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Intrinsically disordered proteins (IDPs) play important roles in disease pathologies; however, their lack of defined stable 3D structures make traditional drug design strategies typically less effective against these targets. Based on promising results of targeted covalent inhibitors (TCIs) on challenging targets, we have developed a covalent design strategy targeting IDPs. As a model system we chose tau, an endogenous IDP of the central nervous system that is associated with severe neurodegenerative diseases via its aggregation. First, we mapped the tractability of available cysteines in tau and prioritized suitable warheads. Next, we introduced the selected vinylsulfone warhead to the non-covalent scaffolds of potential tau aggregation inhibitors. The designed covalent tau binders were synthesized and tested in aggregation models, and inhibited tau aggregation effectively. Our results revealed the usefulness of the covalent design strategy against therapeutically relevant IDP targets and provided promising candidates for the treatment of tauopathies.

Vaccines are one of the most cost-effective tools for improving human health and well-being. The impact of a vaccine on population health is partly determined by its coverage rate, the proportion of eligible individuals vaccinated. Coverage rate is a function of the vaccine presentation and the population in which that presentation is deployed. This population includes not only the individuals vaccinated, but also the logistics and healthcare systems responsible for vaccine delivery. Because vaccine coverage rates remain below targets in many settings, vaccine manufacturers and purchasers have a shared interest in better understanding the relationship between vaccine presentation, population characteristics, and coverage rate. While there have been some efforts to describe this relationship, existing research and tools are limited in their ability to quantify coverage rate changes across a broad set of antigens, vaccine presentations, and geographies. In this article, we present a method for estimating the impact of improved vaccine technologies on vaccination coverage rates. It is designed for use with low- and middle-income country vaccination programs. This method uses publicly available data and simple calculations based on probability theory to generate coverage rate values. We first present the conceptual framework and mathematical approach. Using a Microsoft Excel-based implementation, we then apply the method to a vaccine technology in early-stage development: micro-array patch for a measles-rubella vaccine (MR-MAP). Example outputs indicate that a complete switch from the current subcutaneous presentation to MR-MAP in the 73 countries ever eligible for Gavi support would increase overall vaccination coverage by 3.0–4.9 percentage points depending on the final characteristics of the MR-MAP. This change equates to an additional 2.6–4.2 million children vaccinated per year. Our method can be readily extended to other antigens and vaccine technologies to provide quick, low-cost estimates of coverage impact. As vaccine manufacturers and purchasers face increasingly complex decisions, such estimates could facilitate objective comparisons between options and help these decision makers obtain the most value for money.

Over the past decade intrinsically disordered proteins (IDPs) have emerged as a biologically important class of proteins, many of which are of therapeutic relevance. Here, we investigated the interactions between a model IDP system, tau K18, and nine literature compounds that have been reported as having an effect on tau in order to identify a robust IDP&ndash, ligand system for the optimization of a range of biophysical methods. We used NMR, surface plasmon resonance (SPR) and microscale thermophoresis (MST) methods to investigate the binding of these compounds to tau K18, only one showed unambiguous interaction with tau K18. Several near neighbors of this compound were synthesized and their interactions with tau K18 characterized using additional NMR methods, including 1D ligand-observed NMR, diffusion-ordered spectroscopy (DOSY) and 19F NMR. This study demonstrates that it is possible to detect and characterize IDP&ndash, ligand interactions using biophysical methods. However, care must be taken to account for possible artefacts, particularly the impact of compound solubility and where the protein has to be immobilized.

Detecting regularities in the sensory environment licenses predictions that enable adaptive behaviour. However, it is unclear whether predictions about object category, location, or both dimensions are mediated by overlapping systems, and relatedly, whether constructing predictions about both category and location is associated with processing bottlenecks. To examine this issue, in an fMRI study, we presented participants with image-series in which non-deterministic transition probabilities enabled predictions about either the location of the next image, its semantic category, both dimensions, or neither (the latter forming a “no-regularity” random baseline condition). Speaking to a common system, all three predictable conditions resulted in reduced BOLD activity in four clusters: left rostral anterior cingulate cortex; bilateral putamen, caudate and thalamus; right precentral gyrus, and left visual cortex. Pointing to a processing bottleneck, in some regions, a significant interaction between the two factors was found whereby category-predictable series were associated with lower activity – but only when location regularity was absent. Finally, category regularity decreased activation in areas of the ventral visual stream and semantic areas of lateral temporal cortex, and location regularity decreased activation in a dorsal fronto-parietal cluster, long implicated in the endogenous control of spatial attention. Our findings confirm and expand a role for dACC/dmPFC and striatum in monitoring or responding to uncertainty in the environment and point to a limited capacity bottleneck when multiple predictions are concurrently licensed.

