Comparing pedestal structure in JET-ILW H-mode plasmas with a model for stiff ETG turbulent heat transport.

A predictive model for the electron temperature profile of the H-mode pedestal is described, and its results are compared with the pedestal structure of JET-ILW plasmas. The model is based on a scaling for the gyro-Bohm normalized, turbulent electron heat flux [Formula: see text] resulting from electron temperature gradient (ETG) turbulence, derived from results of nonlinear gyrokinetic (GK) calculations for the steep gradient region. By using the local temperature gradient scale length [Formula: see text] in the normalization, the dependence of [Formula: see text] on the normalized gradients [Formula: see text] and [Formula: see text] can be represented by a unified scaling with the parameter [Formula: see text], to which the linear stability of ETG turbulence is sensitive when the density gradient is sufficiently steep. For a prescribed density profile, the value of [Formula: see text] determined from this scaling, required to maintain a constant electron heat flux [Formula: see text] across the pedestal, is used to calculate the temperature profile. Reasonable agreement with measurements is found for different cases, the model providing an explanation of the relative widths and shifts of the [Formula: see text] and [Formula: see text] profiles, as well as highlighting the importance of the separatrix boundary conditions. Other cases showing disagreement indicate conditions where other branches of turbulence might dominate. This article is part of a discussion meeting issue 'H-mode transition and pedestal studies in fusion plasmas'.

Pembrolizumab as first-line therapy for patients with PD-L1-positive advanced non-small cell lung cancer: a phase 1 trial.

Background: Pembrolizumab improved survival as first- and second-line therapy compared with chemotherapy in patients with highly programmed death ligand 1 (PD-L1) expressing advanced non-small cell lung cancer (NSCLC). We report the long-term safety and clinical activity of pembrolizumab as first-line therapy for patients with advanced NSCLC and the correlation between PD-L1 expression and efficacy. Patients and methods: In the open-label phase 1b KEYNOTE-001 trial, treatment-naive patients with advanced NSCLC whose tumors expressed PD-L1 (≥1% staining, assessed using a prototype assay) were randomly assigned to intravenous pembrolizumab 2 or 10 mg/kg every 3 (Q3W) or 2 (Q2W) weeks. Response was assessed per central RECIST v1.1 every 9 weeks in all patients who received ≥1 pembrolizumab dose. Using pre-treatment tumor tissue, a clinical assay quantified the percentage of tumor cells expressing PD-L1 as tumor proportion score (TPS). Results: Between 1 March 2013 and 18 September 2015, 101 patients received pembrolizumab 2 mg/kg Q3W (n = 6), 10 mg/kg Q3W (n = 49), or 10 mg/kg Q2W (n = 46). Of these, 27 (26.7%) had TPS ≥50%, 52 (51.5%) had TPS 1%-49%, and 12 (11.9%) had TPS <1%. The objective response rate (ORR) was 27% (27/101, 95% CI 18-37) and median overall survival was 22.1 months (95% CI 17.1-27.2). In patients with PD-L1 TPS ≥50%, ORR, 12-month PFS, and 12-month OS were higher [14/27 (51.9%; 95% CI 32%-71%), 54%, and 85%, respectively] than the overall population [27/101 (26.7%; 95% CI 18.4%-36.5%), 35%, 71%]. Pembrolizumab was well tolerated, with only 12 (11.9%) patients experiencing grade 3/4 treatment-related adverse events and no treatment-related deaths. Conclusions: Pembrolizumab provides promising long-term OS benefit with a manageable safety profile for PD-L1-expressing treatment-naive advanced NSCLC, with greatest efficacy observed in patients with TPS ≥50%. Clinical trial name and number: KEYNOTE-001 (ClinicalTrials.gov, NCT01295827).

Temporary alterations to postpartum milking frequency affect whole-lactation milk production and the energy status of pasture-grazed dairy cows.

