 0.931, respectively), and the scores exhibited a strong intercorrelation. Both score-P and score-C were negatively correlated with cognitive performance, and both were excellent in discriminating demented patients from those with normal cognition or MCI (areas under the receiver operating characteristic curve of 0.83 and 0.88, respectively). CONCLUSIONS: The SCCQ-PD is a reliable tool for assessing SCCs in patients with PD. SCCs measured using the SCCQ-PD are correlated with objective cognitive decline and useful for discriminating demented patients from nondemented patients.

Association between sleep disturbance and occupational injury among Korean employees.

BACKGROUND: Occupational injury has been a serious social problem steadily. Sleep disturbance is a risk factor for occupational injury. However, there were few researches studied on the linking between sleep disturbance and occupational injury in general working population of Korea. Therefore, we explored the association between sleep disturbance and occupational injury among Korean workers. METHODS: This study used data from the 5th Korean Working Conditions Survey. Occupational injury was assessed by asking work related injury for 12 months prior to the point of the survey. Sleep disturbance was assessed using the Minimal Insomnia Symptoms Scale (MISS). To analyze the association between sleep disturbance and occupational injury, multiple logistic regression analysis was conducted. RESULTS: The odds ratio (OR) of sleep disturbance group for occupational injury was 2.57 (95% confidence interval [CI]: 1.68-3.93) in the fully adjusted model. For the association between MISS score and occupational injury, it showed increasing trend that ORs increase from the 1st to the 4th quartiles. The OR of 2nd quartile was 1.10 (95% CI: 0.60-2.01), the OR of 3rd quartile was 2.27 (95% CI: 1.53-3.38) and the OR of 4th quartile was 2.80 (95% CI: 1.84-4.26). CONCLUSIONS: Sleep disturbance was associated with occupational injury. In addition, increasing trend was observed between MISS score and occupational injury. These findings imply that developing of intervention programs to manage sleep disturbance and fatigue may be necessary to prevent occupational injury.

Applying Hip Stiffness With an Exoskeleton to Compensate Gait Kinematics.

Neurological disorders and aging induce impaired gait kinematics. Despite recent advances, effective methods using lower-limb exoskeleton robots to restore gait kinematics are as yet limited. In this study, applying virtual stiffness using a hip exoskeleton was investigated as a possible method to guide users to change their gait kinematics. With a view to applications in locomotor rehabilitation, either to provide assistance or promote recovery, this study assessed whether imposed stiffness induced changes in the gait pattern during walking; and whether any changes persisted upon removal of the intervention, which would indicate changes in central neuro-motor control. Both positive and negative stiffness induced immediate and persistent changes of gait kinematics. However, the results showed little behavioral evidence of persistent changes in neuro-motor control, not even short-lived aftereffects. In addition, stride duration was little affected, suggesting that at least two dissociable layers exist in the neuro-motor control of human walking. The lack of neuro-motor adaptation suggests that, within broad limits, the central nervous system is surprisingly indifferent to the details of lower limb kinematics. The lack of neuro-motor adaptation also suggests that alternative methods may be required to implement a therapeutic technology to promote recovery. However, the immediate, significant, and reproducible changes in kinematics suggest that applying hip stiffness with an exoskeleton may be an effective assistive technology for compensation.

Frequency-dependent force direction elucidates neural control of balance.

BACKGROUND: Maintaining upright posture is an unstable task that requires sophisticated neuro-muscular control. Humans use foot-ground interaction forces, characterized by point of application, magnitude, and direction to manage body accelerations. When analyzing the directions of the ground reaction forces of standing humans in the frequency domain, previous work found a consistent pattern in different frequency bands. To test whether this frequency-dependent behavior provided a distinctive signature of neural control or was a necessary consequence of biomechanics, this study simulated quiet standing and compared the results with human subject data. METHODS: Aiming to develop the simplest competent and neuromechanically justifiable dynamic model that could account for the pattern observed across multiple subjects, we first explored the minimum number of degrees of freedom required for the model. Then, we applied a well-established optimal control method that was parameterized to maximize physiologically-relevant insight to stabilize the balancing model. RESULTS: If a standing human was modeled as a single inverted pendulum, no controller could reproduce the experimentally observed pattern. The simplest competent model that approximated a standing human was a double inverted pendulum with torque-actuated ankle and hip joints. A range of controller parameters could stabilize this model and reproduce the general trend observed in experimental data; this result seems to indicate a biomechanical constraint and not a consequence of control. However, details of the frequency-dependent pattern varied substantially across tested control parameter values. The set of parameters that best reproduced the human experimental results suggests that the control strategy employed by human subjects to maintain quiet standing was best described by minimal control effort with an emphasis on ankle torque. CONCLUSIONS: The findings suggest that the frequency-dependent pattern of ground reaction forces observed in quiet standing conveys quantitative information about human control strategies. This study's method might be extended to investigate human neural control strategies in different contexts of balance, such as with an assistive device or in neurologically impaired subjects.

