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The first kinase inhibitor drug approval in 2001 initiated a remarkable decade of tyrosine kinase inhibitor drugs for oncology indications, but a void exists for serine/threonine protein kinase inhibitor drugs and central nervous system indications. Stress kinases are of special interest in neurological and neuropsychiatric disorders due to their involvement in synaptic dysfunction and complex disease susceptibility. Clinical and preclinical evidence implicates the stress related kinase p38αMAPK as a potential neurotherapeutic target, but isoform selective p38αMAPK inhibitor candidates are lacking and the mixed kinase inhibitor drugs that are promising in peripheral tissue disease indications have limitations for neurologic indications. Therefore, pursuit of the neurotherapeutic hypothesis requires kinase isoform selective inhibitors with appropriate neuropharmacology features. Synaptic dysfunction disorders offer a potential for enhanced pharmacological efficacy due to stress-induced activation of p38αMAPK in both neurons and glia, the interacting cellular components of the synaptic pathophysiological axis, to be modulated. We report a novel isoform selective p38αMAPK inhibitor, MW01-18-150SRM (=MW150), that is efficacious in suppression of hippocampal-dependent associative and spatial memory deficits in two distinct synaptic dysfunction mouse models. A synthetic scheme for biocompatible product and positive outcomes from pharmacological screens are presented. The high-resolution crystallographic structure of the p38αMAPK/MW150 complex documents active site binding, reveals a potential low energy conformation of the bound inhibitor, and suggests a structural explanation for MW150's exquisite target selectivity. As far as we are aware, MW150 is without precedent as an isoform selective p38MAPK inhibitor or as a kinase inhibitor capable of modulating in vivo stress related behavior.

Context: No consensus definition exists for excess adiposity during infancy. After age 2 years, high body mass index (BMI) is related to adverse cardiometabolic outcomes. Before age 2 years, the utility of BMI as a metric of excess adiposity is unknown., Objectives: The objective of the study was to characterize infant BMI trajectories in a diverse, longitudinal cohort and investigate the relationship between the infancy BMI trajectory and childhood obesity., Subjects: Healthy, nonpreterm infants (n = 2114) in the Genetic Causes for Complex Pediatric Disorders study (The Children's Hospital of Philadelphia) with six or more BMI measurements in the first 13.5 months participated in the study., Design: For each infant, the BMI trajectory was modeled using polynomial regression. Independent effects of clinical factors on magnitude and timing of peak BMI were assessed. The relationship between infancy BMI and early childhood BMI (age 4 y) was examined (n = 1075)., Results: The cohort was 53% male and 61% African-American. Peak BMI was 18.6 ± 1.7 kg/m(2) and occurred at 8.6 ± 1.4 months. In multivariate analysis, boys had a higher (0.50 kg/m(2), P < .001) peak BMI than girls. The peak was higher (0.53 kg/m(2), P ≤ .001) and occurred earlier (by 12 d, P < .001) in African-American vs white children. The odds of obesity at age 4 years increased among children with higher (odds ratio 2.02; P < .001) and later (odds ratio 1.26; P = .02) infancy peak BMI., Conclusions: We demonstrate sex- and ancestry-specific differences in infancy BMI and an association of infancy peak BMI with childhood BMI. These findings support the potential utility of infancy BMI to identify children younger than age 2 years with increased risk for later obesity.

The time is ripe to usher in a new paradigm in infection control and to move beyond our sole reliance on broad-spectrum antibiotics whose use results in extensive collateral damage to our microbiome and, in so doing, exerts significant selective pressures for resistance to emerge. We propose to supplement the existing pharmacy of conventional antibiotics, with a new drug family, the pheromonicins. These bacteriocin-based antimicrobials are stable, nontoxic proteins that possess potent antibacterial activities, and which can be easily and rapidly retargeted against any bacteria desired. Here we discuss colicin Ia, a pore forming bacteriocin, as the base of a novel drug development platform, the pheromonicins. Recent work suggests this versatile drug development platform can be used to generate pheromonicins active against enveloped viruses, fungi and human cancer cells. Pheromonicins provide a less toxic, more ecologically sound alternative to conventional antibiotics, and their use will help limit our sole reliance on broad-spectrum drugs.

