Alterations of the Mdm2 C-Terminus Differentially Impact Its Function In Vivo.
Murine double minute 2 (Mdm2) is the principal E3-ubiquitin ligase for p53 and contains a C2H2C4 type RING domain wherein the last cysteine residue is followed by an evolutionarily conserved 13 amino acid C-terminal tail. Previous studies have indicated that integrity of the C-terminal tail is critical for Mdm2 function. Recently, a mutation extending the MDM2 length by five amino acids was identified and associated with enhanced p53 response in fibroblasts and premature aging in a human patient. To investigate the importance of the conserved Mdm2 C-terminal length on p53 regulatory function in vivo, we engineered three novel mouse alleles using CRISPR-Cas9 technology. Genetic studies with these murine models showed that curtailing Mdm2 C-terminal length by even a single amino acid leads to p53-dependent embryonic lethality. Extension of the Mdm2 C-terminal length by five amino acids (QLTCL) yielded viable mice that are smaller in size, exhibit fertility problems, and have a shortened life span. Analysis of early passage mouse embryonic fibroblasts indicated impaired Mdm2 function correlates with enhanced p53 activity under stress conditions. Furthermore, analysis in mice showed tissue-specific alterations in p53 target gene expression and enhanced radiosensitivity. These results confirm the physiological importance of the evolutionarily conserved Mdm2 C-terminus in regulating p53 functions. This in vivo study highlights that alterations to the C-terminus of Mdm2 perturb its regulation of the tumor suppressor p53.


Synchronization by injection of common chaotic signal in semiconductor lasers with optical feedback.
We investigate the dynamics of two semiconductor lasers with separate optical feedback when they are driven by a common signal injected from a chaotic laser under the condition of non-identical drive and response. We experimentally and numerically show conditions under which the outputs of the two lasers can be highly correlated with each other even though the correlation with the drive signal is low. In particular, the effects of the phase of the feedback light on the correlation characteristics are described. The maximum correlation between the two response lasers is obtained when the phase of the feedback light is matched between the two response lasers, while the minimum correlation is observed when the difference in the optical phase is pi. On the other hand, the correlation between the drive and response is not sensitive to the phase of the feedback light, unlike the previously studied case of identical drive and response. We numerically examine the difference between the maximum and minimum cross correlations over a wide range of parameters, and show that it is largest when there is a balance between the injection strength and the feedback strength.


Resolving coiled shapes reveals new reorientation behaviors in C. elegans.
We exploit the reduced space of C. elegans postures to develop a novel tracking algorithm which captures both simple shapes and also self-occluding coils, an important, yet unexplored, component of 2D worm behavior. We apply our algorithm to show that visually complex, coiled sequences are a superposition of two simpler patterns: the body wave dynamics and a head-curvature pulse. We demonstrate the precise Ω-turn dynamics of an escape response and uncover a surprising new dichotomy in spontaneous, large-amplitude coils; deep reorientations occur not only through classical Ω-shaped postures but also through larger postural excitations which we label here as δ-turns. We find that omega and delta turns occur independently, suggesting a distinct triggering mechanism, and are the serpentine analog of a random left-right step. Finally, we show that omega and delta turns occur with approximately equal rates and adapt to food-free conditions on a similar timescale, a simple strategy to avoid navigational bias.


Mechanisms of stochastic onset and termination of atrial fibrillation studied with a cellular automaton model.
Mathematical models of cardiac electrical excitation are increasingly complex, with multiscale models seeking to represent and bridge physiological behaviours across temporal and spatial scales. The increasing complexity of these models makes it computationally expensive to both evaluate long term (more than 60 s) behaviour and determine sensitivity of model outputs to inputs. This is particularly relevant in models of atrial fibrillation (AF), where individual episodes last from seconds to days, and interepisode waiting times can be minutes to months. Potential mechanisms of transition between sinus rhythm and AF have been identified but are not well understood, and it is difficult to simulate AF for long periods of time using state-of-the-art models. In this study, we implemented a Moe-type cellular automaton on a novel, topologically equivalent surface geometry of the left atrium. We used the model to simulate stochastic initiation and spontaneous termination of AF, arising from bursts of spontaneous activation near pulmonary veins. The simplified representation of atrial electrical activity reduced computational cost, and so permitted us to investigate AF mechanisms in a probabilistic setting. We computed large numbers (approx. 105) of sample paths of the model, to infer stochastic initiation and termination rates of AF episodes using different model parameters. By generating statistical distributions of model outputs, we demonstrated how to propagate uncertainties of inputs within our microscopic level model up to a macroscopic level. Lastly, we investigated spontaneous termination in the model and found a complex dependence on its past AF trajectory, the mechanism of which merits future investigation.


Neuronal plasticity after a human spinal cord injury: positive and negative effects.
In patients suffering an incomplete spinal cord injury (SCI) an improvement in walking function can be achieved by providing a functional training with an appropriate afferent input. In contrast, in immobilized incomplete and complete subjects a negative neuroplasticity leads to a neuronal dysfunction. After an SCI, neuronal centers below the level of lesion exhibit plasticity that either can be exploited by specific training paradigms or undergo a degradation of function due to the loss of appropriate input. Load- and hip-joint-related afferent inputs seem to be of crucial importance for the generation of a locomotor pattern and, consequently, the effectiveness of the locomotor training. In severely affected SCI subjects rehabilitation robots allow for a longer and more intensive training and can provide feedback information. Conversely, in severely affected chronic SCI individuals without functional training the locomotor activity in the leg muscles exhausts rapidly during assisted locomotion. This is accompanied by a shift from early to dominant late spinal reflex components. The exhaustion of locomotor activity is also observed in non-ambulatory patients with an incomplete SCI. It is assumed that in chronic SCI the patient's immobility results in a reduced input from supraspinal and peripheral sources and leads to a dominance of inhibitory drive within spinal neuronal circuitries underlying locomotor pattern and spinal reflex generation. A training with an enhancement of an appropriate proprioceptive input early after an SCI might serve as an intervention to prevent neuronal dysfunction.


Assessing exposure, uptake and toxicity of silver and cerium dioxide nanoparticles from contaminated environments.
The aim of this project was to compare cerium oxide and silver particles of different sizes for their potential for uptake by aquatic species, human exposure via ingestion of contaminated food sources and to assess their resultant toxicity. The results demonstrate the potential for uptake of nano and larger particles by fish via the gastrointestinal tract, and by human intestinal epithelial cells, therefore suggesting that ingestion is a viable route of uptake into different organism types. A consistency was also shown in the sensitivity of aquatic, fish cell and human cell models to Ag and CeO2 particles of different sizes; with the observed sensitivity sequence from highest to lowest as: nano-Ag > micro Ag > nano CeO2 = micro CeO2. Such consistency suggests that further studies might allow extrapolation of results between different models and species.


Comparison of density functionals for nitrogen impurities in ZnO.
Hybrid functionals and empirical correction schemes are compared to conventional semi-local density functional theory (DFT) calculations in order to assess the predictive power of these methods concerning the formation energy and the charge transfer level of impurities in the wide-gap semiconductor ZnO. While the generalized gradient approximation fails to describe the electronic structure of the N impurity in ZnO correctly, methods that widen the band gap of ZnO by introducing additional non-local potentials yield the formation energy and charge transfer level of the impurity in reasonable agreement with hybrid functional calculations. Summarizing the results obtained with different methods, we corroborate earlier findings that the formation of substitutional N impurities at the oxygen site in ZnO from N atoms is most likely slightly endothermic under oxygen-rich preparation conditions, and introduces a deep level more than 1 eV above the valence band edge of ZnO. Moreover, the comparison of methods elucidates subtle differences in the predicted electronic structure, e.g., concerning the orientation of unoccupied orbitals in the crystal field and the stability of the charged triplet state of the N impurity. Further experimental or theoretical analysis of these features could provide useful tests for validating the performance of DFT methods in their application to defects in wide-gap materials.


The structure and genetic control of a new class of disulphide-linked proteins in wheat endosperm.
Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of unreduced total protein extracts from the endosperm of hexaploid wheat revealed three high molecular weight protein bands (triplet bands) in a zone of heavy background streaking. Electrophoretic examination of 135 hexaploid cultivars showed at least five different patterns of these triplet bands. Nine durum wheat cultivars showed a single band only. Analysis of nullisomic-tetrasomic and ditelocentric lines of 'Chinese Spring' wheat revealed that the slowest moving band (Tri-1) of the triplet was controlled by gene(s) on chromosome arm 1DS and the fastest moving band (Tri-3) by 1AS. The band with intermediate mobility (Tri-2) was found to be a hybrid aggregate of the subunits controlled by 1DS and 1AS. Using a non-reducing/reducing form of 2-dimensional (2-D) electrophoresis, these triplet bands were shown to be heterotetramers of four subunits designated D (M.W. 58,000), δ (22,000), A (52,000) and α (23,000) where Tri-1=DδDδ, Tri-2 = DδAα and Tri-3 = AαAα. With very low concentrations of 2-mercaptoethanol (ME), the tetramers dissociated into dimeric subunit pairs (Dδ, Aα), the monomers being observed with higher concentrations of ME. The structure of these subunit pairs resembles that of the subunit pairs in the globulin storage proteins of oats and some legumes. The 2-D method employed in this study was useful also for separating low molecular weight (LMW) subunits of glutenin from the monomeric gliadins which have similar electrophoretic mobility in 1-D separation. It was shown that at least four of these LMW glutenin subunits are controlled by genes on 1DS and 1AS and at least one subunit is controlled by gene(s) on 1BS. This electrophoretic separation method has proven useful in understanding the aggregation behaviour of the seed proteins of wheat.


Infant acceptance of breast milk after maternal exercise.
Previous research reported that breast milk lactic acid (LA) levels increase after lactating women complete a bout of exhaustive exercise, resulting in poor infant acceptance of the postexercise breast milk. This highly publicized finding may not apply to more practical, everyday exercise conditions of lactating women. The purpose of the present study was to reexamine the composition and infant acceptance of postexercise breast milk while controlling maternal diet, exercise intensity, and the method, timing, and assessment of infant feeding. Twenty-four women, 2 to 4 months' postpartum, completed 3 test sessions: a maximal oxygen uptake test, a 30-minute bout of moderate exercise, and a resting control session. One hour before and 1 hour after each session, participants fully expressed their milk, placed it in a bottle familiar to the infant, fed their infant, and rated their infant's acceptance of the milk. Each feeding was videotaped and viewed individually by 3 lactation consultants who rated infant acceptance; consultants were blinded to the test sessions. Milk was analyzed for LA and infant milk consumption was measured. There were no differences in presession versus postsession values for maternal skin temperature, breast milk temperature, and infant milk acceptance as judged by either the mothers or lactation consultants. These results prevailed despite a small but significant increase in breast milk LA premaximal versus postmaximal exercise (0.09 vs 0.21 mM, respectively); there was no difference in milk LA premoderate versus postmoderate exercise, or prerest versus postrest. These data support the hypothesis that moderate or even high-intensity exercise during lactation does not impede infant acceptance of breast milk consumed 1 hour postexercise.