This study investigated the immediate and long-term effects of temporary alterations to postpartum milking frequency (MF) on milk production, body condition score (BCS), and indicators of energy status in pasture-grazed cows supplemented with concentrates. Multiparous Holstein-Friesian cows (n = 150) were randomly assigned to 1 of 5 groups at calving: milked twice daily (2 ×) throughout lactation (control), or milked either once daily (1 ×) or 3 times daily (3 ×) for 3 or 6 wk immediately postpartum, and then 2 × for the remainder of lactation. During wk 1 to 3 postpartum, cows milked 1 × produced 15% less milk and 17% less energy-corrected milk (ECM) than cows milked 2 ×. This immediate production loss increased to 20% less milk and 22% less ECM during wk 4 to 6 postpartum for cows that remained on 1 × milking; these animals also produced less than 1 × cows switched to 2 × milking after 3 wk. During wk 8 to 32, when all cows were milked 2 ×, those previously milked 1 × had sustained reductions in milk (-6%) and ECM (-8%) yields, which were not affected by the duration of reduced postpartum MF. In contrast, cows milked 3 × postpartum had 7% greater milk yields during wk 1 to 6 compared with 2 × controls, irrespective of the duration of increased MF. Milk yields also remained numerically greater (+5%) during wk 8 to 32 in cows previously milked 3 ×. Nevertheless, yields of ECM were not increased by 3 × milking, because of lower milk fat and protein contents that persisted for the rest of lactation. In addition, indicators of cow energy status reflected an increasing state of negative energy balance with increasing MF. Cows milked 1 × postpartum had greater plasma glucose and lower plasma nonesterified fatty acid concentrations during the reduced MF, and plasma glucose remained lower for 2 wk after cows had switched to 2 × milking. Moreover, BCS was improved relative to 2 × controls from wk 5 to 6. In contrast, cows milked 3 × had lower plasma glucose concentrations, greater plasma nonesterified fatty acid concentrations, and greater BCS loss during wk 1 to 3; however, greater body fat mobilization was not sustained, indicating that additional energy supplements may be required to achieve better milk production responses. In conclusion, temporary 1 × milking had lactation-long negative effects on milk and milk component yields but improved cow energy status and BCS, whereas temporary 3 × milking immediately increased milk yield but did not improve milk fat and protein yields in pasture-grazed cows.

Kinetic instabilities that limit ß in the edge of a tokamak plasma: a picture of an H-mode pedestal.

Plasma equilibria reconstructed from the Mega-Amp Spherical Tokamak have sufficient resolution to capture plasma evolution during the short period between edge-localized modes (ELMs). Immediately after the ELM, steep gradients in pressure, P, and density, n(e), form pedestals close to the separatrix, and they then expand into the core. Local gyrokinetic analysis over the ELM cycle reveals the dominant microinstabilities at perpendicular wavelengths of the order of the ion Larmor radius. These are kinetic ballooning modes in the pedestal and microtearing modes in the core close to the pedestal top. The evolving growth rate spectra, supported by gyrokinetic analysis using artificial local equilibrium scans, suggest a new physical picture for the formation and arrest of this pedestal.

Zero-turbulence manifold in a toroidal plasma.

Sheared toroidal flows can cause bifurcations to zero-turbulent-transport states in tokamak plasmas. The maximum temperature gradients that can be reached are limited by subcritical turbulence driven by the parallel velocity gradient. Here it is shown that q/ϵ (magnetic field pitch/inverse aspect ratio) is a critical control parameter for sheared tokamak turbulence. By reducing q/ϵ, far higher temperature gradients can be achieved without triggering turbulence, in some instances comparable to those found experimentally in transport barriers. The zero-turbulence manifold is mapped out, in the zero-magnetic-shear limit, over the parameter space (γ(E), q/ϵ, R/L(T)), where γ(E) is the perpendicular flow shear and R/L(T) is the normalized inverse temperature gradient scale. The extent to which it can be constructed from linear theory is discussed.

Turbulent transport in tokamak plasmas with rotational shear.

Nonlinear gyrokinetic simulations are conducted to investigate turbulent transport in tokamak plasmas with rotational shear. At sufficiently large flow shears, linear instabilities are suppressed, but transiently growing modes drive subcritical turbulence whose amplitude increases with flow shear. This leads to a local minimum in the heat flux, indicating an optimal E×B shear value for plasma confinement. Local maxima in the momentum fluxes are observed, implying the possibility of bifurcations in the E×B shear. The critical temperature gradient for the onset of turbulence increases with flow shear at low flow shears; at higher flow shears, the dependence of heat flux on temperature gradient becomes less stiff. The turbulent Prandtl number is found to be largely independent of temperature and flow gradients, with a value close to unity.

Whole-brain dynamic CT angiography and perfusion imaging.