Identifying human postural dynamics and control from unperturbed balance.

BACKGROUND: Upright standing requires control of an inherently unstable multi-joint human body within a small base of support, despite biological motor and / or sensory noise which challenge balance. Without applying perturbations, system identification methods have been regarded as inadequate, because the relevant internal biological noise processes are not accessible to direct measurement. As a result, unperturbed balance studies have been limited to investigation of behavioral patterns rather than possible underlying control strategies. METHODS: In this paper, we present a mathemathically rigorous system identification method that is applicable to study the dynamics and control of unperturbed balance. The method is derived from autocorrelation matrices with non-zero time lags and identifies the system matrix of a discrete-time dynamic system in the presence of unknown noise processes, without requiring any information about the strength of the noise. RESULTS: Unlike reasonable 'least-squares' approaches, the performance of the new method is consistent across a range of different combinations of internal and measurement noise strengths, even when measurement noise is substantial. We present a numerical example of a model that simulates human upright balancing and show that its dynamics can be identified accurately. With a biomechanically reasonable choice of state and input variables, a state feedback controller can also be identified. CONCLUSIONS: This study provides a new method to correctly identify the dynamics of human standing without the need for known external perturbations. The method was numerically validated using simulation that included realistic features of human balance. This method avoids potential issues of adaptation or possible reflex responses evoked by external perturbations, and does not require expensive in-lab, high-precision measurement equipment. It may eventually enable diagnosis and treatment of individuals with impaired balance, and the development of safe and effective assistive and / or rehabilitative technologies.

What's new on EEG monitoring in the ICU.

Continuous video-EEG (cEEG, lasting hours to several days) is increasingly used in ICU patients, as it is more sensitive than routine video-EEG (rEEG, lasting 20-30 min) to detect seizures or status epilepticus, and allows more frequent changes in therapeutic regimens. However, cEEG is more resource-consuming, and its relationship to outcome compared to repeated rEEG has only been formally assessed very recently in a randomized controlled trial, which did not show any significant difference in terms of long-term mortality or functional outcome. Awaiting more refined trials, it seems therefore that using repeated rEEG in ICU patients may represent a reasonable alternative in resource-limited settings. Prolonged EEG has been used recently in patients with severe COVID-19 infection, the proportion of seizures seems albeit relatively low, and similar to ICU patients with medical conditions. As a timely EEG recording is recommended in the ICU in any case, recent technical developments may ease its use in clinical practice.

Association between employment status and sickness presenteeism among Korean employees: a cross-sectional study.

BACKGROUND: Sickness presenteeism (SP) indicates "going to work while being ill." The importance of SP has only recently been investigated, and the association between SP and employment status has been inconsistent across studies. Therefore, we conducted this study to explore the association between SP and employment status by using presenteeism propensity (PP), which can reflect the individual decision-making process. METHODS: The study population included employees participating in the 5th Korean Working Condition Survey. We analyzed data of only employees with at least one health event, which was calculated as the sum of SP and sickness absenteeism days. Employment status was grouped into 3 categories: stable employment, unstable employment (contract period ≥ 1 year), and unstable employment (contract period < 1 year). Survey-weighted logistic regression analysis was conducted to assess the association between employment status and PP (dichotomized as "≤ 0.5" and "> 0.5"). RESULTS: Unstable employees (contract period ≥ 1 year) had higher odds of PP than stable employees (odds ratio [OR]: 1.23, 95% confidence interval [CI]: 1.03-1.47), whereas unstable employees (contract period < 1 year) had lower odds of PP than stable employees (OR: 0.82, 95% CI: 0.71-0.96). CONCLUSIONS: Employment status was associated with SP. Given the negative health impact of SP, social efforts, such as paid sick leave, are required to reduce SP and enhance the health status of unstable workers.

High-Speed 3D Printing of Millimeter-Size Customized Aspheric Imaging Lenses with Sub 7 nm Surface Roughness.

Advancements in three-dimensional (3D) printing technology have the potential to transform the manufacture of customized optical elements, which today relies heavily on time-consuming and costly polishing and grinding processes. However the inherent speed-accuracy trade-off seriously constrains the practical applications of 3D-printing technology in the optical realm. In addressing this issue, here, a new method featuring a significantly faster fabrication speed, at 24.54 mm3 h-1 , without compromising the fabrication accuracy required to 3D-print customized optical components is reported. A high-speed 3D-printing process with subvoxel-scale precision (sub 5 µm) and deep subwavelength (sub 7 nm) surface roughness by employing the projection micro-stereolithography process and the synergistic effects from grayscale photopolymerization and the meniscus equilibrium post-curing methods is demonstrated. Fabricating a customized aspheric lens 5 mm in height and 3 mm in diameter is accomplished in four hours. The 3D-printed singlet aspheric lens demonstrates a maximal imaging resolution of 373.2 lp mm-1 with low field distortion less than 0.13% across a 2 mm field of view. This lens is attached onto a cell phone camera and the colorful fine details of a sunset moth's wing and the spot on a weevil's elytra are captured. This work demonstrates the potential of this method to rapidly prototype optical components or systems based on 3D printing.