Regulatory effects of phospholamban on cardiac function in intact mice.
Phospholamban (PLB) regulates Ca(2+)-adenosinetriphosphatase activity in cardiac sarcoplasmic reticulum and participates in the regulation of myocardial performance. Animal models with altered levels of PLB permit in vivo evaluation of the physiological role of PLB. This study examined left ventricular (LV) performance in intact PLB heterozygous and homozygous mice under basal and stimulated conditions. A Millar Mikro-Tip transducer was inserted into the right carotid artery and advanced into the LV for direct measurement of ventricular pressure and the first derivative of intraventricular pressure (dP/dt). Baseline blood pressures were increased in PLB heterozygotes and even more so in PLB homozygotes compared with wild types (WT), and there were no differences in heart rate or LV end-diastolic pressure. The increase in pressure was primarily caused by an increase in systolic pressure. Baseline values for positive and negative dP/dt were linearly correlated with PLB levels. In PLB heterozygotes, contractile response to isoproterenol (Iso) was blunted compared with WT, but maximum rates of contraction were similar between the two groups. Contractile performance in PLB homozygous mice, which under baseline conditions was similar to maximum levels seen in WT, showed a blunted response to Iso, and maximum rates of contraction were significantly greater than in either of the other groups, indicating an essential but perhaps not exclusive role for PLB in mediating the inotropic effects of beta-adrenergic agonists. The effects of Iso on negative dP/dt were also blunted in both PLB heterozygous and PLB homozygous animals. Our results demonstrate that myocardial function in highly dependent on PLB level and suggest that the cardiovascular effects of PLB perturbations are largely uncompensated for in the intact mouse.


Cold defense mechanisms in vagotomized rats.
Subdiaphragmatically vagotomized rats cannot mount a febrile response to pyrogens and are believed to have severe thermoregulatory deficiencies. We addressed the issue of thermoeffector competence of vagotomized rats by asking three questions. In Expt. 1 we asked, can vagotomized rats readily recruit tail skin vasoconstriction in the course of a moderate cold exposure? In Expt. 2 the question was, can brown adipose tissue (BAT) thermogenesis readily be activated in vagotomized rats (e.g., in response to a tail pinch)? In Expt. 3, we investigated the question: can vagotomized rats elevate their body temperature in response to ephedrine (a drug of high hyperthermizing potential) to the same extent as sham-operated controls? Rats were vagotomized or sham operated and implanted with a catheter into the jugular vein and a thermocouple into the interscapular BAT. To prevent the common complications of vagotomy, special perioperative care was given. During experiments, colonic, tail skin, and BAT temperatures (Tc, Tsk, and TBAT, respectively) were measured. The vagotomized animals were well nourished and had a body mass (325 +/- 6 g) similar to that of the controls (338 +/- 6 g). In Expt. 1, in response to external cooling (15 degrees C, 1 h), the vagotomized (n = 30) and sham-operated (n = 31) rats recruited tail skin vasoconstriction at close values of both Tc (37.84 +/- 0.08 and 37.97 +/- 0.07 degrees C) and Tsk (33.16 +/- 0.17 and 33.18 +/- 0.18 degrees C, respectively). In Expt. 2, tail pinch-associated stress in vagotomized rats resulted in a sharp rise in the TBAT-Tc gradient by 0.3-1.0 degree C. In Expt. 3, ephedrine administered intravenously (whether in a 5 or 35 mg/kg dose) evoked similar hyperthermic responses in the vagotomized and sham-operated rats: a moderate (approximately 2.5 degrees C) Tc rise in the low dose and a "supramaximal" (approximately 5.0 degrees C) rise in the high dose. In sum, the answer to all three questions asked is yes. Vagotomized rats, at least when well nourished, exhibit no signs of thermoeffector deficiency. It is, therefore, not effector incompetence but rather vagal deafferentation per se that can explain the febrile irresponsiveness of vagotomized rats.


Cysteine conjugate beta-lyase in the gastrointestinal bacterium Eubacterium limosum.
A cysteine conjugate beta-lyase (beta-lyase) from the gastrointestinal bacterium Eubacterium limosum has been isolated and characterized. This organism has the highest specific activity for cysteine conjugate beta-lyase of the gastrointestinal bacteria studied. The beta-lyase was found to cleave the thioether linkage of S-alkyl- and S-aryl-L-cysteine conjugates. Stoichiometric amounts of 2-mercaptobenzothiazole, pyruvic acid, and ammonia were produced from the beta-lyase cleavage of S-(2-benzothiazolyl)-L-cysteine. The enzyme activity was inhibited by hydroxylamine, iodoacetic acid, or KCN. The enzyme appears to be a 75,000-Da dimer of two 38,000-Da subunits. A natural substrate, cystathionine, was cleaved by this enzyme, indicating that this beta-lyase has beta-cystathionase activity. These data suggest that a beta-cystathionase from E. limosum may be an important enzyme in the metabolism of a wide range of cysteine conjugates of xenobiotics to thiol-containing products.


The Clinical Stages of Sporadic Creutzfeldt-Jakob Disease with Met/Met Genotype in Korean Patients.
Clinical diagnosis of sporadic Creutzfeldt-Jakob disease (sCJD) is currently based on changes occurring in the late disease stages, which limits early-stage detection. Therefore, we investigated the disease course from the vague symptomatic to the terminal phase. We retrospectively reviewed 36 sCJD patient records, classifying the disease progression into 4 stages based on clinical manifestations: vague symptomatic, possible CJD, probable CJD and chronic vegetative state. We analyzed findings from diffusion-weighted imaging (DWI), electroencephalography (EEG) and cerebrospinal fluid (CSF) 14-3-3 protein testing performed at each stage. In stage 1, the most distinctive feature was DWI hyperintensities in the neocortex, even with negative CSF 14-3-3 protein and EEG results. In stage 2, DWI hyperintensities in the limbic cortex were more remarkable. CSF 14-3-3 protein testing yielded positive results in >80% of patients; EEG showed sensitivity in <30% of patients. With progression toward stage 3, DWI hyperintensities in the subcortical nucleus increased, with a sustained higher rate of hyperintensities in the limbic and neocortical regions. With gradual progression to stage 4, the sensitivity of CSF 14-3-3 protein testing and EEG decreased and increased, respectively within limited data. Understanding disease stage-dependent differences in clinical symptoms and laboratory test results will facilitate early and accurate diagnosis.


Unknown partner for USP6 and unusual SS18 rearrangement detected by fluorescence in situ hybridization in a solid aneurysmal bone cyst.
USP6 rearrangement is the most common genetic abnormality in primary aneurysmal bone cyst, and SS18 rearrangement has not been previously described in any type of tumor where synovial sarcoma was excluded from the differential diagnosis. We report a case of solid aneurysmal bone cyst in which fluorescence in situ hybridization (FISH) analysis indicated rearrangements of both USP6 and SS18, but histologic features were consistent with aneurysmal bone cyst throughout the lesion. Reverse-transcription polymerase chain reaction (RT-PCR) for the SS18-SSX1 and SS18-SSX2 translocations, identity testing, and SS18 FISH were performed on cytogenetic monolayer cultures and formalin-fixed paraffin-embedded (FFPE) tissue. Genomic microarray, FISH, and immunohistochemistry were performed on follow-up studies of the FFPE specimen. The karyotype was 45,X,add(X)(p11.2),add(4)(q13),add(8)(p21),-13,add(17)(p11.2),add(18)(q11.2) in all 20 cells analyzed from monolayer cultures. The karyotype showed no cytogenetically visible alterations of chromosomal regions harboring known partners for USP6. Metaphase FISH with a commercial SS18 break-apart probe showed translocation of the 5' portion of the SS18 probe to the short arm of the derivative X, as is observed in synovial sarcoma. RT-PCR showed no evidence of a SS18-SSX fusion, and immunohistochemistry was negative for TLE1, EMA, and cytokeratin AE1/3 expression. FISH on FFPE sections with a custom break-apart probe flanking USP6 showed evidence for a USP6 rearrangement throughout the tumor (25-50%). FISH on FFPE sections with a commercial SS18 break-apart FISH probe showed more variable results (0-50% split signals). There was no evidence of a SS18-USP6 fusion by FISH or RT-PCR. A molecular inversion probe array revealed a deletion encompassing the entire SS18 gene and its promoter, as well as portions of the region targeted by the commercial SS18 FISH probe. In conclusion, results obtained from commercially available FISH probes may occasionally yield misleading results. In this case, the SS18 rearrangement by FISH resulted from a complex rearrangement of 18q11.2 with a deletion of the SS18 gene. The translocation partner for USP6 remains unknown in this case.


[Pain and painkillers. 1. Neurophysiologic component].
A pain impulse is transmitted in the thin nerves from nerve cell to nerve cell, by means of neurotransmitters and -modulators, shed in the synapses. In the dorsal horn of the spinal medulla the impulse is either or not allowed to travel to the brain by a 'gate control' mechanism, and pain may be felt. The gate is influenced by cells in the nearby situated substantia gelatinosa, thick nerves and efferent nerves from higher situated centra.


Effect of long-term intensive lipid-lowering therapy with rosuvastatin on progression of carotid intima-media thickness--Justification for Atherosclerosis Regression Treatment (JART) extension study.
Recently, it was reported from the Justification for Atherosclerosis Regression Treatment (JART) Study that intensive therapy with rosuvastatin significantly slowed progression of carotid intima-media thickness (IMT) compared with conventional therapy with pravastatin at 12 months. To assess the long-term efficacy of intensive therapy, the present extension study was conducted. Subjects in the intensive therapy group of the JART Study were asked to participate in the extension study and to continue rosuvastatin treatment. A total of 113 subjects were enrolled into the extension study and were included in the analysis. At 24 months, the mean daily dose of rosuvastatin (±SD) was 7.9±2.9 mg. Mean change in mean IMT was -0.005 mm (range, -0.024 to 0.015 mm) at 24 months (P=0.633, compared with baseline). Rosuvastatin lowered low-density lipoprotein cholesterol (mean±SD) by 46.4±13.8% and elevated high-density lipoprotein cholesterol (mean±SD) by 8.9±24.0% at 24 months compared with baseline. Gray scale median was measured in 25 subjects. It increased by 16.93±33.12 (mean±SD) % at 12 months and by 22.50±52.83% at 24 months from baseline (P=0.017, P=0.044, respectively). Two-year treatment with rosuvastatin inhibited progression of carotid IMT. Rosuvastatin also improved the plaque composition, and this qualitative change occurred relatively early after starting therapy.


