Engineering Improved Photoswitches for the Control of Nucleocytoplasmic Distribution

Optogenetic techniques use light-responsive proteins to study dynamic processes in living cells and organisms. These techniques typically rely on repurposed naturally occurring light-sensitive proteins to control subcellular localization and activity. We previously engineered two optogenetic systems, the light activated nuclear shuttle (LANS) and the light-inducible nuclear exporter (LINX), by embedding nuclear import or export sequence motifs into the C-terminal helix of the light-responsive LOV2 domain of Avena sativa phototropin 1, thus enabling light-dependent trafficking of a target protein into and out of the nucleus. While LANS and LINX are effective tools, we posited that mutations within the LOV2 hinge-loop, which connects the core PAS domain and the C-terminal helix, would further improve the functionality of these switches. Here, we identify hinge-loop mutations that favorably shift the dynamic range (the ratio of the on- to off-target subcellular accumulation) of the LANS and LINX photoswitches. We demonstrate the utility of these new optogenetic tools to control gene transcription and epigenetic modifications, thereby expanding the optogenetic "tool kit" for the research community

Multisensor data fusion algorithm development

This report presents a two-year LDRD research effort into multisensor data fusion. We approached the problem by addressing the available types of data, preprocessing that data, and developing fusion algorithms using that data. The report reflects these three distinct areas. First, the possible data sets for fusion are identified. Second, automated registration techniques for imagery data are analyzed. Third, two fusion techniques are presented. The first fusion algorithm is based on the two-dimensional discrete wavelet transform. Using test images, the wavelet algorithm is compared against intensity modulation and intensity-hue-saturation image fusion algorithms that are available in commercial software. The wavelet approach outperforms the other two fusion techniques by preserving spectral/spatial information more precisely. The wavelet fusion algorithm was also applied to Landsat Thematic Mapper and SPOT panchromatic imagery data. The second algorithm is based on a linear-regression technique. We analyzed the technique using the same Landsat and SPOT data

Human Cord Blood Stem Cell-Modulated Regulatory T Lymphocytes Reverse the Autoimmune-Caused Type 1 Diabetes in Nonobese Diabetic (NOD) Mice

Background: The deficit of pancreatic islet b cells caused by autoimmune destruction is a crucial issue in type 1 diabetes (T1D). It is essential to fundamentally control the autoimmunity for treatment of T1D. Regulatory T cells (Tregs) play a pivotal role in maintaining self-tolerance through their inhibitory impact on autoreactive effector T cells. An abnormality of Tregs is associated with initiation of progression of T1D. Methodology/Principal Findings: Here, we report that treatment of established autoimmune-caused diabetes in NOD mice with purified autologous CD4 + CD62L + Tregs co-cultured with human cord blood stem cells (CB-SC) can eliminate hyperglycemia, promote islet b-cell regeneration to increase b-cell mass and insulin production, and reconstitute islet architecture. Correspondingly, treatment with CB-SC-modulated CD4 + CD62L + Tregs (mCD4CD62L Tregs) resulted in a marked reduction of insulitis, restored Th1/Th2 cytokine balance in blood, and induced apoptosis of infiltrated leukocytes in pancreatic islets. Conclusions/Significance: These data demonstrate that treatment with mCD4CD62L Tregs can reverse overt diabetes

Acute Cardiovascular and Metabolic Responses to Three Modes of Treadmill Exercise in Older Adults with Parkinson’s Disease

Parkinson’s disease (PD) is a neurodegenerative condition characterized by muscle tremors, rigidity and dyskinesis leading to balance and gait abnormalities that could alter physiologic responses during exercise. Locomotion on an aquatic treadmill (ATM) or anti-gravity treadmill (AGTM) may be a safe alternative to exercise on a traditional land treadmill (LTM) in those with PD. PUPROSE: To determine the acute cardiovascular and metabolic responses to three different modes of treadmill exercise in older adults diagnosed with Parkinson’s disease. METHODS: Eight adults diagnosed with PD (68 ± 3 years of age) completed one exercise session on an LTM, one session on an ATM, and one session on an AGTM at 50% body weight. Participants walked from 1 to 3 mph in 0.5 mph increments at 0% grade during each exercise session. Heart rate (HR), energy expenditure (EE), systolic blood pressure (SBP), and diastolic blood pressure (DBP) were measured at rest and during steady-state exercise at each speed on each treadmill. Rate of perceived exertion was also measured during steady-state exercise. Rate pressure product (RPP) was calculated. RESULTS: All variables, with the exception of DBP, increased as speed increased across all treadmill modes (p \u3c 0.001). Between treadmill modes across all speeds, EE was statistically different (p = 0.025). There was a significant interaction effect for mode and speed for HR (p \u3c 0.001) and RPP (p = 0.003). At all speeds except 1.5 mph, HR was higher on the LTM versus the AGTM (p \u3c 0.05). CONCLUSION: Exercising on an ATM or an AGTM elicits similar physiologic responses to exercise on an LTM in adults with P

Changes in Balance, Gait and Motor Skills Following Treadmill Exercise in Adults with Parkinson’s Disease

Locomotion on an aquatic treadmill or anti-gravity treadmill may be a safe and effective alternative to exercise on a traditional land treadmill in those with Parkinson’s disease as the removal of body weight in these environments may allow the participant to exercise with less concern of falling at higher speeds before reaching volitional fatigue. PURPOSE: To determine the training effects of three different treadmill modalities on dynamic balance, gait, and fine motor control in older adults diagnosed with Parkinson’s disease. METHODS: Ten adults diagnosed with Parkinson’s disease (70 ± 5 years of age) completed 8 exercise sessions (4 weeks, 2x/week) each separately on a land treadmill, aquatic treadmill, and anti-gravity treadmill at 50% body weight. Two weeks separated each intervention and the order was randomized. A 4-week control period occurred at the start of the study in which no treadmill exercise was performed. Each exercise session included a 2-minute warm-up and 30 minutes at a moderate intensity. Before and after each intervention, balance, gait and fine motor control were measured. Dynamic balance and gait were assessed using a Timed-Up-and-Go test and Performance Oriented Mobility Assessment (POMA). Fine motor control was assessed with the Purdue Pegboard Test. RESULTS: The gait assessment of the POMA was significant across all time points (p = 0.028). All other variables were statistically similar (p \u3e 0.05) across all time points. CONCLUSION: Exercising on a traditional land treadmill, aquatic treadmill, or anti-gravity treadmill for 60 min/week for 4 weeks at a moderate intensity did not alter balance, gait or fine motor control in adults with Parkinson’s disease