Miracle-dz/UniFAR-FTUs huggingface dataset:
Oxidant‐Induced Regulation of Glutathione Synthesis

This unit describes protocols for characterizing the expression of two glutathione biosynthesis enzymes: gamma‐glutamylcysteine synthase (GCS) and gamma‐glutamyl transpeptidase (GGT) in response to oxidants. GCS catalyzes the first and rate‐limiting step of glutathione synthesis, while GGT degrades extracellular glutathione (GSH) to provide the amino acids required for intracellular synthesis of GSH. Northern blot hybridization is used to quantitatively assess the mRNAs for the two enzymes in response to oxidant. Nuclear run‐on is used to determine the rate of transcription.
Cataracts and Hypertension in Salt-Sensitive Rats

In previous unrelated studies, we observed a 35 to 50% incidence of cataract formation in several groups of Dahl salt-sensitive hypertensive rats (DS) over a 4-year period. In the present study we evaluated longitudinal changes in blood pressure In DS in which cataracts eventually developed and those in which cataracts did not develop when all animals were maintained on a high sodium diet. Lenses were evaluated by slit-lamp microscopy to determine if cataractous lesions were similar among rats, to classify lesion types, and to define the age at which cataracts were detectable in DS. The possible participation of several cataractogenic risk factors as major Influences on cataract formation also was evaluated. Finally, aqueous humor concentrations and lenticular content of sodium and potassium were determined to evaluate the possibility that a defect in ion transport at the lens epithelium and ciliary body might play a role in cataractogenesis in DS, since ion transport defects have been shown to lead to lens opaciflcatlon in other models of genetic and experimental cataracts. Parallel studies were performed in Dahl salt-resistant control rats (DR). A high incidence of cataract formation was found in DS. Although systolic blood pressure was not consistently greater in adult DS with cataracts compared with values in age-matched DS without cataracts, the initial pressor response to a high salt diet was greatest in weanling DS in which cataractous lesions later developed. Slit-lamp analysis revealed that cataracts in this genetic model were cortical, with one mixed cortical, nuclear lesion. Posterior subcapsular lesions were not observed, suggesting that lesions were not steroidinduced. Serum ultrafiltrable calcium concentration was similar among DS with and without cataracts and normotensive DR, indicating that hypocalcemia was not involved in cataractogenesis. Analysis of aqueous humor showed high potassium concentration and low sodium concentration in DS with cataracts, suggesting a possible ion transport defect in the ciliary processes or lens epithelium (or both). In addition, increased lenticular water and sodium content, as well as decreased potassium content, indicated that mature cataracts in DS were associated with altered sodium and potassium transport. Although aqueous humor sodium concentration and lenticular sodium content were not altered in DS without cataracts, lenticular potassium content was decreased while aqueous humor potassium concentration was increased compared with values in control DR. These data suggesting a decreased transmembrane potassium gradient in DS without cataracts also suggest that a specific defect in potassium transport may precede the development of cataracts in this genetic model. The finding of a potential genetic model of cataracts in which cataractogenesis is associated with hypertension suggests a possible link between hypertension and cataract formation. (Hypertension 9: 304-308, 1987)
Mutational Analysis of the L1 Neuronal Cell Adhesion Molecule Identifies Membrane-Proximal Amino Acids of the Cytoplasmic Domain That Are Required for Cytoskeletal Anchorage

The preferential localization of the L1 cell adhesion molecule in the axons and growth cones of differentiating neurons suggests the existence of a mechanism for targeting or anchoring the molecule to these locations. We have used B28 glioma cells, which have an extremely flattened morphology, as a model system to study the organization of L1 on the cell structure. Transfection of L1 cDNA into B28 cells results in expression of the L1 protein in organized linear cell surface arrays which are codistributed with cytoskeletal stress fibers, but not with microtubles or intermediate filaments. Transfection studies with L1 deletion mutants identify the juxtamembrane segment of the cytoplasmic domain as the critical entity for arrangement of L1 into ordered cell surface arrays. The seventh cytoplasmic amino acid of L1, lysine 1150, and to a lesser extent the fourth cytoplasmic amino acid, lysine 1147, appear to be critical residues for maintaining normal L1 anchorage and distribution.
Energy balance and obesity: a UK perspective on the gluttony v. sloth debate