Fruiting bodies of the social amoeba Dictyostelium discoideum increase spore transport by Drosophila.
Many microbial phenotypes are the product of cooperative interactions among cells, but their putative fitness benefits are often not well understood. In the cellular slime mold Dictyostelium discoideum, unicellular amoebae aggregate when starved and form multicellular fruiting bodies in which stress-resistant spores are held aloft by dead stalk cells. Fruiting bodies are thought to be adaptations for dispersing spores to new feeding sites, but this has not been directly tested. Here we experimentally test whether fruiting bodies increase the rate at which spores are acquired by passing invertebrates. Drosophila melanogaster accumulate spores on their surfaces more quickly when exposed to intact fruiting bodies than when exposed to fruiting bodies physically disrupted to dislodge spore masses from stalks. Flies also ingest and excrete spores that still express a red fluorescent protein marker. Multicellular fruiting bodies created by D. discoideum increase the likelihood that invertebrates acquire spores that can then be transported to new feeding sites. These results thus support the long-hypothesized dispersal benefits of altruism in a model system for microbial cooperation.


[The functional status of the small intestine in rats in the dynamics of a bone marrow form of acute radiation sickness].
A study was made of 75Se-methionine absorption rate and contraction and choline-reactivity of smooth muscle cells of rat jejunum longitudinal layer. Whole-body X-irradiation with a dose of 5.8 Gy (LD50/30) caused a sharp decrease in the rate of absorption of 75Se-methionine by the blood from the intestine on days 3--4 after irradiation followed by an incomplete recovery of absorption by the 14th day. At the height of acute radiation sickness there was an 8- to 10-fold decrease in the sensitivity of the smooth muscle cell membrane to acetylcholine the contractile capacity being retained. This was perhaps the cause of the intestine motor activity inhibition. It is concluded that rat intestine functions are suppressed considerably with the bone marrow form of acute radiation sickness.


Investigation of Position Sensing and Energy Harvesting of a Flexible Triboelectric Touch Pad.
Triboelectric nanogenerator (TENG) is a promising technology because it can harvest energy from the environment to enable self-sustainable mobile and wearable electronic devices. In this work, we present a flexible touch pad capable of detecting the contact location of an object and generating substantial energy simultaneously based on the coupling of triboelectric effects and electrostatic induction. The touch pad consists of Polytetrafluoroethylene (PTFE) thin film, multiple Aluminum (Al) electrodes and Polyethylene terephthalate (PET) layers, which can be achieved through low cost, simplified and scalable fabrication process. Different from the conventional multi-pixel-based positioning sensor (i.e., large array of sensing elements and electrodes), the analogue method proposed here is used to implement the positioning function with only four electrodes. Position location can achieve a detecting resolution of as small as 1.3 mm (the size of locating layer is 7.5 cm × 7.5 cm). For the energy harvesting part, a multilayer structure is designed to provide higher current output. The open circuit voltage of the device is around 420 V and the short circuit current can reach up to 6.26 µA with current density of 0.25 µA/cm². The maximum output power obtained is approximately 10 mW, which is 0.4 mW/cm². The flexibility and significantly reduced number of electrodes enable the proposed touch pad to be readily integrated into portable electronic devices, such as intelligent robots, laptops, healthcare devices, and environmental surveys, etc.


ING2 (inhibitor of growth protein-2) plays a crucial role in preimplantation development.
ING2 (inhibitor of growth protein-2) is a member of the ING-gene family and participates in diverse cellular processes involving tumor suppression, DNA repair, cell cycle regulation, and cellular senescence. As a subunit of the Sin3 histone deacetylase complex co-repressor complex, ING2 binds to H3K4me3 to regulate chromatin modification and gene expression. Additionally, ING2 recruits histone methyltransferase (HMT) activity for gene repression, which is independent of the HDAC class I or II pathway. However, the physiological function of ING2 in mouse preimplantation embryo development has not yet been characterized previously. The expression, localization and function of ING2 during preimplantation development were investigated in this study. We showed increasing expression of ING2 within the nucleus from the 4-cell embryo stage onwards; and that down-regulation of ING2 expression by endoribonuclease-prepared small interfering RNA (esiRNA) microinjection results in developmental arrest during the morula to blastocyst transition. Embryonic cells microinjected with ING2-specific esiRNA exhibited decreased blastulation rate compared to the negative control. Further investigation of the underlying mechanism indicated that down-regulation of ING2 significantly increased expression of p21, whilst decreasing expression of HDAC1. These results suggest that ING2 may play a crucial role in the process of preimplantation embryo development through chromatin regulation.


SYT12 plays a critical role in oral cancer and may be a novel therapeutic target.
Synaptotagmin12 (SYT12) has been well characterized as the regulator of transmitter release in the nervous system, however the relevance and molecular mechanisms of SYT12 in oral squamous cell carcinoma (OSCC) are not understood. In the current study, we investigated the expression of SYT12 and its molecular biological functions in OSCC by quantitative reverse transcriptase polymerase chain reaction, immunoblot analysis, and immunohistochemistry. SYT12 were up-regulated significantly in OSCC-derived cell lines and primary OSCC tissue compared with the normal counterparts (P<0.05) and the SYT12 expression levels were correlated significantly with clinical indicators, such as the primary tumoral size, lymph node metastasis, and TNM stage (P<0.05). SYT12 knockdown OSCC cells showed depressed cellular proliferation, migration, and invasion with cell cycle arrest at G1 phase. Surprisingly, we found increased calcium/calmodulin-dependent protein kinase 2 (CAMK2) inhibitor 1 (CAMK2N1) and decreased CAMK2-phosphorylation in the knockdown cells. Furthermore, treatment with L-3, 4-dihydroxyphenylalanine (L-dopa), a drug approved for Parkinson's disease, led to down-regulation of SYT12 and similar phenotypes to SYT12 knockdown cells. Taken together, we concluded that SYT12 plays a significant role in OSCC progression via CAMK2N1 and CAMK2, and that L-dopa would be a new drug for OSCC treatment through the SYT12 expression.


Understanding the cell behavior on nano-/micro-patterned surfaces.
This article reports on studies conducted in the same laboratory on interactions between patterned substrates with different pattern dimensions and chemistries, and various types of cells. In order to compare the influence of various parameters, bone marrow stromal cells, retinal pigment epithelial cells, human corneal stromal cells (keratocytes), Saos-2 (human osteosarcoma cells), human microvascular endothelial cells and vascular smooth muscle cells were tested on surfaces with different physical patterns and chemical properties. It was observed that cell type and surface topography are more influential than surface chemistry in determining the alignment tendency of a cell on a substrate surface. Low walls (several microns high) could not confine cells into the microgrooves of the films but alignment was still possible if the cells had a natural alignment property. This information is very useful in designing tissue engineering scaffolds and in the long-term success of implants.


Imaging performance of finite uniaxial metamaterials with large anisotropy.
Metamaterials with extreme anisotropy overcome the diffraction limit by supporting the propagation of otherwise evanescent waves. Recent experiments in slabs of wire media have shown that images deteriorate away from the longitudinal Fabry-Perot resonances of the slab. Existing theoretical models explain this using nonlocality, surface waves, and additional boundary conditions. We show that image aberrations can be understood as originating from cavity resonances of uniaxial media with large local axial permittivity. We apply a simple cavity resonator model and a transfer matrix approach to replicate salient experimental features of wire media hyperlenses. These results offer avenues to reduce observed imaging artefacts, and are applicable to all uniaxial media with large magnitude of the axial permittivity, e.g., wire media and layered media.


Cutaneous epitheliotropic T-cell lymphoma in the cat: a review of the literature and five new cases.
Cutaneous epitheliotropic T-cell lymphoma (CETL) is characterized by cutaneous infiltration of neoplastic T lymphocytes with a specific tropism for the epidermis and adnexal epithelium. This disease is reported very rarely in the cat. Clinical data were collected from an informal discussion with veterinary dermatologists through the Vetdermlist (firstname.lastname@example.com). In parallel, case archives of two European diagnostic histopathology laboratories (Institut de Pathologie et Génétique/Bio.be Gosselies, Belgium and the School of Veterinary Sciences, University of Bristol, UK) were reviewed. Fifteen cases with a good clinical description were selected, and five sets of skin biopsies were available for review. Cutaneous epitheliotropic T-cell lymphoma generally affects older cats with no apparent sex or breed predisposition. Solitary or multiple lesions were reported without predilection for any particular location. The lesions consisted of erythematous plaques or patches, scaly alopecic patches and nonhealing ulcers or nodules, which sometimes mimicked an eosinophilic plaque. Pruritus was rarely reported. No lesions affecting the oral mucosa were observed. Clinical diagnosis of CETL is more challenging in cats than in dogs. Final diagnosis must be based on histopathological examination of skin biopsy samples. The characteristic lesions of feline CETL are similar to those reported in the dog, but involvement of the adnexal glands was not observed in this series (n=5). The neoplastic T cells were generally small to medium in size. The survival time of cats with CETL seems to be more variable than that of affected dogs. Too few cases have been evaluated to permit clear recommendations to be made with respect to treatment.


A multicenter cohort study of treatments and hospital outcomes in neonatal abstinence syndrome.
To compare pharmacologic treatment strategies for neonatal abstinence syndrome (NAS) with respect to total duration of opioid treatment and length of inpatient hospital stay. We conducted a cohort analysis of late preterm and term neonates who received inpatient pharmacologic treatment of NAS at one of 20 hospitals throughout 6 Ohio regions from January 2012 through July 2013. Physicians managed NAS using 1 of 6 regionally based strategies. Among 547 pharmacologically treated infants, we documented 417 infants managed using an established NAS weaning protocol and 130 patients managed without protocol-driven weaning. Regardless of the treatment opioid chosen, when we accounted for hospital variation, infants receiving protocol-based weans experienced a significantly shorter duration of opioid treatment (17.7 vs. 32.1 days, P < .0001) and shorter hospital stay (22.7 vs. 32.1 days, P = .004). Among infants receiving protocol-based weaning, there was no difference in the duration of opioid treatment or length of stay when we compared those treated with morphine with those treated with methadone. Additionally, infants treated with phenobarbital were treated with the drug for a longer duration among those following a morphine-based compared with methadone-based weaning protocol. (P ≤ .002). Use of a stringent protocol to treat NAS, regardless of the initial opioid chosen, reduces the duration of opioid exposure and length of hospital stay. Because the major driver of cost is length of hospitalization, the implications for a reduction in cost of care for NAS management could be substantial.