Understanding how humans code for and respond to environmental uncertainty/regularity is a question shared by current computational and neurobiological approaches to human cognition. To date, studies investigating neurobiological systems that track input uncertainty have examined responses to uni-sensory streams. It is not known, however, whether there exist brain systems that combine information about the regularity of input streams presented to different senses. We report an fMRI study that aimed to identify brain systems that relate statistical information across sensory modalities. We constructed temporally extended auditory and visual streams, each of which could be random or highly regular, and presented them concurrently. We found strong signatures of “regularity matching” in visual cortex bilaterally; responses were higher when the level of regularity in the auditory and visual streams mismatched than when it matched, [(AudHigh/VisLow and AudLow/VisHigh) >(AudLow/VisLow and AudHigh/VisHigh)]. In addition, several frontal and parietal regions tracked regularity of the auditory or visual stream independently of the other stream's regularity. An individual-differences analysis suggested that signatures of single-modality-focused regularity tracking in these fronto-parietal regions are inversely related to signatures of regularity-matching in visual cortex. Our findings suggest that i) visual cortex is a junction for integration of temporally-extended auditory and visual inputs and that ii) multisensory regularity-matching depends on balanced processing of both input modalities. We discuss the implications of these findings for neurobiological models of uncertainty and for understanding computations that underlie multisensory interactions in occipital cortex.

Myeloid cell leukemia 1 (Mcl-1), an antiapoptotic member of the Bcl-2 family of proteins, whose upregulation when observed in human cancers is associated with high tumor grade, poor survival, and resistance to chemotherapy, has emerged as an attractive target for cancer therapy. Here, we report the discovery of selective small molecule inhibitors of Mcl-1 that inhibit cellular activity. Fragment screening identified thienopyrimidine amino acids as promising but nonselective hits that were optimized using nuclear magnetic resonance and X-ray-derived structural information. The introduction of hindered rotation along a biaryl axis has conferred high selectivity to the compounds, and cellular activity was brought on scale by offsetting the negative charge of the anchoring carboxylate group. The obtained compounds described here exhibit nanomolar binding affinity and mechanism-based cellular efficacy, caspase induction, and growth inhibition. These early research efforts illustrate drug discovery optimization from thienopyrimidine hits to a lead compound, the chemical series leading to the identification of our more advanced compounds S63845 and S64315.

Aim: The objective was to investigate factors influencing patients’ willingness to pay (WTP) and physician’s views on the cost of therapy for two contrasted chronic diseases, chronic heart failure and psoriasis. Methods: A cross-sectional survey was conducted in ten developing countries, using a stated WTP contingent valuation method. Multivariate analyses were performed by linear regression. Results: Independent factors influencing patient WTP were income (+0.04 $PPP [purchasing power parity] in WTP per $PPP in monthly income; p Conclusion: Disease-specific factors may affect WTP for treatment that should be accounted for to support effective public health programs in developing countries.

Sleep has been shown to subtly disrupt the spatial organization of functional connectivity networks in the brain, but in a way that largely preserves the connectivity within sensory cortices. Here we evaluated the hypothesis that sleep does impact sensory cortices, but through alteration of activity dynamics. We therefore examined the impact of sleep on hemodynamics using a method for quantifying non-random, high frequency signatures of the blood-oxygen-level dependent (BOLD) signal (amplitude variance asymmetry; AVA). We found that sleep was associated with the elimination of these dynamics in a manner that is restricted to auditory, motor and visual cortices. This elimination was concurrent with increased variance of activity in these regions. Functional connectivity between regions showing AVA during wakefulness maintained a relatively consistent hierarchical structure during wakefulness and N1 and N2 sleep, despite a gradual reduction of connectivity strength as sleep progressed. Thus, sleep is related to elimination of high frequency non-random activity signatures in sensory cortices that are robust during wakefulness. The elimination of these AVA signatures conjointly with preservation of the structure of functional connectivity patterns may be linked to the need to suppress sensory inputs during sleep while still maintaining the capacity to react quickly to complex multimodal inputs., Highlights • We examined time-domain dynamics of BOLD activity during sleep. • We used a method previously showing unique dynamics in sensory cortices during wakefulness. • We find elimination of wakefulness dynamics during N2 and N3 sleep, but not N1.

There is limited data pertaining to the triage and transportation of patients with penetrating trauma in rural states. Large urban trauma centers have found rapid transport to be beneficial even when done by nonemergency medical staff. However, there is limited application to a rural state with only a single level 1 trauma center.This a retrospective observational study of 854 trauma patients transported by helicopter emergency services between 2009 and 2015 to the state's only level 1 trauma center.After excluding patients with other injuries or lack of data, 854 patients underwent final analysis. Compared with penetrating trauma, blunt trauma had a significantly different chance of survival (92.0% versus 81.2%, P = 0.002) and a significantly different injury severity score (17 ± 12 versus 12 ± 9, P = 0.002). After controlling for blunt injuries, age, gender, injury severity score, tachycardia, tachypnea, hypotension, glasgow coma scale, and dispatch to hospital arrival time in multivariate analysis, blunt trauma had higher odds of survival than penetrating trauma (OR, 5.97; 95% CI, 2.52-14.12; P = 0.001 = 1). Gender, tachycardia, tachypnea, and dispatch to arrival time did not impact a patient's likelihood of survival.Penetrating trauma has a higher mortality when compared with blunt trauma in Helicopter Emergency Services transported patients in a rural state. Perhaps a new algorithm in the management of penetrating trauma would include hemorrhage control at a locoregional hospital before definitive care. Further study is required to understand the exact variables that lead to a higher mortality in penetrating trauma in a rural state.