Obesity in the UK was assumed to have developed against a population decline in physical activity, with health messages focused on diet and exercise prevention strategies. Doubly-labelled water (DLW) studies of energy expenditure have indicated the alternative scenario that the increased obesity prevalence reflects excessive food energy intake with physical activity levels unchanged. This analysis is questionable, deriving in part from a weakness of the DLW methodology in identifying changing physical activity levels within populations of increasing body weight. This has resulted in an underestimation of the reduction in physical activity in the overweight and obese, as revealed by direct studies of such behaviour. Furthermore, a close examination of food energy supply, household food purchases and individual food energy consumption since 1955, in relation to likely estimates of current intakes indicated by simple modelling of predicted energy expenditure, identifies: (a) food energy supply as markedly overestimating energy intakes; (b) individual food energy consumption as markedly underestimating energy intakes; and (c) household food purchase data as the closest match to predicted current food energy intakes. Energy intakes indicated by this latter method have fallen by between 20 to 30 %, suggesting comparable falls in physical activity. Although unequivocal evidence for a matching UK trend in falling physical activity is limited, as is evidence that obesity follows reductions in physical activity, such a link has been recently suggested in a large prospective study in adolescents. Thus, for the UK, obesity has developed within a ‘move less–eat somewhat less but still too much’ scenario. A focus on both diet and exercise should remain the appropriate public health policy.
Mash1-dependent Notch Signaling Pathway Regulates GABAergic Neuron-Like Differentiation from Bone Marrow-Derived Mesenchymal Stem Cells

GABAergic neuronal cell grafting has promise for treating a multitude of neurological disorders including epilepsy, age-related memory dysfunction, Alzheimer’s disease and schizophrenia. However, identification of an unlimited source of GABAergic cells is critical for advancing such therapies. Our previous study implied that reprogramming of bone marrow-derived mesenchymal stem cells (BMSCs) through overexpression of the Achaete-scute homolog 1 (Ascl1, also called Mash1) could generate GABAergic neuron-like cells. Here, we investigated mechanisms underlying the conversion of BMSCs into GABAergic cells. We inhibited γ-secretase (an enzyme that activates Notch signaling) with N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT) or manipulated the expression of Notch signaling components such as the recombination signal binding protein for immunoglobulin kappa J region (RBPJ), hairy and enhancer of split-1 (Hes1) or Mash1. We demonstrate that inhibition of γ-secretase through DAPT down-regulates RBPJ and Hes1, up-regulates Mash1 and results in an enhanced differentiation of BMSCs into GABAergic cells. On the other hand, RBPJ knockdown in BMSCs has no effect on Mash1 gene expression whereas Hes1 knockdown increases the expression of Mash1. Transduction of Mash1 in BMSCs also increases the expression of Hes1 but not RBPJ. Moreover, increased GABAergic differentiation in BMSCs occurs with concurrent Mash1 overexpression and Hes1-silencing. Thus, the Mash1-dependent Notch signaling pathway regulates GABAergic neuron-like differentiation of BMSCs. These results also suggest that genetic engineering of BMSCs is a useful avenue for obtaining GABAergic neuron-like donor cells for the treatment of neurological disorders.
THE CONVERSION OF NEGATIVES TO POSITIVES IN “SLOW” ADAPTING POPULATIONS OF YEAST,

Winge and Roberts (1948) discovered that Saccharomyces chevalieri was a "slow" adapting type to galactose and demonstrated that the character was determined by a single recessive allele (g.) which segregates normally in crosses to the wild type. Spiegelman, Sussman, and Pinska (1950) analyzed strains of the g. -genotype and revealed the existence of phenotypic heterogeneity in clones carrying this allele. The heterogeneity is exhibited easily and simply by plating glucose grown g8cells on purified galactose agar test plates upon which two kinds of colonies develop. The predominant type is small, light colored, and negative in appearance. Present, in the proportion of about one in a thousand, are cells which yield large, dark colored, galactose positive colonies. Such positives retain and transmit their character for an indefinite number of generations provided galactose represents the sole or primary source of carbon and energy. If, however, positive cells are allowed to grow in the abbence of galactose, a mass reversion to the original negative phenotype occurs quite abruptly between the sixth and seventh generations. The data obtained eliminated the possibility that mutation was operative in the process by means of which a positive population is derived from a negative one in the presence of galactose. The experimental results led to the conclusion that the inheritance of the capacity to synthesize galactozymase in this strain is a predominantly cytoplasmic event and involves the discrete transmission of elements necessary for enzyme formation. On the basis of the behavior of positive cells in the pres-
A de-blocking algorithm and a blockiness metric for highly compressed images

