Search

Anti-IL17A therapies have proven effective for numerous inflammatory diseases including psoriasis, axial spondylitis and psoriatic arthritis. Modulating and/or antagonizing protein-protein interactions of IL17A cytokine binding to its cell surface receptors with oral therapies offers the promise to bring forward biologics-like efficacy in a pill to patients. We used an NMR-based fragment screen of recombinant IL17A to uncover starting points for small molecule IL17A antagonist discovery. By examining chemical shift perturbations in 2D [ 1 H, 13 C-HSQC] spectra of isotopically labeled IL17A, we discovered fragments binding the cytokine at a previously undescribed site near the IL17A C-terminal region, albeit with weak affinity (> 250 µM). Importantly this binding location was distinct from previously known chemical matter modulating cytokine responses. Subsequently through analog screening, we identified related compounds that bound symmetrically in this novel site with two copies. From this observation we employed a linking strategy via structure-based drug design and obtained compounds with increased binding affinity (< 50 nM) and showed functional inhibition of IL17A-induced cellular signaling (IC 50 ~1 µM). We also describe a fluorescence-based probe molecule suitable to discern/screen for additional molecules binding in this C-terminal site., (© 2022. The Author(s).)

Commercially available hydroxypropyl methylcellulose capsules are employed as a fast, safe, and user-friendly chemical delivery system containing all reagents (catalyst, ligand, and base) for three important transition-metal-catalyzed reactions: Buchwald-Hartwig, Suzuki-Miyaura, and metallophotoredox C-N cross-coupling reactions. This encapsulation methodology simplifies the screening of reaction conditions and the preparation of compound libraries using parallel synthesis in organic solvents or aqueous media. These reagents-containing HPMC capsules are easy to prepare, come in different sizes, and can be stored on the bench under noninert conditions.

Tumor necrosis factor α (TNFα) is a soluble cytokine that is directly involved in systemic inflammation through the regulation of the intracellular NF-κB and MAPK signaling pathways. The development of biologic drugs that inhibit TNFα has led to improved clinical outcomes for patients with rheumatoid arthritis and other chronic autoimmune diseases; however, TNFα has proven to be difficult to drug with small molecules. Herein, we present a two-phase, fragment-based drug discovery (FBDD) effort in which we first identified isoquinoline fragments that disrupt TNFα ligand-receptor binding through an allosteric desymmetrization mechanism as observed in high-resolution crystal structures. The second phase of discovery focused on the de novo design and optimization of fragments with improved binding efficiency and drug-like properties. The 3-indolinone-based lead presented here displays oral, in vivo efficacy in a mouse glucose-6-phosphate isomerase (GPI)-induced paw swelling model comparable to that seen with a TNFα antibody.

Background: Food authenticity is demanded by the consumer at all times. The consumer places trust in the manufacturer that the food product is genuine in terms of what is recorded on the packaging label. Objective: Recent advancements in LC-tandem MS methodology in the detection of allergens, meat, and gelatin speciation in raw food products and processed foods are detailed in this paper. Method: For each of the three methods, initial proteome analysis and the screening leading to the determination of unique tryptic peptides were conducted using a high-resolution, accurate tandem mass spectrometer. Having identified the unique markers, the method was transferred to a tandem quadrupole mass spectrometer for a higher-sensitivity quantitative study, multiple reaction monitoring transition analysis. Results: For the allergens method a detection limit of at least 10 ppm was attained across the 12 allergen peptides in this workflow. In the gluten workflow the resulting chromatograms show good detection down to 5 ppm, with no interference from the food matrices. The meat speciation method details that signature peptides could be readily identified at 1% w/w with no matrix interference. Conclusions: These single-injection workflows with cycle-time optimization enable wide coverage of analytes to identify multiple species within challenging matrix samples.

Apolipoprotein E is a 299-residue lipid carrier protein produced in both the liver and the brain. The protein has three major isoforms denoted apoE2, apoE3, and apoE4 which differ at positions 112 and 158 and which occur at different frequencies in the human population. Genome-wide association studies indicate that the possession of two apoE4 alleles is a strong genetic risk factor for late-onset Alzheimer's disease (LOAD). In an attempt to identify a small molecule stabilizer of apoE4 function that may have utility as a therapy for Alzheimer's disease, we carried out an NMR-based fragment screen on the N-terminal domain of apoE4 and identified a benzyl amidine based fragment binder. In addition to NMR, binding was characterized using various other biophysical techniques, and a crystal structure of the bound core was obtained. Core elaboration ultimately yielded a compound that showed activity in an IL-6 and IL-8 cytokine release assay.

