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Background: Recent findings suggest that the plasma axonal structural protein, neurofilament light (NFL) chain, may serve as a potential blood biomarker for early signs of neurodegenerative diseases, such as Alzheimer's disease (AD). Given the need for early detection of neurodegenerative disorders, the current study investigated the associations between regional cerebral blood flow (rCBF) in brain regions associated with neurodegenerative disorders and memory function with plasma NFL in AD, mild cognitive impairment (MCI), and healthy controls (HCs). Methods: We recruited 29 AD, 76 MCI, and 39 HCs from the Alzheimer's Disease Neuroimaging Initiative (ADNI) database in the current cross-sectional study. We used Pearson's correlation models adjusted for the effect of age, sex, and APOE genotype to investigate the association between plasma NFL and rCBF. Results: We found non-significant differences in age (F(2, 141) = 1.304; P = 0.275) and years of education (F(2, 141) = 0.013; P = 0.987). Additionally, we found significant differences between groups in terms of MMSE scores (F(2, 141) = 100.953; P < 0.001). Despite the observation of significantly reduced rCBF in AD and MCI groups versus HCs, we did not detect significant differences in plasma NFL between these groups. We found significant negative associations between plasma NFL and rCBF in various AD-related regions, these findings were only observed after analyses in all participants, and were observed in HCs alone and no significant associations were observed in the AD or MCI groups. Conclusion: These outcomes add to our current understanding surrounding the use of rCBF and plasma NFL biomarkers as tools for early detection and diagnosis of neurodegenerative diseases. A conclusion might be that the association between NFL and impaired rCBF exists before the clinical symptoms appear. Further longitudinal studies with a large sample size should be performed to examine the correlation between plasma NFL and rCBF in order to understand these complex relationships. [ABSTRACT FROM AUTHOR]

Between the first recorded execution at Tyburn in 1196 and the abolition of public executions in 1868, tens of thousands of offenders were publicly executed in London. In Section 6, for example, there was no discussion of the extent to which debates about punishment in the nineteenth century were divided along political lines, and in particular that the abolition of public executions in 1868 was at least in part a move by those who wanted to retain capital punishment as a whole. Some of the research represents genuinely original contributions to our historical knowledge of public execution in London, such as the exhibition's exhaustive identification of past execution sites. [Extracted from the article]

'Dementia-friendly communities' herald a shift toward the neighbourhood as a locus for the care and support of people with dementia, sparking growing interest in the geographies of dementia care and raising questions over the shifting spatial and social experience of the condition. Existing research claims that many people with dementia experience a 'shrinking world' whereby the boundaries to their social and physical worlds gradually constrict over time, leading to a loss of control and independence. This paper reports a five-year, international study that investigated the neighbourhood experience of people with dementia and those who care for and support them. We interrogate the notion of a shrinking world and in so doing highlight an absence of attention paid to the agency and actions of people with dementia themselves. The paper draws together a socio-relational and embodied-material approach to question the adequacy of the shrinking world concept as an explanatory framework and to challenge reliance within policy and practice upon notions of place as fixed or stable. We argue instead for the importance of foregrounding 'lived place' and attending to social practices and the networks in which such practices evolve. Our findings have implications for policy and practice, emphasising the need to bolster the agency of people living with dementia as a route to fostering accessible and inclusive neighbourhoods. [ABSTRACT FROM AUTHOR]

Dementia is a global health challenge and currently the focus of a coordinated international response articulated through the notion of 'dementia‐friendly communities and initiatives' (DFCIs). Yet, while increasing research attention has been paid to the social and spatial dimensions to life with dementia in a neighbourhood setting, the temporalities of dementia have been largely overlooked. This article sets out different aspects of the lived experience of time for people with dementia and unpaid carers, before exploring the temporal politics of formal dementia care and support. The authors show that time is a site for material struggle and a marker of unequal relations of power. People with dementia and unpaid carers are disempowered through access to formal care, and this is illustrated in their loss of (temporal) autonomy and limited options for changing the conditions of the care received. The authors advocate for a time‐space configured understanding of the relationship with neighbourhood and foreground a tempo‐material understanding of dementia. Set against the backdrop of austerity policy in the UK, the findings reveal that ongoing budgetary restrictions have diminished the capacity for social care to mediate in questions of social justice and inequality, at times even compounding inequity. [ABSTRACT FROM AUTHOR]

In today's society, we are constantly absorbing information via text (e.g., news, social media), much of which may be affectively charged. However, to date, little is known about how the affective framing of the text itself may give rise to various affective experiences during reading. We examined how subtle changes to wording (negatively or positively valenced framing of the same content) influenced affective experiences during reading and subsequent comprehension. Results show that (a) affective framing in text elicited congruent subjective affective states throughout reading, (b) subjective affective valence throughout reading impacted comprehension at various levels, and (c) subjective affective valence mediated the relationship between affective framing and how participants integrated information from the text in their summaries, as well as if they included affect in their summaries. This indicates that even subtle affectively valenced content in text can indirectly influence some aspects of comprehension through eliciting subjective affective experiences in the reader. [ABSTRACT FROM AUTHOR]

Approximately 20% of individuals who experience a traumatic injury will subsequently develop posttraumatic stress disorder (PTSD). Physical pain following traumatic injury has received increasing attention as both a distinct, functionally debilitating disorder and a comorbid symptom related to PTSD. Studies have demonstrated that both clinician‐assessed injury severity and patient pain ratings can be important predictors of nonremitting PTSD; however, few have examined pain and PTSD alongside socioenvironmental factors. We postulated that both area‐ and individual‐level socioeconomic circumstances and lifetime trauma history would be uniquely associated with PTSD symptoms and interact with the pain–PTSD association. To test these effects, pain and PTSD symptoms were assessed at four visits across a 1‐year period in a sample of 219 traumatically injured participants recruited from a Level 1 trauma center. We used a hierarchal linear modeling approach to evaluate whether (a) patient‐reported pain ratings were a better predictor of PTSD than clinician‐assessed injury severity scores and (b) socioenvironmental factors, specifically neighborhood socioeconomic disadvantage, individual income, and lifetime trauma history, influenced the pain–PTSD association. Results demonstrated associations between patient‐reported pain ratings, but not clinician‐assessed injury severity scores, and PTSD symptoms, R2(fvm) =.65. There was a significant interaction between neighborhood socioeconomic disadvantage and pain such that higher disadvantage decreased the strength of the pain–PTSD association but only among White participants, R2(fvm) =.69. Future directions include testing this question in a larger, more diverse sample of trauma survivors (e.g., geographically diverse) and examining factors that may alleviate both pain and PTSD symptoms. [ABSTRACT FROM AUTHOR]