Blockiness is a typical artifact in reconstructed images that have been coded by a block-based discrete cosine transform (BDCT). In highly compressed images and video, the blocking artifacts are easily noticeable as the discontinuities between relatively homogeneous regions. Many current noniterative de-blocking algorithms attempt to remove the blocking artifacts by smoothing a few pixels around the block boundaries; however, the results are not satisfactory, especially at very low bit rates. We propose a de-blocking algorithm based on the number of connected blocks in a relatively homogeneous region, the magnitude of abrupt changes between neighboring blocks, and the quantization step size of DCT coefficients. Due to its adaptability, the proposed algorithm can smooth out the blocking artifacts while keeping the strong edges and texture areas untouched. Since this algorithm is noniterative and only identifies those block pairs that actually need de-blocking, its computation cost is low. In addition, we have developed a new metric to measure the blocking artifacts in images. Through analyzing the 2N-point (N is the block size) one-dimensional DCT coefficients of the two neighboring blocks with blocking artifacts, we show that all of the even DCT coefficients of the combined 2N points are zeros (except frequency k=0). The odd DCT coefficients are proportional to the pixel value difference between these two blocks with their magnitudes almost inversely proportional to frequency k. We selected the first DCT coefficient (frequency k=1) as an indicator for the strength of blocking artifacts in the reconstructed images. For the postprocessed images, we used a weighted summation of the squared first DCT coefficient to measure their blocking artifacts. Experimental results demonstrate that our proposed de-blocking algorithm produces better results than other methods, both visually and quantitatively, while the proposed blocking artifact metric is more consistent with subjective evaluation than the peak signal-to-noise ratio.
Make Gestures to Learn: Reproducing Gestures Improves the Learning of Anatomical Knowledge More than Just Seeing Gestures

Manual gestures can facilitate problem solving but also language or conceptual learning. Both seeing and making the gestures during learning seem to be beneficial. However, the stronger activation of the motor system in the second case should provide supplementary cues to consolidate and re-enact the mental traces created during learning. We tested this hypothesis in the context of anatomy learning by naïve adult participants. Anatomy is a challenging topic to learn and is of specific interest for research on embodied learning, as the learning content can be directly linked to learners' body. Two groups of participants were asked to look at a video lecture on the forearm anatomy. The video included a model making gestures related to the content of the lecture. Both groups see the gestures but only one also imitate the model. Tests of knowledge were run just after learning and few days later. The results revealed that imitating gestures improves the recall of structures names and their localization on a diagram. This effect was however significant only in long-term assessments. This suggests that: (1) the integration of motor actions and knowledge may require sleep; (2) a specific activation of the motor system during learning may improve the consolidation and/or the retrieval of memories.
Suppressed visual looming stimuli are not integrated with auditory looming signals: Evidence from continuous flash suppression

Previous studies using binocular rivalry have shown that signals in a modality other than the visual can bias dominance durations depending on their congruency with the rivaling stimuli. More recently, studies using continuous flash suppression (CFS) have reported that multisensory integration influences how long visual stimuli remain suppressed. In this study, using CFS, we examined whether the contrast thresholds for detecting visual looming stimuli are influenced by a congruent auditory stimulus. In Experiment 1, we show that a looming visual stimulus can result in lower detection thresholds compared to a static concentric grating, but that auditory tone pips congruent with the looming stimulus did not lower suppression thresholds any further. In Experiments 2, 3, and 4, we again observed no advantage for congruent multisensory stimuli. These results add to our understanding of the conditions under which multisensory integration is possible, and suggest that certain forms of multisensory integration are not evident when the visual stimulus is suppressed from awareness using CFS.
What does the bispectral EEG index monitor?

