Molecular targeting of protein arginine deiminases to suppress colitis and prevent colon cancer

Ulcerative colitis (UC) is a chronic disease, in which the lining of the colon becomes inflamed and develops ulcers leading to abdominal pain, diarrhea, and rectal bleeding. The extent of these symptoms depends on disease severity. The protein arginine deiminase (PAD) family of enzymes converts peptidyl-Arginine to peptidyl-Citrulline through citrullination. PADs are dysregulated, with abnormal citrullination in many diseases, including UC and colorectal cancer (CRC). We have developed the small molecule, pan-PAD inhibitor, Chlor-amidine (Cl-amidine), with multiple goals, including treating UC and preventing CRC. Building off our recent results showing that: 1) Cl-amidine suppresses colitis in vivo in a dextran sulfate sodium (DSS) mouse model; and 2) Cl-amidine induces microRNA (miR)-16 in vitro causing cell cycle arrest, we tested the hypothesis that Cl-amidine can prevent tumorigenesis and that miR-16 induction, by Cl-amidine, may be involved in vivo. Consistent with our hypothesis, we present evidence that Cl-amidine, delivered in the drinking water, prevents colon tumorigenesis in our mouse model of colitis-associated CRC where mice are given carcinogenic azoxymethane (AOM), followed by multiple cycles of 2% DSS to induce colitis. To begin identifying mechanisms, we examined the effects of Cl-amidine on miR-16. Results show miR-16 suppression during the colitis-to-cancer sequence in colon epithelial cells, which was rescued by drinking Cl-amidine. Likewise, Ki67 and cellular proliferation targets of miR-16 (Cyclins D1 and E1) were suppressed by Cl-amidine. The decrease in cell proliferation markers and increase in tumor suppressor miRNA expression potentially define a mechanism of how Cl-amidine is suppressing tumorigenesis in vivo

The effect of relationship status on communicating emotions through touch

Research into emotional communication to date has largely focused on facial and vocal expressions. In contrast, recent studies by Hertenstein, Keltner, App, Bulleit, and Jaskolka (2006) and Hertenstein, Holmes, McCullough, and Keltner (2009) exploring nonverbal communication of emotion discovered that people could identify anger, disgust, fear, gratitude, happiness, love, sadness and sympathy from the experience of being touched on either the arm or body by a stranger, without seeing the touch. The study showed that strangers were unable to communicate the self-focused emotions embarrassment, envy and pride, or the universal emotion surprise. Literature relating to touch indicates that the interpretation of a tactile experience is significantly influenced by the relationship between the touchers (Coan, Schaefer, & Davidson, 2006). The present study compared the ability of romantic couples and strangers to communicate emotions solely via touch. Results showed that both strangers and romantic couples were able to communicate universal and prosocial emotions, whereas only romantic couples were able to communicate the self-focused emotions envy and pride

Comparison of Body Composition Measurements using a New Caliper, Two Established Calipers, Hydrostatic Weighing, and BodPod

Purposes: (1) To compare the Lafayette Instruments (LI) skinfold caliper to the Lange (L) and Harpenden (H) calipers using a diverse subject population. (2) To determine the validity of the LI caliper in a subset of subjects by comparing body compositions from skinfold thicknesses to those measured by hydrostatic weighing (HW) and air displacement plethysmography (ADP). (3) To compare measurements obtained by experienced (EX) and inexperienced (IX) technicians using all three calipers. Methods: Skinfold measurements were performed by both EX and IX technicians using three different calipers on 21 younger (21.2 ± 1.5 yrs) and 20 older (59.2 ± 4 yrs) subjects. Body compositions were calculated using the Jackson-Pollock seven-site and three-site formulas. HW and ADP tests were performed on a subset of subjects (10 younger, 10 older). Results: No significant differences existed between LI and L or H when measurements were made by EX. Further, the LI-EX measurements were highly correlated to both H-EX and L-EX. No significant differences existed in the subgroup between LI-EX and HW or ADP. Skinfold determinations made by EX and IX were similar. Conclusions: Similar body compositions determined using LI, H, and L suggest that LI determines body composition as effectively as H and L. High correlations between the three calipers support this notion. Similar results between LI and HW/ADP subgroup suggest that the LI caliper may be a valid method of measuring body composition. Overall, performance by IX was similar to EX and suggests similar ease of use for all three calipers

The induction of microRNA-16 in colon cancer cells by protein arginine deiminase inhibition causes a p53-dependent cell cycle arrest.

Protein Arginine Deiminases (PADs) catalyze the post-translational conversion of peptidyl-Arginine to peptidyl-Citrulline in a calcium-dependent, irreversible reaction. Evidence is emerging that PADs play a role in carcinogenesis. To determine the cancer-associated functional implications of PADs, we designed a small molecule PAD inhibitor (called Chor-amidine or Cl-amidine), and tested the impact of this drug on the cell cycle. Data derived from experiments in colon cancer cells indicate that Cl-amidine causes a G1 arrest, and that this was p53-dependent. In a separate set of experiments, we found that Cl-amidine caused a significant increase in microRNA-16 (miRNA-16), and that this increase was also p53-dependent. Because miRNA-16 is a putative tumor suppressor miRNA, and others have found that miRNA-16 suppresses proliferation, we hypothesized that the p53-dependent G1 arrest associated with PAD inhibition was, in turn, dependent on miRNA-16 expression. Results are consistent with this hypothesis. As well, we found the G1 arrest is at least in part due to the ability of Cl-amidine-mediated expression of miRNA-16 to suppress its\u27 G1-associated targets: cyclins D1, D2, D3, E1, and cdk6. Our study sheds light into the mechanisms by which PAD inhibition can protect against or treat colon cancer

Comparing serial X-ray crystallography and microcrystal electron diffraction (MicroED) as methods for routine structure determination from small macromolecular crystals.

