Miracle-dz/UniFAR-FTUs huggingface dataset:
SEC-MALLS ANALYSIS OF HYALURONAN SIZE DISTRIBUTIONS MADE BY MEMBRANE-BOUND HYALURONAN SYNTHASE

SEC-MALLS analyses of E. coli membranes expressing Streptococcus equisimilis hyaluronan synthase (seHAS) demonstrated an inherent artifact (10–100 MDa) that co-eluted with HA, and skewed the apparent weight-average mass of HA to erroneously high values. Briefly heating samples to 65–75°C eliminated this artifact and increased the yield of recovered HA, due to the release of HA chains that were attached to membrane-bound HAS. Inclusion of alkaline phosphatase, which removed UDP produced during the reaction, improved the linearity of HA synthesis - even at high substrate utilization. Surprisingly, addition of EDTA, to chelate Mg +2 ions, did not completely stop the HAS reaction at 30°C or at 4°C. The best conditions for stopping the reaction without altering SEC-MALLS profiles of the product HA were to chill samples on ice in the presence of both EDTA and UDP. Even with excess substrate, the maximum size of product HA decreased as the enzyme concentration increased. Therefore, the maximum HA size made by HAS was determined by extrapolation to zero enzyme concentration. Using the above conditions, membrane-bound seHAS synthesized a cohort of HA products that steadily increased in weight-average molar mass, reaching a final maximal steady-state size of 4–6 MDa within 2–4 hours. by UDP and EDTA to final concentrations of 10 mM and 40 mM, respectively, chilling on ice, and then briefly to inactivate the phosphatase. (iii) The stored samples are heated to 65–75 °C just prior to injection for SEC-MALLS analysis. (iv) The time-dependence for the increase in average HA size is determined to ensure that a steady-state size distribution is obtained. (v) The kinetics of HA product size increase is determined for 3–4 different concentrations of HAS-containing membranes in the presence of sufficient alkaline phosphatase to maintain linearity of HA synthesis. (vi) Finally, the M w (or M n ) values obtained from SEC-MALLS analyses of the 3– 4 samples are plotted versus membrane protein (HAS content does not need to be known), and the zero-intercept then calculated to determine the maximum M w of HA products that can be made by the HAS or HAS variants being studied. A key point is that in an analysis of HA size, investigators need to consider and separately assess both rate of HA synthesis i.e. total amount of sugar incorporated into HA and rate HA size
Differential Contribution of Kainate Receptors to EPSCs in Superficial Layer Neurons of the Rat Medial Entorhinal Cortex

Although in situ hybridization studies have revealed the presence of kainate receptor (KAR) mRNA in neurons of the rat medial entorhinal cortex (mEC), the functional presence and roles of these receptors are only beginning to be examined. To address this deficiency, whole cell voltage clamp recordings of locally evoked EPSCs were made from mEC layer II and III neurons in combined entorhinal cortex -hippocampal brain slices. Three types of neurons were identified by their electroresponsive membrane properties, locations, and morphologies: stellate-like “Sag” neurons in layer II (S), pyramidal-like “No Sag” neurons in layer III (NS), and “Intermediate Sag” neurons with varied morphologies and locations (IS). Non-NMDA EPSCs in these neurons were composed of two components, and the slow decay component in NS neurons had larger amplitudes and contributed more to the combined EPSC than did those observed in S and IS neurons. This slow component was mediated by KARs and was characterized by its resistance to either GYKI 52466 (100 μ M) or NBQX (1 μ M), relatively slow decay kinetics, and sensitivity to CNQX (10–50 μ M). KAR mediated EPSCs in pyramidal-like NS neurons contributed significantly more to the combined non-NMDA EPSC than did those from S and IS neurons. Layer III neurons of the mEC are selectively susceptible to degeneration in human temporal lobe epilepsy (TLE) and animal models of TLE such as kainate-induced status epilepticus. Characterizing differences in the complement of postsynaptic receptors expressed in injury prone versus injury resistant mEC neurons represents an important step toward understanding the vulnerability of layer III neurons seen in TLE. and and NS neurons after the (pA) IS, and NS neurons, between S and IS neurons p S and NS neurons p , the relative amplitude of the EPSC at as a of the and NS are and On a neuron-by-neuron the of the EPSC (N=24) peak for IS, and as as IS and
Phosphorylated MAPK/ERK1/2 may not always represent its kinase activity in a rat model of focal cerebral ischemia with or without ischemic preconditioning