Background Artemisinin-based combination therapy (ACT) is recommended as the first-line anti-malarial treatment strategy in sub-Saharan African countries. WHO policy recommends parasitological confirmation by microscopy or rapid diagnostic test (RDT) in all cases of suspected malaria prior to treatment. Gaps remain in understanding the factors that influence patient treatment-seeking behaviour and anti-malarial drug purchase decisions in the private sector. The objective of this study was to identify patient treatment-seeking behaviour in Ghana, Kenya, Nigeria, Tanzania, and Uganda. Methods Face-to-face patient interviews were conducted at a total of 208 randomly selected retail outlets in five countries. At each outlet, exit interviews were conducted with five patients who indicated they had come seeking anti-malarial treatment. The questionnaire was anonymous and standardized in the five countries and collected data on different factors, including socio-demographic characteristics, history of illness, diagnostic practices (i.e. microscopy or RDT), prescription practices and treatment purchase. The price paid for the treatment was also collected from the outlet vendor. Results A total of 994 patients were included from the five countries. Location of malaria diagnosis was significantly different in the five countries. A total of 484 blood diagnostic tests were performed, (72.3% with microscopy and 27.7% with RDT). ACTs were purchased by 72.5% of patients who had undergone blood testing and 86.5% of patients without a blood test, regardless of whether the test result was positive or negative (p

Complex systems are described according to two central dimensions: (a) the randomness of their output, quantified via entropy; and (b) their complexity, which reflects the organization of a system's generators. Whereas some approaches hold that complexity can be reduced to uncertainty or entropy, an axiom of complexity science is that signals with very high or very low entropy are generated by relatively non-complex systems, while complex systems typically generate outputs with entropy peaking between these two extremes. In understanding their environment, individuals would benefit from coding for both input entropy and complexity; entropy indexes uncertainty and can inform probabilistic coding strategies, whereas complexity reflects a concise and abstract representation of the underlying environmental configuration, which can serve independent purposes, e.g., as a template for generalization and rapid comparisons between environments. Using functional neuroimaging, we demonstrate that, in response to passively processed auditory inputs, functional integration patterns in the human brain track both the entropy and complexity of the auditory signal. Connectivity between several brain regions scaled monotonically with input entropy, suggesting sensitivity to uncertainty, whereas connectivity between other regions tracked entropy in a convex manner consistent with sensitivity to input complexity. These findings suggest that the human brain simultaneously tracks the uncertainty of sensory data and effectively models their environmental generators., Highlights • Complexity science holds that highly ordered and random signals have low complexity. • We examine whether effective connectivity tracks both stimulus entropy and complexity. • Connectivity tracked entropy both linearly and via an inverse U-shaped profile. • Both profiles were identified for hippocampal and anterior cingulate networks.

Current models of brain function assign a central role to predictive processes calibrated to the structure of the environment. Although several neuroimaging studies have examined how the human brain encodes the uncertainty of incoming stimuli, most have relied exclusively on experimental manipulations of uncertainty in which stimuli were presented in a single sensory modality, and further assumed that neural responses vary monotonically with uncertainty. This has left a gap in theoretical development with respect to two core issues: i) are there cross-modal brain systems that encode input uncertainty in way that generalizes across sensory modalities, and ii) are there brain systems that track input uncertainty in a non-monotonic fashion? Here we directly addressed the issues of cross-modal and non-monotonic processing by quantifying neural sensitivity to uncertainty in auditory, visual and audiovisual inputs using multivariate pattern analysis. We found signatures of cross-modal encoding in frontoparietal, orbitofrontal, and association cortices using a searchlight cross-classification analysis where classifiers trained to discriminate levels of uncertainty in one modality were tested in another modality. Additionally, we found widespread systems encoding uncertainty non-monotonically using classifiers trained to discriminate intermediate levels of uncertainty from both the highest and lowest uncertainty levels. These findings comprise the first comprehensive report of cross-modal and non-monotonic neural sensitivity to statistical regularities in the environment, and suggest that conventional paradigms testing for monotonic responses to uncertainty in a single sensory modality may have limited generalizability.

Libraries of nonpurified resorcinol amide derivatives were screened by surface plasmon resonance (SPR) to determine the binding dissociation constant (off-rate, kd) for compounds binding to the pyruvate dehydrogenase kinase (PDHK) enzyme. Parallel off-rate measurements against HSP90 and application of structure-based drug design enabled rapid hit to lead progression in a program to identify pan-isoform ATP-competitive inhibitors of PDHK. Lead optimization identified selective sub-100-nM inhibitors of the enzyme which significantly reduced phosphorylation of the E1α subunit in the PC3 cancer cell line in vitro.