Bipolar depression: criteria for treatment selection, definition of refractoriness, and treatment options.
This paper reviews controlled studies of bipolar depression, outlines criteria for choosing treatment, defines refractoriness in bipolar depression, and provides options for treatment of refractory bipolar depression. Controlled studies that examined the efficacy of treatments for acute and long-term treatment of bipolar depression were located through electronic searches of several databases and by manual crosssearch of references and proceedings of international meetings. Lithium comes close to fulfilling the proposed criteria for first-line treatment for bipolar depression, and those not responding to lithium should be considered to have refractory bipolar depression. Options for such patients include addition of lamotrigine or a second mood stabilizer, or a newer-generation antidepressant such as a serotonin re-uptake inhibitor or bupropion, or the atypical antipsychotic olanzapine. Although there is a paucity of research in the treatment of refractory bipolar depression, available data could be used for providing rational treatment options for such patients. However, further studies are urgently needed to determine which options are most appropriate for which type of patients.


Anti-inflammatory and immunomodulatory effects of bortezomib in various in vivo models.
Bortezomib (Velcade®) is a proteasome inhibitor that has been approved for the treatment of multiple myeloma and mantle cell lymphoma. It has been shown to inhibit the expression of cell adhesion molecules, co-stimulatory molecules, and NFκB activation, to deplete alloreactive T lymphocytes, and to decrease Th1 cytokine production. The anti-inflammatory effects of bortezomib were further investigated in this current set of studies. Systemic treatment with bortezomib was efficacious in the thioglycolate-induced MCP-1 production model, and the dinitrofluorobenzene-induced delayed-type hypersensitivity model. Psoriasis is an autoimmune disease that affects about 2% of the world population. Many treatments have been reported with varying degrees of efficacy. A topical bortezomib formulation was developed to minimize systemic exposure. Its tolerability was investigated in a topical imiquimod (IMQ)-induced psoriasis model. Daily application of IMQ on mouse skin induced inflamed scaly skin lesions resembling plaque-type psoriasis. Fatality was observed in the 1-mg/ml dose group. At 0.1 and 0.01 mg/ml, bortezomib potentiated IMQ-induced erythema, scaling, skin thickening, and caused necrotic lesions. Lower doses had no effect on the clinical observations. Histologically, bortezomib dose-dependently increased parakeratosis, hyperkeratosis, acanthosis, and inflammatory cell infiltration. This study demonstrated that topical bortezomib is not suitable for the treatment of psoriasis.


Limulus alpha 2-macroglobulin. First evidence in an invertebrate for a protein containing an internal thiol ester bond.
Intra-chain thiol ester bonds are present in a limited number of proteins. The thiol ester class of proteins includes vertebrate alpha 2-macroglobulin and the complement proteins C3 and C4. We report here the first instance of a thiol ester protein from an invertebrate, the alpha 2-macroglobulin proteinase-inhibitor homologue present in the plasma of the American horseshoe crab Limulus polyphemus. Our evidence is of three kinds: (1) the proteinase-binding activity of Limulus alpha 2-macroglobulin is inactivated by the low-molecular-mass primary amine methylamine; (2) the native protein is subject to autolytic fragmentation during mild thermal denaturation, yielding fragments of approx. 125 kDa and 55 kDa, whereas the methylamine-treated protein is stable under these conditions of thermal treatment; (3) new thiol groups are generated rapidly during reaction of the protein with trypsin. The demonstration of the thiol ester bond in a protein from an ancient invertebrate provides evolutionary evidence for the importance of this bond in the function of plasma forms of the alpha 2-macroglobulin-like proteinase inhibitors.


Development and Emergence of Individual Variability in the Functional Connectivity Architecture of the Preterm Human Brain.
Individual variability in human brain networks underlies individual differences in cognition and behaviors. However, researchers have not conclusively determined when individual variability patterns of the brain networks emerge and how they develop in the early phase. Here, we employed resting-state functional MRI data and whole-brain functional connectivity analyses in 40 neonates aged around 31-42 postmenstrual weeks to characterize the spatial distribution and development modes of individual variability in the functional network architecture. We observed lower individual variability in primary sensorimotor and visual areas and higher variability in association regions at the third trimester, and these patterns are generally similar to those of adult brains. Different functional systems showed dramatic differences in the development of individual variability, with significant decreases in the sensorimotor network; decreasing trends in the visual, subcortical, and dorsal and ventral attention networks, and limited change in the default mode, frontoparietal and limbic networks. The patterns of individual variability were negatively correlated with the short- to middle-range connection strength/number and this distance constraint was significantly strengthened throughout development. Our findings highlight the development and emergence of individual variability in the functional architecture of the prenatal brain, which may lay network foundations for individual behavioral differences later in life.


Dual modulation of osteoclast differentiation by lipopolysaccharide.
Lipopolysaccharide (LPS) modulates bone resorption by augmentation of osteoclastogenesis. It increases in osteoblasts the production of RANKL, interleukin (IL)-1, prostaglandin E2 (PGE2), and TNF-alpha, each known to induce osteoclast activity, viability, and differentiation. We examined the role of direct interactions of LPS with osteoclast precursors in promoting their differentiation. To this end, we have used bone marrow mononuclear cell preparations in the absence of osteoblasts or stromal cells. We found that LPS does not induce osteoclast differentiation in these cells. Moreover, the inclusion of LPS blocked the osteoclastogenic activity of RANKL. However, LPS is a potent inducer of osteoclastogenesis in RANKL-pretreated cells, even if present in the absence of exogenous RANKL. Osteoprotegerin (OPG), does not affect the stimulatory phase of LPS modulation of osteoclastogenesis, ruling out involvement of endogenous RANKL. LPS induces the expression of TNF-a and IL-1beta in osteoclast precursors, regardless if they were or were not pretreated with RANKL. These two cytokines induced osteoclast differentiation in RANKL-pretreated cells. To examine if these cytokines mediate LPS effect in an autocrine mechanism, we measured the effect of their neutralization on LPS osteoclastogenic activity. Although neutralization of IL-1beta did not affect LPS activity, a marked inhibition was observed when TNF-alpha was neutralized. However, TNF-a expression was increased also in conditions in which LPS inhibited RANKL osteoclastogenic activity. We found that LPS reduces the expression of RANK and macrophage colony-stimulating factor (M-CSF) receptor. In summary, LPS impacts on osteoclastogenesis also via its interactions with the precursor cells. LPS inhibits RANKL activity by reducing the expression of RANK and M-CSF receptor and stimulates osteoclastogenesis in RANKL-pretreated cells via TNF-alpha.


Syrinx reduction due to spontaneous spinal cord tear: demonstration on 3 T MRI and review of the literature.
A holocord syringomyelia due to Chiari 1.5 malformation (CM) in a 12-year-old girl was serially imaged with 3 T MRI over 4 years. The serial MRI showed reduction in size of the syrinx, without any surgical intervention or CM improvement, but rather due to spontaneous spinal cord tear. The tear was clearly demonstrated by evidence of flow signal across the tear between syrinx and subarachnoid space at the upper thoracic level. The tear showed spontaneous closure at follow-up. A medullary tear has been described in the adult population as one of the putative causes of spontaneous syringomyelia reduction, but its clear demonstration with modern high-resolution MRI has not been reported in the paediatric population. Moreover, this is the first reported case of syrinx reduction due to spontaneous fissuration in a paediatric patient.


Associations between rotating night shifts, sleep duration, and telomere length in women.
Telomere length has been proposed as a marker of aging. However, our knowledge of lifestyle risk factors determining telomere length is limited. We evaluated the associations between years of rotating night shifts, self-reported sleep duration, and telomere length in 4,117 female participants from the Nurses' Health Study. Telomere length in peripheral blood leukocytes was determined by Real-Time PCR assay. Information on rotating night shifts and sleep duration was collected via questionnaires prior to blood collection. We used multivariable linear regression to investigate the associations between rotating night shifts, sleep duration, and telomere length. Compared with women in the category (9 hours), those in the lowest category of sleep duration (≤6 hours) had a 0.12 unit decrease in z score after adjustment for age, BMI and cigarette smoking (equivalent to 9-year telomere attrition, P for trend = 0.05). Significant positive association between sleep duration and telomere length was seen among women under age of 50 (P for trend = 0.004), but not among those over 50 (P for trend = 0.33) (P for interaction = 0.005). In addition, we observed that women with a longer history of rotating night shifts tended to have shorter telomere length, but this relation was not statistically significant (P for trend = 0.36). We found that sleep duration was positively associated with telomere length among women under 50 years old. Further research is needed to confirm the observed associations.


Identification of the Obstetric Factors Increasing Tendency to Smoking Cessation During Pregnancy.
Cigarette, one of the various forms of tobacco, is the most commonly used tobacco product in Turkey and in the world. Cigarette has several negative effects on general health and is a major problem in the pregnancy period. This study aims to identify pregnancy-related factors that affect the tendency to smoking cessation and their effect levels in pregnant women who continue to smoke during pregnancy. The sample of this descriptive and cross-sectional study was 430 pregnant women who smoked. Data were collected through an original interview form and face-to-face interviews. Data analysis was performed using chi-square, independent t test, and multiple logistic regression model in terms of demographic, obstetric, maternal, and fetal features. The average number of cigarettes smoked before pregnancy was 13.28 ± 8.62, whereas the average number of cigarettes smoked during pregnancy was 6.60 ± 7.64. The tendency to decrease smoking increases 2.8 times ( OR = 2.825, 95% CI [1.631, 4.895]) with the number of pregnancies and approximately 2 times in case of a planned pregnancy ( OR = 1.946, 95% CI [1.076, 3.520]). The frequency of having prenatal visits showed a weak but significant relationship with the number of abortuses and the number of living children. Developing a risk map in line with the findings of the study and considering the obstetric features of smoking women could enable to hypothesize about the types of behaviors in smoking in the following processes of pregnancy. With the precautions to be taken, the negative effects of smoking on maternal and fetal health could be prevented or minimized.


