Miracle-dz/UniFAR-FTUs huggingface dataset:
In Vivo Adenoviral Delivery of Recombinant Human Protein Kinase C-ζ Stimulates Glucose Transport Activity in Rat Skeletal Muscle*

An in vivo adenoviral gene delivery system was utilized to assess the effect of overexpressing protein kinase C (PKC)-ζ on rat skeletal muscle glucose transport activity. Female lean Zucker rats were injected with adenoviral/human PKC-ζ (hPKC-ζ) and adenoviral/LacZ in opposing tibialis anterior muscles. One week subsequent to adenoviral/gene delivery rats were subjected to hind limb perfusion. The hPKC-ζ protein was expressed at the same level (fast-twitch white) or at ∼80% of the level (fast-twitch red) of endogenous PKC-ζ, thus approximately doubling the amount of PKC-ζ in tibialis anterior. Basal glucose transport activity was elevated ∼3.4- and 2-fold, respectively, in fast-twitch white and red hPKC-ζ muscle relative to control. Submaximal insulin-stimulated glucose transport activity, corrected for basal transport, was ∼90 and 40% over control values, respectively, in fast-twitch white and red hPKC-ζ muscle. The enhancement of glucose transport activity in muscle expressing hPKC-ζ occurred in the absence of any change in GLUT1 or GLUT4 protein levels, suggesting a redistribution of existing transporters to the cell surface. These results demonstrate that an adenoviral vector can be used to deliver expressible hPKC-ζ to adult rat skeletal muscle in vivo and also affirm a role for PKC-ζ in the regulation of glucose transport activity.
Physicochemical and mechanical properties of carbamazepine cocrystals with saccharin

The aim of present research was to investigate the physicochemical, mechanical properties, and stability characteristics of cocrystal of carbamazepine (CBZ) using saccharin (SAC) as a coformer. The cocrystals were prepared by solubility method and characterized by pH-solubility profile, intrinsic dissolution by static disk method, and surface morphology by scanning electron microscopy (SEM), crystallinity by differential scanning calorimetry (DSC) and powder X-ray diffraction (PXRD), and mechanical properties by Heckel analysis. Stability of the cocrystals were assessed by storing them at 60 °C for two weeks, 25 °C/60%RH, 40 °C/75%RH and 40 °C/94%RH for one month and compared with the stability of CBZ. The solubility profile of cocrystal was similar to CBZ. The cocrystal and CBZ have shown the same stability profile at all the conditions of studies except at 40 °C/94%RH. Unlike the CBZ, cocrystal was stable against dihydrate transformation. The compacts of cocrystal have a greater tensile strength and more compressibility. The Heckel analysis suggested that plastic deformation started at low compression pressure in the cocrystal than CBZ. In summary, the cocrystal form of carbamazepine provides another avenue for product development which is more stable than the parent drug.
Immobilization of Lipase to Chitosan Beads using a Natural Cross‐Linker

Abstract Genipin, a reagent of plant origin was used for the immobilization of lipase by cross‐linking to chitosan beads. The catalytic properties and operational and storage stabilities of the immobilized lipase were compared with the soluble lipase. Under optimum conditions, 198 µg protein was bound per g chitosan with a protein‐coupling yield of 35%. The hydrolytic activity was 10.8 U/g chitosan and the relative specific activity was 108%. The immobilized lipase showed better thermal and pH stabilities compared to the soluble form. The immobilized enzyme exhibited mass transfer limitations as reflected by a higher apparent Km value and a lower energy of activation. The immobilized enzyme retained about 74% of its initial activity after five hydrolytic cycles.
Validation and modification of a diagnostic scoring system to predict microscopic colitis