Innovative new crystallographic methods are facilitating structural studies from ever smaller crystals of biological macromolecules. In particular, serial X-ray crystallography and microcrystal electron diffraction (MicroED) have emerged as useful methods for obtaining structural information from crystals on the nanometre to micrometre scale. Despite the utility of these methods, their implementation can often be difficult, as they present many challenges that are not encountered in traditional macromolecular crystallography experiments. Here, XFEL serial crystallography experiments and MicroED experiments using batch-grown microcrystals of the enzyme cyclophilin A are described. The results provide a roadmap for researchers hoping to design macromolecular microcrystallography experiments, and they highlight the strengths and weaknesses of the two methods. Specifically, we focus on how the different physical conditions imposed by the sample-preparation and delivery methods required for each type of experiment affect the crystal structure of the enzyme

Comparison of Methods for Modeling Fractional Cover Using Simulated Satellite Hyperspectral Imager Spectra

Remotely sensed data can be used to model the fractional cover of green vegetation (GV), non-photosynthetic vegetation (NPV), and soil in natural and agricultural ecosystems. NPV and soil cover are difficult to estimate accurately since absorption by lignin, cellulose, and other organic molecules cannot be resolved by broadband multispectral data. A new generation of satellite hyperspectral imagers will provide contiguous narrowband coverage, enabling new, more accurate, and potentially global fractional cover products. We used six field spectroscopy datasets collected in prior experiments from sites with partial crop, grass, shrub, and low-stature resprouting tree cover to simulate satellite hyperspectral data, including sensor noise and atmospheric correction artifacts. The combined dataset was used to compare hyperspectral index-based and spectroscopic methods for estimating GV, NPV, and soil fractional cover. GV fractional cover was estimated most accurately. NPV and soil fractions were more difficult to estimate, with spectroscopic methods like partial least squares (PLS) regression, spectral feature analysis (SFA), and multiple endmember spectral mixture analysis (MESMA) typically outperforming hyperspectral indices. Using an independent validation dataset, the lowest root mean squared error (RMSE) values were 0.115 for GV using either normalized difference vegetation index (NDVI) or SFA, 0.164 for NPV using PLS, and 0.126 for soil using PLS. PLS also had the lowest RMSE averaged across all three cover types. This work highlights the need for more extensive and diverse fine spatial scale measurements of fractional cover, to improve methodologies for estimating cover in preparation for future hyperspectral global monitoring missions

Selection for Forage and Avoidance of Risk by Woodland Caribou (Rangifer Tarandus Caribou) at Coarse andLocal Scales

The relationship between selection at coarse and fine spatiotemporal spatial scales is still poorly understood. Some authors claim that, to accommodate different needs at different scales, individuals should have contrasting selection patterns at different scales of selection, while others claim that coarse scale selection patterns should reflect fine scale selection decisions. Here we examine site selection by 110 woodland caribou equipped with GPS radio‐collars with respect to forage availability and predation risk across a broad gradient in availability of both variables in boreal forests of Northern Ontario. We tested whether caribou selection for forage and avoidance of risk was consistent between coarse (seasonal home range) and fine scales of selection. We found that local selection patterns predicted coarse scale selection patterns, indicating a close relationship between the drivers of selection at both spatial scales

Near-Infrared Monitoring of Volatiles in Frozen Lunar Simulants While Drilling

In Situ Resource Utilization (ISRU) focuses on using local resources for mission consumables. The approach can reduce mission cost and risk. Lunar polar volatiles, e.g. water ice, have been detected via remote sensing measurements and represent a potential resource for both humans and propellant. The exact nature of the horizontal and depth distribution of the ice remains to be documented in situ. NASA's Resource Prospector mission (RP) is intended to investigate the polar volatiles using a rover, drill, and the RESOLVE science package. RP component level hardware is undergoing testing in relevant lunar conditions (cryovacuum). In March 2015 a series of drilling tests were undertaken using the Honeybee Robotics RP Drill, Near-Infrared Volatile Spectrometer System (NIRVSS), and sample capture mechanisms (SCM) inside a 'dirty' thermal vacuum chamber at the NASA Glenn Research Center. The goal of these tests was to investigate the ability of NIRVSS to monitor volatiles during drilling activities and assess delivery of soil sample transfer to the SCMs in order to elucidate the concept of operations associated with this regolith sampling method

Neutron Scattering Cross Section Measurements for \u3csup\u3e56\u3c/sup\u3eFe

Elastic and inelastic differential cross sections for neutron scattering from 56Fe have been measured for several incident energies from 1.30 to 7.96 MeV at the University of Kentucky Accelerator Laboratory. Scattered neutrons were detected using a C6D6 liquid scintillation detector using pulse-shape discrimination and time-of-flight techniques. The deduced cross sections have been compared with previously reported data, predictions from evaluation databases ENDF, JENDL, and JEFF, and theoretical calculations performed using different optical model potentials using the TALYS and EMPIRE nuclear reaction codes. The coupled-channel calculations based on the vibrational and soft-rotor models are found to describe the experimental (n,n0) and (n,n1) cross sections well