Hot and cold water infusion aroma profiles of Hibiscus sabdariffa: fresh compared with dried.
Calyxes from the Roselle plant (Hibiscus sabdariffa L.) were used to prepare cold (22 °C for 4 h) and hot (98 °C for 16 min) infusions/teas from both fresh and dried forms. Aroma volatiles were extracted using static headspace SPME and analyzed using GC-MS and GC-O with 2 different columns (DB-5 and DB-Wax). Totals of 28, 25, 17, and 16 volatiles were identified using GC-MS in the dried hot extract (DHE), dried cold extract (DCE), fresh hot extract (FHE), and fresh cold extract (FCE) samples, respectively. In terms of total GC-MS peak areas DHE ≫ DCE > FHE ≫ FCE. Nonanal, decanal, octanal, and 1-octen-3-ol were among the major volatiles in all 4 beverage types. Thirteen volatiles were common to all 4 teas. Furfural and 5-methyl furfural were detected only in dried hibiscus beverages whereas linalool and 2-ethyl-1-hexanol were detected only in beverages from fresh hibiscus. In terms of aroma active volatiles, 17, 16, 13, and 10 aroma active volatiles were detected for DHE, DCE, FHE, and FCE samples, respectively. The most intense aroma volatiles were 1-octen-3-one and nonanal with a group of 4 aldehydes and 3 ketones common to all samples. Dried samples contained dramatically higher levels of lipid oxidation products such as hexanal, nonanal, and decanal. In fresh hibiscus extracts, linalool (floral, citrus) and octanal (lemon, citrus) were among the highest intensity aroma compounds but linalool was not detected in any of the dried hibiscus extracts. Hibiscus teas/infusions are one of the highest volume specialty botanical products in international commerce. The beverage is consumed for both sensory pleasure and health attributes and is prepared a number of ways throughout the world. Although color and taste attributes have been examined, little information is known about its aroma volatiles and no other study has compared extractions from both fresh and dried as well as extraction temperature differences. This is also, apparently, the first study to identify the aroma active volatiles in hibiscus beverages using GC-olfactometry. Manufacturers and consumers will now have a better understanding of why hibiscus teas prepared in different ways from either fresh or dried forms have a different flavor quality and intensity.


Changing Inequalities in Health-Adjusted Life Expectancy by Income and Race in South Africa.
Trends in socioeconomic-related health inequalities is a particularly pertinent topic in South Africa where years of systematic discrimination under apartheid bequeathed a legacy of inequalities in health outcomes. We use three nationally representative datasets to examine trends in income- and race-related inequalities in life expectancy (LE) and health-adjusted life expectancy (HALE) since the beginning of the millennium. We find that, in aggregate, (HA)LE at age five fell substantially between 2001 and 2007, but then increased to above 2001 levels by 2016, with the largest changes observed among prime age adults. Income- and race-related inequalities in both LE and HALE favor relatively well-off and non-Black South Africans in all survey years. Both income- and race-related inequalities in (HA)LE grew between 2001 and 2007, and then narrowed between 2007 to 2016. However, while race-related inequalities in (HA)LE in 2016 were smaller than in 2001, income-related inequalities in (HA)LE were greater in 2016 than in 2001. Based on the patterns and timing observed, these trends in income- and race-related inequalities in (HA)LE are most likely related to the delayed initial policy response to the HIV epidemic, the subsequent rapid and effective rollout of anti-retroviral therapy, and the changes in the overall income distribution among Black South Africans. In particular, the growth of the Black middle class narrowed the HA(LE) gap with the non-Black population but reinforced income-related inequalities.


Diversity of yeast and mold species from a variety of cheese types.
To generate a comprehensive profile of viable fungi (yeasts and molds) on cheese as it is purchased by consumers, 44 types of cheese were obtained from a local grocery store from 1 to 4 times each (depending on availability) and sampled. Pure cultures were obtained and identified by DNA sequence of the ITS region, as well as growth characteristics and colony morphology. The yeast Debaryomyces hansenii was the most abundant fungus, present in 79 % of all cheeses and 63 % of all samples. Penicillium roqueforti was the most common mold, isolated from a variety of cheeses in addition to the blue cheeses. Eighteen other fungal species were isolated, ten from only one sample each. Most fungi isolated have been documented from dairy products; a few raise potential food safety concerns (i.e. Aspergillus flavus, isolated from a single sample and capable of producing aflatoxins; and Candida parapsilosis, an emerging human pathogen isolated from three cheeses). With the exception of D. hansenii (present in most cheese) and P. roqueforti (a necessary component of blue cheese), no strong correlation was observed between cheese type, manufacturer, or sampling time with the yeast or mold species composition.


A Review of the Epigenetic Contributions to Endometriosis.
Epigenetic influences have been postulated to affect the course of endometriosis. DNA methyltransferases, estrogen and progesterone receptors, micro-RNAs, and histone deacetylators, have shown differential expression in endometriosis compared with normal endometrium. Others such as aromatase, Steroid Factor-1, COX-2, and Homeobox A10 also have epigenetic modifications in endometriosis. Limitations in this area of research include heterogeneity in study design, patient populations, and methods of analysis. Larger, controlled studies are needed. Future targeted uses of this work may include using methylomes to noninvasively diagnose endometriosis, or targeting histone-deacetylase inhibitors for treatment.


Glutamate-induced currents in acutely dissociated guinea pig locus coeruleus neurons.
To investigate the properties of L-glutamate (Glu) receptors in single locus coeruleus (LC) neurons at mature stage, we recorded the Glu-induced currents in the LC neurons acutely dissociated from adult guinea pigs, using the whole-cell patch-clamp technique. The concentration-current relationships show that Glu-induced currents in LC neurons might be predominantly mediated by non-NMDA receptors rather than NMDA receptors. The current-voltage relationship of NMDA or non-NMDA receptor-mediated currents indicated that both subtypes of the Glu-receptors operate non-selective cation channels, and that the NMDA receptor-operated channels but not the non-NMDA receptor-operated channels are permeable to Ca2+. These results suggest that in LC neurons, the normal excitatory neurotransmission may be mediated through non-NMDA receptors, and that NMDA receptors may be involved in intracellular signal transduction by their Ca2+ permeability.


[Spectrophotometric study of DNA complexes with dodecyltrimethylammonium bromide].
Multistep nature of complex formation of bromide dodecyltrimethylammonium (DTMA) with thymus DNA is shown by spectrophotometric method application in the range of 220-600 nm. While adsorbing DTMA DNA undergoes some structural changes. DTMA concentration raise from 0.4 to 0.8 mM per DNA provokes hydrophobic layer formation leading to following aggregation of the complexes. The analysis of light scattering in the range of 340-600 nm estimated the size of these aggregates. Assuming that these particles are spherical, their diameter appeared to be 95-110 nm.


Characterization of histamine receptors in isolated rabbit veins.
Veins were isolated from 16 sites of the rabbit venous tree and responses to histamine and histamine receptor agonists were studied to characterize the histamine receptors. Isometric contraction and relaxation of ring segment preparations were recorded. Histamine produced concentration-dependent contractions in all veins in the resting state. Both the maximum response and pD2 value varied remarkably from vein to vein and regional differences in sensitivities to histamine varied considerably from previous findings in dog veins. Also in the precontracted state with a vasoconstricting agent, histamine predominantly contracted the veins. The contractile responses to histamine, in both resting and precontracted states, were antagonized competitively by the histamine H1-receptor antagonist, mepyramine. On the other hand, histamine relaxed the precontracted veins, in the presence of mepyramine. Selective H2-receptor agonists, dimaprit and impromidine, relaxed the precontracted veins even in the absence of mepyramine. These responses to histamine were antagonized competitively by the H2-receptor antagonist, cimetidine or ranitidine. The present study provides quantitative and systematic data regarding histamine receptors in rabbit veins. We propose that: 1) there are both vasoconstrictor H1-receptors and vasodilator H2-receptors, 2) histamine generally contracts rabbit veins through predominant H1-receptors and that 3) the H2-receptor-mediated relaxation does not depend on the presence of the endothelium.


Differential mapping of the amino acids mediating agonist and antagonist coordination with the human thromboxane A2 receptor protein.
Despite the well documented involvement of thromboxane A(2) receptor (TPR) signaling in the pathogenesis of thrombotic diseases, there are currently no rationally designed antagonists available for clinical use. To a large extent, this derives from a lack of knowledge regarding the topography of the TPR ligand binding pocket. On this basis, the purpose of the current study was to identify the specific amino acid residues in the TPR protein that regulate ligand coordination and binding. The sites selected for mutation reside within or in close proximity to a region we previously defined as a TPR ligand binding region (i.e. the C terminus of the second extracellular loop and the leading edge of the fifth transmembrane domain). Mutation of these residues caused varying effects on the TPR-ligand coordination process. Specifically, the D193A, D193Q, and D193R mutants lost SQ29,548 (antagonist) binding and exhibited a dramatically reduced calcium response, which could not be restored by elevated U46619 (agonist) doses. The F184Y mutant lost SQ29,548 binding and exhibited a reduced calcium response (which could be restored by elevated U46619); and the T186A and S191T mutants lost SQ29,548 binding and retained a normal U46619-induced calcium response. Furthermore, these last three mutants also revealed a divergence in the binding of two structurally different antagonists, SQ29,548 and BM13.505. Two separate mutants that exhibited SQ29,548 binding yielded either a normal (F196Y) or reduced (S201T) U46619 response. Finally, mutation of other residues directly adjacent to those described above (e.g. E190A and F200A) produced no detectable effects on either SQ29,548 binding or the U46619-induced response. In summary, these results identify key amino acids (in particular Asp(193)) involved in TPR ligand coordination. These findings also demonstrate that TPR-specific ligands interact with different residues in the ligand-binding pocket.


Site-directed mutagenesis of the phosphate-binding consensus sequence in Escherichia coli adenylosuccinate synthetase.
Adenylosuccinate synthetases from different sources contain an N-terminal glycine-rich sequence GDEGKGK, which is homologous to the conserved sequence GXXXXGK found in many other guanine nucleotide-binding proteins or enzymes. To determine the role of this sequence in the structure and function of Escherichia coli adenylosuccinate synthetase, site-directed mutagenesis was performed to generate five mutant enzymes: G12V (Gly12----Val), G15V (Gly15----Val), G17V (Gly17----Val), K18R (Lys18----Arg), and I19T (Ile19----Thr). Comparison of the kinetic properties of the wild-type enzyme and those of the mutant enzymes revealed that the sequence is critical for enzyme activity. Replacement of Gly12, Gly15, or Gly17 with Val, or replacement of Lys18 with Arg, resulted in significant decreases in the kcat/Km values of the enzyme. Because the consensus sequence GXXXXGK(T/S) has been found in many GTP-binding proteins, isoleucine at position 19 in the E. coli adenylosuccinate synthetase was changed to threonine to produce the sequence GDEGKGKT. This mutation, which more closely resembles the consensus sequence, resulted in a 160-fold increase in the Km value for substrate GTP; however, there were no great changes for the other two substrates, IMP and aspartate. Based on these data, we suggest that the N-terminal glycinerich sequence in E. coli adenylosuccinate synthetase plays a more important role in enzyme catalysis than in substrate binding. In addition, a hydrophobic amino acid residue such as isoleucine, leucine, or valine, rather than threonine, may play a critical role in GTP binding in adenosuccinate synthetase. These findings suggest that the glycine-rich sequence in adenylosuccinate synthetase functions differently relative to those in other GTP binding proteins or enzymes.