Novel conformationally constrained BET bromodomain inhibitors have been developed. These inhibitors were optimized in two similar, yet distinct chemical series, the 6-methyl-1H-pyrrolo[2,3-c]pyridin-7(6H)-ones (A) and the 1-methyl-1H-pyrrolo[2,3-c]pyridin-7(6H)-ones (B). Each series demonstrated excellent activity in binding and cellular assays, and lead compounds from each series demonstrated significant efficacy in in vivo tumor xenograft models., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Herein we disclose SAR studies of a series of dimethylamino pyrrolidines which we recently reported as novel inhibitors of the PRC2 complex through disruption of EED/H3K27me3 binding. Modification of the indole and benzyl moieties of screening hit 1 provided analogs with substantially improved binding and cellular activities. This work culminated in the identification of compound 2, our nanomolar proof-of-concept (PoC) inhibitor which provided on-target tumor growth inhibition in a mouse xenograft model. X-ray crystal structures of several inhibitors bound in the EED active-site are also discussed., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Timing of precipitation is equally important as amount for determining ecosystem function, especially aboveground net primary productivity (ANPP), in a number of ecosystems. In tallgrass prairie of the Central Plains of North America, grass flowering stalks of dominant C4 grasses, such as Andropogon gerardii, can account for more than 70 % of ANPP, or almost none of it, as the number of flowering stalks produced is highly variable. Although growing season precipitation amount is important for driving variation in flowering stalk production, it remains unknown whether there are critical periods within the growing season in which sufficient rainfall must occur to allow for flowering. The effect of timing of rainfall deficit (drought) on flowering of A. gerardii, was tested by excluding rainfall during three periods within the growing season (starting in mid-April, mid-May and mid-June). Mid-summer drought (starting in mid-June) strongly reduced the flowering rate (e.g., density and biomass) of A. gerardii (e.g., as high as 94 % compared to the control), suggesting flowering is highly sensitive to precipitation at this time. This effect appeared to be related to plant water status at the time of flowering stalk initiation, rather than an indirect consequence of reduced C assimilation. Our results suggest that increased frequency of growing season drought forecast with climate change could reduce sexual reproduction in this dominant grass species, particularly if it coincides with timing of flowering stalk initiation, with important implications for ecosystem functioning.

Long-lasting synaptic plasticity is often accompanied by morphological changes as well as formation and/or loss of dendritic spines. Since the spine cytoskeleton mainly consists of actin filaments, morphological changes are primarily controlled by actin binding proteins (ABPs). Inositol-1,4,5-trisphosphate-3-kinase-A (ITPKA) is a neuron-specific, actin bundling protein concentrated at dendritic spines. Here, we demonstrate that ITPKA depletion in mice increases the number of hippocampal spine-synapses while reducing average spine length. By employing actin to ABP ratios similar to those occurring at post synaptic densities, in addition to cross-linking actin filaments, ITPKA strongly inhibits Arp2/3-complex induced actin filament branching by displacing the complex from F-actin. In summary, our data show that in vivo ITPKA negatively regulates formation and/or maintenance of synaptic contacts in the mammalian brain. On the molecular level this effect appears to result from the ITPKA-mediated inhibition of Arp2/3-complex F-actin branching activity., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Background: Recently, high-frequency repetitive transcranial magnetic stimulation (rTMS) is reported to evaluating the corticospinal pathway and improving both cortical excitability and motor function significantly in subjects. According to some previous reports, the maximum voluntary muscle contraction (MVC) of target muscle can reinforce the influence by rTMS. The aim of this study was to confirm 5 Hz rTMS with MVC in healthy individuals is an effective method to facilitate motor neuron excitability and the efficiency can last at least 30 min post stimulation., Objective: To compare the motor evoked potentials (MEPs) elicited by 5Hz rTMS and 5Hz rTMS combined with MVC., Methods: In this randomized, controlled, assessor-blinded, crossover trial, 40 healthy right-handed subjects were divided into group A (n=20) and group B (n=20). All subjects received rTMS over the primary motor cortex area (M1) in the left hemisphere. The parameters of rTMS were 5 Hz, 90.of the resting motor threshold (RMT), for a total of 500 pulses in 100 trains (1-sec inter-stimulus and 8- sec inter-interval). Method 1: All subjects received rTMS over the hand area of left M1. Method 2: All subjects received rTMS at the same stimulated point, combined with maximum voluntary hand griping in each 10 trains. Test 1: group A underwent method 1, while group B underwent method 2. Test 2: A week later, group B underwent method 1, while group A underwent method 2. In each test, the MEP amplitude and latency was measured before (P-rTMS), 5 min after (Post 1) and 30 min after (Post 2) the rTMS intervention., Results: MEP amplitude increased significantly from baseline at 5 minutes post intervention under both treatment regimes. However for both sequences, it decreased towards baseline under the rTMS intervention at 30 minutes post intervention but remained relatively high when rTMS was combined with MVC. MEP latency decreased significantly from baseline at 5 minutes post intervention under both treatment regimes. For both sequences, it then increased again towards baseline under both treatment regimes at 30 minutes post intervention. Although there was a trend for a less pronounced increase under the combined treatment, this effect was not significant., Conclusion: Both 5 Hz rTMS and 5 Hz rTMS combined with MVC facilitate motor cortical excitability, but the enhancement in rTMS with MVC is more pronounced and maintained longer than simple rTMS.