Tissue homeostasis is one of the central requirements for the existence of multicellular organisms, and is maintained by complex feedback regulatory processes. Homeostasis can be disturbed by diseases such as viruses and tumors. Here, we use mathematical models to investigate how tissue architecture influences the ability to maintain tissue homeostasis during viral infections. In particular, two different tissue designs are considered. In the first scenario, stem cells secrete negative feedback factors that influence the balance between stem cell self-renewal and differentiation. In the second scenario, those feedback factors are not produced by stem cells but by differentiated cells. The model shows a tradeoff. If feedback factors are produced by stem cells, then a viral infection will lead to a significant reduction in the number of differentiated cells leading to tissue pathology, but the number of stem cells is not affected at equilibrium. In contrast, if the feedback factors are produced by differentiated cells, a viral infection never reduces the number of tissue cells at equilibrium because the feedback mechanism compensates for virus-induced cells death. The number of stem cells, however, becomes elevated, which could increase the chance of these stem cells to accumulate mutations that can drive cancer. Interestingly, if the virus interferes with feedback factor production by cells, uncontrolled growth can occur in the presence of the virus even in the absence of genetic lesions in cells. Hence, the optimal design would be to produce feedback factors by both stem and differentiated cells in quantities that strike a balance between protecting against tissue destruction and stem cell elevation during infection., (© 2013 Elsevier Ltd. All rights reserved.)

Splenectomy is considered the treatment of choice for patients with "symptomatic" hereditary spherocytosis (HS). Published guidelines offering recommendations for splenectomy in HS categorize patients primarily based on hemoglobin concentration. We performed a retrospective review of 64 children having splenectomy for HS at Children's Medical Center Dallas. On the basis of hemoglobin concentration alone, 16 children (25%) had mild, 38 (59%) moderate, and 10 (16%) severe HS. However, when reticulocyte count was used to categorize disease severity, only 3 patients (5%) having splenectomy would be considered mild, 17 (27%) moderate, and 42 (66%) severe. Despite otherwise having "mild" disease defined by near-normal hemoglobin levels, many children with nontraditional or subjective signs and symptoms related to hemolytic rate received a splenectomy, and, therefore, reticulocyte count might be a more reliable laboratory marker suggesting consideration of splenectomy. A validated assessment tool incorporating quality of life indicators in addition to the traditional medical indications for splenectomy in HS would be valuable in assessing indications for the procedure.

Serine-threonine protein kinases are critical to CNS function, yet there is a dearth of highly selective, CNS-active kinase inhibitors for in vivo investigations. Further, prevailing assumptions raise concerns about whether single kinase inhibitors can show in vivo efficacy for CNS pathologies, and debates over viable approaches to the development of safe and efficacious kinase inhibitors are unsettled. It is critical, therefore, that these scientific challenges be addressed in order to test hypotheses about protein kinases in neuropathology progression and the potential for in vivo modulation of their catalytic activity. Identification of molecular targets whose in vivo modulation can attenuate synaptic dysfunction would provide a foundation for future disease-modifying therapeutic development as well as insight into cellular mechanisms. Clinical and preclinical studies suggest a critical link between synaptic dysfunction in neurodegenerative disorders and the activation of p38αMAPK mediated signaling cascades. Activation in both neurons and glia also offers the unusual potential to generate enhanced responses through targeting a single kinase in two distinct cell types involved in pathology progression. However, target validation has been limited by lack of highly selective inhibitors amenable to in vivo use in the CNS. Therefore, we employed high-resolution co-crystallography and pharmacoinformatics to design and develop a novel synthetic, active site targeted, CNS-active, p38αMAPK inhibitor (MW108). Selectivity was demonstrated by large-scale kinome screens, functional GPCR agonist and antagonist analyses of off-target potential, and evaluation of cellular target engagement. In vitro and in vivo assays demonstrated that MW108 ameliorates beta-amyloid induced synaptic and cognitive dysfunction. A serendipitous discovery during co-crystallographic analyses revised prevailing models about active site targeting of inhibitors, providing insights that will facilitate future kinase inhibitor design. Overall, our studies deliver highly selective in vivo probes appropriate for CNS investigations and demonstrate that modulation of p38αMAPK activity can attenuate synaptic dysfunction.