Bayesian methods for estimating pathogen prevalence within groups of animals from faecal-pat sampling.
Pathogens such as Escherichia coli O157:H7 and Campylobacter spp. have been implicated in outbreaks of food poisoning in the UK and elsewhere. Domestic animals and wildlife are important reservoirs for both of these agents, and cross-contamination from faeces is believed to be responsible for many human outbreaks. Appropriate parameterisation of quantitative microbial-risk models requires representative data at all levels of the food chain. Our focus in this paper is on the early stages of the food chain-specifically, sampling issues which arise at the farm level. We estimated animal-pathogen prevalence from faecal-pat samples using a Bayesian method which reflected the uncertainties inherent in the animal-level prevalence estimates. (Note that prevalence here refers to the percentage of animals shedding the bacteria of interest). The method offers more flexibility than traditional, classical approaches: it allows the incorporation of prior belief, and permits the computation of a variety of distributional and numerical summaries, analogues of which often are not available through a classical framework. The Bayesian technique is illustrated with a number of examples reflecting the effects of a diversity of assumptions about the underlying processes. The technique appears to be both robust and flexible, and is useful when defecation rates in infected and uninfected groups are unequal, where population size is uncertain, and also where the microbiological-test sensitivity is imperfect. We also investigated the determination of the sample size necessary for determining animal-level prevalence from pat samples to within a pre-specified degree of accuracy.


The unique features of Th9 cells and their products.
Although it was discovered more than two decades ago, new information concerning the biological activities of IL-9 has been provided in recent years, after the isolation of cells that selectively produce this cytokine, designated "Th9." Th9 cells are generated in vitro by polarization, mainly by TGF-β and IL-4, during activation with the specific antigen, or with anti-CD3/CD28 antibodies. This review deals mainly with Th9 generated by the former, "physiological" mode of activation. Of particular interest is the unique production kinetics of IL-9: the cytokine is produced very rapidly, but after reaching its peak (day 3 in our studies), it declines sharply to trace levels. In addition to IL-9, Th9 cells also produce similar amounts of another cytokine, IL-10, but the production kinetics of these two cytokines are strikingly different. Antigen-activated Th9 in our studies also developed pathogenic capacity, but only during the short time period of peak IL-9 production. Interestingly, no IL-9-producing cells were detected in sites of inflammation induced by Th9, in contrast to Thl and Thl7. The unique features of Th9 cells and their products are discussed with regards to the known and assumed functions of the cytokine.


Leukaemia reactivates mouse cytomegalovirus.
An established model for latency by mouse cytomegalovirus (MCMV) was used to assess the effects of leukaemia on reactivation of MCMV. After inoculation of 400 spleen focus-forming units of Friend leukaemia virus (FLV), a significant number of mice yielded MCMV in salivary glands and kidneys 4 to 5 weeks later. Administration of cyclophosphamide during FLV infection increased the percentage of MCMV-positive animals. These results suggest that cytomegalovirus infections in leukaemic patients may reactivate as a result of the leukaemia itself and may be exacerbated by chemotherapy.


Role of FGF receptors as an emerging therapeutic target in lung squamous cell carcinoma.
Recent advances in molecular medicine and high-throughput sequencing technologies have achieved major cancer strategies and therapeutics over the past decades. For example, identification of oncogenic EGF receptor mutations that are present in up to 20% of lung adenocarcinoma patients confer exquisite sensitivity to EGF receptor inhibitors. However, currently known 'druggable' targets are enriched in the subgroup of adenocarcinomas and individuals who have never smoked. We present an overview of FGFs and FGF receptor (FGFR) signaling in cancer, and the role of FGFR1 as a novel druggable target in lung squamous cell carcinoma. FGFR1 amplification in lung squamous cell carcinoma is required for the survival of FGFR1-amplified cell lines. Currently, clinical reagents that target the FGFs and FGFRs are being developed accordingly. This review focuses on the emerging role of FGFR1 as a therapeutic target in lung squamous cell carcinoma and reviews current agents that are in clinical development for the treatment of FGFR-dependent cancer.


[Monosynaptic pathway responsible for generation of burst activity in neuron PPa1 of Helix pomatia].
The connections between an interneuron in the visceral ganglion initiating the pacemaker activity in RPa1 neuron and RPa1 neuron itself have been analyzed in snail. The interneuron stimulation led to initiation of the bursting activity in the RPa1 neuron. Replacement of extracellular Ca2+ by Mg2+ ions, as well as addition of CdCl2 to the external solution reversibly inhibited the effect of the interneuronal stimulation on the RPa1 neuron. An increase of extracellular Ca2+ ions concentration to 70 mM did not prevent the effect. Intracellular Cs+ and TEA injection in the interneuron increasing the action potential duration in the interneuron increased also the effectiveness of connection between the neurons. It is concluded that the connection between the neurons under investigation is monosynaptic and peptidergic.


Case Report: Paliperidone Palmitate in the Management of Bipolar I Disorder With Non-compliance.
Background: Medication non-adherence is prevalent in patients with bipolar disorder (BD). Long-acting injectable antipsychotics (LAIAs) are widely used to improve compliance with treatment. This study aimed to illustrate the effectiveness, compliance, and safety profile of once-monthly paliperidone palmitate (PP1M), a novel therapeutic LAIA, in the management of bipolar I disorder (BDI). Method: A prospective follow-up was arranged to 11 BDI patients who were prescribed PP1M as monotherapy or adjunctive treatment. Severity of symptoms, disturbing behavior, status of employment, 17-item Hamilton Depression Rating Scale (HAMD-17), and Young Mania Rating Scale (YMRS) were evaluated at the baseline and the endpoint of follow-up. Clinical Global Impression-Bipolar Disorder-Severity of Illness Scale (CGI-BP) and Treatment Emergent Symptom Scale (TESS) were measured at each injection of PP1M. Compliance, relapse or switch, and new hospitalization were monitored through the period of follow-up. Results: The median duration of treatment was 14 months, ranging from 5 to 22 months. The scores (mean ± standard deviation) of HAMD-17, YMRS, and CGI-BP generally decreased from the baseline (16.1 ± 10.3, 30.9 ± 12.6, 5.3 ± 0.7) to the endpoint (7.4 ± 5.7, 3.7 ± 3.2, 2.3 ± 0.7). No disturbing behavior was detected at the endpoint. Neither new hospitalization nor manic/mixed episode occurred during treatment, whereas mild to moderate depressive episodes were reported in three cases. The status of employment of 10 participants (90.9%) was improved, and no new safety concern was detected. Conclusion: PP1M might offer a new valid treatment option in the long-term management of BDI, especially for those with poor compliance with oral medication. However, more studies are needed to further justify such role.


Who Needs Longer Tolvaptan Treatment?
The vasopressin receptor 2 (V2) receptor antagonist tolvaptan is an aquaretic agent that has been approved for heart failure patients with volume overload in Japan. In this study (SMILE study), we investigated patient characteristics and effectiveness in both a 14 days and shorter treated group (14DS) and 15 days and longer treated group (15DL). The results showed that the patients in the 15DL group had low cardiac output with intensive diuretic administration (ie, diuretic resistance). The congestive symptoms were greatly improved within 14 days of treatment in both the 14DS and 15DL groups. Further improvements in lower limb edema, pulmonary congestion, dyspnea, third sound, and rales after 2 weeks were statistically significant in the 15DL group, but the amount of improvement was subtle and the 15DL group might have consisted of a considerable number of "non-responders". Therefore, identifying "responders" by biomarkers and conducting a prospective randomized study is required to validate our findings.


Treatment of humeral condylar fractures and humeral intracondylar fissures in cats with patellar fracture and dental anomaly syndrome.
The aim of this study is to describe the treatment and outcome of humeral condylar fractures and humeral intracondylar fissures in cats with patellar fracture and dental anomaly syndrome (PADS) and to provide advice on how to manage these cases in practice. Data were collated on cats with PADS that were reported to have sustained humeral fractures or had fractures or fissures of the humerus identified on radiographs. The details of the fractures were recorded in addition to any treatment and outcome information. Of the 207 cases reported with PADS, 18 cats (8.7%) were found to have humeral condylar fractures, none of which was known to have resulted from significant trauma. Where treatment occurred, it involved the placement of transcondylar positional or lag screws. In some cases additional implants, including supracondylar bone plates and screws or Kirschner wires (K-wires), were used. Follow-up data revealed that only two cats were euthanased owing to the presence of the humeral fractures, with at least eight achieving some degree of recovery of function. These humeral fractures all have the characteristics of stress insufficiency fractures, being simple isolated fractures that are short oblique, with increased radio-density at the fracture line and occurring following minimal or no trauma. Humeral intracondylar fissures were identified in two cats and it is possible that some of the other fractures may have occurred secondary to pre-existing fissures. To our knowledge, no prior reports exist of fissures in cats that do not meet the criteria for PADS. Surgical repair primarily consisted of the placement of transcondylar lag or positional screws with, in some cases, adjunct implants such as bone plates and screws or K-wires. Though there were insufficient data to determine the prognosis for these fractures in the long term, unlike patellar fractures, many of these fractures will heal if treated appropriately.