GPR84 is an immune cell-expressed, proinflammatory receptor currently being assessed as a therapeutic target in conditions including fibrosis and inflammatory bowel disease. Although it was previously shown that the orthosteric GPR84 activators 2-HTP and 6-OAU promoted its interactions with arrestin-3, a G protein-biased agonist DL-175 did not. Here, we show that replacement of all 21 serine and threonine residues within i-loop 3 of GPR84, but not the two serines in the C-terminal tail, eliminated the incorporation of [32P] and greatly reduced receptor-arrestin-3 interactions promoted by 2-HTP. GPR84 was phosphorylated constitutively on residues Ser221 and Ser224, while various other amino acids are phosphorylated in response to 2-HTP. Consistent with this, an antiserum able to identify pSer221/pSer224 recognized GPR84 from cells treated with and without activators, whereas an antiserum able to identify pThr263/pThr264 only recognized GPR84 after exposure to 2-HTP and not DL-175. Two distinct GPR84 antagonists as well as inhibition of G protein-coupled receptor kinase 2/3 prevented phosphorylation of pThr263/pThr264, but neither strategy affected constitutive phosphorylation of Ser221/Ser224. Furthermore, mutation of residues Thr263 and Thr264 to alanine generated a variant of GPR84 also limited in 2-HTP-induced interactions with arrestin-2 and -3. By contrast, this mutant was unaffected in its capacity to reduce cAMP levels. Taken together, these results define a key pair of threonine residues, regulated only by subsets of GPR84 small molecule activators and by GRK2/3 that define effective interactions with arrestins and provide novel tools to monitor the phosphorylation and functional status of GPR84. [ABSTRACT FROM AUTHOR]

The nineteenth century witnessed the creation of a 'paper Panopticon' designed to capture information about offenders in England, especially those who were transported to Australia. This article considers the effectiveness of this new record-keeping system and asks whether convicts had some agency within it. These questions are explored through a macroscopic analysis of the recording of convict ages in nineteenth-century England and Australia, made possible by the Digital Panopticon project. By using the methodological opportunities opened up by digital technologies, we can test the accuracy of historical records in new ways, and in the process develop a better understanding of the encounter between state authority and convict agency. [ABSTRACT FROM AUTHOR]

Background: Trait anxiety is characterized by impaired gating of threat from working memory (WM), allowing unnecessary maintenance of anxious cognitions. Improving filtering efficiency of threatening information through computerized WM training might reduce intrusive, worrisome thoughts. Methods: We randomized high-trait anxious individuals to 9 sessions of high-potency n-back (n = 19) or low-potency 1-back (n = 18) training to examine their effects on various neurocognitive indices of WM functioning and emotional symptoms. Results: Following the interventions, multilevel modeling analyses revealed both training conditions resulted in significant improvements in attentional (Flanker), WM capacity (operation and spatial span tasks), and WM filtering efficiency (change detection task) measures, and increased N2 amplitudes. However, the high-potency training produced more favorable results at post-training, indexed by larger ERN amplitudes. We also observed significant reductions in trait anxiety and worry symptoms for the high-potency training following the intervention, although, low-potency training caught up at follow-up with comparably reduced symptoms. Conclusions: These results show that emotional WM training can improve neurocognitive processes of attention and WM as well as symptoms of worrying. Overall, this study encourages the development of a standalone or adjunctive cognitive intervention focused on WM for vulnerable populations with high trait anxiety or worry symptoms. [ABSTRACT FROM AUTHOR]

CXCR4, a member of the family of chemokine-activated G protein-coupled receptors, is widely expressed in immune response cells. It is involved in both cancer development and progression as well as viral infection, notably by HIV-1. A variety of methods, including structural information, have suggested that the receptor may exist as a dimer or an oligomer. However, the mechanistic details surrounding receptor oligomerization and its potential dynamic regulation remain unclear. Using both biochemical and biophysical means, we confirm that CXCR4 can exist as a mixture of monomers, dimers, and higher-order oligomers in cell membranes and show that oligomeric structure becomes more complex as receptor expression levels increase. Mutations of CXCR4 residues located at a putative dimerization interface result in monomerization of the receptor. Additionally, binding of the CXCR4 antagonist IT1t--a small drug-like isothiourea derivative--rapidly destabilizes the oligomeric structure, whereas AMD3100, another well-characterized CXCR4 antagonist, does not. Although a mutation that regulates constitutive activity of CXCR4 also results in monomerization of the receptor, binding of IT1t to this variant promotes receptor dimerization. These results provide novel insights into the basal organization of CXCR4 and how antagonist ligands of different chemotypes differentially regulate its oligomerization state. [ABSTRACT FROM AUTHOR]

Current theories propose that anxiety adversely impacts working memory (WM) by restricting WM capacity and interfering with efficient filtering of task‐irrelevant information. The current study investigated the effect of shock‐induced state anxiety on WM capacity and the ability to filter task‐irrelevant neutral stimuli. We measured the contralateral delay activity (CDA), an event‐related potential that indexes the number of items maintained in WM, while participants completed a lateralized change detection task. The task included low and high WM loads, as well as a low load plus distracter condition. This design was used to assess WM capacity for low and high loads and investigate an individual's ability to filter neutral task‐irrelevant stimuli. Participants completed the task under two conditions, threat of shock and safe. We observed a reduced CDA in the threat compared to the safe condition that was specific for high memory load. However, we did not find any differences in CDA filtering cost between threat and safe conditions. In addition, we did not find any differences in behavioral performance between the threat and safe conditions. These findings suggest that being in an anxious state reduces the neural representation for large amounts of information in WM, but have little effect on the filtering of neutral distracters. Anxiety is believed to restrict working memory capacity and impact the ability to filter task‐irrelevant information, but little is known how an anxious state impacts these processes. Here, we provide electrophysiological evidence of reduced working memory capacity, but intact filtering abilities for neutral task‐irrelevant distracters in state anxious individuals. [ABSTRACT FROM AUTHOR]