Postresection Outcomes of Combined Hepatocellular Carcinoma-Cholangiocarcinoma, Hepatocellular Carcinoma and Intrahepatic Cholangiocarcinoma.

INTRODUCTION: Combined hepatocellular carcinoma and cholangiocarcinoma (cHCC-CC) is rare. This study investigated the clinicopathological features of cHCC-CC and compared the postresection survival outcomes of cHCC-CC, hepatocellular carcinoma (HCC), and intrahepatic cholangiocarcinoma (IHC). METHODS: Between January 2000 and September 2012, 53 patients with cHCC-CC underwent tumor resection, accounting for 1.1 % of surgeries for primary liver malignancies. Control groups included patients with HCC (n = 1452) and IHC (n = 149) who underwent R0 resection of stage I/II tumors ≤5 cm. RESULTS: Mean tumor diameter of cHCC-CC group was 5.5 ± 2.9 cm, and single tumor was identified in 50. Pathological classification included combined (n = 41), mixed (n = 11), and double (n = 1) tumors. The 1-, 3-, 5-, and 10-year tumor recurrence rates were 60.8, 71.8, 80.7, and 80.7 %, respectively. The 1-, 3-, 5-, and 10-year overall survival rates were 73.3, 35.6, 30.5, and 11.1 %, respectively. Tumor recurrence and patient survival did not differ significantly according to AJCC tumor staging and histological type (all p ≥ 0.2). Tumor recurrence rates did not differ significantly between the cHCC-CC, HCC, and IHC groups (p = 0.43), whereas differences in survival rates were significant (p = 0.000), with a median survival after tumor recurrence of 8, 51, and 6 months, respectively (p = 0.000). CONCLUSIONS: Patients with cHCC-CC showed similar recurrence rates to those of control patients with HCC and IHC, whereas their survival outcomes were worse than those of control HCC patients because of poor responses to recurrence treatment. Further evaluation of differences in tumor characteristics and tumor biology is necessary to accurately predict the prognosis of patients with cHCC-CC.

Negative regulation of the novel norpA(P24) suppressor, diehard4, in the endo-lysosomal trafficking underlies photoreceptor cell degeneration.

Rhodopsin has been used as a prototype system to investigate G protein-coupled receptor (GPCR) internalization and endocytic sorting mechanisms. Failure of rhodopsin recycling upon light activation results in various degenerative retinal diseases. Accumulation of internalized rhodopsin in late endosomes and the impairment of its lysosomal degradation are associated with unregulated cell death that occurs in dystrophies. However, the molecular basis of rhodopsin accumulation remains elusive. We found that the novel norpA(P24) suppressor, diehard4, is responsible for the inability of endo-lysosomal rhodopsin trafficking and retinal degeneration in Drosophila models of retinal dystrophies. We found that diehard4 encodes Osiris 21. Loss of its function suppresses retinal degeneration in norpA(P24), rdgC(306), and trp(1), but not in rdgB(2), suggesting a common cause of photoreceptor death. In addition, the loss of Osiris 21 function shifts the membrane balance between late endosomes and lysosomes as evidenced by smaller late endosomes and the proliferation of lysosomal compartments, thus facilitating the degradation of endocytosed rhodopsin. Our results demonstrate the existence of negative regulation in vesicular traffic between endosomes and lysosomes. We anticipate that the identification of additional components and an in-depth description of this specific molecular machinery will aid in therapeutic interventions of various retinal dystrophies and GPCR-related human diseases.

Multifunctional epidermal electronics printed directly onto the skin.

Materials and designs are presented for electronics and sensors that can be conformally and robustly integrated onto the surface of the skin. A multifunctional device of this type can record various physiological signals relevant to health and wellness. This class of technology offers capabilities in biocompatible, non-invasive measurement that lie beyond those available with conventional, point-contact electrode interfaces to the skin.

Role of type II protein arginine methyltransferase 5 in the regulation of Circadian Per1 gene.