Grade IV glioblastoma is characterized by increased kinase activity of epidermal growth factor receptor (EGFR); however, EGFR kinase inhibitors have failed to improve survival in individuals with this cancer because resistance to these drugs often develops. We showed that tumor necrosis factor-α (TNFα) produced in the glioblastoma microenvironment activated atypical protein kinase C (aPKC), thereby producing resistance to EGFR kinase inhibitors. Additionally, we identified that aPKC was required both for paracrine TNFα-dependent activation of the transcription factor nuclear factor κB (NF-κB) and for tumor cell-intrinsic receptor tyrosine kinase signaling. Targeting aPKC decreased tumor growth in mouse models of glioblastoma, including models of EGFR kinase inhibitor-resistant glioblastoma. Furthermore, aPKC abundance and activity were increased in human glioblastoma tumor cells, and high aPKC abundance correlated with poor prognosis. Thus, targeting aPKC might provide an improved molecular approach for glioblastoma therapy., (Copyright © 2014, American Association for the Advancement of Science.)

The mammalian target of rapamycin (mTOR) and mitogen-activated protein kinase signaling cascades have been implicated in a number of human cancers. The tumor suppressor gene tuberous sclerosis-2 (Tsc-2) functions as a negative regulator of mTOR. Critical proteins in both pathways are activated following treatment of Eker rats (Tsc-2(EK/+)) with the nephrocarcinogen 2,3,5-tris-(glutathion-S-yl)hydroquinone (TGHQ), which also results in loss of the wild-type allele of Tsc-2 in renal preneoplastic lesions and tumors. Western blot analysis of kidney tumors formed following treatment of Tsc-2(EK/+) rats with TGHQ for 8 months revealed increases in B-Raf, Raf-1, pERK, cyclin D1, 4EBP1, and p-4EBP1-Ser65, -Thr70, and -Thr37/46 expression. Similar changes are observed following TGHQ-mediated transformation of primary renal epithelial cells derived from Tsc-2(EK/+) rats (quinol-thioether rat renal epithelial [QTRRE] cells) that are also null for tuberin. These cells exhibit high ERK, B-Raf, and Raf-1 kinase activity and increased expression of all p-4EBP1s and cyclin D1. Treatment of the QTRRE cells with the Raf kinase inhibitor, sorafenib, or the MEK1/2 kinase inhibitor, PD 98059, produced a significant decrease in the protein expression of all p-4EBP1s and cyclin D1. Following siRNA knockdown of Raf-1, Western blot analysis revealed a significant decrease in Raf-1, cyclin D1, and all p-4EBP1 forms noted above. In contrast, siRNA knockdown of B-Raf resulted in a nominal change in these proteins. The data indicate that Raf-1/MEK/ERK participates in crosstalk with 4EBP1, which represents a novel pathway interaction leading to increased protein synthesis, cell growth, and kidney tumor formation.