Purpose: The purpose of this study was to evaluate patient satisfaction and clinical outcomes among subjects with mandibular overdentures supported by one or two immediately placed dental implants 5 years after loading. Materials and Methods: Thirty-six subjects (16 men and 20 women) received one or two dental implants in the anterior mandible, and all implants were loaded the day of surgery. Subjects were scheduled for follow-up 3-, 6-, and 12 months after implant placement and thereafter annually for 4 more years. Patient satisfaction scores were measured with the Oral Health Impact Profile-EDENT (OHIPEDENT) questionnaire. Results: Seventeen subjects (7 male and 10 female) with a mean age of 59.4 years (range, 44 to 74 years) were available for the 5-year follow-up examination. Nine subjects with 10 failing implants were excluded during the first year and nine subjects were lost to follow-up. No implants failed between the 12- and 60-month follow-up examinations, and the need for denture maintenance was low. Mean peri-implant bone change was 0.92 mm, and the Spearman test failed to show correlation between the insertion torque value and implant stability quotient. Patient satisfaction scores increased significantly when compared with baseline values and continued to be high for both groups, with no significant differences. Conclusion: Ten implants in nine subjects failed early, but no failures were observed after the 12-month examination. No significant differences were found between subjects in the two groups with respect to implant survival rates and peri-implant bone loss, and patient satisfaction scores continued to be high. Although patient satisfaction and implant success were high during the 12- to 60-month period, the results should be interpreted with caution because of the high number of failing implants and patients lost to followup. More research is needed to study outcomes of treatment with immediately loaded mandibular implant overdentures.

Pyruvate dehydrogenase kinase (PDK) is a pivotal enzyme in cellular energy metabolism that has previously been implicated in cancer through both RNAi based studies and clinical correlations with poor prognosis in several cancer types. Here, we report the discovery of a novel and selective ATP competitive pan-isoform inhibitor of PDK, VER-246608. Consistent with a PDK mediated MOA, VER-246608 increased pyruvate dehydrogenase complex (PDC) activity, oxygen consumption and attenuated glycolytic activity. However, these effects were only observed under D-glucose-depleted conditions and required almost complete ablation of PDC E1α subunit phosphorylation. VER-246608 was weakly anti-proliferative to cancer cells in standard culture media; however, depletion of either serum or combined D-glucose/L-glutamine resulted in enhanced cellular potency. Furthermore, this condition-selective cytostatic effect correlated with reduced intracellular pyruvate levels and an attenuated compensatory response involving deamination of L-alanine. In addition, VER-246608 was found to potentiate the activity of doxorubicin. In contrast, the lipoamide site inhibitor, Nov3r, demonstrated sub-maximal inhibition of PDK activity and no evidence of cellular activity. These studies suggest that PDK inhibition may be effective under the nutrient-depleted conditions found in the tumour microenvironment and that combination treatments should be explored to reveal the full potential of this therapeutic strategy.

Condyle fractures are a common injury, but only a few of these injuries require immediate or late reconstruction. The complications that most frequently necessitate condylar reconstruction include proximal segment degeneration, malunion, and ankylosis. Costochondral grafts and total joint prostheses, both stock and custom, remain the most common methods of reconstruction. Reconstruction plates with condylar extensions should only be used temporarily as an unacceptable number cause serious complications. Distraction osteogenesis may have an occasional role in reconstructing the posttraumatic condyle.

There is a pressing need for new technologies that improve the efficacy and efficiency of drug discovery. Structure-based methods have contributed towards this goal but they focus on predicting the binding affinity of protein-ligand complexes, which is notoriously difficult. We adopt an alternative approach that evaluates structural, rather than thermodynamic, stability. As bioactive molecules present a static binding mode, we devised dynamic undocking (DUck), a fast computational method to calculate the work necessary to reach a quasi-bound state at which the ligand has just broken the most important native contact with the receptor. This non-equilibrium property is surprisingly effective in virtual screening because true ligands form more-resilient interactions than decoys. Notably, DUck is orthogonal to docking and other 'thermodynamic' methods. We demonstrate the potential of the docking-undocking combination in a fragment screening against the molecular chaperone and oncology target Hsp90, for which we obtain novel chemotypes and a hit rate that approaches 40%.

Inhibitors of the Hsp90 molecular chaperone are showing promise as anti-cancer agents. Here we describe a series of 4-aryl-5-cyanopyrrolo[2,3-d]pyrimidine ATP competitive Hsp90 inhibitors that were identified following structure-driven optimization of purine hits revealed by NMR based screening of a proprietary fragment library. Ligand-Hsp90 X-ray structures combined with molecular modeling led to the rational displacement of a conserved water molecule leading to enhanced affinity for Hsp90 as measured by fluorescence polarization, isothermal titration calorimetry and surface plasmon resonance assays. This displacement was achieved with a nitrile group, presenting an example of efficient gain in binding affinity with minimal increase in molecular weight. Some compounds in this chemical series inhibit the proliferation of human cancer cell lines in vitro and cause depletion of oncogenic Hsp90 client proteins and concomitant elevation of the co-chaperone Hsp70. In addition, one compound was demonstrated to be orally bioavailable in the mouse. This work demonstrates the power of structure-based design for the rapid evolution of potent Hsp90 inhibitors and the importance of considering conserved water molecules in drug design.

Pin1 is an emerging oncology target strongly implicated in Ras and ErbB2-mediated tumourigenesis. Pin1 isomerizes bonds linking phospho-serine/threonine moieties to proline enabling it to play a key role in proline-directed kinase signalling. Here we report a novel series of Pin1 inhibitors based on a phenyl imidazole acid core that contains sub-μM inhibitors. Compounds have been identified that block prostate cancer cell growth under conditions where Pin1 is essential.