A hallmark of human immunodeficiency virus is its ability to infect CD4+ T helper cells, thus impairing helper cell responses and consequently effector responses whose maintenance depends on help (such as killer T cells and B cells). In particular, the virus has been shown to infect HIV-specific helper cells preferentially. Using mathematical models, we investigate the consequence of this assumption for the basic dynamics between HIV and its target cells, assuming the existence of two independently regulated helper cell clones, directed against different epitopes of the virus. In contrast to previous studies, we examine a relatively simple scenario, only concentrating on the interactions between the virus and its target cells, not taking into account any helper-dependent effector responses. Further, there is no direct competition for space or antigenic stimulation in the model. Yet, a set of interesting outcomes is observed that provide further insights into factors that shape helper cell responses. Despite the absence of competition, a stronger helper cell clone can still exclude a weaker one because the two clones are infected by the same pathogen, an ecological concept called "apparent competition". Moreover, we also observe "facilitation": if one of the helper cell clones is too weak to become established in isolation, the presence of a stronger clone can provide enhanced antigenic stimulation, thus allowing the weaker clone to persist. The dependencies of these outcomes on parameters is explored. Factors that reduce viral infectivity and increase the death rate of infected cells promote coexistence, which is in agreement with the observation that stronger immunity correlates with broader helper cell responses. The basic model is extended to explicitly take into account helper-dependent CTL responses and direct competition. This study sheds further light onto the factors that can influence the clonal composition of HIV-specific helper cell responses, which has implications for the overall pattern of disease progression., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Diaquasuccinatocadmium(II) hemihydrate (DSCH) single crystals were grown successfully by single gel diffusion technique at room temperature. Block shaped crystals of size 11 mm × 6 mm × 3 mm have been obtained in 24 days. The grown crystals were colorless and transparent. The crystalline nature and reflection planes of the sample were confirmed by the powder X-ray diffraction technique. The crystal structure of the DSCH crystal was determined by single crystal X-ray diffraction technique. According to microanalytical results the formula of the crystal is [Cd(C(4)H(4)O(4))H(2)O](2)·0.5H(2)O. Fourier Transform Infrared (FT-IR) and Fourier Transform Raman (FT-Raman) studies were used to confirm the presence of various functional groups in the crystal. The thermal behavior of the crystal was studied by simultaneous TGA-DTA methods in nitrogen atmosphere. The mechanical strength of the grown crystal was estimated by Vickers microhardness test. The dielectric behavior of the sample was also studied., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Engineering robust protein production and purification of correctly folded biotherapeutic proteins in cell-based systems is often challenging due to the requirements for maintaining complex cellular networks for cell viability and the need to develop associated downstream processes that reproducibly yield biopharmaceutical products with high product quality. Here, we present an alternative Escherichia coli-based open cell-free synthesis (OCFS) system that is optimized for predictable high-yield protein synthesis and folding at any scale with straightforward downstream purification processes. We describe how the linear scalability of OCFS allows rapid process optimization of parameters affecting extract activation, gene sequence optimization, and redox folding conditions for disulfide bond formation at microliter scales. Efficient and predictable high-level protein production can then be achieved using batch processes in standard bioreactors. We show how a fully bioactive protein produced by OCFS from optimized frozen extract can be purified directly using a streamlined purification process that yields a biologically active cytokine, human granulocyte-macrophage colony-stimulating factor, produced at titers of 700 mg/L in 10 h. These results represent a milestone for in vitro protein synthesis, with potential for the cGMP production of disulfide-bonded biotherapeutic proteins., (Copyright © 2011 Wiley Periodicals, Inc.)