Objectives: The purpose of this study was to compare clinical decision-making in iron overload patients using FerriScan and an R2*-based approach.Methods: One-hundred and six patients were imaged at two consecutive timepoints (454 ± 158 days) on a 1.5-T Siemens MAGNETOM Avanto Fit scanner. For both timepoints, patients underwent the standard FerriScan MRI protocol. During the second exam, each patient additionally underwent R2*-MRI mapping. For each patient, a retrospective (simulated) decision was made to increase, decrease, or maintain chelator levels. Two different decision models were considered: The fixed threshold model assumed that chelator adjustments are based strictly on fixed liver iron concentration (LIC) thresholds. Decisions made with this model depend only on the most recent LIC value and do not require any clinician input. The second model utilized decisions made by two hematologists retrospectively based on trends between two consecutive LIC values. Agreement (κA) between hematologists (i.e., interobserver variability) was compared with the agreement (κB) between a single hematologist using the two different LIC techniques.Results: Good agreement between R2*- and FerriScan-derived decisions was achieved for the fixed threshold model. True positive/negative rates were greater than 80%, and false positive/negative rates were less than 10%. ROC analysis yielded areas under the curve greater than 0.95. In the second model, the agreement in clinical decision-making for the two scenarios (κA vs. κB) was equal at the 95% confidence level.Conclusions: Switching to R2*-based LIC estimation from FerriScan has the same level of agreement in patient management decisions as does switching from one hematologist to another.Key Points: • Good agreement between R2*- and FerriScan-derived decisions in liver iron overload patient management • Switching to R2*-based LIC estimation from FerriScan has the same level of agreement in patient management decisions as does switching from one hematologist to another. [ABSTRACT FROM AUTHOR]

Objective: While doses of deferiprone up to 75 mg/kg/d have been demonstrated to be effective in cardiac iron removal, their efficacy in the reduction of liver iron has been equivocal. The aim of this study was to evaluate the effect of deferiprone dose on liver iron concentrations in adult iron overload patients. Methods: A single‐centered, retrospective, cohort observational study was conducted involving 71 patients exposed to deferiprone doses up to 113 mg/kg/d between January 2009 and June 2015 for a median of 33 months. Results: At the end of the study period, liver iron measured by R2 MRI was reduced by a mean 1.7 mg/g dw. A dose effect was observed, with incremental reductions of 2.8 mg/g dw in end of study LIC for every 10 mg/kg/d higher dose of deferiprone (P < 0.001). A dose effect was also observed in end of study ferritin and cardiac iron concentration measured by T2* MRI (P < 0.0001 and P = 0.048, respectively). No associations between adverse effects and deferiprone dose were observed, but there was a trend toward higher rates of agranulocytosis at higher doses and two of three hereditary hemochromatosis patients developed this complication. Conclusion: The present study failed to demonstrate that the use of deferiprone at >90 mg/kg/d was associated with increased risk of agranulocytosis or neutropenia, but did demonstrate more effective liver iron control in iron overload patients. [ABSTRACT FROM AUTHOR]

Hemodiafiltration (HDF) increases the removal of middle-molecular-weight uremic toxins and may improve outcomes in patients with end-stage kidney disease (ESKD), but it requires complex equipment and comes with risks associated with infusion of large volumes of substitution solution. New high-flux hemodialysis membranes with improved diffusive permeability profiles do not have these limitations and offer an attractive alternative to HDF. However, both strategies are associated with increased albumin loss into the dialysate, raising concerns about the potential for decreased serum albumin concentrations that have been associated with poor outcomes in ESKD. Many factors can contribute to hypoalbuminemia in ESKD, including protein energy wasting, inflammation, volume expansion, renal loss and loss into the dialysate; of these factors, loss into the dialysate is not necessarily the most important. Furthermore, recent studies suggest that mild hypoalbuminemia per se is not an independent predictor of increased mortality in dialysis patients, but in combination with inflammation it is a poor prognostic sign. Thus, whether hypoalbuminemia predisposes to increased morbidity and mortality may depend on the presence or absence of inflammation. In this review we summarize recent findings on the role of dialysate losses in hypoalbuminemia and the importance of concomitant inflammation on outcomes in patients with ESKD. Based on these findings, we discuss whether hypoalbuminemia may be a price worth paying for increased dialytic removal of middle-molecular-weight uremic toxins. [ABSTRACT FROM AUTHOR]

Cartoon depictions of (a) Dynamic interchange between receptor monomers, (homo, orange GPCR1, and hetero, orange GPCR1 and green GPCR2) dimers and oligomers which may be ligand regulated (orange sphere). (b) Signalling cross-talk between protomers of a dimer. Left: ligand (orange sphere) binds to one protomer and the conformational change transmitted to the second protomer results in signalling through activation of a Gα subunit (blue). Right: ligand (green sphere) binding to the second protomer may modulate signalling (large or small yellow flash). G protein-coupled receptors (GPCRs) are an extensive class of trans-plasma membrane proteins that function to regulate a wide range of physiological functions. Despite a general perception that GPCRs exist as monomers an extensive literature has examined whether GPCRs can also form dimers and even higher-order oligomers, and if such organization influences various aspects of GPCR function, including cellular trafficking, ligand binding, G protein coupling and signalling. Here we focus on recent studies that employ approaches ranging from computational methods to single molecule tracking and both quantal brightness and fluorescence fluctuation measurements to assess the organization, stability and potential functional significance of dimers and oligomers within the class A, rhodopsin-like GPCR family. [ABSTRACT FROM AUTHOR]

Prior research suggests a relationship between metacognitive awareness and academic success, in which individuals with greater metacognitive awareness perform better in academic settings compared to those with a lesser degree of metacognitive awareness. In the current study, we sought to generalize this finding to the population of college freshmen students by examining the relationship between metacognitive awareness and academic performance. In addition, we aimed to explore the potential value of metacognitive awareness training in college freshmen students to encourage retention in higher academic settings. Results revealed a significant positive correlation between metacognitive awareness and college freshmen students' academic performance, as indicated by cumulative grade point average (GPA). This implies that students with a higher degree of metacognitive awareness tend to also succeed academically when compared to those with a lesser degree of metacognitive awareness. In addition, these results generalize the association between academic performance and metacognitive awareness to the college freshmen student population. Since the degree of metacognitive awareness can be increased with instruction, these findings suggest a route in assisting at risk freshmen succeed in academic settings. [ABSTRACT FROM AUTHOR]