Effects of carnosine on contractile apparatus Ca²⁺ sensitivity and sarcoplasmic reticulum Ca²⁺ release in human skeletal muscle fibers.
There is considerable interest in potential ergogenic and therapeutic effects of increasing skeletal muscle carnosine content, although its effects on excitation-contraction (EC) coupling in human muscle have not been defined. Consequently, we sought to characterize what effects carnosine, at levels attained by supplementation, has on human muscle fiber function, using a preparation with all key EC coupling proteins in their in situ positions. Fiber segments, obtained from vastus lateralis muscle of human subjects by needle biopsy, were mechanically skinned, and their Ca(2+) release and contractile apparatus properties were characterized. Ca(2+) sensitivity of the contractile apparatus was significantly increased by 8 and 16 mM carnosine (increase in pCa(50) of 0.073 ± 0.007 and 0.116 ± 0.006 pCa units, respectively, in six type I fibers, and 0.063 ± 0.018 and 0.103 ± 0.013 pCa units, respectively, in five type II fibers). Caffeine-induced force responses were potentiated by 8 mM carnosine in both type I and II fibers, with the potentiation in type II fibers being entirely explicable by the increase in Ca(2+) sensitivity of the contractile apparatus caused by carnosine. However, the potentiation of caffeine-induced responses caused by carnosine in type I fibers was beyond that expected from the associated increase in Ca(2+) sensitivity of the contractile apparatus and suggestive of increased Ca(2+)-induced Ca(2+) release. Thus increasing muscle carnosine content likely confers benefits to muscle performance in both fiber types by increasing the Ca(2+) sensitivity of the contractile apparatus and possibly also by aiding Ca(2+) release in type I fibers, helping to lessen or slow the decline in muscle performance during fatiguing stimulation.


Serum type IV collagen concentrations in diabetic patients with microangiopathy as determined by enzyme immunoassay with monoclonal antibodies.
In long-term diabetes mellitus, thickening of basement membrane in capillaries and small vessels is a characteristic lesion and plays an important role in the progression of diabetic microangiopathy. We have developed a sandwich enzyme immunoassay for human serum type IV collagen peptide with monoclonal antibodies. Previous studies suggested that collagen levels reflect the activity of fibrogenesis in basement membrane. Serum type IV collagen levels were measured in 137 non-insulin-dependent diabetic patients (aged 50-75 yr) with or without clinical signs of retinopathy, nephropathy, and/or neuropathy and 110 healthy subjects (aged 50-75 yr) without serological abnormality. Serum concentrations of type IV collagen were significantly higher (P less than 0.01) in diabetic patients (mean +/- SE 124.1 +/- 4.1 ng/ml) than in healthy subjects (73.9 +/- 2.2 ng/ml) and were increased with the prevalence or incidence of diabetic complications. In the patients with diabetic microangiopathy, serum type IV collagen levels became higher as clinical signs worsened. Especially in the patients with diabetic nephropathy, serum type IV collagen levels became higher with elevation of blood urea nitrogen, serum creatinine, and serum beta 2-microglobulin but not urinary excretion of beta 2-microglobulin and N-acetyl-beta-glucosaminidase. These observations indicated that elevation of serum type IV collagen in diabetic nephropathy was related to glomerular filtration dysfunction rather than renal tubular dysfunction. However, the antigen, which can be detected by our assay system, did not exist in urine specimens of healthy subjects, and an intimate relationship was not observed between serum type IV collagen level and serum creatinine level.(ABSTRACT TRUNCATED AT 250 WORDS)


Put a (Limbal) Ring on It: Women Perceive Men's Limbal Rings as a Health Cue in Short-Term Mating Domains.
Limbal rings are dark annuli encircling the iris that fluctuate in visibility based on health and age. Research also indicates their presence augments facial attractiveness. Given individuals' prioritization of health cues in short-term mates, those with limbal rings may be implicated as ideal short-term mates. Three studies tested whether limbal rings serve as veridical health cues, specifically the extent to which this cue enhances a person's value as a short-term mating partner. In Study 1, targets with limbal rings were rated as healthier, an effect that was stronger for female participants and male targets. In Study 2, temporally activated short-term mating motives led women to report a heightened preference for targets with limbal rings. In Study 3, women rated targets with limbal rings as more desirable short-term mates. Results provide evidence for limbal rings as veridical cues to health, particularly in relevant mating domains.


Endometriosis and infertility: epidemiology and evidence-based treatments.
Endometriosis is an estrogen-dependent disorder defined as the presence of endometrial tissue outside of the uterine cavity. A leading cause of infertility, endometriosis has a prevalence of 0.5-5% in fertile and 25-40% in infertile women. The optimal choice of management for endometriosis-associated infertility remains obscure. Removal or suppression of endometrial deposits by medical or surgical means constitutes the basis of endometriosis management. Current evidence indicates that suppressive medical treatment of endometriosis does not benefit fertility and should not be used for this indication alone. Surgery is probably efficacious for all stages of the disease. Controlled ovarian hyperstimulation with intrauterine insemination is recommended in early-stage and surgically corrected endometriosis when pelvic anatomy is normal. In advanced cases, in vitro fertilization is a treatment of choice, and its success may be augmented with prolonged gonadotropin-releasing hormone analog treatment. Further randomized clinical trials focusing on diverse etiopathogenic mechanisms and therapeutic innovation are necessary to find more conclusive, evidence-based answers regarding this enigmatic disease.


Phytoremediation of organomercurial compounds via chloroplast genetic engineering.
Mercury (Hg), especially in organic form, is a highly toxic pollutant affecting plants, animals, and man. In plants, the primary target of Hg damage is the chloroplast; Hg inhibits electron transport and photosynthesis. In the present study, chloroplast genetic engineering is used for the first time to our knowledge to enhance the capacity of plants for phytoremediation. This was achieved by integrating a native operon containing the merA and merB genes (without any codon modification), which code for mercuric ion reductase (merA) and organomercurial lyase (merB), respectively, into the chloroplast genome in a single transformation event. Stable integration of the merAB operon into the chloroplast genome resulted in high levels of tolerance to the organomercurial compound, phenylmercuric acetate (PMA) when grown in soil containing up to 400 micro M PMA; plant dry weights of the chloroplast transformed lines were significantly higher than those of wild type at 100, 200, and 400 micro M PMA. That the merAB operon was stably integrated into the chloroplast genome was confirmed by polymerase chain reaction and Southern-blot analyses. Northern-blot analyses revealed stable transcripts that were independent of the presence or absence of a 3'-untranslated region downstream of the coding sequence. The merAB dicistron was the more abundant transcript, but less abundant monocistrons were also observed, showing that specific processing occurs between transgenes. The use of chloroplast transformation to enhance Hg phytoremediation is particularly beneficial because it prevents the escape of transgenes via pollen to related weeds or crops and there is no need for codon optimization to improve transgene expression. Chloroplast transformation may also have application to other metals that affect chloroplast function.


Effects of Lipopolysaccharide Core Sugar Deficiency on Colanic Acid Biosynthesis in Escherichia coli.
When 10 Escherichia coli mutant strains with defects in lipopolysaccharide (LPS) core biosynthesis were grown on agar medium at 30°C, four of them, the ΔwaaF, ΔwaaG, ΔwaaP, and ΔwaaB strains, formed mucoid colonies, while the other six, the ΔwaaU, ΔwaaR, ΔwaaO, ΔwaaC, ΔwaaQ, and ΔwaaY strains, did not. Using light microscopy with tannin mordant staining, the presence of exopolysaccharide around the cells of the mutants that formed mucoid colonies could be discerned. The ΔwaaF mutant produced the largest amounts of exopolysaccharide, regardless of whether it was grown on agar or in liquid medium. The exopolysaccharide was isolated from the liquid growth medium of ΔwaaF cells, hydrolyzed, and analyzed by high-performance liquid chromatography with an ion-exchange column, and the results indicated that the exopolysaccharide was consistent with colanic acid. When the key genes related to the biosynthesis of colanic acid, i.e., wza, wzb, wzc, and wcaA, were deleted in the ΔwaaF background, the exopolysaccharide could not be produced any more, further confirming that it was colanic acid. Colanic acid could not be produced in strains in which rcsA, rcsB, rcsD, or rcsF was deleted in the ΔwaaF background, but a reduced level of colanic acid production was detected when the rcsC gene was deleted, suggesting that a change of lipopolysaccharide structure in ΔwaaF cells might be sensed by the RcsCDB phosphorelay system, leading to the production of colanic acid. The results demonstrate that E. coli cells can activate colanic acid production through the RcsCDB phosphorelay system in response to a structural deficiency of lipopolysaccharide. Lipopolysaccharide and colanic acid are important forms of exopolysaccharide for Escherichia coli cells. Their metabolism and biological significance have been investigated, but their interrelation with the cell stress response process is not understood. This study demonstrates, for the first time, that E. coli cells can activate colanic acid production through the RcsCDB phosphorelay system in response to a structural change of lipopolysaccharide, suggesting that bacterial cells can monitor the outer membrane integrity, which is essential for cell survival and damage repair.


Sickle cell anemia with moyamoya disease: outcomes after EDAS procedure.
Moyamoya disease is a relatively uncommon neurovascular complication of sickle cell anemia. We report a case series of six patients with sickle cell anemia who developed moyamoya disease and underwent encephaloduroarteriosynangiosis procedures. These six patients presented with either cerebrovascular accidents, transient ischemic attacks, or seizures, and subsequent magnetic resonance imaging scans were suggestive of moyamoya-like changes in the cerebral vasculature. Conventional cerebral angiography was used to confirm the diagnosis in all six patients. Four of six patients manifested a cerebrovascular accident before surgery, and two of these patients were compliant on a transfusion protocol at the time of their cerebrovascular accident. Bilateral (n = 4) or unilateral (n = 2) encephaloduroarteriosynangiosis procedures were performed without any complications. The patient who was stroke-free preoperatively had a cerebrovascular accident 2 weeks after the procedure; otherwise, all patients have remained free of neurovascular complications with an average follow-up of 33 months. Collateral anastomoses between external and internal carotid arteries were established by magnetic resonance angiography in three patients. The encephaloduroarteriosynangiosis procedure is a safe and effective treatment option in patients with sickle cell anemia who develop moyamoya disease.


Islamic sunni mainstream opinions on compensation to unrelated live organ donors.
This article focuses on contemporary Islamic attitudes towards the question of compensation to a non-relative live organ donor. This article presents the history of the debate on organ transplantation in Islam since the 1950s and the key ethical questions. It continues by presenting the opinions of the mainstream ulema such as Tantawi and Qaradawi. The article ends with a conclusion that there must be no compensation made to a non-related live organ donor, not even a symbolic gift of honor (ikramiyya).