The availability of whole brain computed tomography (CT) perfusion has expanded the opportunities for analysing the haemodynamic parameters associated with varied neurological conditions. Examples demonstrating the clinical utility of whole-brain CT perfusion imaging in selected acute and chronic ischaemic arterial neurovascular conditions are presented. Whole-brain CT perfusion enables the detection and focused haemodynamic analyses of acute and chronic arterial conditions in the central nervous system without the limitation of partial anatomical coverage of the brain.

Transport bifurcation in a rotating tokamak plasma.

The effect of flow shear on turbulent transport in tokamaks is studied numerically in the experimentally relevant limit of zero magnetic shear. It is found that the plasma is linearly stable for all nonzero flow shear values, but that subcritical turbulence can be sustained nonlinearly at a wide range of temperature gradients. Flow shear increases the nonlinear temperature gradient threshold for turbulence but also increases the sensitivity of the heat flux to changes in the temperature gradient, except over a small range near the threshold where the sensitivity is decreased. A bifurcation in the equilibrium gradients is found: for a given input of heat, it is possible, by varying the applied torque, to trigger a transition to significantly higher temperature and flow gradients.

Adapting smoking relapse-prevention materials for pregnant and postpartum women: formative research.

OBJECTIVE: To decrease smoking relapse among pregnant and postpartum women by adapting existing, validated relapse-prevention materials to meet the unique needs of pregnant and postpartum women. METHODS: A series of semi-structured interviews and learner verification activities were conducted with pregnant abstinent, postpartum abstinent, and postpartum relapsed women. Results were used to create new relapse-prevention materials, specific to the needs of pregnant and postpartum women, which are currently being used in a randomized clinical trial. RESULTS: Findings are consistent with the recurrent themes in the literature regarding smoking cessation among pregnant and postpartum women and revealed exceptional needs for coping and stress reduction strategies related to remaining abstinent postpartum. Conflict levels were also high in areas of identity, social support, and reasons for quitting. CONCLUSION: By interviewing women about their cessation related needs, the current study was able to produce smoking relapse-prevention materials specific to this population. Having pregnant and postpartum women review the modified program materials before starting the clinical trial enhanced the quality, dependability, and validity of the materials. We await the results of the clinical trial to determine if this intervention is indeed more efficacious than previous attempts to intervene with this population.

L-H transition in the mega-amp spherical tokamak.

H-mode plasmas have been achieved on the MAST spherical tokamak at input power considerably higher than predicted by conventional threshold scalings. Following L- H transition, a clear improvement in energy confinement is obtained, exceeding recent international scalings even at densities approaching the Greenwald density limit. Transition is accompanied by an order-of-magnitude increase in edge-density gradient, a marked decrease in turbulence, the efficient conversion of internal electron Bernstein waves into free space waves, and the onset and saturation of edge poloidal rotation.

An unexpected extended conformation for the third TPR motif of the peroxin PEX5 from Trypanosoma brucei.

A number of helix-rich protein motifs are involved in a variety of critical protein-protein interactions in living cells. One of these is the tetratrico peptide repeat (TPR) motif that is involved, amongst others, in cell cycle regulation, chaperone function and post-translation modifications. So far, these helix-rich TPR motifs have always been observed to be a compact unit of two helices interacting with each other in antiparallel fashion. Here, we describe the structure of the first three TPR-motifs of the peroxin PEX5 from Trypanosoma brucei, the causative agent of sleeping sickness. Peroxins are proteins involved in peroxisome, glycosome and glyoxysome biogenesis. PEX5 is the receptor of the proteins targeted to these organelles by the "peroxisomal targeting signal-1", a C-terminal tripeptide called PTS-1. The first two of the three TPR-motifs of T. brucei PEX5 appear to adopt the canonical antiparallel helix hairpin structure. In contrast, the third TPR motif of PEX5 has a dramatically different conformation in our crystals: the two helices that were supposed to form a hairpin are folded into one single 44 A long continuous helix. Such a conformation has never been observed before for a TPR motif. This raises interesting questions including the potential functional importance of a "jack-knife" conformational change in TPR motifs.

Exploration of the GM1 receptor-binding site of heat-labile enterotoxin and cholera toxin by phenyl-ring-containing galactose derivatives.