Circadian clocks are the endogenous oscillators that regulate rhythmic physiological and behavioral changes to correspond to daily light-dark cycles. Molecular dissections have revealed that transcriptional feedback loops of the circadian clock genes drive the molecular oscillation, in which PER/CRY complexes inhibit the transcriptional activity of the CLOCK/BMAL1 heterodimer to constitute a negative feedback loop. In this study, we identified the type II protein arginine methyltransferase 5 (PRMT5) as an interacting molecule of CRY1. Although the Prmt5 gene was constitutively expressed, increased interaction of PRMT5 with CRY1 was observed when the Per1 gene was repressed both in synchronized mouse liver and NIH3T3 cells. Moreover, rhythmic recruitment of PRMT5 and CRY1 to the Per1 gene promoter was found to be associated with an increased level of histone H4R3 dimethylation and Per1 gene repression. Consistently, decreased histone H4R3 dimethylation and altered rhythmic Per1 gene expression were observed in Prmt5-depleted cells. Taken together, these findings provide an insight into the link between histone arginine methylation by PRMT5 and transcriptional regulation of the circadian Per1 gene.

Drosophila arf72A acts as an essential regulator of endoplasmic reticulum quality control and suppresses autosomal-dominant retinopathy.

The eukaryotic endoplasmic reticulum operates multiple quality control mechanisms to ensure that only properly folded proteins are exported to their final destinations via the secretory pathway and those that are not are destroyed via the degradation pathway. However, molecular mechanisms underlying such regulated exportation to these distinct routes are unknown. In this article, we report the role of Drosophila arf72A--the fly homologue of the mammalian Arl1 - in the quality checks of proteins and in the autosomal-dominant retinopathy. ARF72A localizes to the Golgi membranes of Drosophila photoreceptor cells, consistent with mammalian Arl1 localization in cell culture systems. A loss of arf72A function changes the membrane character of the endoplasmic reticulum and shifts the membrane balance between the endoplasmic reticulum and the Golgi complex toward the Golgi complex, resulting in over-proliferated Golgi complexes and accelerated protein secretion. Interestingly, our study indicated that more ARF72A localized on the endoplasmic reticulum in the ninaE(D1) photoreceptor cell, a Drosophila model of autosomal-dominant retinitis pigmentosa, compared to that in the wild-type. In addition, arf72A loss was shown to rescue the ninaE(D1)-related membrane accumulation and the rhodopsin maturation defect, and suppress ninaE(D1)-triggered retinal degeneration, indicating that rhodopsin accumulated in the endoplasmic reticulum bypasses the quality checks. While previous studies of ARF small GTPases have focused on their roles in vesicular budding and transport between the specific organelles, our findings establish an additional function of arf72A in the quality check machinery of the endoplasmic reticulum distinguishing the cargoes for secretion from those for degradation.

Spatiotemporal regulation of fibroblast growth factor signal blocking for endoderm formation in Xenopus laevis.

We have previously shown that the inhibition of fibroblast growth factor (FGF) signaling induced endodermal gene expression in the animal cap and caused the expansion of the endodermal mass in Xenopus embryos. However, we still do not know whether or not the alteration of FGF signaling controls embryonic cell fate, or when FGF signal blocking is required for endoderm formation in Xenopus. Here, we show that FGF signal blocking in embryonic cells causes their descendants to move into the endodermal region and to express endodermal genes. It is also interesting that blocking FGF signaling between fertilization and embryonic stage 10.5 promotes endoderm formation, but persistent FGF signaling blocking after stage 10.5 restricts endoderm formation and differentiation.

The cis-regulatory logic of the mammalian photoreceptor transcriptional network.

The photoreceptor cells of the retina are subject to a greater number of genetic diseases than any other cell type in the human body. The majority of more than 120 cloned human blindness genes are highly expressed in photoreceptors. In order to establish an integrative framework in which to understand these diseases, we have undertaken an experimental and computational analysis of the network controlled by the mammalian photoreceptor transcription factors, Crx, Nrl, and Nr2e3. Using microarray and in situ hybridization datasets we have produced a model of this network which contains over 600 genes, including numerous retinal disease loci as well as previously uncharacterized photoreceptor transcription factors. To elucidate the connectivity of this network, we devised a computational algorithm to identify the photoreceptor-specific cis-regulatory elements (CREs) mediating the interactions between these transcription factors and their target genes. In vivo validation of our computational predictions resulted in the discovery of 19 novel photoreceptor-specific CREs near retinal disease genes. Examination of these CREs permitted the definition of a simple cis-regulatory grammar rule associated with high-level expression. To test the generality of this rule, we used an expanded form of it as a selection filter to evolve photoreceptor CREs from random DNA sequences in silico. When fused to fluorescent reporters, these evolved CREs drove strong, photoreceptor-specific expression in vivo. This study represents the first systematic identification and in vivo validation of CREs in a mammalian neuronal cell type and lays the groundwork for a systems biology of photoreceptor transcriptional regulation.