Traditionally, the left frontal and parietal lobes have been associated with language production while regions in the temporal lobe are seen as crucial for language comprehension. However, recent evidence suggests that the classical language areas constitute an integrated network where each area plays a crucial role both in speech production and perception. We used functional MRI to examine whether observing speech motor movements (without auditory speech) relative to non-speech motor movements preferentially activates the cortical speech areas. Furthermore, we tested whether the activation in these regions was modulated by task difficulty. This dissociates between areas that are actively involved with speech perception from regions that show an obligatory activation in response to speech movements (e.g. areas that automatically activate in preparation for a motoric response). Specifically, we hypothesized that regions involved with decoding oral speech would show increasing activation with increasing difficulty. We found that speech movements preferentially activate the frontal and temporal language areas. In contrast, non-speech movements preferentially activate the parietal region. Degraded speech stimuli increased both frontal and parietal lobe activity but did not differentially excite the temporal region. These findings suggest that the frontal language area plays a role in visual speech perception and highlight the differential roles of the classical speech and language areas in processing others' motor speech movements.

Heat shock protein (HSP) 60 is a mitochondrial and cytosolic protein. Previously, we reported that HSP60 doubled in end-stage heart failure, even though levels of the protective HSP72 were unchanged. Furthermore, we observed that acute injury in adult cardiac myocytes resulted in movement of HSP60 to the plasma membrane. We hypothesized that the inflammatory state of heart failure would cause translocation of HSP60 to the plasma membrane and that this would provide a pathway for cardiac injury. Two models were used to test this hypothesis: 1) a rat model of heart failure and 2) human explanted failing hearts. We found that HSP60 localized to the plasma membrane and was also found in the plasma early in heart failure. Plasma membrane HSP60 localized to lipid rafts and was detectable on the cell surface with the use of both flow cytometry and confocal microscopy. Localization of HSP60 to the cell surface correlated with increased apoptosis. In heart failure, HSP60 is in the plasma membrane fraction, on the cell surface, and in the plasma. Membrane HSP60 correlated with increased apoptosis. Release of HSP60 may activate the innate immune system, promoting a proinflammatory state, including an increase in TNF-α. Thus abnormal trafficking of HSP60 to the cell surface may be an early trigger for myocyte loss and the progression of heart failure.

The targeting of RNA for the design of novel anti-viral compounds represents an area of vast potential. We have used NMR and computational methods to model the interaction of a series of synthetic inhibitors of the in vitro RNA binding activities of a peptide derived from the transcriptional activator protein, Tat, from human immunodeficiency virus type 1. Inhibition has been measured through the monitering of fluorescence resonance energy transfer between fluorescently labeled peptide and RNA components. A series of compounds containing a bi-aryl heterocycle as one of the three substituents on a benzylic scaffold, induce a novel, inactive TAR conformation by stacking between base-pairs at the site of a three-base bulge within TAR. The development of this series resulted in an enhancement in potency (with Ki < 100 nM in an in vitro assay) and the removal of problematic guanidinium moieties. Ligands from this series can act as inhibitors of Tat-induced transcription in a cell-free system. This study validates the drug design strategy of using a ligand to target the RNA receptor in a non-functional conformation.

Case: A traumatic nondisplaced periprosthetic trochanteric fracture around a well-fixed and functioning Birmingham Hip Resurfacing prosthesis was managed with a reconstruction intramedullary nail. The surgical procedure provided excellent pain relief, and the patient was able to mobilize with crutches the day after surgery. Follow-up at 1 year confirmed excellent clinical and radiographic results. Conclusion: In a patient with a previously well-functioning hip resurfacing prosthesis, internal fixation rather than revision hip arthroplasty can offer good clinical and radiographic outcomes. The use of a reconstruction intramedullary nail may be a good option for fixing a periprosthetic trochanteric fracture around a Birmingham Hip Resurfacing implant.

Saturable radioligand binding of [3H]L-689,560 to the glycine site of the N-methyl-D-aspartate (NMDA) glutamate receptor complex was employed to determine the density of this receptor in putamen, caudate and nucleus accumbens taken post mortem from schizophrenic patients and matched controls. Receptor density was found to be significantly increased in putamen of schizophrenics (P=0.012), although no significant change was found in the other two areas studied. Further experiments following 21 days administration of haloperidol to rats provided no evidence that the increase was related to prior drug treatment.

Neuroimaging studies have shown that task demands affect connectivity patterns in the human brain not only during task performance but also during subsequent rest periods. Our goal was to determine whether ongoing connectivity patterns during rest contain information about both the current rest state, as well as the recently terminated task. Our experimental design consisted of two types of active tasks that were followed by two types of low‐demand rest states. Using this design, we examined whether hippocampal functional connectivity during wakeful rest reflects both features of a recently terminated task and those of the current resting‐state condition. We identified four types of networks: (i) one whose connectivity with the hippocampus was determined only by features of a recently terminated task, (ii) one whose connectivity was determined only by features of the current resting‐state, (iii) one whose connectivity reflected aspects of both the recently terminated task and ongoing resting‐state features, and (iv) one whose connectivity with the hippocampus was strong, but not affected by any external factor. The left and right hippocampi played distinct roles in these networks. These findings suggest that ongoing hippocampal connectivity networks mediate information integration across multiple temporal scales, with hippocampal laterality moderating these connectivity patterns. Hum Brain Mapp 36:519–537, 2015. © 2014 Wiley Periodicals, Inc.