Health Risks Associated with Oil Pollution in the Niger Delta, Nigeria.
Although there is considerable public concern about the environmental impacts of oil pollution in the Niger Delta of Nigeria, actual evidence on the pathological and psychological effects in the health of local communities is minimally known. We sought to associate the perspective measures of exposure to oil pollution with health outcomes (inventory of health symptoms and functional capacity limitations) and determine how emotional reactions to environmental risks moderate these health outcomes. The study was conducted with 600 participants selected from five local government areas in Akwa Ibom State where oil pollution is rampant. A structured questionnaire was used to collect the data on the respondents' exposure to oil pollution, self-rated health and disease symptoms, perception of risk of exposure and emotional reactions to local oil pollution. Most of the participants lived in areas with visible oil pollution and/or near gas flaring facilities and regularly suffered direct exposure to oil in their environment. High level of emotional distress was a part of everyone's life for the study population. Risk perception in the study area was mediated, to a large extent, by dreaded hazards (catastrophic fears of pipeline explosions and oil spill fire), visual cues (gas flares and smoke stacks) and chemosensory cues (off-flavor in drinking water). The exposure metrics were found to be significant predictors of the health effects and influencing factors (emotional reactions). Multi-levels models suggest that at the individual level, the demographic variables and direct contact with oil pollution were important mediators of functional capacity limitation. At the community level, emotional distress from fear of the sources of exposure was an important mediator of the health symptoms. This study documents high levels of disease symptoms and environmental distress (worry, annoyance and intolerance) associated with oil pollution in the Niger Delta areas of Nigeria. It highlights the need for some intervention to ameliorate the psychological distress associated with living under such environmental adversity.


[Effect of ionol on superoxide radical metabolism in murine liver].
The influence of ionol (100mg/kg) on the rate of superoxide generation (V) and activities of antioxidant enzymes: CuZn- and Mn-SOD, glutathione peroxidase (GSH-Px), glutathione S-transferase (GST) in different subcellular organelles of mice liver was studied. Ionol is shown to result in realiable a synchronous changes of all studied antioxidant enzyme activities in cytosol and whole blood. On the first day the level of these enzymes increased by 1.5 times and on the third day it returned to normal. The obtained data indicate retention of regulatory relation in antioxidant system in liver cytosol within the sector SOD-GSH-Px. In the mitochondria the Mn-SOD activity changes in antibate manner as compared CuZn-SOD activity, on the first day Mn-SOD activity decreases and remains on lowered level during the whole period investigated. In microsomes the value of V is found to be reduced. In the case of SMP on the first day after the administration of ionol V value didn't increase significantly. However, owing to Mn-SOD activity decrease the ratio V/A, showing the level of superoxide radicals in subcellular organelles grows 3-fold. In nuclei V value increases 4-6-fold during 1-3 hours after ionol injection. The data obtained show that administration of high dose of ionol to intact mice suppresses antioxidant enzyme system of mitochondria, induces abrupt production of superoxide radicals in nuclei and reduces of functioning of electron transport chaine in microsomes. The observed disturbances have short-lived character and are normalized during 3 days after administration of ionol. The toxic effects of ionol may be connected with the action of oxidative modification products formed in organism.


Integrating Fiber Optic Data in Numerical Reservoir Simulation Using Intelligent Optimization Workflow.
A novel workflow is presented for integrating fiber optic Distributed Temperature Sensor (DTS) data in numerical simulation model for the Cyclic Steam Stimulation (CSS) process, using an intelligent optimization routine that automatically learns and improves from experience. As the steam-oil relationship is the main driver for forecasting and decision-making in thermal recovery operations, knowledge of downhole steam distribution across the well over time can optimize injection and production. This study uses actual field data from a CSS operation in a heavy oil field in California, and the value of integrating DTS in the history matching process is illustrated as it allows the steam distribution to be accurately estimated along the entire length of the well. The workflow enables the simultaneous history match of water, oil, and temperature profiles, while capturing the reservoir heterogeneity and the actual physics of the injection process, and ultimately reducing the uncertainty in the predictive models. A novel stepwise grid-refinement approach coupled with an evolutionary optimization algorithm was implemented to improve computational efficiency and predictive accuracy. DTS surveillance also made it possible to detect a thermal communication event due to steam channeling in real-time, and even assess the effectiveness of the remedial workover to resolve it, demonstrating the value of continuous fiber optic monitoring.


[Effect of cryopowder Rhodiola rosae L. on cardiorespiratory parameters and physical performance of humans].
Seven-hour continuous physical loading test (bicycle ergometry) was used to assess effects of Rhodiola rosae L. (cryopowder) on the human cardiorespiratory system. Comparing with the control, the preparation facilitated activation of the energy-supplying mechanisms in human organism during physical work. In addition, it increased efficiency of the cardiovascular and respiration systems and prevented fatigue growth. The medical herb powder demonstrated explicitly the protective action and can be recommended for use in extreme conditions.


Empowering law enforcement officers to engage effectively with individuals in crisis.
As first responders with a duty to protect the safety of civilians, law enforcement officers are required to carry firearms. Over the course of a career, officers are exposed to multiple traumatic events and are required to make rapid decisions regarding safety. In the last few years, the use of deadly force by law enforcement officers has become a public concern. Persons with untreated mental illness have a comparatively higher risk than most for deadly outcomes in crisis situations. The purpose of this manuscript is to describe policies and programs that have been developed and implemented by teams that include psychiatric mental health nurse practitioners to help law enforcement officers recognize and interact effectively with persons in crisis. The same programs help officers understand the effects of severe stress and trauma on emotional, behavioral, and cognitive functioning, and provide support, encouragement, and access for officers who need help.


Allophycocyanin from the filamentous cyanophyte, Phormidium luridum.
Allophycocyanin from the filamentous cyanophyte, Phormidium luridum, was purified by ammonium sulfate fractionation and ion exchange chromatography on brushite columns. The specific absorption coefficient (E 0.1% 1cm) of purified allophycocyanin was 6.1 in distilled water and 7.3 in 0.05 M potassium phosphate buffer (pH 7). Absorption maxima of allophycocyanin occurred at 650, 618 (shoulder), 350, and 275 nm. Circular dichroic spectra displayed positive ellipticity bands at 655 and 625 nm, and a major negative ellipticity band at 340 nm. Computer analysis of the circular dichroic spectrum of allophycocyanin from 207 to 243 nm indicated that the secondary structure contained 60% alpha helix and 40% beta form. The estimated molecular weight of allophycocyanin on Sephadex G-200 columns at pH 7.0 was 155,000. Electrophoretic examination of allophycocyanin on sodium dodecyl sulfate polyacrylamide gels revealed two subunits, alpha and beta, with apparent molecular weights of 17,300 and 19,000, respectively. Densitometric analysis of unstained gels at 600 nm indicated that one phycocyanobilin chromophore was associated with each subunit. Treatment of allophycocyanin with 12% formic acid or 8 M urea and subsequent removal of the denaturant yielded a derivative with spectroscopic characteristics similar to phycocyanin. Subsequent incubation in phosphate buffer (pH 7), but not in acetate buffer (pH 5) or in water, was accompanied by a progressive reappearance of absorption maxima at 650 and 618 nm (shoulder), and positive ellipticity bands at 655 and 617 nm. Automated sequence analysis of allophycocyanin (a) showed that the sequence of amino acids at the amino terminus of the alpha and beta subunits is different, (b) showed that the subunits occur in a ratio of 1:1, and (c) demonstrated sequence homology at the amino terminus of allophycocyanin, phycocyanin, and phycoerythrin.


Mitochondrial control of acute glutamate excitotoxicity in cultured cerebellar granule cells.
Mitochondria within cultured rat cerebellar granule cells have a complex influence on cytoplasmic free Ca2+ ([Ca2+]c) responses to glutamate. A decreased initial [Ca2+]c elevation in cells whose mitochondria are depolarized by inhibition of the ATP synthase and respiratory chain (conditions which avoid ATP depletion) was attributed to enhanced Ca2+ extrusion from the cell rather than inhibited Ca2+ entry via the NMDA receptor. Even in the presence of elevated extracellular Ca2+, when [Ca2+]c responses were restored to control values, such cells showed resistance to acute excitotoxicity, defined as a delayed cytoplasmic Ca2+ deregulation (DCD) during glutamate exposure. DCD was a function of the duration of mitochondrial polarization in the presence of glutamate rather than the total period of glutamate exposure. Once initiated, DCD could not be reversed by NMDA receptor inhibition. In the absence of ATP synthase inhibition, respiratory chain inhibitors produced an immediate Ca2+ deregulation (ICD), ascribed to an ATP deficit. In contrast to DCD, ICD could be reversed by subsequent ATP synthase inhibition with or without additional NMDA receptor blockade. DCD could not be ascribed to the failure of an ATP yielding metabolic pathway. It is concluded that mitochondria can control Ca2+ extrusion from glutamate-exposed granule cells by the plasma membrane in three ways: by competing with efflux pathways for Ca2+, by restricting ATP supply, and by inducing a delayed failure of Ca2+ extrusion. Inhibitors of the mitochondrial permeability transition only marginally delayed the onset of DCD.


Biological activities of commercial bee pollens: antimicrobial, antimutagenic, antioxidant and anti-inflammatory.
Bee pollen is considered, since memorable times, a good source of nourishing substances and energy. The present study aimed to evaluate the biological activities of eight commercial bee pollens purchased from the market. The origin of sample A was not specified in the labeling; samples B, C, D and G were from Portugal and the remaining were from Spain. The sample E presented the highest value of phenolics (32.15±2.12 mg/g) and the H the lowest (18.55±095 mg/g). Sample C had the highest value of flavonoids (10.14±1.57 mg/g) and sample H the lowest (3.92±0.68 mg/g). All the samples exhibited antimicrobial activity, being Staphylococcus aureus the most sensitive and Candida glabrata the most resistant of the microorganisms studied. All the samples exhibited antimutagenic activity, even though some samples were more effective in decreasing the number of gene conversion colonies and mutant colonies. Regarding the antioxidant activity, assessed using two methods, the more effective was sample B. The anti-inflammatory activity, assessed using the hyaluronidase enzyme, was highest in samples B and D. Pearson's correlation coefficients between polyphenols, flavonoids, antioxidant activity and antimicrobial activity were computed. It was also performed a discriminant analysis.


Benzodiazepine use in a small community hospital. Appropriate prescribing or not?
Benzodiazepine (BDZ)-prescribing patterns in relation to indications and to drug characteristics in a small short-stay hospital were examined. In a sample of 800 patients, 183 were prescribed BDZs during their stay. Female patients received more BDZs (BDZ:female patients 1.31:1.0) than males (BDZs:male patients 1.02:1.0), particularly in the 21-40-year age group in which polypharmacy was highest. BDZs were classified according to their elimination half-lives. Our data showed that the majority fell into the long-acting (half-life greater than 24 h) (55%) and intermediate-acting (half-life 12-24 h) (20.7%) categories, despite the fact that most indications (pre-anaesthetic and night-time sedation; total 71.6%) called for the shorter-acting drugs. The merits or otherwise of this situation are discussed, and a number of questions put regarding the validity of this approach. Overall, BDZs accounted for 5.8% of the calculated medications given to the 800-patient sample.