The ERK 1/2 protein require a dual phosphorylation at conserved threonine and tyrosine residues to be fully activated under normal physiological conditions. Thus, ERK1/2 kinase activity is often defined by the quantity of phosphorylated kinase. However, this may not accurately represent its true activity under certain pathological conditions. We investigated whether ERK1/2 kinase activity is proportional to its phosphorylation state in a rat focal ischemia model with and without rapid ischemic preconditioning. We showed that phosphorylated-ERK1/2 protein levels were increased 2.6±0.07 fold, and ERK1/2 kinase activity was increased 10.6±1.9 fold in animals receiving ischemic preconditioning alone without test ischemia compared with sham group ( P <0.05, n=6/group), suggesting that phosphorylated-ERK1/2 protein levels represent its kinase activity under these conditions. However, preconditioning plus test ischemia robustly blocked ERK1/2 kinase activity, while it increased phosphorylated-ERK1/2 protein levels beyond those receiving test ischemia alone, suggesting that phosphorylated-ERK1/2 protein levels were not representative of actual kinase activity in this pathological condition. In conclusion, protein phosphorylation levels of ERK1/2 do not always correspond to kinase activity, thus, measuring the true kinase activity is essential. and test ischemia plus RIPC. Our results were reliable and reproducible in each experimental setting and elucidated valuable information regarding the relationship between ERK1/2 phosphorylation and kinase activity.
The Development and Validation of the Memory Support Rating Scale (MSRS)

Patient memory for treatment information is poor, and worse memory for treatment information is associated with poorer clinical outcomes. Memory support techniques have been harnessed to improve patient memory for treatment. However, a measure of memory support used by treatment providers during sessions has yet to be established. The present study reports on the development and psychometric properties of the Memory Support Rating Scale (MSRS) – an observer-rated scale designed to measure memory support. Forty-two adults with major depressive disorder (MDD) were randomized to either cognitive therapy plus memory support (CS+MS; n = 22) or cognitive therapy as-usual (CT-as-usual; n = 20). At post-treatment, patients freely recalled treatment points via the Patient Recall Task. Sessions (n = 171) were coded for memory support using the MSRS, 65% of which were also assessed for the quality of cognitive therapy via the Cognitive Therapy Rating Scale (CTRS). A unidimensional scale composed of 8 items was developed using exploratory factor analysis, though a larger sample is needed to further assess the factor structure of MSRS scores. High inter-rater and test-retest reliabilities of MSRS scores were observed across seven MSRS coders. MSRS scores were higher in the CT+MS condition compared to CT-as-usual, demonstrating group differentiation ability. MSRS scores were positively associated with Patient Recall Task scores but not associated with CTRS scores, demonstrating convergent and discriminant validity, respectively. Results indicate that the MSRS yields reliable and valid scores for measuring treatment providers’ use of memory support while delivering cognitive therapy.
Highly stretchable and transparent dielectric gels for high sensitivity tactile sensors