Depth of anaesthesia is an old concept originally based on the depressant effects of volatile anaesthetics on the central nervous system. As the dose of anaesthetic is increased, there is progressive loss of consciousness and suppression of somatic and autonomic responses to noxious stimuli. It seemed logical to search for a measure of central nervous system depression that would serve as a monitor of anaesthesia. The electroencephalogram (EEG) was much studied and a number of signal processing techniques were developed to facilitate interpretation [1]. Unfortunately, none of the early processed EEG derivatives (95% spectral edge frequency or median power frequency) were able to correlate reliably with consciousness, movement or autonomic responses. In the European Journal of Anaesthesiology, Driessen et al. concluded that the bispectral Index (BIS), the latest EEG derivative, also did not predict the haemodynamic response associated with endotracheal intubation and stenotomy [2]. To understand Driessen et al.’s findings, one must review the evolution of BIS. Unlike early processed EEG variables, BIS is derived from the bifrontal EEG recordings collected from a huge group of subjects (n × 5000) sedated with different anaesthetics [1,3]. These EEG waveforms were then analysed off-line according to three features: (a) Burst suppression ratio. This is a time-domain feature that quantifies the extent of electrical silence during deep anaesthesia. (b) Relative alpha/beta ratio. This is a frequencydomain feature and contributions from these frequency bands (alpha 8–13 Hz, beta 13–30 Hz) are often seen during light sedation. (c) Bicoherence of the EEG. This describes the phase coupling relations between individual waves. In simple terms, a signal with strong phase relations and a high bicoherence value implies a common generator and may be associated with moderate sedation.
Complementation analysis of pseudorabies virus gE and gI mutants in retinal ganglion cell neurotropism

Pseudorabies virus glycoproteins gE and gI are required to infect some, but not all, regions of the rodent central nervous system after peripheral injection. After infection of the retina, pseudorabies virus mutants lacking either gE or gI can subsequently infect neural centers involved in the control of circadian function but cannot infect visual circuits mediating visual perception or the reflex movement of the eyes. In this study, we used genetic complementation to test the hypothesis that gE and gI are required for entry into the specific retinal ganglion cells that project to visual centers. These data strongly suggest that gE and gI must function after the viruses enter primary neurons in the retina.
Thermal Denaturation Profiles of Tuna Myoglobin

Myoglobin (Mb) purified from fast skeletal muscle of bluefin tuna Thunnus thynnus orientalis was subjected to thermal treatment, and the denaturation profiles were examined by thermodynamic analysis. Based on the ellipticity or helical content obtained by circular dichroism (CD) spectrometry, it was found that denaturation of tuna Mb consisted of three steps, and that slight structural changes of Mb started below 20 °C. However, major structural changes were observed at around 58 and 72 °C. Differential scanning calorimetry (DSC) analysis revealed a similar but somewhat different thermal denaturation profile of Mb. In comparison with the denaturing profiles of whale Mb under the same conditions, the thermal stability of tuna Mb was found to be much lower. In the modeled tertiary structures of these Mbs, they were roughly similar to each other, though minor conformational differences were recognized and the total energy was found to be lower for tuna Mb.
Holographic microscopy reconstruction in both object and image half-spaces with an undistorted three-dimensional grid.

We propose a holographic microscopy reconstruction method that propagates the hologram in the object half-space in the vicinity of the object. The calibration yields reconstructions with an undistorted reconstruction grid, i.e., with orthogonal x, y, and z axes and constant pixel pitch. The method is validated with a USAF target imaged by a ×60 microscope objective (MO), whose holograms are recorded and reconstructed for different USAF locations along the longitudinal axis: -75 to +75 μm. Since the reconstruction numerical phase mask, the reference phase curvature, and the MO form an afocal device, the reconstruction can be interpreted as occurring equivalently in the object or image half-space.
Extended Release System of Hypoglycemic Agent Containing Solid Dispersions: Strategies and Mechanisms