Objective: The aim of this pilot study was to describe problems in functioning and associated rehabilitation needs in persons with spinal cord injury after the 2010 earthquake in Haiti by applying a newly developed tool based on the International Classification of Functioning, Disability and Health (ICF)., Design: Pilot study., Subjects: Eighteen persons with spinal cord injury (11 women, 7 men) participated in the needs assessment. Eleven patients had complete lesions (American Spinal Injury Association Impairment Scale; AIS A), one patient had tetraplegia., Methods: Data collection included information from the International Spinal Cord Injury Core Data Set and a newly developed needs assessment tool based on ICF Core Sets. This tool assesses the level of functioning, the corresponding rehabilitation need, and required health professional. Data were summarized using descriptive statistics., Results: In body functions and body structures, patients showed typical problems following spinal cord injury. Nearly all patients showed limitations and restrictions in their activities and participation related to mobility, self-care and aspects of social integration. Several environmental factors presented barriers to these limitations and restrictions. However, the availability of products and social support were identified as facilitators. Rehabilitation needs were identified in nearly all aspects of functioning. To address these needs, a multidisciplinary approach would be needed., Conclusion: This ICF-based needs assessment provided useful information for rehabilitation planning in the context of natural disaster. Future studies are required to test and, if necessary, adapt the assessment.

This special report presents the role of the World Health Organization (WHO) Liaison Sub-Committee on Rehabilitation Disaster Relief (CRDR) of the International Society of Physical and Rehabilitation Medicine (ISPRM) in developing an enhanced physical rehabilitation relief response to large-scale natural disasters. The CRDR has stated that disaster rehabilitation is an emerging subspecialty within physical and rehabilitation medicine (PRM). In reviewing the existing literature it was found that large natural disasters result in many survivors with disabling impairments, that these survivors may have better clinical outcomes when they are treated by PRM physicians and teams of rehabilitation professionals, that the delivery of these rehabilitation services to disaster sites is complicated, and that their absence can result in significant negative consequences for individuals, communities and society. To advance its agenda, the CRDR sponsored an inaugural Symposium on Rehabilitation Disaster Relief as a concurrent scientific session at the 2011 ISPRM 6th World Congress in San Juan, Puerto Rico. The symposium included oral and poster presentations on a range of relevant topics and concluded with an international non-governmental organization panel discussion that addressed the critical question "How can rehabilitation actors coordinate better in disaster?" Building upon the symposium, the CRDR is developing a disaster rehabilitation evidence-base, which will inform and educate the global professional rehabilitation community about needs and best practices in disaster rehabilitation. The Journal of Rehabilitation Medicine (JRM) has commissioned this special report to announce a series of papers on disaster rehabilitation from the symposium's scientific programme. Authors are invited to submit papers on the topic for inclusion in this special series. JRM also encourages expert commentary in the form of Letters to the Editor.

Purpose: Visual cues from persons with impairments may trigger stereotypical generalisations that lead to prejudice and discrimination. The main objective of this pilot study is to examine whether visual stimuli of impairment activate latent prejudice against disability and whether this connection can be counteracted with priming strategies., Methods: In a field experiment, participants were asked to rate photographs showing models with mental impairments, wheelchair users with paraplegia, and persons without any visible impairment. Participants should appraise the models with regard to several features (e.g. communicativeness, intelligence). One hundred participants rated 12 photo models yielding a total of 1183 observations. One group of participants was primed with a cover story introducing visual perception of impairment as the study's gist, while controls received neutral information., Results: Photo models with mental impairments were rated lowest and models without visible impairment highest. In participants who did not have prior contacts with persons with impairments, priming led to a levelling of scores of models with and without impairment. Prior contacts with persons with impairments created similar effects as the priming. Unexpectedly, a pattern of converse double discrimination to the disadvantage of men with mental impairments was revealed., Conclusion: Signs of stereotypical processing of visual cues of impairment have been found in participants of the Swiss general population. Personal contact with persons with impairments as well as priming participants seems to reduce stereotyping.