Abstract Objective: Many patients with diarrhoea undergo colonoscopy. If this is macroscopically normal, random biopsies are obtained to rule out microscopic colitis (MC), but most patients have functional disease. Accurate predictors of MC could avoid the need to take biopsies in a substantial proportion of patients, saving money for the health service. We validated a previously described diagnostic scoring system for MC, and incorporated further variables to assess whether this improved performance. Material and methods: Consecutive adults with loose stools undergoing colonoscopy in Leeds, UK were included. Demographic and symptom data were collected prospectively. The diagnostic scoring system described previously was applied. In addition, the incorporation of further variables, including drugs associated with MC, number of stools, nocturnal passage of stools, and duration of loose stools, into the scoring system was assessed. Sensitivities, specificities, and positive and negative predictive values were calculated. Results: Among 242 patients (mean age 51.0 years, 163 (67.4%) female), 26 (10.7%) of whom had MC, a cut off of ≥4 on the original scoring system had a sensitivity of 92.3% and specificity of 35.2%. Nocturnal passage of stools and duration of loose stools <6 months were significant predictors of MC. Incorporating these variables in a new scoring system with a cut off of ≥6 identified MC with 95.7% sensitivity and 46.0% specificity. Conclusions: Incorporating nocturnal passage of stools and duration of loose stools into the scoring system may improve ability to predict MC, and avoid random colonic biopsies in a greater proportion of patients with loose stools.
The influence of physical activity level on climacteric symptoms in menopausal women

Objective To evaluate the relationship between physical activity level and climacteric symptoms in menopausal women. Methods A study was conducted on 336 healthy women aged 45–55 years. They were asked to complete a questionnaire consisting of two parts: (1) a socioeconomic and obstetric/gynecological survey, and (2) the International Physical Activity Questionnaire long form and the Blatt–Kupperman Index. Results The women assigned to the high physical activity group felt better and had less severe climacteric symptoms; 52.08% of the women had no climacteric symptoms. The majority of women in this group were assigned to the total high physical activity group. Significant differences were observed in the relationship between physical activity level and climacteric symptoms. In each domain, women with no climacteric symptoms were classified into either the high or the moderate physical activity level group. In the work domain, 25.30% of the women had a moderate, 20.53% a high, and 6.25% a low physical activity level. In the transportation domain, 32.72% of the women had a moderate, and 19.36% a low physical activity level. In the domestic chores domain, 30.06% of the women had a moderate, 16.07% a high and 5.95% a low physical activity level. In the leisure-time domain, 25.61% of the women had a moderate, 16.95% a high and 9.52% a low physical activity level. Conclusions Physical activity is associated with less menopausal symptoms. It is important to encourage regular physical activity as it is beneficial to health and relieves menopausal symptoms.
Frontal eye field as defined by intracortical microstimulation in squirrel monkeys, owl monkeys, and macaque monkeys II. cortical connections

Physiological (intracortical microstimulation) and anatomical (transport of horseradish peroxidase conjugated to wheat germ agglutinin as shown by tetramethyl benzidine) approaches were combined in the same animals to reveal the locations, extents, and cortical connections of the frontal eye fields (FEF) in squirrel, owl, and macaque monkeys. In some of the same owl and macaque monkeys, intracortical microstimulation was also used to evoke eye movements from dorsomedial frontal cortex (the supplementary motor area). In addition, in all of the owl and squirrel monkeys, intracortical microstimulation was also used to evoke body movements from the premotor and motor cortex situated between the central dimple and the FEF. These microstimulation data were directly compared to the distribution of anterogradely and retrogradely transported label resulting from injections of tracer into the FEF in each monkey. Since the injection sites were limited to the physiologically defined FEF, the demonstrated connections were solely those of the FEF. To aid in the interpretation of areal patterns of connections, the relatively smooth cortex of owl and squirrel monkeys was unfolded, flattened, and cut parallel to the flattened surface. Cortex of macaque monkeys, which has numerous deep sulci, was cut coronally.
Effect of neuroactive nutritional supplementation on body weight and composition in growing puppies