Membrane fusion is a key event in many biological processes. The fusion process, both in vivo and in vitro, is induced by different agents which include mainly proteins and peptides. For protein- and peptide-mediated membrane fusion, conformational reorganization serves as a driving force. Small drug molecules do not share this advantage; hence, drug induced membrane fusion occurring in absence of any other fusogenic agent and at physiologically relevant concentration of the drugs is a very rare event. To date, only three drugs, namely, meloxicam (Mx), piroxicam (Px), and tenoxicam (Tx), belonging to the oxicam group of non steroidal anti-inflammatory drugs (NSAIDs), have been shown by us to induce fusion at very low drug to lipid ratio without the aid of any other fusogenic agent. In our continued effort to understand the interplay of different physical and chemical parameters of both the participating drugs and the membrane on the mechanism of this drug induced membrane fusion, we present here the effect of increase in orientational order of the lipid chains and increase in head group spacing. This is achieved by studying the effect of low concentration cholesterol (<10 mol %) at temperatures above the chain-melting transition. Low concentration cholesterol (<10 mol %), above the gel to fluid transition temperature, is mainly known to increase orientational order of the lipid chains and increase head group spacing. To isolate the effect of these parameters, small unilameller vesicles (SUVs) formed by dimyristoylphosphatidylcholine (DMPC) with an average diameter of 50-60 nm were used as simple model membranes. Fluorescence assays were used to probe the time dependence of lipid mixing, content mixing, and leakage and also used to determine the partitioning of the drugs in the membrane bilayer. Differential scanning calorimetry (DSC) was used to study the effect of drugs in the presence of cholesterol on the chain-melting temperature which reflects the fluidization effect of the hydrophobic tail region of the bilayer. Our results show contradictory effect of low concentration cholesterol on the fusion induced by the three drugs, which has been explained by parsing the effect of orientational order and increase in head group spacing on the fusion process.

Purpose: We investigated the ability to perform a clinical proteomic study using samples collected at different times from two independent clinical sites., Experimental Design: Label-free 2-D-LC-MS proteomic analysis was used to differentially quantify tens of thousands of peptides from human plasma. We have asked whether samples collected from two sites, when analyzed by this type of peptide profiling, reproducibly contain detectable peptide markers that are differentially expressed in the plasma of disease (advanced renal cancer) patients relative to healthy normals., Results: We have demonstrated that plasma proteins enriched in disease patients are indeed detected reproducibly in both clinical collections. Regression analysis, unsupervised hierarchical clustering and PCA detected no systematic bias in the data related to site of sample collection and processing. Using a genetic algorithm, support vector machine classification method, we were able to correctly classify disease samples at 88% sensitivity and 94% specificity using the second site as an independent validation set., Conclusions and Clinical Relevance: We conclude that multiple site collection, when analyzed by label-free 2-D-LC-MS, generates data that are sufficiently reproducible to guide reliable biomarker discovery., (Copyright © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background and Objectives: This in-vitro study was conducted to evaluate the strength and properties of Type III and Type IV gypsum mixed with disinfectant solutions., Materials and Methods: Type III and Type IV gypsum were used for the study. Three different mixing solutions namely waterqueous solutions of 0.525% sodium hypochlorite and 2% glutaraldehyde were used. Gypsum materials were subjected to further modification by adding a mixture of 1.0% gum arabic and 0.132% calcium hydroxide before mixing with the disinfectant solutions, at two different liquid/powder (L/P) ratios for each. Both, the unmodified and the modified gypsum were tested for compressive and tensile strength after one hour and one week from the start of the mix. The crystalline configuration of the fracture fragments of the unmodified and modified set gypsum were studied under the scanning electron microscope., Results: The disinfectant solutions reduced the strength of both Type III and Type IV gypsum. Water showed higher-strength, which was followed by 0.525% sodium hypochlorite and 2% glutaraldehyde. The modified Type III and Type IV gypsum with reduced L/P ratio also showed strength values less than that of the control groups., Interpretation and Conclusion: Chemical disinfectants reduced the strength of gypsum when used as water substitutes. Gum Arabic and calcium hydroxide additives permitted lower L/P ratio, however, there was still excess water retained in the set gypsum that lowered the strength values of Type III and Type IV gypsum. Hence, further reduction of L/P ratio may increase the properties of the modified Type III and Type IV gypsum.