Coping with limited resources is one of the key challenges in patient care. Rationalisation reserves have already been exhausted in many areas. Particularly in the health sector, the question needs to be clarified as to how the limited resources can be allocated fairly from both the ethical and legal point of view. The medical community proposes that health care services be prioritised. In other words, priorities are defined in order to increase fair distribution in the health sector. In this context prioritisation does not mean the exclusion of medically necessary services, but grading the granting of services in accordance with the principles of priority. The advantages of prioritisation lie primarily in the transparency of the procedure and in the chance of achieving consistent distribution decisions. Protection of the patient-physician relationship as a confidential relation would remain unaffected since prioritisation is not the same thing as rationing., (Copyright © 2010. Published by Elsevier GmbH.)

With the International Classification of Functioning, Disability and Health (ICF) the World Health Organization (WHO) has prepared the ground for a comprehensive understanding of Human Functioning and Rehabilitation Research, integrating the biomedical perspective on impairment with the social model of disability. This poses a number of old and new challenges regarding the enhancement of adequate research capacity. Here we will summarize approaches to address these challenges with respect to 3 areas: the organization of Human Functioning and Rehabilitation Research into distinct scientific fields, the development of suitable academic training programmes and the building of university centres and collaboration networks.

Up to the early 1990's the health care system was essentially characterised through:--the insured' right of choice of therapist,--therapeutic freedom of patients and physicians, and--the freedom of establishment for medical doctors.--In accordance with the Hospital Funding Act the hospital system was--in compliance with federal constitutional law using capacity requirements--based on the "fire-fighting" principle, i.e. that if required, every patient should have access to a suitable hospital bed within about 15 minutes.--The responsibility for ensuring the provision of general and specialist health care services had been conferred by the government to the National Association of Statutory Health Insurance Physicians (1955) in the legal form of a public corporation. In the face of a foreseeable rise in expenses as a result of advances in medicine and a higher demand for health care services because of the demographic development (long-life society) the Advisory Council for Concerted Action in Health Care concludes in its Annual Report that maintaining this level of health care for all people is not financially viable any longer. This is why the state--on the basis of the Health Care Reform Act of 2002 and the Statutory Health Insurance System Modernisation Act of 2004--retreated from the provision of services in the ambulatory and inpatient setting by privatising these sectors and by proclaiming competition (introduction of diagnosis-related groups). Presently, the once liberal performance tradition is more and more turning into a centrally planned system in the spirit of "From Therapeutic Freedom to Therapeutic Programmes". The guidelines that on the basis of the methods of evidence-based medicine were developed by the international community of physicians for the treatment of patients with defined diagnoses and intended to be decision aids for individual treatment decisions are now used to implement disease management programmes for the provision of health care to people with certain chronic diseases. Another steering instrument of the above mentioned Modernisation Act is the definition of a list stating all the benefits available under the statutory health insurance system. The assessment of the health gain of new medical drugs and other medical procedures has been assigned to the Institute for Quality and Efficiency in Health Care (IQWiG) specifically founded to that purpose. With regard to the assessment of diagnostic and therapeutic procedures the Joint Federal Committee (Gemeinsamer Bundesausschuss, G-BA) in the summer of 2005 gave itself a Code of Procedures that defines uniform cross-sector criteria for the appraisal of diagnosis and treatment. In Germany the principle of evidence-based health care has by law--and this is unique as compared to other countries--fully penetrated everyday health care where the decisions of the Joint Federal Committee in the form of directives have mandatory effect for health care providers and hence for the insured, too. This is why the German Medical Association and the National Association of Statutory Health Insurance Physicians have embarked on the implementation of the "National Programme for Disease Management Guidelines" and the "Health Services Research" Project as a means of continuously evaluating health care provision which are intended to guide the future political control of the system of statutory health insurance in terms of target-performance comparisons and for the purpose of identifying health care deficits.

Lege artis procedures, medical standards as well as guideline contents ought to be defined by health care professionals themselves. Therefore the medical profession carries a great responsibility to both continuously optimise diagnostic and therapeutic methods as well as their results and adapt them to medical progress. Standards and guidelines cannot replace a doctor's responsibility towards his patients. They can help doctors to find the right answer but they are not check-off lists. The practical application of an evidence-based guideline must result from a doctor's identification of a certain patient problem. This stage of identification is predominated by the individual doctor's internal evidence which reflects his medical knowledge, practical experience and the information arising from the individual relation between patient and doctor.