Abstract Nutritional factors can dramatically affect development of young animals during the early stage of life. The objective of the present study was to examine the effects of a neuroactive nutritional supplement (NNS) containing DHA, taurine, carotenoids and vitamins on the body weight and body composition of growing puppies. A total of twenty-four 2-month-old Beagles were fed a nutritionally complete and balanced base diet and a control supplement daily during an initial 1-month baseline assessment, after which they were divided into control and treatment groups. They were fed daily either control or treatment supplements in addition to the base diet from 3 to 12 months of age. Lean body mass and fat mass were assessed using quantitative magnetic resonance scans at 0 (baseline), 3, 6 and 9 months of treatment. Total body weight and lean body mass did not differ between groups over time. The puppies in the treatment group showed a trend of reduced fat gain compared with those in the control group, and with a marginally significant difference at 6 months (P = 0·05). At 3 months, insulin-like growth factor 1 was higher (P = 0·02) in the treatment group compared with the control group. At 9 months, fasting lipid levels were lower (P < 0·05) and fat-oxidation metabolite 3-hydroxybutyrate was higher (P < 0·05) in the treatment group compared with the control group. These results may indicate that NNS has an impact on puppy growth and development, possibly by promoting fat metabolism; further investigation would be necessary to determine the full impact of this supplement on growth and development.
Ethanol Cytotoxicity to a Transfected HepG2 Cell Line Expressing Human Cytochrome P4502E1*

The effect of ethanol on the viability of a HepG2 cell model which was developed to constitutively express human CYP2E1 was studied in an attempt to establish a linkage between CYP2E1, reactive oxygen intermediates, and ethanol toxicity. Assays of toxicity included leakage of lactate dehydrogenase, trypan blue uptake, morphology, and formazan production. Ethanol was toxic to HepG2 E9 cells, which express CYP2E1, but not to HepG2 MV5 cells, which do not express CYP2E1. The ethanol toxicity was dependent on the concentration of ethanol, starting with 10 mM ethanol, and on the time of incubation with ethanol. Phorbol 12-myristate 13-acetate, which increases the expression of CYP2E1 in this model, increased the toxicity by ethanol. Ethanol toxicity was prevented by 4-methylpyrazole and by diallyl sulfide, inhibitors of CYP2E1. The ethanol toxicity was also prevented by radical trapping agents such as N-acetylcysteine and N-t-butyl-α-phenylnitrone, antioxidative agents such as catalase, superoxide dismutase, thiourea, and uric acid, and inhibitors of lipid peroxidation, such as vitamin E phosphate, Trolox, and diphenylphenylenediamine. Besides ethanol, other substrates such as Me2SO, CCl4, isoniazid, and N,N-dimethylnitrosamine were cytotoxic to cells expressing CYP2E1 but not to control cells. These results indicate that ethanol was toxic to HepG2 cells which express human CYP2E1 by a pathway sensitive to inhibitors of CYP2E1 and to a variety of antioxidative agents. This model appears to be useful in efforts to establish a CYP2E1-dependent ethanol hepatotoxicity system and to evaluate the role of oxidative stress and reactive radical species in the toxicity by ethanol.
The Representation of Snow in Land Surface Schemes: Results from PILPS 2(d)

Twenty-one land surface schemes (LSSs) performed simulations forced by 18 yr of observed meteorological data from a grassland catchment at Valdai, Russia, as part of the Project for the Intercomparison of Land-Surface Parameterization Schemes (PILPS) Phase 2(d). In this paper the authors examine the simulation of snow. In comparison with observations, the models are able to capture the broad features of the snow regime on both an intra- and interannual basis. However, weaknesses in the simulations exist, and early season ablation events are a significant source of model scatter. Over the 18-yr simulation, systematic differences between the models’ snow simulations are evident and reveal specific aspects of snow model parameterization and design as being responsible. Vapor exchange at the snow surface varies widely among the models, ranging from a large net loss to a small net source for the snow season. Snow albedo, fractional snow cover, and their interplay have a large effect on energy available for ablation, with differences among models most evident at low snow depths. The incorporation of the snowpack within an LSS structure affects the method by which snow accesses, as well as utilizes, available energy for ablation. The sensitivity of some models to longwave radiation, the dominant winter radiative flux, is partly due to a stability-induced feedback and the differing abilities of models to exchange turbulent energy with the atmosphere. Results presented in this paper suggest where weaknesses in macroscale snow modeling lie and where both theoretical and observational work should be focused to address these weaknesses.
Proteins Homologous to p47phox and p67phox Support Superoxide Production by NAD(P)H Oxidase 1 in Colon Epithelial Cells*