G protein-coupled receptors (GPCRs) have been classically described as monomeric entities that function by binding in a 1:1 stoichiometric ratio to both ligand and downstream signalling proteins. However, in recent years, a growing number of studies has supported the hypothesis that these receptors can interact to form dimers and higher order oligomers although the molecular basis for these interactions, the overall quaternary arrangements and the functional importance of GPCR oligomerization remain topics of intense speculation. Muscarinic acetylcholine receptors belong to class A of the GPCR family. Each muscarinic receptor subtype has its own particular distribution throughout the central and peripheral nervous systems. In the central nervous system, muscarinic receptors regulate several sensory, cognitive, and motor functions while, in the peripheral nervous system, they are involved in the regulation of heart rate, stimulation of glandular secretion and smooth muscle contraction. Muscarinic acetylcholine receptors have long been used as a model for the study of GPCR structure and function and to address aspects of GPCR dimerization using a broad range of approaches. In this review, the prevailing knowledge regarding the quaternary arrangement for the various muscarinic acetylcholine receptors has been summarized by discussing work ranging from initial results obtained using more traditional biochemical approaches to those generated with more modern biophysical techniques. This article is part of the Special Issue entitled ‘Neuropharmacology on Muscarinic Receptors’. [ABSTRACT FROM AUTHOR]

Covalent inhibition is a rapidly growing discipline within drug discovery. Many historical covalent inhibitors were discovered by serendipity, with such a mechanism of action often regarded as undesirable due to potential toxicity issues. Recent progress has seen a major shift in this outlook, as covalent inhibition shows promise for targets where previous efforts to identify non-covalent small molecule inhibitors have failed. Targeted covalent inhibitors (TCIs) can offer drug discovery scientists the ability to increase the potency and/or selectivity of small molecule inhibitors, by attachment of reactive functional groups designed to covalently bind to specific sites in a target. In this tutorial review we introduce the broader concept of covalent inhibition, discuss the potential benefits and challenges of such an approach, and provide an overview of the current status of the field. We also describe some strategies and computational tools to enable successful covalent drug discovery. [ABSTRACT FROM AUTHOR]

Spatial intensity distribution analysis (SpIDA) is a recently developed approach for determining quaternary structure information on fluorophore-labelled proteins of interest in situ . It can be applied to live or fixed cells and native tissue. Using confocal images, SpIDA generates fluorescence intensity histograms that are analysed by super-Poissonian distribution functions to obtain density and quantal brightness values of the fluorophore-labelled protein of interest. This allows both expression level and oligomerisation state of the protein to be determined. We describe the application of SpIDA to investigate the oligomeric state of G protein-coupled receptors (GPCRs) at steady state and following cellular challenge, and consider how SpIDA may be used to explore GPCR quaternary organisation in pathophysiology and to stratify medicines. [ABSTRACT FROM AUTHOR]

A paramagnetic nanocomposite coated with chitosan and N-(5-Amino-1-carboxy-pentyl) iminodiacetic acid (NTA) that is suitable for protein immobilization applications has been prepared and characterized. The nanoparticle core was synthesized by controlled aggregation of Fe 3 O 4 under alkaline conditions, and Transmission Electron Microscopy revealed a size distribution of 10–50 nm. The nanoparticle core was coated with chitosan and derivatized with glutaraldehyde and NTA, as confirmed by Fourier Transform Infrared Spectroscopy. The final nanoparticles were used as a metal affinity matrix to separate a recombinant polyhistidine-tagged β-galactosidase from Bacillus subtilis directly from E. coli cell lysates with high purity (>95%). After loading with Ni 2+ , nanoparticles demonstrated a binding capacity of 250 μg of a polyhistidine-tagged β-galactosidase per milligram of support. The immobilized enzyme retained 80% activity after 9 cycles of washing, and the immobilized recombinant protein could be eluted with high purity with imidazole. The applications for these nanomagnetic composites extend beyond protein purification, and can also be used for immobilizing enzymes, where the β-galactosidase immobilized on the nanomagnetic support was used in multiple cycles of catalytic reactions with no significant loss of catalytic activity. [ABSTRACT FROM AUTHOR]

This article seeks to understand why detailed personal information about accused criminals and convicts was recorded from the late 18th century in England, and why some of this information was converted into statistics from the 1820s, such that by 1860, extensive information about criminals' physical characteristics and backgrounds was regularly collected and tabulated. These developments in record-keeping and statistics were mostly the result of local initiatives and imperatives, revealing a grass-roots information-gathering culture, with limited central government direction. Rather than primarily driven by efforts at control or the practical demands of judicial administration, the substantial amount of information recorded reveals a strong and widely held desire to understand the criminal, long before the self-conscious enterprise of 'criminology' was invented. [ABSTRACT FROM AUTHOR]

Previous studies have indicated that the G-protein-coupled secretin receptor is present as a homodimer, organized through symmetrical contacts in transmembrane domain IV, and that receptor dimerization is critical for high-potency signalling by secretin. However, whether all of the receptor exists in the dimeric form or if this is regulated is unclear. We used measures of quantal brightness of the secretin receptor tagged with monomeric enhanced green fluorescent protein (mEGFP) and spatial intensity distribution analysis to assess this. Calibration using cells expressing plasma membrane-anchored forms of mEGFP initially allowed us to demonstrate that the epidermal growth factor receptor is predominantly monomeric in the absence of ligand and while wild-type receptor was rapidly converted into a dimeric form by ligand, a mutated form of this receptor remained monomeric. Equivalent studies showed that, at moderate expression levels, the secretin receptor exists as a mixture of monomeric and dimeric forms, with little evidence of higher-order complexity. However, sodium butyrate-induced up-regulation of the receptor resulted in a shift from monomeric towards oligomeric organization. In contrast, a form of the secretin receptor containing a pair of mutations on the lipid-facing side of transmembrane domain IV was almost entirely monomeric. Down-regulation of the secretin receptor- interacting G-protein Gαsdid not alter receptor organization, indicating that dimerization is defined specifically by direct protein–protein interactions between copies of the receptor polypeptide, while short-term treatment with secretin had no effect on organization of the wild-type receptor but increased the dimeric proportion of the mutated receptor variant. [ABSTRACT FROM AUTHOR]

There are a number of small-molecule inhibitors targeting the RAS/RAF/MEK/ERK signaling pathway that have either been approved or are in clinical development for oncology across a range of disease indications. The inhibition of ERK1/2 is of significant current interest, as cell lines with acquired resistance to BRAF and MEK inhibitors have been shown to maintain sensitivity to ERK1/2 inhibition in preclinical models. This article reports on our recent work to identify novel, potent, and selective reversible ERK1/2 inhibitors from a low-molecular-weight, modestly active, and highly promiscuous chemical start point, compound 4. To guide and inform the evolution of this series, inhibitor binding mode information from X-ray crystal structures was critical in the rapid exploration of this template to compound 35, which was active when tested in in vivo antitumor efficacy experiments. [ABSTRACT FROM AUTHOR]