Soft tactile sensors have been widely used in wearable electronics and soft robotics. Among different sensing technologies, soft capacitive sensors receive much attention due to good signal repeatability, temperature insensitivity, relative simplicity of fabrication and adaptability of configuration. Many methods such as adding nano-fillers have been developed to increase the sensitivity of capacitive sensors. However, the existing methods usually sacrifice the transparency and reduces the flexibility. In this work, we propose to use dielectric gels to fabricate capacitive sensors for the first time. We synthesize dielectric gels by using ethylene carbonate and propylene carbonate as solvents and 2-Ethylhexyl acrylate and acryloyl morpholine as polymer networks. The synthesized dielectric gels are extremely soft, highly stretchable, and fully transparent to visible lights. The relative permittivity is as high as 30. We demonstrate that the sensitivity of the capacitive sensor made of the new dielectric gel increases about 6 times compared to the sensors made of VHB, PDMS, or Ecoflex. We design a touch sensor with the dielectric gel to control the LED, and also demonstrate the use of the dielectric gels as the transparent cover layer of a cell phone which may displace the traditional protective layer made of glass in future wearable electronics.
Animal Model Transforming Growth Factor- b 1 Fails to Stimulate Wound Healing and Impairs Its Signal Transduction in an Aged Ischemic Ulcer Model Importance of Oxygen and Age

Clinical trials of exogenous growth factors in treating chronic wounds have been less successful than ex-pected. One possible explanation is that most studies used animal models of acute wounds in young animals, whereas most chronic wounds occur in elderly patients with tissue ischemia. We described an animal model of age- and ischemia-impaired wound healing and analyzed the wound-healing response as well as the transforming growth factor (TGF)- b 1 effect in this model. Rabbits of increasing ages were made ischemic in the ear where dermal ulcers were created. Histological analysis showed that epithelium in-growth and granulation tissue deposition were signif-icantly impaired with increased age under ischemia. TGF- b 1 stimulated wound repair under both ischemic and non-ischemic conditions in young animals, although it showed no statistical difference in aged animals. Procollagen mRNA expression decreased under ischemic conditions and with aging. Neither TGF- b 1 nor procollagen a 1(I) mRNA expression increased in response to TGF- b 1 treatment under ischemia in aged animals. Therefore, the wound-healing process is impaired additively by aging and ischemia. The lack of a wound-healing response to TGF- b 1 in aged ischemic wounds may play a role in the chronic wounds. (Am J Pathol 1999, 154:301–309) Many animal appear promising therapeutic agents promoting wound healing. However, chronic wounds have been less encouraging. 2,3 We rea-soned that preclinical studies using young animals may not
Deposition and Characterization of ZnSnSe2 Thin-Films Deposited by Using Sintered Stoichiometric Powder

In this work, ZnSnSe2 (ZTSe) thin films were deposited using crystalline powder grown by vertical Bridgman-Stockbarger technique. The deposition process was carried out by means of e-beam evaporation on the well-cleaned soda lime glass substrates and keeping them at the substrate temperature of 200°C. The structural, optical and electrical properties of ternary ZTSe thin films were investigated depending on the annealing temperature at 250 and 300°C. X-ray diffraction analysis showed that as-grown films were in amorphous structure, however annealing at 250°C triggered the crystallization on the preferred ternary structure and annealing at 300°C resulted in the changes from amorphous to the polycrystalline structure. Using the compositional analysis, the detail information about the stoichiometry and the segregation mechanisms of the constituent elements in the structure were determined for both as-grown and annealed samples. In addition, they were morphologically characterized using scanning electron microscopy technique. The electrical properties were analyzed using temperature dependent dark- and photo-conductivity measurements. From the variation of electrical conductivity as a function of the ambient temperature, the current transport mechanisms and corresponding activation energies at specific temperature intervals for each sample were determined. The optical properties for the ZTSe thin films were studied depending on the structural changes with annealing.
Heparin-derived heparan sulfate mimics that modulate inflammation and cancer