Superoxide production by phagocytes involves activation of a multi-component NADPH oxidase. Recently, several homologues of the catalytic component of the phagocyte oxidase, gp91phox, were identified in various tissues. Here we describe two proteins, p41 and p51, with significant homology to two cytosolic components of the phagocytic oxidase, p47phox and p67phox. Like p47phox, p41 contains an amino-terminal Phox homology domain, two SH3 domains, and a conserved carboxyl-terminal, proline-rich motif. Similarly, p51 is homologous to p67phox, containing four amino-terminal tetratrico-peptide repeats, a conserved “activation domain” motif, a PB1 domain, and a carboxyl-terminal SH3 domain. The highest levels of p41 transcript are detected in the colon and in other gastrointestinal tissues that express Nox1, the predominant gp91phox homologue in these tissues. In contrast, the p51 transcript showed a more widespread expression pattern, suggesting that it may support other tissue-specific oxidases. Mouse colon in situ hybridization detected both transcripts in the epithelial cells of colon crypts. Heterologous co-expression of p41 and p51 significantly enhances the superoxide-generating activity of Nox1-expressing cells; thus, p41 and p51 appear to be novel regulators of Nox1. These proteins also support the activity of gp91phox, albeit at much lower levels than the cytosolic phox counterparts. Our results suggest colon epithelial cells contain a multi-component NAD(P)H oxidase with a molecular architecture similar to the phagocytic oxidase.
Batch fabrication of carbon nanotube bearings

Relative displacements between the atomically smooth, nested shells in multiwalled carbon nanotubes (MWNTs) can be used as a robust nanoscale motion enabling mechanism. Here, we report on a novel method suited for structuring large arrays of MWNTs into such nanobearings in a parallel fashion. By creating MWNT nanostructures with nearly identical electrical circuit resistance and heat transport conditions, uniform Joule heating across the array is used to simultaneously engineer the shell geometry via electric breakdown. The biasing approach used optimizes process metrics such as yield and cycle-time. We also present the parallel and piecewise shell engineering at different segments of a single nanotube to construct multiple, but independent, high density bearings. We anticipate this method for constructing electromechanical building blocks to be a fundamental unit process for manufacturing future nanoelectromechanical systems (NEMS) with sophisticated architectures and to drive several nanoscale transduction applications such as GHz-oscillators, shuttles, memories, syringes and actuators.
Learning to Perceive with a Visuo — Auditory Substitution System: Localisation and Object Recognition with ‘The Voice’

We investigated to what extent participants can acquire the mastery of an auditory-substitution-of-vision device (‘The vOICe’) using dynamic tasks in a three-dimensional environment. After extensive training, participants took part in four experiments. In the first experiment we explored locomotion and localisation abilities. Participants, blindfolded and equipped with the device, had to localise a target by moving a hand-held camera, walk towards the target, and point at it. In the second experiment, we studied the localisation ability in a constrained pointing task. In the third experiment we explored participants' ability to recognise natural objects via their auditory rendering. In the fourth experiment we tested the ability of participants to discriminate objects belonging to the same category. We analysed participants' performance from both an objective and a subjective point of view. The results showed that participants, through sensorimotor interactions with the perceptual scene while using the hand-held camera, were able to make use of the auditory stimulation to obtain the information necessary for locomotor guidance, localisation, and pointing, as well as for object recognition. Furthermore, analysis from a subjective perspective yielded insights into participants' qualitative experience and into the strategies they used to master the device, and thus to pass from a kind of deductive reasoning to a form of immediate apprehension of what is being perceived.
Stromal Cells Confer Lymph Node-Specific Properties by Shaping a Unique Microenvironment Influencing Local Immune Responses1