The main perspective of the present study aims at overcoming solubility problems by using the technique of solid dispersion. Repaglinide is a BCS Class II drug, having low aqueous solubility and therefore, low bioavailability. Solid dispersions of repaglinide with different carriers Polyvinyl Pyrrolidone (PVP) and Ethyl Cellulose (EC) in different ratios were prepared by suspending method and Dissolving methods. In vitro release studies revealed that the F7 formulation showed extended drug release. So, the dissolution profile of solid dispersion containing EC and PVP K30 (1: 3) was selected as the best formulation because of its extended drug release among all formulations. In conclusion, solid dispersions of Repaglinide in PVP have shown to be a promising approach to improve the bioavailability of Repaglinide. Keywords—Ethyl Cellulose, Glibenclamide, Polyvinyl Pyrrolidone, Repaglinide, Solid Dispersion.
Different Mechanisms of Syndecan-1 Activation through a Fibroblast-growth-factor-inducible Response Element (FiRE) in Mucosal and Cutaneous Wounds

Syndecan-1 expression is enhanced in cutaneous and mucosal wounds. We have previously demonstrated that wounding-induced syndecan-1 expression in the skin occurs transcriptionally, through a fibroblast-growth-factor-inducible element (FiRE). Here, we show that FiRE is also activated in mucosal wounds. However, both the expression patterns and the activation mechanisms of FiRE are different from those in the skin. In the mucosa in vivo, the activation starts and ends earlier than in cutaneous wounds. FiRE is first detected at around 12 hours in keratinocytes, and the activation declines by the third day after wounding occurs. The activation is seen on the migrating sheet of epithelial mucosa, as in the case of cutaneous wounding. In contrast to the situation in vivo, organ-cultured mucosal wounds exhibit no FiRE activity, while organ-cultured cutaneous wounds show robust activity. Activation in mucosal wounds is enhanced, however, by the application of epidermal growth factor. This suggests that exogenous growth factor activity is required for activation of syndecan-1 in mucosal wounds but not in cutaneous wounds.
6-{5-Amino-3-tert-butyl-4-[(E)-(3-methyl-1,2,4-thiadiazol-5-yl)diazenyl]-1H-pyrazol-1-yl}-1,3,5-triazine-2,4(1H,3H)-dione–1-methylpyrrolidin-2-one–water (1/1/1)

In the title compound, C13H16N10O2S·C5H9NO·H2O, the entire 1-methylpyrrolidin-2-one (NMP) molecule is disordered over two sites with occupancies of 0.488 (5) and 0.512 (5). The six-membered triazine ring and the two five-membered pyrazole and thiadiazole rings, together with the diazene (–N=N–) linkage are almost coplanar (r.m.s. deviation for the non-H atoms = 0.0256 Å) with methyl groups from the tert-butyl substituent on the pyrazole ring located above and below the plane. Three intramolecular N—H⋯N hydrogen bonds contribute to the planarity of the system. The O atom of the NMP molecule is hydrogen bonded to an O—H group of water. In turn, the water molecule is hydrogen bonded to the mono-azo skeleton through intermolecular N—H⋯O and O—H⋯N hydrogen bonds. At both ends of the long molecular axis of the main molecule there are intermolecular N—H⋯N hydrogen bonds, arranged in a head-to-tail fashion, between the N—H group of the triazine ring of one molecule and the N atom of the thiadiazole ring of a neighboring molecule. These form a polymeric chain along [110] or [10]. The main molecules are stacked alternately along the b axis, which effectively cancels their dipole moments. In addition, pairs of alternate molecules are dimerized via intermolecular hydrogen bonds involving the solvent molecules.
Disorientation-induced disordering at a nematic-liquid-crystal-solid interface.

To explain the observed deviation from the Rapini-Papoular surface energy of nematic liquid crystals, we propose a model of the form Tr[(Q ij -Q 0ij ) 2 ] (Q ij is the quadrupolar order parameter of the nematic liquid crystal and Q 0ij a quadrupolar surface field). For a broken anchoring, this model predicts a surface order decrease ΔS≃3/2 (ξ/d)S B , where ξ and d are the coherence and surface extrapolation lengths of the nematic liquid crystal and S B is the bulk order parameter