The heparan sulfate (HS) chains of heparan sulfate proteoglycans (HSPG) are “ubiquitous” components of the cell surface and the extracellular matrix (EC) and play important roles in the physiopathology of developmental and homeostatic processes. Most biological properties of HS are mediated by interactions with “heparin-binding proteins” and can be modulated by exogenous heparin species (unmodified heparin, low molecular weight heparins, shorter heparin oligosaccharides and various non-anticoagulant derivatives of different sizes). Heparin species can promote or inhibit HS activities to different extents depending, among other factors, on how closely their structure mimics the biologically active HS sequences. Heparin shares structural similarities with HS, but is richer in “fully sulfated” sequences (S domains) that are usually the strongest binders to heparin/HS-binding proteins. On the other hand, HS is usually richer in less sulfated, N-acetylated sequences (NA domains). Some of the functions of HS chains, such as that of activating proteins by favoring their dimerization, often require short S sequences separated by rather long NA sequences. The biological activities of these species cannot be simulated by heparin, unless this polysaccharide is appropriately chemically/enzymatically modified or biotechnologically engineered. This mini review covers some information and concepts concerning the interactions of HS chains with heparin-binding proteins and some of the approaches for modulating HS interactions relevant to inflammation and cancer. This is approached through a few illustrative examples, including the interaction of HS and heparin-derived species with the chemokine IL-8, the growth factors FGF1 and FGF2, and the modulation of the activity of the enzyme heparanase by these species. Progresses in sequencing HS chains and reproducing them either by chemical synthesis or semi-synthesis, and in the elucidation of the 3D structure of oligosaccharide–protein complexes, are paving the way for rational approaches to the development of HS-inspired drugs in the field of inflammation and cancer, as well in other therapeutic fields.
Arginine reduces Cryptosporidium parvum infection in undernourished suckling mice involving both nitric oxide synthase and arginase

Objective— This study investigated the role of L-arginine supplementation to undernourished and Cryptosporidium parvum-infected suckling mice. Methods— The following regimens were initiated on the 4th day of life and given subcutaneously daily: either 200mM of L-arginine or PBS for the C. parvum-infected controls. L-arginine-treated mice were grouped to receive either 20mM of NG-nitroarginine-methyl-ester (L-NAME) or PBS. Infected mice received orally 10 6 excysted-C. parvum oocysts on day 6 and were euthanized on day 14 th at the infection peak. Results— L-arginine improved weight gain compared to the untreated infected controls. L-NAME profoundly impaired body weight gain as compared to all other groups. Cryptosporidiosis was associated with ileal crypt hyperplasia, villus blunting, and inflammation. L-arginine improved mucosal histology following infection. L-NAME abrogated these arginine-induced improvements. Infected control mice showed an intense arginase expression, which was even greater with L-NAME. L-arginine reduced parasite burden, an effect that was reversed by L-NAME. C. parvum infection increased urine NO 3 -/NO 2 - concentration when compared to uninfected controls, which was increased by L-arginine supplementation, an effect that was also reversed by L-NAME. Conclusion— These findings show a protective role of L-arginine during C. parvum infection in undernourished mice with involvement of arginase I and nitric oxide synthase enzymatic actions.
Low Reynolds number flow control through small-amplitude high-frequency motion