Nuclear magnetic resonance (NMR) is well suited to probing the interactions between ligands and macromolecular receptors. It is a truly label-free technique, requiring only the presence of atoms (usually (1)H or (19)F) which give rise to observable resonances on either the ligand or the receptor. A number of parameters associated with these resonances can be used to distinguish rapidly tumbling compounds from ligands which bind to a macromolecular receptor. As such, NMR reports directly on the molecular components involved in the binding interaction whilst avoiding artifacts arising from the addition of an observable label. NMR is also unique amongst biophysical techniques in giving information on the chemical nature of almost all of the constituents present in the sample, thus allowing ready identification of sample, contaminants, degraded material and buffers. Solution phase NMR is also free of artifacts introduced by the presence of a solid support or matrix, although some interesting NMR techniques have been developed to identify ligand-receptor interactions in both solid and heterogeneous phase systems.NMR can readily report on molecular interactions across a wide range of affinities and timescales. Although NMR is not an inherently sensitive technique, the development of cryogenic probeheads over the past decade has dramatically increased the range of applicability of the technique and reduced the stringent sample requirements that used to be regarded as an "Achilles' heel" of NMR. The last, but by no means the least, NMR has the ability to determine structural information at atomic resolution; this has proved to be particularly useful when applied to those protein-ligand systems which cannot be readily crystallized.

Background In 2009, the Global Fund to Fight AIDS, Tuberculosis and Malaria established the Affordable Medicines Facility-malaria (AMFm) in order to increase access to quality-assured artemisinin combination therapy (QAACT). AMFm Phase 1, which includes nine pilot programmes in eight countries, was launched in 2009. The objective of this study was to assess anti-malarial stock and purchase patterns at private outlets in five AMFm Phase 1 countries in regard to three of the core AMFm goals: increase the affordability of QAACT, increase the availability of QAACT, and crowd out artemisinin monotherapies and other substandard therapies. Methods The study was conducted between April and May 2012 and included interviews with personnel in 598 private pharmaceutical outlets in Ghana, Kenya, Nigeria, Tanzania, and Uganda. Questionnaires were administered at private retail outlets and the data were analyzed to assess within- and between-country differences in QAACT price, availability, and popularity. Results AMFm medications were less expensive than their non-AMFm counterparts, yet prices for both types were above country-specific suggested retail prices. Market penetration of AMFm QAACT in both urban and rural areas was high, although stock-outs of both AMFm and non-AMFm products were more common in rural compared with urban outlets in Ghana and Kenya (p = 0.0013). Government recommendation was the most significant factor influencing anti-malarial stock choices in urban (41.5%) and rural (31.9%) outlets. The three top-selling anti-malarials reported for both urban and rural areas in each country were, with the exception of rural Uganda and urban Nigeria, combination therapies. Conclusions Results from this study indicate that the AMFm has not fully achieved its affordability and crowd-out objectives. Still, the final purchase price of AMFm QAACT was substantially lower than non-AMFm equivalents. Moreover, for both urban and rural areas, AMFm QAACT availability was found to be high, and the various forms of QAACT were the best-selling products among all anti-malarials. These findings suggest a continued need for initiatives like the AMFm that improve the affordability and accessibility of QAACT. Similar programmes may be especially effective if employed in combination with rapid diagnostic testing to ensure the appropriate use of these products.

The objective of the study was to evaluate and compare treatments with mandibular overdentures supported by one or two immediately placed implants 3 years after loading.Thirty-six edentulous subjects were eligible for inclusion. Using a random sampling system, one or two implants were placed in the mandible. Separate ball attachments were connected to the implants, and the denture was relined and delivered the day of surgery with the retentive components incorporated in the denture base. At the follow-up examinations peri-implant bone levels, implant and denture stability/retention, and need for maintenance and adjustments were evaluated. Moreover, the OHIP-EDENT questionnaire was used to measure patient satisfaction.Nineteen subjects (10 men and 9 women) with a mean age of 56 years were available for the 3-year follow-up examination. The group with 1 implant (Group 1) consisted of 11 subjects (5 women and 6 men) while the remaining 8 (5 women and 3 men) belonged to Group2. Nine subjects had been excluded during the first year due to failing implants, 6 had moved, 1 had died, and 1 reported severe illness. No implant failures between the 12-and 36 month follow-up were observed. The mean peri-implant bone change was.86 mm and the Implant Stability Quotient showed only minor changes with no significant difference between the groups when compared with the 12-month follow-up. Patient satisfaction scores increased significantly when compared with the baseline values and continued to be high for both groups and need for denture maintenance was low.No significant differences were found between subjects in the two groups with respect to implant survival and peri implant bone loss, and patient satisfaction scores continued to be high for both groups. Need for denture maintenance was low in both groups.