Aims The aim of this study was to investigate the impact of the death of an older member on families. Background The death of a significant other in later life is a dramatic moment. Research has demonstrated that some older persons face negative consequences for their well-being. A majority, however, exhibit resilience in the wake of loss. Nonetheless, the relational process through which older persons come to terms with the loss in interaction with their families is little understood, but vital to support bereaved families. Design Heideggerian hermeneutic phenomenology. Methods A purposive sample of ten older persons with their families, represented by children, grandchildren and in-laws ( n = 30) were interviewed several times in 2013, alone ( n = 16) and in family groups ( n = 21), 6-23 months after their significant other's death (mean age 81 years). Data collection and thematic analysis was informed by van Manen's and Benner's analytical strategies. Findings Three family themes were discerned. First, through meaning-making, bereaved families weaved the death into their family narrative. Second, through sharing-not sharing their feelings and daily moments, family members lived with the loss both together and alone. Third, some families faced upheaval in their family life, which required them to re-create their everyday life, whereas other families continued with little change. Conclusions Findings demonstrate that families hold an inherent capacity to make meaning of the death and enact family thereafter. Family relations arose as interplay of different, contradicting forces. Nurses should facilitate families' meaning-making of the death, attend to their converging and diverging sense of loss and strengthen family caring. [ABSTRACT FROM AUTHOR]

Tomato brown rugose fruit virus (ToBRFV) is a contact-transmitted tobamovirus affecting many tomato growing regions of the world. This study investigated the effects of different glasshouse surfaces on the survival of the virus; the efficacy of different disinfectants; and heat treatment against ToBRFV (surfaces included steel, aluminium, hard plastic, polythene, glass and concrete). A bioassay followed by ELISA was used to check virus viability. ToBRFV survived for at least 7 days on all surfaces tested and on some for at least 6 months. The virus survived for over two hours on hands and gloves. Hand washing was shown to be unreliable for the removal of the virus. Glutaraldehyde and quaternary ammonium compound disinfectants were effective at one hour on all surfaces. Some other disinfectants were effective at one hour of contact time, on all surfaces except concrete. Sodium hypochlorite was partially effective against ToBRFV, even on concrete. A 5 min soak of plastic trays in water at 90 °C was effective at denaturing ToBRFV; however, 5 min at 70 °C was not. Heating infected sap showed the thermal inactivation point to be 90 °C, confirming the hot water treatment results and showing that deactivation was due to the heat treatment and not a washing effect of the water. [ABSTRACT FROM AUTHOR]

This article explores the labour and experiences of a hitherto entirely overlooked section of the dementia care workforce: care-based hairdressers. Reporting on findings from the ESRC-funded 'Hair and Care' project, the analysis and discussion focus upon the 'doing of hair' in the context of dementia care. The authors challenge existing assumptions and approaches to the management of appearance in dementia care, arguing for greater recognition of the subjective and culturally meaningful qualities of a visit to the salon. The article draws upon a wider debate on body work as a framework for the discussion, and considers the employment and working conditions of this largely hidden group of workers in the care system. The article offers an account of the praxis of care-based hairdressing, with particular attention paid to narrative, intercorporeal and place-making practices in the salon, showing how a particular approach to the body shapes the labour, relationships and activities that unfold within it. The authors argue that as an alternative form of body work much can be learned from hairdressing that can inform and enhance the provision of dementia care. [ABSTRACT FROM AUTHOR]

Our aim was to develop and assess the feasibility of an education tool to improve health outcomes of patients with thalassemia. Thirty-five patients attending a Canadian thalassemia clinic were enrolled. Acting in an expert role, they participated in a Delphi method to reach consensus as to what tools and information should be incorporated in the development of a self management Smartphone app. One- and 6-month usability and health impact feedback surveys were built-in. Sixty percent of responders were 18–34 years old, over 50.0% had a college degree. The Delphi method successfully generated a comprehensive list of features important to patients. The app has been downloaded 147 times globally. Between March 2015 and January 2016, 19 responses for the 1-month survey were collected and the trends described. Responders reported improved medication adherence. The personal adherence pledge feature supports gamification of health apps to individualize goals of therapy. The impact of tracking iron levels was highly favorable. The Delphi method was an effective way to introduce a patient education and empowerment tool to the thalassemia population. The long-term impact requires data maturation. Use of validated methodology is essential to ensure ehealth interventions are positively contributing to patient education and disease outcomes. [ABSTRACT FROM AUTHOR]

Although rhodopsin-like G protein-coupled receptors can exist as both monomers and non-covalently associated dimers/ oligomers, the steady-state proportion of each form and whether this is regulated by receptor ligands are unknown. Herein we address these topics for the M1 muscarinic acetylcholine receptor, a key molecular target for novel cognition enhancers, by using spatial intensity distribution analysis. This method can measure fluorescent particle concentration and assess oligomerization states of proteins within defined regions of living cells. Imaging and analysis of the basolateral surface of cells expressing some 50 molecules·μm-2 human muscarinic M1 receptor identified a ~75:25 mixture of receptor monomers and dimers/oligomers. Both sustained and shorter term treatment with the selective M1 antagonist pirenzepine resulted in a large shift in the distribution of receptor species to favor the dimeric/ oligomeric state. Although sustained treatment with pirenzepine also resulted in marked up-regulation of the receptor, simple mass action effects were not the basis for ligand-induced stabilization of receptor dimers/oligomers. The related antagonist telenzepine also produced stabilization and enrichment of the M1 receptor dimer population, but the receptor subtype non-selective antagonists atropine and N-methylscopolamine did not. In contrast, neither pirenzepine nor telenzepine altered the quaternary organization of the related M3 muscarinic receptor. These data provide unique insights into the selective capacity of receptor ligands to promote and/or stabilize receptor dimers/oligomers and demonstrate that the dynamics of ligand regulation of the quaternary organization of G protein-coupled receptors is markedly more complex than previously appreciated. This may have major implications for receptor function and behavior. [ABSTRACT FROM AUTHOR]