CYP2D6 substrate status is a critical Go/No Go decision criteria in central nervous system (CNS) drug discovery efforts because the polymorphic nature of CYP2D6 can lead to variable patient safety and drug efficacy. In addition, CYP2D6 is disproportionately involved in the metabolism of CNS drugs compared with other drug classes. Therefore, identifying trends in small molecule properties of CNS-penetrant compounds that can help discriminate potential CYP2D6 substrates from nonsubstrates would allow additional prioritization in the synthesis and biological evaluation of new therapeutic candidates. We report here the conversion of the CNS drug minaprine from substrate to nonsubstrate, as well as the conversion of the related CNS drug minozac from nonsubstrate to substrate, through the use of analog synthesis and CYP2D6 enzyme kinetic analyses. No single molecular property strongly correlated with substrate status for this 3-amino-4-methyl-6-phenylpyridazine scaffold, although molecular volume and charge appeared to be indirectly related. A parsed database of CYP2D6 substrates across diverse chemical structures was assembled and analyzed for physical property trends correlating with substrate status. We found that a complex interplay of properties influenced CYP2D6 substrate status and that the particular chemical scaffold affects which properties are most prominent. The results also identified an unexpected issue in CNS drug discovery, in that some property trends correlative with CYP2D6 substrates overlap previously reported properties that correlate with CNS penetrance. These results suggest the need for a careful balance in the design and synthesis of new CNS therapeutic candidates to avoid CYP2D6 substrate status while maintaining CNS penetrance.

We show that when a suitable entanglement-generating unitary operator depending on a parameter is applied on N qubits in parallel, a precision of the order of 2(-N) in estimating the parameter may be achieved. This exponentially improves the precision achievable in classical and in quantum nonentangling strategies.

Background: An accumulating body of evidence is consistent with the hypothesis that excessive or prolonged increases in proinflammatory cytokine production by activated glia is a contributor to the progression of pathophysiology that is causally linked to synaptic dysfunction and hippocampal behavior deficits in neurodegenerative diseases such as Alzheimer's disease (AD). This raises the opportunity for the development of new classes of potentially disease-modifying therapeutics. A logical candidate CNS target is p38 alpha MAPK, a well-established drug discovery molecular target for altering proinflammatory cytokine cascades in peripheral tissue disorders. Activated p38 MAPK is seen in human AD brain tissue and in AD-relevant animal models, and cell culture studies strongly implicate p38 MAPK in the increased production of proinflammatory cytokines by glia activated with human amyloid-beta (A beta) and other disease-relevant stressors. However, the vast majority of small molecule drugs do not have sufficient penetrance of the blood-brain barrier to allow their use as in vivo research tools or as therapeutics for neurodegenerative disorders. The goal of this study was to test the hypothesis that brain p38 alpha MAPK is a potential in vivo target for orally bioavailable, small molecules capable of suppressing excessive cytokine production by activated glia back towards homeostasis, allowing an improvement in neurologic outcomes., Methods: A novel synthetic small molecule based on a molecular scaffold used previously was designed, synthesized, and subjected to analyses to demonstrate its potential in vivo bioavailability, metabolic stability, safety and brain uptake. Testing for in vivo efficacy used an AD-relevant mouse model., Results: A novel, CNS-penetrant, non-toxic, orally bioavailable, small molecule inhibitor of p38 alpha MAPK (MW01-2-069A-SRM) was developed. Oral administration of the compound at a low dose (2.5 mg/kg) resulted in attenuation of excessive proinflammatory cytokine production in the hippocampus back towards normal in the animal model. Animals with attenuated cytokine production had reductions in synaptic dysfunction and hippocampus-dependent behavioral deficits., Conclusion: The p38 alpha MAPK pathway is quantitatively important in the A beta-induced production of proinflammatory cytokines in hippocampus, and brain p38 alpha MAPK is a viable molecular target for future development of potential disease-modifying therapeutics in AD and related neurodegenerative disorders.