Neuroimaging research has identified several brain systems sensitive to statistical regularities within environmental input. However, the continuous input impinging on sensory organs is rarely stationary and its degree of regularity may itself change over time. The goals of the current fMRI study were to identify systems sensitive to changes in statistical regularities within an ongoing stimulus, and determine to what extent sensitivity to such changes depends on intentional monitoring of order. We predicted that changes in regularity would be coded for in systems previously associated with statistical coding (hippocampus and middle frontal regions) or event segmentation (posterior medial regions). Participants listened to a rapid train of four different tones whose order levels fluctuated over time. In an active task, participants monitored the tones and indicated when they perceived a change in regularity; in a passive task, they performed a concurrent visuo-motor task and could ignore the auditory input. Behavioral responses in the active task were used to define points of consensus between participants regarding changes in regularity. Activity in 7.5s epochs that preceded these order-change points was contrasted with activity during matched-length epochs where no participant indicated a change in order. We found that brain regions differentiating these two types of epochs matched those identified in prior research as mediating event segmentation in narratives and movies. These consisted mainly of medial posterior parietal and occipital regions, with limited involvement of temporal and lateral frontal cortices and no hippocampal involvement. In both tasks, order-change epochs were associated with a higher BOLD response than stable-order epochs, but the specific regions showing this pattern varied across tasks. We suggest that partitioning an input stream on the basis of statistical shifts constitutes a basic neural function underlying the ability to segment both semantic and non-semantic inputs. We further discuss the implications of these findings for neurobiological theories of statistical coding and event segmentation.

The purpose of the present study was to evaluate and compare treatment with mandibular overdentures supported by one or two implants placed and loaded immediately.Edentulous subjects with maxillary and mandibular complete dentures were recruited to participate in the study. All subjects received new maxillary and mandibular complete dentures, after which they were scheduled for implant surgery. Using a randomized sampling system, one or two implants were placed in the interforaminal area, and separate ball attachments were connected to the implants. The existing mandibular denture was relined, provided with retentive o-rings, and delivered the day of surgery. Subjects were seen for follow-up examinations after 3, 6, and 12 months, during which marginal bone levels and implant stability were measured. Survival rates were calculated and the data were analyzed statistically.Thirty-six subjects (16 men and 20 women with a mean age of 53.2 years [range, 38 to 69 years]) received 55 implants. Seventeen subjects received one implant in the symphyseal area, and the other 19 received two implants in the canine positions. Ten implants in nine subjects failed during the observation period, and three subjects with two implants each withdrew from the study, resulting in a 12-month implant survival rate of 81.8%. Three subjects lost their only implant and one patient lost both implants. The remaining five subjects lost one of their two implants. Six of the failures occurred during the first month after placement.Immediate loading of one or two separate implants by means of ball attachment-supported mandibular overdentures should be performed with caution, as a higher than expected implant failure rate was observed. No statistically significant differences in failure rates between the two groups were observed.

To determine the general utility of clinical (Asthma Control Test) and physiologic (forced expiratory volume in the first second of exhalation [FEV(1)] and fractionated exhaled nitric oxide level [FeNO]) parameters for characterizing asthma patients.Two cross-sectional independent studies simultaneously enrolled 100 patients in the US and 109 patients in Spainor = 18 years of age with a physician-diagnosis of asthma and confirmed by aor = 12% improvement in FEV(1) after bronchodilators or the presence of airway hyperresponsiveness, a central feature of asthma, as measured by methacholine challenge (PC(20)10 mg/mL). There was no restriction on asthma severity or treatment. Patients were excluded if they had a diagnosis of chronic obstructive pulmonary disease and/or were current cigarette smokers. Statistical analyses were performed to compare ACT, FeNO, and spirometry within and between sites.Population characteristics revealed significant differences in distributions of age, percent-predicted FEV(1) (%FEV(1)), FeNO, inhaled corticosteroid usage, and atopy between the two populations. The Spain site enrolled younger patients with milder asthma, based on higher %FEV(1) values and less frequent treatment with inhaled corticosteroids. At each site, mean FeNO levels decreased as asthma control categories increased, and means were lower in the US. There was a negative correlation between ACT and FeNO that was statistically significant for Spain patients not treated with inhaled corticosteroids.The results of this study support the use of FeNO as an adjunctive tool for assessing asthma primarily in mild inhaled corticosteroid (ICS)-naïve asthma patients. The lack of correlation of ACT with FeNO in this and other studies across the entire population appears to reflect the heterogeneity of asthma patients who have an admixture of asthma severity and treatment regimens making it very difficult to appreciate the nuances of sensitive tests like FeNO.

Inhibitors of the Hsp90 molecular chaperone are showing considerable promise as potential molecular therapeutic agents for the treatment of cancer. Here we describe novel 2-aminothieno[2,3-d]pyrimidine ATP competitive Hsp90 inhibitors, which were designed by combining structural elements of distinct low affinity hits generated from fragment-based and in silico screening exercises in concert with structural information from X-ray protein crystallography. Examples from this series have high affinity (IC50 = 50-100 nM) for Hsp90 as measured in a fluorescence polarization (FP) competitive binding assay and are active in human cancer cell lines where they inhibit cell proliferation and exhibit a characteristic profile of depletion of oncogenic proteins and concomitant elevation of Hsp72. Several examples (34a, 34d and 34i) caused tumor growth regression at well tolerated doses when administered orally in a human BT474 human breast cancer xenograft model.