Surface waters are increasingly contaminated by cyanobacteria, which may produce potent cyanotoxins harmful to animals and humans. Hemodialysis patients are at high risk of injury from waterborne contaminants in the water used to prepare dialysate. Episodes of acute illness and death among hemodialysis patients have been reported following exposure to dialysate prepared from drinking water contaminated with elevated concentrations of cyanotoxins. Protecting dialysis patients from these toxins is complicated by a lack of monitoring and regulation of cyanotoxins in drinking water, uncertainty as to their safe levels in dialysate, and incomplete knowledge of how well current dialysate preparation and water treatment practices remove them. Until these issues are adequately addressed, hemodialysis centers should be aware of the potential for cyanotoxins to be present in their potable water supply, particularly when it comes from surface water sources prone to cyanobacterial blooms. [ABSTRACT FROM AUTHOR]

In this paper we explore the experience and implications of getting lost with dementia. While getting lost has become culturally emblematic of dementia, speaking as it does to a widespread fear of losing our place in the world, it is marked by an overall absence of critical attention. We argue that this critical hesitancy is part of a broader unease with 'dementing' that reveals a paradox in dementia scholarship as growing emphasis on strengths-based and capacity-oriented approaches to the condition shift attention away from episodes of disorientation, forgetting and unknowing that commonly arise after onset. We therefore explore getting lost with dementia, not only as a route to better understanding what happens during such events and the meaning it holds but also to consider the implications for a broader politics of the social inclusion of people living with dementia. Reporting findings from a five-year international study of the neighbourhood experiences of people with dementia, we suggest that through such experiences as getting lost, people with dementia have a unique and distinctive contribution to make to the ever-evolving character of public space and civic culture. In particular, we argue that getting lost and the subsequent recovery or reconstitution that ensues can help inform efforts to reimagine public space. This includes looking beyond risk reduction in responses to dementia and public and outdoor settings to consider how freedom of movement for people with dementia might be enhanced rather than curtailed. Our learning points to the value of making the process of dementing more visible and central to the politics and practices of social inclusion. [ABSTRACT FROM AUTHOR]

Chronic red blood cell (RBC) transfusion is employed for a wide range of sickle cell disease complications, ranging from primary and secondary stroke prophylaxis to prevention of painful vaso-occlusive episodes. Currently different methods are employed by centers for chronic transfusion that include simple, automated and partial manual RBC exchange transfusion. A retrospective cohort study of two different manual RBC exchange transfusion methods was conducted between two comprehensive care centers in Toronto, ON, Canada and London, United Kingdom in 19 and 21 sickle cell disease adults, respectively. London used a weight-based protocol, while Toronto used a unit-based method. Our results indicated that sickle cell disease patients utilizing a weight-based method are more often unable to achieve the prescribed Hb S (HBB: c.20A > T) target compared to the unit-based method (90.0vs.53.0% in the weight-based and unit-based methods, respectively,p = 0.0123). On multivariable logistic regression, none of the covariates examined was found to influence the ability to achieve the prescribed Hb S target after accounting for the exchange transfusion method. Mean interval of exchange sessions, session duration, total units of packed RBC, volume of blood used by body weight each year, the mean post exchange hematocrit [or packed cell volume (PCV)] and ferritin change were similar in both cohorts. In conclusion, the unit-based method was more effective at maintaining the prescribed Hb S target. [ABSTRACT FROM AUTHOR]

The article explores the geographical history of capital punishment such as hanging in Great Britain and the implementation of the Bloody Code in peripheral areas. Emphasis is given to topics such as the hegemony of the 18th-century elite, the autonomy of rural areas in the implementation of social policy, and execution rates by county.

The article offers a comparison of chronic manual and automated red blood cell exchange transfusion among patients with sickle cell disease. Topics discussed include treatment and prevention of the multisystem complications in sickle cell disease (SCD), reduction of sickle haemoglobin (HbS) without raising final hematocrit (Hct) by Red blood cell (RBC) exchange transfusion (RBCX), and use of manual RBCX for patients undergoing sequential phlebotomies and transfusions.

The questions of whether G protein-coupled receptors exist as monomers, dimers, and/or oligomers and if these species interconvert in a ligand-dependent manner are among the most contentious current issues in biology. When employing spatial intensity distribution analysis to laser scanning confocal microscope images of cells stably expressing either a plasma membrane- associated form of monomeric enhanced green fluorescent protein (eGFP) or a tandem version of this fluorophore, the eGFP tandem was identified as a dimer. Similar studies on cells stably expressing an eGFP-tagged form of the epidermal growth factor receptor demonstrated that, although largely a monomer in the basal state, this receptor rapidly became predominantly dimeric upon the addition of its ligand epidermal growth factor. In cells induced to express an eGFP-tagged form of the serotonin 5-hydroxytryptamine 2C (5-HT2C) receptor, global analysis of construct quantal brightness was consistent with the predominant form of the receptor being dimeric. However, detailed spatial intensity distribution analysis demonstrated the presence of multiple forms ranging from monomers to higher-order oligomers. Furthermore, treatment with chemically distinct 5-HT2C) receptor antagonists resulted in a time-dependent change in the quaternary organization to one in which there was a preponderance of receptor monomers. This antagonist-mediated effect was reversible, because washout of the ligand resulted in the regeneration of many of the oligomeric forms of the receptor. [ABSTRACT FROM AUTHOR]

There is a growing demand for enzymes with improved catalytic performance or tolerance to process-specific parameters, and biotechnology plays a crucial role in the development of biocatalysts for use in industry, agriculture, medicine and energy generation. Metagenomics takes advantage of the wealth of genetic and biochemical diversity present in the genomes of microorganisms found in environmental samples, and provides a set of new technologies directed towards screening for new catalytic activities from environmental samples with potential biotechnology applications. However, biased and low level of expression of heterologous proteins in E scherichia coli together with the use of non-optimal cloning vectors for the construction of metagenomic libraries generally results in an extremely low success rate for enzyme identification. The bottleneck arising from inefficient screening of enzymatic activities has been addressed from several perspectives; however, the limitations related to biased expression in heterologous hosts cannot be overcome by using a single approach, but rather requires the synergetic implementation of multiple methodologies. Here, we review some of the principal constraints regarding the discovery of new enzymes in metagenomic libraries and discuss how these might be resolved by using synthetic biology methods. [ABSTRACT FROM AUTHOR]