There is currently growing interest in the field of Micro Air Vehicles (MAVs). A MAV is characterized by its low Reynolds numbers flight regime which makes lift and thrust creation a significant challenge. One possible solution inspired by nature is flapping flight, but instead of the large-amplitude low-frequency motion suited to the muscular actuators of nature, small-amplitude high-frequency motion may be more suitable for electrical actuators. In this thesis the effect of small-amplitude high-frequency motion is experimentally investigated focusing on three aspects: general performance improvement, deflected jets, and the effect of geometry Results presented herein demonstrate that using small-amplitude high-frequency plunging motion on a NACA 0012 airfoil at a post-stall angle of attack of 15° can lead to significant thrust production accompanying a 305% increase in lift coefficient. At low Strouhal numbers vortices form at the leading-edge during the downward motion and then convect into the wake. This ‘mode 1’ flow field is associated with high lift but low thrust. The maximum lift enhancement was due to resonance with the natural shedding frequency, its harmonics and subharmonics. At higher Strouhal numbers the vortex remains over the leading-edge area for a larger portion of the cycle and therefore loses its coherency through impingement with the upward moving airfoil. This ‘mode 2’ flowfield is associated with low lift and high thrust. At angles of attack below 12.5° very large force bifurcations are observed. These are associated with the formation of upwards or downwards deflected jets with the direction determined by initial conditions. The upwards deflected jet is associated with the counter-clockwise Trailing Edge Vortex (TEV) loitering over the airfoil and thereby pairing with the clockwise TEV to form a dipole that convects upwards. It therefore draws fluid from the upper surface enhancing the upper surface vortex leading to high lift. The downwards deflected jet is associated with the inverse. Deflected jets were not observed at larger angles of attack as the asymmetry in the strength of the TEVs was too great; nor at smaller amplitudes as the TEV strength was insufficient. To understand the effect of geometry comparable experiments were performed for a flat plate geometry. At zero degrees angle of attack deflected jets would form, as for the NACA 0012 airfoil, however their direction would switch sinusoidally with a period on the order of 100 cycles. The lift coefficient therefore also switched. At 15° angle of attack for Strouhal numbers up to unity the performance of the flat plate was comparable to the NACA 0012 airfoil. Above unity, the upper surface and lower surface leading-edge vortices form a dipole which convects away from the upper surface resulting in increased time-averaged separation and reduced lift.
A Unique B-cell Regulome Links Notch to Downstream Oncogenic Pathways in Small B-cell Lymphomas

Gain-of-function Notch mutations are recurrent in mature small B-cell lymphomas such as mantle cell lymphoma (MCL) and chronic lymphocytic leukemia (CLL), but the Notch target genes that contribute to B-cell oncogenesis are largely unknown. We performed integrative analysis of Notch-regulated transcripts, genomic binding of Notch transcription complexes, and genome conformation data to identify direct Notch target genes in MCL cell lines. This unique B-cell Notch regulome is largely controlled through Notch-bound distal enhancers and includes genes involved in B-cell receptor and cytokine signaling and the oncogene MYC, which sustains proliferation of Notch-dependent MCL cell lines via a Notch-regulated lineage-restricted enhancer complex. Expression of direct Notch target genes is associated with Notch activity in an MCL xenograft model and in CLL lymph node biopsies. Our findings provide key insights into the role of Notch in MCL and other B-cell malignancies and have important implications for therapeutic targeting of Notch-dependent oncogenic pathways. rationale targeting Notch
Endocannabinoid Signaling, Glucocorticoid-Mediated Negative Feedback and Regulation of the HPA Axis

The hypothalamic-pituitary-adrenal (HPA) axis regulates the outflow of glucocorticoid hormones under basal conditions and in response to stress. Within the last decade, a large body of evidence has mounted indicating that the endocannabinoid system is involved in the central regulation of the stress response; however, the specific role endocannabinoid signalling plays in phases of HPA axis regulation, or the neural sites of action mediating this regulation, was not mapped out until recently. This review aims to collapse the current state of knowledge regarding the role of the endocannabinoid system in the regulation of the HPA axis to put together a working model of how and where endocannabinoids act within the brain to regulate outflow of the HPA axis. Specifically, we discuss the role of the endocannabinoid system in the regulation of the HPA axis under basal conditions, activation in response to acute stress and glucocorticoid-mediated negative feedback. Interestingly, there appears to be some anatomical specificity to the role of the endocannabinoid system in each phase of HPA axis regulation, as well as distinct roles of both anandamide and 2-arachidonoylglycerol in these phases. Ultimately, the current level of information indicates that endocannabinoid signalling acts to suppress HPA axis activity through concerted actions within the prefrontal cortex, amygdala and hypothalamus. a constellation of physiological systems designed to increase the survival of the organism. The common stress response to both physiological and psychological stressors can be summarized generally as the concerted activation of a two-pronged physiological defense mechanism, an autonomic response and a neuroendocrine response. The autonomic response involves stimulation of sympathetic motor and hormonal outputs via descending neural circuits originating in hypothalamic preautonomic control centers. The neuroendocrine stress response is mediated by activation