Lymph nodes (LN) consist not only of highly motile immune cells coming from the draining area or from the systemic circulation, but also of resident stromal cells building the backbone of the LN. These two cell types form a unique microenvironment which is important for initiating an optimal immune response. The present study asked how the unique microenvironment of the mesenteric lymph node (mLN) is influenced by highly motile cells and/or by the stromal cells. A transplantation model in rats and mice was established. After resecting the mLN, fragments of peripheral lymph node (pLN) or mLN were inserted into the mesentery. The pLN and mLN have LN-specific properties, resulting in differences of, for example, the CD103+ dendritic cell subset, the adhesion molecule mucosal addressin cell adhesion molecule 1, the chemokine receptor CCR9, the cytokine IL-4, and the enzyme retinal dehydrogenase 2. This new model clearly showed that during regeneration stromal cells survived and immune cells were replaced. Surviving high endothelial venules retained their site-specific expression (mucosal addressin cell adhesion molecule 1). In addition, the low expression of retinal dehydrogenase 2 and CCR9 persisted in the transplanted pLN, suggesting that stromal cells influence the lymph node-specific properties. To examine the functional relevance of this different expression pattern in transplanted animals, an immune response against orally applied cholera toxin was initiated. The data showed that the IgA response against cholera toxin is significantly diminished in animals transplanted with pLN. This model documents that stromal cells of the LN are active players in shaping a unique microenvironment and influencing immune responses in the drained area.
Metal‐dependent SV40 viruses containing inducible enhancers from the upstream region of metallothionein genes.

We have isolated SV40 recombinant viruses which are dependent on heavy metal ions for efficient propagation. They were obtained after‐co‐transfection of enhancerless SV40 DNA (the so‐called enhancer trap) with sonicated DNA from the mouse metallothionein‐I (mMT‐I) or human metallothionein‐IIA (hMT‐IIA) upstream regions. To substitute for the SV40 enhancer, these viruses have incorporated a segment of the immediate upstream region of the metallothionein genes. Two recombinant viruses of the SVMT‐I type carry segments of the mMT‐I gene from positions ‐73 to ‐187 and ‐39 to ‐194 inverted with respect to their natural configuration. The overlapping segment contains two of the four metal‐responsive elements involved in the induction of the mMT‐I gene by heavy metal ions. The SVMT‐II recombinant virus contains a segment of the hMT‐IIA gene from position ‐39 to ‐366 which harbors the metal‐ and hormone‐responsive elements of the hMT‐IIA gene. Insertion of the mMT‐I segment downstream of a rabbit beta‐globin test gene enhances beta‐globin transcription upon metal ion stimulation. This shows that the immediate upstream region of the mouse metalliothionein‐I gene, when detached from its TATA box, can act as an inducible enhancer. It may be generally true that the enhancer/promoters of inducible genes are composed of several regulatory sequence elements which are interspersed with constitutive elements. The number and spatial arrangement of these elements probably determines the basal versus induced level of expression.
Two Functionally Distinct Steps Mediate High Affinity Binding of U1A Protein to U1 Hairpin II RNA*

Binding of the U1A protein to its RNA target U1 hairpin II has been extensively studied as a model for a high affinity RNA/protein interaction. However, the mechanism and kinetics by which this complex is formed remain largely unknown. Here we use real-time biomolecular interaction analysis to dissect the roles various protein and RNA structural elements play in the formation of the U1A·U1 hairpin II complex. We show that neutralization of positive charges on the protein or increasing the salt concentration slows the association rate, suggesting that electrostatic interactions play an important role in bringing RNA and protein together. In contrast, removal of hydrogen bonding or stacking interactions within the RNA/protein interface, or reducing the size of the RNA loop, dramatically destabilizes the complex, as seen by a strong increase in the dissociation rate. Our data support a binding mechanism consisting of a rapid initial association based on electrostatic interactions and a subsequent locking step based on close-range interactions that occur during the induced fit of RNA and protein. Remarkably, these two steps can be clearly distinguished using U1A mutants containing single amino acid substitutions. Our observations explain the extraordinary affinity of U1A for its target and may suggest a general mechanism for high affinity RNA/protein interactions.