Cholera toxin (CT) and the closely related heat-labile enterotoxin of Escherichia coli (LT) are responsible for numerous cases of diarrhea worldwide, leading to considerable morbidity and mortality. The B subunits of these heterohexameric AB(5) toxins form a pentameric arrangement which is responsible for binding to the receptor GM1 of the target epithelial cells of the host. Blocking these B pentamer-receptor interactions forms an avenue for therapeutic intervention. Here, the structural characterization of potential receptor-blocking compounds are described based on the previously identified inhibitor m-nitrophenyl-alpha-D-galactoside (MNPG). The structure of a CTB-MNPG complex confirms that the binding mode of this inhibitor is identical in the two homologous toxins CT and LT and is characterized by a glycosyl linkage geometry that leads to displacement of a well ordered water molecule near the amide group of Gly33 by the O1-substituent of MNPG. This glycosyl geometry is not maintained in the absence of a substituent that can displace this water, as shown by a complex of LTB with p-aminophenyl-alpha-D-galactoside (PAPG). New compounds were synthesized to investigate the feasibility of maintaining the favorable binding interactions exhibited by MNPG while gaining increased affinity through the addition of hydrophobic substituents complementary to either of two hydrophobic regions of the receptor-binding site. The structural characterization of complexes of LTB with two of these compounds, 3-benzylaminocarbonylphenyl-alpha-D-galactoside (BAPG) and 2-phenethyl-7-(2,3-dihydrophthalazine-1,4-dione)-alpha-D-galactoside (PEPG), demonstrates a partial success in this goal. Both compounds exhibit a mixture of binding modes, some of which are presumably influenced by the local packing environment at multiple crystallographically independent binding sites. The terminal phenyl ring of BAPG associates either with the phenyl group of Tyr12 or with the hydrophobic patch formed by Lys34 and Ile58. The latter interaction is also made by the terminal phenyl substituent of PEPG, despite a larger ring system linking the galactose moiety to the terminal phenyl. However, neither BAPG nor PEPG displaces the intended target water molecule. Both of the designed compounds exhibit increased affinity relative to the galactose and to PAPG notwithstanding the failure to displace a bound water, confirming that additional favorable hydrophobic interactions can be gained by extending the starting inhibitor by a hydrophobic tail. The insight gained from these structures should allow the design of additional candidate inhibitors that retain both the glycosyl geometry and water displacement exhibited by MNPG and the favorable hydrophobic interactions exhibited by BAPG and PEPG.

Lovastatin biosynthesis in Aspergillus terreus: characterization of blocked mutants, enzyme activities and a multifunctional polyketide synthase gene.

BACKGROUND: Lovastatin, an HMG-CoA reductase inhibitor produced by the fungus Aspergillus terreus, is composed of two polyketide chains. One is a nonaketide that undergoes cyclization to a hexahydronaphthalene ring system and the other is a simple diketide, 2-methylbutyrate. Fungal polyketide synthase (PKS) systems are of great interest and their genetic manipulation should lead to novel compounds. RESULTS: An A. terreus mutant (BX102) was isolated that could not synthesize the nonaketide portion of lovastatin and was missing a approximately 250 kDa polypeptide normally present under conditions of lovastatin production. Other mutants produced lovastatin intermediates without the methylbutyryl sidechain and were missing a polypeptide of approximately 220 kDa. The PKS inhibitor cerulenin reacted covalently with both polypeptides. Antiserum raised against the approximately 250 kDa polypeptide was used to isolate the corresponding gene, which complemented the BX102 mutation. The gene encodes a polypeptide of 269 kDa containing catalytic domains typical of vertebrate fatty acid and fungal PKSs, plus two additional domains not previously seen in PKSs: a centrally located methyltransferase domain and a peptide synthetase elongation domain at the carboxyl terminus. CONCLUSIONS: The results show that the nonaketide and diketide portions of lovastatin are synthesized by separate large multifunctional PKSs. Elucidation of the primary structure of the PKS that forms the lovastatin nonaketide, as well as characterization of blocked mutants, provides new details of lovastatin biosynthesis.

Structure-based exploration of the ganglioside GM1 binding sites of Escherichia coli heat-labile enterotoxin and cholera toxin for the discovery of receptor antagonists.