Pulsed electron–electron double resonance (PELDOR) coupled with site-directed spin labeling is a powerful technique for the elucidation of protein or nucleic acid, macromolecular structure and interactions. The intrinsic high sensitivity of electron paramagnetic resonance enables measurement on small quantities of bio-macromolecules, however short relaxation times impose a limit on the sensitivity and size of distances that can be measured using this technique. The persistence of the electron spin-echo, in the PELDOR experiment, is one of the most crucial limitations to distance measurement. At a temperature of around 50 K one of the predominant factors affecting persistence of an echo, and as such, the sensitivity and measurable distance between spin labels, is the electron spin echo dephasing time ( T m ). It has become normal practice to use deuterated solvents to extend T m and recently it has been demonstrated that deuteration of the underlying protein significantly extends T m . Here we examine the spatial effect of segmental deuteration of the underlying protein, and also explore the concentration and temperature dependence of highly deuterated systems. [ABSTRACT FROM AUTHOR]

Appearance and the work invested in it by and for people with dementia are a neglected issue within dementia studies. In policy and practice there exists an assumption that the role of supporting a person to manage their appearance is easily assumed by another within caring encounters, only to be subsumed within the daily task-oriented provision of care. This paper reports on interviews conducted as part of the Hair and Care project, which explored questions of appearance and the meanings it holds with people with dementia. The research used ‘appearance biographies’, a method which allows for a range of topics to be considered about appearance throughout the life course, acting as a conduit for reminiscence and life story work. The paper reports on the key themes and findings from these interviews, discussing them in the context of a wider debate on dementia, self-expression and agency. A key question posed by the authors is whether appearance and the work invested in it are legitimate considerations for dementia care policy and practice. And if so, how should we make sense of this work and what significance should we attach to it? In seeking to answer these questions the authors position the perspectives and experiences of people with dementia as central to their analysis. A narrative framework is suggested as a useful basis on which to understand the work of managing appearance over the life course. The implications for policy and practice are outlined. [ABSTRACT FROM AUTHOR]

Robo (Roundabout) receptors and their Slit polypeptide ligands are known to play key roles in neuronal development and have been implicated in both angiogenesis and cancer. Like the other family members, Robo1 is a large single transmembrane domain polypeptide containing a series of well-defined extracellular elements. However, the intracellular domain lacks structural definition and little is known about the quaternary structure of Robo receptors or how binding of a Slit might affect this. To address these questions combinations of both autofluorescent protein-based FRET imaging and time-resolved FRET were employed. Both approaches identified oligomeric organization of Robo1 that did not require the presence of the intracellular domain. SpIDA (spatial intensity distribution analysis) of eGFPtagged forms of Robo1 indicated that for a C-terminally deleted version approximately two-thirds of the receptor was present as a dimer and one-third as a monomer. By contrast, full-length Robo1 was present almost exclusively as a dimer. In each case this was unaffected by the addition of Slit2, although parallel studies demonstrated the biological activity of Slit2 and its interaction with Robo1. Deletion of both the immunoglobulin and fibronectin type III extracellular repeats prevented dimer formation, with the immunoglobulin repeats providing the bulk of the protein-protein interaction affinity. [ABSTRACT FROM AUTHOR]

A series of 2,5-bis(arylethynyl)rhodacyclopentadienes has been prepared by a rare example of regiospecific reductive coupling of 1,4-( p-R-phenyl)-1,3-butadiynes (RH, Me, OMe, SMe, NMe2, CF3, CO2Me, CN, NO2, −CC-( p-C6H4NHex2), −CC( p-C6H4CO2Oct)) at [RhX(PMe3)4] ( 1) (X=−CCSiMe3 ( a), −CC-( p-C6H4NMe2) ( b), −CCCC( p-C6H4NPh2) ( c) or CC{ p-C6H4-CC( p-C6H4-N(C6H13)2)} ( d) or Me ( e)), giving the 2,5-bis(arylethynyl) isomer exclusively. The rhodacyclopentadienes bearing a methyl ligand in the equatorial plane (compound 1 e) have been converted into their chloro analogues by reaction with HCl etherate. The rhodacycles thus obtained are stable to air and moisture in the solid state and the acceptor-substituted compounds are even stable to air and moisture in solution. The photophysical properties of the rhodacyclopentadienes are highly unusual in that they exhibit, exclusively, fluorescence between 500-800 nm from the S1 state, with quantum yields of Φ=0.01-0.18 and short lifetimes ( τ=0.45-8.20 ns). The triplet state formation ( ΦISC=0.57 for 2 a) is exceptionally slow, occurring on the nanosecond timescale. This is unexpected, because the Rh atom should normally facilitate intersystem crossing within femto- to picoseconds, leading to phosphorescence from the T1 state. This work therefore highlights that in some transition-metal complexes, the heavy atom can play a more subtle role in controlling the photophysical behavior than is commonly appreciated. [ABSTRACT FROM AUTHOR]

Abstract: The interaction between ligands and the G protein-coupled receptors (GPCRs) to which they bind has long been the focus of intensive investigation. The signalling cascades triggered by receptor activation, due in most cases to ligand binding, are of great physiological and medical importance; indeed, GPCRs are targeted by in excess of 30% of small molecule therapeutic medicines. Attempts to identify further pharmacologically useful GPCR ligands, for receptors with known and unknown endogenous ligands, continue apace. In earlier days direct assessment of such interactions was restricted largely to the use of ligands incorporating radioactive isotope labels as this allowed detection of the ligand and monitoring its interaction with the GPCR. This use of such markers has continued with the development of ligands labelled with fluorophores and their application to the study of receptor–ligand interactions using both light microscopy and resonance energy transfer techniques, including homogenous time-resolved fluorescence resonance energy transfer. Details of ligand–receptor interactions via X-ray crystallography are advancing rapidly as methods suitable for routine production of substantial amounts and stabilised forms of GPCRs have been developed and there is hope that this may become as routine as the co-crystallisation of serine/threonine kinases with ligands, an approach that has facilitated widespread use of rapid structure-based ligand design. Conformational changes involved in the activation of GPCRs, widely predicted by biochemical and biophysical means, have inspired the development of intramolecular FRET-based sensor forms of GPCRs designed to investigate the events following ligand binding and resulting in a signal propagation across the cell membrane. Finally, a number of techniques are emerging in which ligand–GPCR binding can be studied in ways that, whilst indirect, are able to monitor its results in an unbiased and integrated manner. This article is part of a Special Issue entitled: Structural and biophysical characterisation of membrane protein–ligand binding. [Copyright &y& Elsevier]