We report the prevalence and incidence of methicillin-resistant Staphylococcus aureus (MRSA) colonisation during the patient journey for patients admitted to orthopaedic and trauma wards. Patients were swabbed for MRSA colonisation on admission, transfer, and discharge from hospital. Elective patients undergoing major joint surgery were also swabbed at a pre-operative assessment clinic. Of the 559 patients admitted, 323 (101 elective, 192 trauma and 30 non-orthopaedic) were included in the study. Of these, 27 elective (27%), 41 trauma (21%), and seven non-orthopaedic (23%) patients were colonised with MRSA at any time during the audit period. There is a high prevalence of MRSA colonisation in patients admitted to the orthopaedic and trauma wards in our setting. A policy of pre-admission screening, though able to identify MRSA carriage, does not guarantee that patients are not colonised in the period between screening and admission. We suggest to screen for MRSA all patients admitted to an orthopaedic ward.

Perceptual temporal order judgments require an individual to determine the relative timing of two spatially separate events. Here we reveal the brain regions involved with this task. We had participants observe perceptually identical visual stimuli while conducting two different tasks: discriminating temporal order or discriminating spatial properties. By contrasting the functional magnetic resonance imaging signals during these tasks, we were able to isolate regions specifically engaged by each task. Participants observed two briefly presented rectangles. In one task, participants were instructed to report which appeared first, and, in the other, they were requested to report which rectangle was squarer. A potential confound of this study is that the temporal order judgment (TOJ) task required processing of brief events (onsets), whereas the shape task did not require temporal selectivity. To address this, we conducted a second study in which both tasks required discriminating brief events concurrent with the object onsets. The stimuli were similar to the first experiment, except a gray line was briefly superimposed on each rectangle at onset. Participants reported either which rectangle appeared first (TOJ) or which rectangle had a slightly wider gray line (shape). The first study found that the TOJ task resulted in greater bilateral activation of the temporal parietal junction (TPJ). The second revealed TOJ activation in the TPJ of the left hemisphere. This suggests that TPJ activation increases when we need to temporally sequence information. This finding supports the notion that the TPJ may be a crucial component of the “when” pathway.

The decision to manage an organized hemarthrosis conservatively or surgically is difficult and still controversial, especially in the postoperative patient compared with the trauma patient. We describe a cheap, effective, and readily available technique for the surgeon who elects for a surgical solution to this problem using a Karman catheter (Rocket Medical PLC, Watford, UK) during arthroscopy to complete an irrigation and vacuum circuit. Arthroscopy: The Journal of Arthroscopic and Related Surgery, Vol 18, No 5 (May-June), 2002: E26

We studied a group of verbal memory specialists to determine whether intensive oral text memory is associated with structural features of hippocampal and lateral-temporal regions implicated in language processing. Professional Vedic Sanskrit Pandits in India train from childhood for around 10years in an ancient, formalized tradition of oral Sanskrit text memorization and recitation, mastering the exact pronunciation and invariant content of multiple 40,000–100,000 word oral texts. We conducted structural analysis of gray matter density, cortical thickness, local gyrification, and white matter structure, relative to matched controls. We found massive gray matter density and cortical thickness increases in Pandit brains in language, memory and visual systems, including i) bilateral lateral temporal cortices and ii) the anterior cingulate cortex and the hippocampus, regions associated with long and short-term memory. Differences in hippocampal morphometry matched those previously documented for expert spatial navigators and individuals with good verbal working memory. The findings provide unique insight into the brain organization implementing formalized oral knowledge systems.

CD5 is a type-I transmembrane glycoprotein found on thymocytes, T-cells and a subset of B-cells. The extracellular region consists of three domains belonging to the scavenger receptor cysteine-rich (SRCR) superfamily for which the three-dimensional polypeptide fold is as yet unknown. Glycosylated CD5 domain 1 (CD5d1) has been obtained by expression by secretion from both Chinese hamster ovary (CHO) cells and Pichia pastoris. Recombinant CD5d1 expressed in this manner was shown to be correctly folded by binding to anti-CD5 L17F12/Leu1 monoclonal antibody. Preliminary nuclear magnetic resonance (NMR) spectra obtained for CD5d1 (residues 1-118) had spectral dispersion typical of a folded protein, but otherwise of such poor quality that NMR structural studies were not feasible. The analysis of glycoproteins by NMR is frustrated by sample heterogeneity and poor spectral quality associated with glycan resonance overlap and the potential for increased line-widths due to the large hydrodynamic volume. In order to pursue NMR structural studies of CD5d1 it was necessary to optimize the quality of NMR spectra of CD5d1. A range of constructs of varying length and carbohydrate content were expressed in CHO cells and in P. pastoris. In addition the P. pastoris CD5d1 proved susceptible to N-glycan cleavage with endoglycosidase H. The protein products were characterised using size exclusion chromatography, NMR measurement of translational self-diffusion coefficients and two-dimensional 1H nuclear Overhauser effect spectroscopy experiments. Removal of an eight residue O-glycosylated C-terminal peptide, in particular, resulted in significant improvements in the quality of the CD5d1 NMR data, while retaining native protein structure. Two-dimensional heteronuclear NMR spectroscopy of nitrogen-15 isotope labelled deglycosylated CD5d1 (residues 1-110) prepared from P. pastoris suggests that this protein product is now amenable to solution structure determination.