We report the development of a novel, aqueous-soluble, safe, small molecule, experimental therapeutic that suppresses injury-induced, proinflammatory cytokine increases in the brain, with resultant attenuation of synaptic protein biomarker loss and improvement in hippocampus-dependent behavioral deficits. A GMP production scheme for the active pharmaceutical ingredient, compound 17, is presented. The development and large-scale availability of this novel compound allow exploration of new, potentially disease-modifying, therapeutic approaches to CNS disorders.

We demonstrate the quantification capability and robustness of a new integrated liquid chromatography-mass spectrometry (LC-MS) approach for large-scale profiling of proteins and metabolites. This approach to determine differential expression relies on linearity of signal vs molecular concentration using electrospray ionization LC-MS, reproducibility of sample processing, a novel normalization strategy and associated data analysis software. No isotopic tagging or spiking of internal standards is required. The method is general and applicable to the proteome and metabolome from all biological fluids and tissues. Small or large numbers of samples can be profiled in a single experiment. Differential profiling of 6000 molecular ions per sample by one-dimensional chromatography LC-MS and 30,000 molecular ions per sample by two-dimensional chromatography LC-MS is demonstrated using rheumatoid arthritis patient samples compared with control samples. A new approach to peptide identification is described that involves building libraries of previously identified peptides, circumventing the need to acquire MS/MS data during profiling. Robustness of the platform was tested by repeating sample preparation and LC-MS differential expression analysis after 10 mo, using independent serum aliquots stored at -80 degrees C. To the best of our knowledge, this is the first demonstration of long-term robustness of a platform for quantitative proteomics and metabolomics.

We construct an algorithm for suppressing the transitions of a quantum mechanical system, initially prepared in a subspace P of the full Hilbert space of the system, to outside this subspace by subjecting it to a sequence of unequally spaced short-duration pulses. Each pulse multiplies the amplitude of the vectors in the subspace by -1. The number of pulses required by the algorithm to limit the leakage probability to epsilon in time increases as T exp[square root log(T(2)/epsilon)], compared to T(2)epsilon(-1) in the standard quantum Zeno effect.

Currently, there is no single test for multiple sclerosis (MS). Diagnosis is confirmed through clinical evaluation, abnormalities revealed by magnetic resonance imaging (MRI), and analysis of cerebrospinal fluid (CSF) chemistry. The early and accurate diagnosis of the disease, monitoring of progression, and gauging of therapeutic intervention are important but elusive elements of patient care. Moreover, a deeper understanding of the disease pathology is needed, including discovery of accurate biomarkers for MS. Herein we review putative biomarkers of MS relating to neurodegeneration and contributions to neuropathology, with particular focus on autoimmunity. In addition, novel assessments of biomarkers not driven by hypotheses are discussed, featuring our application of advanced proteomics and metabolomics for comprehensive phenotyping of CSF and blood. This strategy allows comparison of component expression levels in CSF and serum between MS and control groups. Examination of these preliminary data suggests that several CSF proteins in MS are differentially expressed, and thus, represent putative biomarkers deserving of further evaluation.

I show that separability of N-partite quantum states implies new inequalities on Bell correlations which are stronger than the corresponding Mermin-Roy-Singh local reality inequalities by a factor of 2((N-1)/2).

We establish an exact differential equation for the operator describing time-dependent measurements continuous in time and obtain a series solution. Suppose the projection operator E(t) = U(t)EUdagger(t) is measured continuously from t = 0 to T, where E is a projector leaving the initial state unchanged and U(t) a unitary operator obeying U(0) = 1. We prove that the probability of always finding E(t) = 1 from t = 0 to T is unity. If U(t) not equal1, the watched kettle is sure to "boil."

The Gaza Strip now has limited self-rule. Its people inherit a health-care system provided by four sectors--the Israeli government, the UN Relief and Works Agency, non-governmental organisations, and private facilities. They also inherit a legacy of health indices comparing unfavourably with those of Israel, huge numbers of refugees, a looming water crisis, civil unrest, and a health care expenditure bill (as a proportion of gross domestic product) that will be unsustainable. Four plans have been drawn up for health services in autonomous Gaza but none is exactly right.