Nanometre distance measurements by pulsed electron−electron double resonance (PELDOR) spectroscopy have become an increasingly important tool in structural biology. The theoretical underpinning of the experiment is well defined for systems containing two nitroxide spin-labels (spin pairs); however, recently experiments have been reported on homo-oligomeric membrane proteins consisting of up to eight spin-labelled monomers. We have explored the theory behind these systems by examining model systems based on multiple spins arranged in rotationally symmetric polygons. The results demonstrate that with a rising number of spins within the test molecule, increasingly strong distortions appear in distance distributions obtained from an analysis based on the simple spin pair approach. These distortions are significant over a range of system sizes and remain so even when random errors are introduced into the symmetry of the model. We present an alternative approach to the extraction of distances on such systems based on a minimisation that properly treats multi-spin correlations. We demonstrate the utility of this approach on a spin-labelled mutant of the heptameric Mechanosensitive Channel of Small Conductance ofE. coli. [ABSTRACT FROM PUBLISHER]

A novel series of small-moleculeinhibitors has been developedto target the double mutant form of the epidermal growth factor receptor(EGFR) tyrosine kinase, which is resistant to treatment with gefitiniband erlotinib. Our reported compounds also show selectivity over wild-typeEGFR. Guided by molecular modeling, this series was evolved to targeta cysteine residue in the ATP binding site via covalent bond formationand demonstrates high levels of activity in cellular models of thedouble mutant form of EGFR. In addition, these compounds show significantactivity against the activating mutations, which gefitinib and erlotinib target and inhibition of which gives rise totheir observed clinical efficacy. A glutathione (GSH)-based assaywas used to measure thiol reactivity toward the electrophilic functionalityof the inhibitor series, enabling both the identification of a suitablereactivity window for their potency and the development of a reactivityquantitative structure-property relationship (QSPR) to support design. [ABSTRACT FROM AUTHOR]

Background: Spousal loss can be a dramatic life event for older persons, and a difficult experience that affects everyday life. Research shows that bereavement influences health and well-being in old age and involves changes in social networks. However, the nature of the bereavement experience for older widows/ers remains unclear. Objectives: To determine key characteristics of the bereavement experience of older widowed persons. Design: An integrative review of the literature was performed using the framework developed by Whittemore and Knafl (2005). Data sources: Research studies investigating aspects of the bereavement experiences of widowed older (>65) persons (n = 39) were included. The electronic databases Medline, CINAHL, PsychInfo, Sociological Abstracts, and Cochrane Library and relevant journals were searched up to February 2011. Review methods: The health sciences literature was searched systematically and iteratively to determine relevant studies. Data analysis and synthesis of primary sources of qualitative and quantitative research studies occurred through the processes of data reduction, display, comparison, conclusion drawing and verification (Whittemore and Knafl, 2005). Results: A majority of the reviewed studies included more women than men as participants. The studies' findings suggest that daily activities and routines are disrupted in bereavement, while certain coping strategies seem to sustain older persons in their everyday life. Many older persons face a pervasive sense of loneliness and difficult daily and yearly times. Health concerns prevail or intensify, and older persons have to negotiate their independence and new identity as widow/ers in a social context. Findings show changes in relationships with close others, which are both a challenge and resource for older widow/ers. Moreover, many older persons maintain connections with the deceased spouse. Conclusions: This review suggests that constructing a new identity as widow/er and striving for independence in the face of disrupted everyday activities and routines, loneliness, health concerns and changed relationships within the family and social network are essential features of older persons' bereavement experience. While many of the identified characteristics of the bereavement experience may not be specific to later life, they need to be considered when working with this population given the concurrent challenges of the ageing process and declining health. [ABSTRACT FROM AUTHOR]

The general objective assigned to the EUropean DIALlysis (EUDIAL) Working Group by the European Renal Association-European Dialysis and Transplant Association (ERA-EDTA) was to enhance the quality of dialysis therapies in Europe in the broadest possible sense. Given the increasing interest in convective therapies, the Working Group has started by focusing on haemodiafiltration (HDF) therapies. Several reports suggest that those therapies potentially improve the outcomes for end-stage renal disease patients. Europe is the leader in the field, having introduced the concept of ultra-purity for water and dialysis fluids and with notified bodies of the European Community having certified water treatment systems and online HDF machines. The prevalence of online HDF-treated patients is steadily increasing in Europe, averaging 15%. A EUDIAL consensus conference was held in Paris on 13 October 2011 to revisit terminology, safety and efficacy of online HDF. This is the first report of the expert group arising from that conference. [ABSTRACT FROM PUBLISHER]

Intramolecular fluorescence resonance energy transfer (FRET) sensors able to detect changes in distance or orientation between the 3rd intracellular loop and C-terminal tail of the human orexin OX1 and OX2 G protein-coupled receptors following binding of agonist ligands were produced and expressed stably. These were directed to the plasma membrane and, despite the substantial sequence alterations introduced, in each case were able to elevate [Ca2+]i, promote phosphorylation of the ERK1/2 MAP kinases and become internalized effectively upon addition of the native orexin peptides. Detailed characterization of the OX1 sensor demonstrated that it was activated with rank order of potency orexin A > orexin B > orexin A 16-33, that it bound antagonist ligands with affinity similar to the wild-type receptor, and that mutation of a single residue, D203A, greatly reduced the binding and function of orexin A but not antagonist ligands. Addition of orexin A to individual cells expressing an OX1 sensor resulted in a time- and concentration- dependent reduction in FRET signal consistent with mass-action and potency/affinity estimates for the peptide. Compared with the response kinetics of a muscarinicM3 acetylcholine receptor sensor upon addition of agonist, response of theOX1 andOX2 sensors to orexinAwas slow, consistent with a multistep binding and activation process. Such sensors provide means to assess the kinetics of receptor activation and how this may be altered by mutation and sequence variation of the receptors. [ABSTRACT FROM AUTHOR]

Lactate dehydrogenase A (LDHA) catalyzes the conversion of pyruvate to lactate, utilizing NADH as a cofactor. It has been identified as a potential therapeutic target in the area of cancer metabolism. In this manuscript we report our progress using fragment-based lead generation (FBLG), assisted by X-ray crystallography to develop small molecule LDHA inhibitors. Fragment hits were identified through NMR and SPR screening and optimized into lead compounds with nanomolar binding affinities via fragment linking. Also reported is their modification into cellular active compounds suitable for target validation work. [ABSTRACT FROM AUTHOR]