Ganglioside GM1 is the natural receptor for cholera toxin (CT) and heat-labile enterotoxin (LT), which are the causative agents of cholera and traveler's diarrhea, respectively. This observation suggests that small molecules interfering with this recognition process may prevent entry of the toxins into intestinal cells, thereby averting their devastating effects. Here, the terminal sugar of ganglioside GM1, galactose, was chosen as a lead in designing such receptor antagonists. Guided by the experimentally determined binding mode of galactose, we selected a "substructure" for searching the Available Chemicals Database, which led to the purchase of 35 galactose derivatives. Initial screening of these compounds in an LT ELISA revealed that 22 of them have a higher affinity for LT than galactose itself. A structurally diverse subset of these galactose derivatives was selected for determination of IC50 values in the LT ELISA and IC50 values in a CT assay, as well as for the determination of Kd's using the intrinsic fluorescence of LT. The best receptor antagonist found in this study was m-nitrophenyl alpha-galactoside with an IC50 of 0.6 (2) mM in the LT ELISA and 0.72 (4) mM in the CT assay, 100-fold lower than both IC50 values of galactose. Careful analysis of our binding data and comparison with crystal structures led to the derivation of correlations between the structure and affinity of the galactose derivatives. These characteristics will be used in the design of a second round of LT and CT receptor antagonists.

Crystal structure of heat-labile enterotoxin from Escherichia coli with increased thermostability introduced by an engineered disulfide bond in the A subunit.

Cholera toxin (CT) produced by Vibrio cholerae and heat-labile enterotoxin (LT-I), produced by enterotoxigenic Escherichia coli, are AB5 heterohexamers with an ADP-ribosylating A subunit and a GM1 receptor binding B pentamer. These toxins are among the most potent mucosal adjuvants known and, hence, are of interest both for the development of anti-diarrheal vaccines against cholera or enterotoxigenic Escherichia coli diarrhea and also for vaccines in general. However, the A subunits of CT and LT-I are known to be relatively temperature sensitive. To improve the thermostability of LT-I an additional disulfide bond was introduced in the A1 subunit by means of the double mutation N40C and G166C. The crystal structure of this double mutant of LT-I has been determined to 2.0 A resolution. The protein structure of the N40C/G166C double mutant is very similar to the native structure except for a few local shifts near the new disulfide bond. The introduction of this additional disulfide bond increases the thermal stability of the A subunit of LT-I by 6 degrees C. The enhancement in thermostability could make this disulfide bond variant of LT-I of considerable interest for the design of enterotoxin-based vaccines.

Cooperative stroke management project by a peer-review organization.

To improve the care of patients in Mississippi through increased adherence to nationally accepted ischemic stroke management guidelines, patterns for ischemic stroke services were determined from hospital chart review. Hospital-specific education and data feedback were performed to encourage international systems improvements. The Mississippi Foundation for Medical Care, Inc, reviewed records of Medicare beneficiaries discharged with the principal diagnosis of acute ischemic stroke from four hospitals over a 1-year period. Records were analyzed for compliance with stroke management guidelines. Hospital-specific and aggregate data were presented to the staffs of each hospital and the hospitals were encouraged to develop internal quality improvement projects. The Foundation reviewed 427 records of acute stroke patients, of whom 375 (87.8%) had ischemic stroke. Among the 427 stroke patients, there were 76 (17.8%) in-hospital deaths. Notable variances from the ischemic stroke management guidelines included those for emergent hypertension management, deep vein thrombosis prophylaxis, evaluation for cause of ischemic stroke, and use of antithrombotic therapy on discharge of ischemic stroke patients. Thus, the management of acute stroke patients in these four regional hospitals in Mississippi often differed from nationally accepted guidelines. We hope to improve the care of stroke patients by using the expertise of academic stroke physicians and hospital-specific analyses that are personally meaningful but not personally threatening to treating physicians.

Mutants of Streptomyces cattleya defective in the synthesis of a factor required for thienamycin production.

Thienamycin non-producing mutants of Streptomydes cattleya were identified that displayed a cross-feeding relationship. A diffusible product from one of these mutants (RK-11) resulted in restoration of thienamycin production when fed to cultures of another mutant (RK-4). In vivo radiolabeling experiments were conducted to test whether the RK-11 mutant produced a late biosynthetic intermediate which contained a carbapenem ring and a cysteaminyl and/or a hydroxyethyl side chain. Both [35S]cystine and [methyl-3H]methionine were used to label the RK-11 product which was then fed to RK-4 cultures. None of the thienamycin subsequently produced by RK-4 converter cells was labeled, implying the lack of either side chain of the thienamycin molecule in the RK-11 product. Further stability studies suggested that the RK-11 product does not contain a carbapenem ring. Additional feeding experiments with RK-4 cells also ruled out the possibility that the RK-11 product is a co-factor necessary for thienamycin production. It is concluded that the RK-11 product may regulate expression of the thienamycin gene cluster.