Miracle-dz/UniFAR-FTUs huggingface dataset:
JC virus persistence following progressive multifocal leukoencephalopathy in multiple sclerosis patients treated with natalizumab

JC virus (JCV) DNA in the cerebrospinal fluid (CSF) provides the laboratory confirmatory diagnosis of progressive multifocal leukoencephalopathy (PML) in patients whose clinical symptoms and magnetic resonance imaging findings are consistent with PML.The Laboratory of Molecular Medicine and Neuroscience (LMMN), National Institute of Neurological Disorders and Stroke (NINDS), National Institutes of Health (NIH), made the confirmatory laboratory diagnosis in 35 multiple sclerosis (MS) patients treated with natalizumab. Thirteen patients had 3 or more CSF samples taken from weeks to months following PML diagnosis. Seven of the 13 patients demonstrated persistence of JCV DNA in the CSF even though all patients experienced immune reconstitution inflammatory syndrome (IRIS), 11 patients had plasma exchange, and 2 had immunoabsorption. Specific anti‐JCV antibody was measured in plasma/sera samples from 25 of the 35 patients. Most of the samples showed moderate to high or rising antibody levels from the time of PML diagnosis. However, plasma from 1 patient at or near the time of PML diagnosis had a titer considered seronegative and 2 other plasma samples from patients had titers considered at baseline for seropositivity. In several PML cases, viral persistence and neurological deficits have continued for several years, indicating that once initiated, JCV infection may not entirely clear, even with IRIS. Ann Neurol 2010;68:384–391
Activation of CXCR7 receptor promotes oligodendroglial cell maturation

Differentiation of oligodendroglial precursor cells is crucial for central nervous system (re)myelination and is influenced by multiple extrinsic and intrinsic factors. Chemokines, a group of small proteins, are highly conserved among mammals and have been implicated in a variety of biological processes during development, tissue homeostasis, and repair. We investigated whether the chemokine CXCL12 influences oligodendrocytes and what cellular differentiation/maturation processes are controlled by this molecule.
Arf6 Guanine Nucleotide Exchange Factor Cytohesin-2 Binds to CCDC120 and Is Transported Along Neurites to Mediate Neurite Growth*

Background: The Arf6 activator, cytohesin-2, is involved in neurite growth. Results: Cytohesin-2 binds to CCDC120 and is transported along growing neurites. Conclusion: This interaction is required for Arf6 activation and neurite growth. Significance: The previously unknown functional CCDC120 is a new cytohesin adaptor protein, which regulates neurite growth. The mechanism of neurite growth is complicated, involving continuous cytoskeletal rearrangement and vesicular trafficking. Cytohesin-2 is a guanine nucleotide exchange factor for Arf6, an Arf family molecular switch protein, controlling cell morphological changes such as neuritogenesis. Here, we show that cytohesin-2 binds to a protein with a previously unknown function, CCDC120, which contains three coiled-coil domains, and is transported along neurites in differentiating N1E-115 cells. Transfection of the small interfering RNA (siRNA) specific for CCDC120 into cells inhibits neurite growth and Arf6 activation. When neurites start to extend, vesicles containing CCDC120 and cytohesin-2 are transported in an anterograde manner rather than a retrograde one. As neurites continue extension, anterograde vesicle transport decreases. CCDC120 knockdown inhibits cytohesin-2 localization into vesicles containing CCDC120 and diffuses cytohesin-2 in cytoplasmic regions, illustrating that CCDC120 determines cytohesin-2 localization in growing neurites. Reintroduction of the wild type CCDC120 construct into cells transfected with CCDC120 siRNA reverses blunted neurite growth and Arf6 activity, whereas the cytohesin-2-binding CC1 region-deficient CCDC120 construct does not. Thus, cytohesin-2 is transported along neurites by vesicles containing CCDC120, and it mediates neurite growth. These results suggest a mechanism by which guanine nucleotide exchange factor for Arf6 is transported to mediate neurite growth.
3D visualization and quantification of rat cortical bone porosity using a desktop micro‐CT system: a case study in the tibia

Although micro‐computed tomography (micro‐CT) has become the gold standard for assessing the 3D structure of trabecular bone, its extension to cortical bone microstructure has been relatively limited. Desktop micro‐CT has been employed to assess cortical bone porosity of humans, whereas that of smaller animals, such as mice and rats, has thus far only been imaged using synchrotron‐based micro‐CT. The goal of this study was to determine if it is possible to visualize and quantify rat cortical porosity using desktop micro‐CT. Tibiae (n = 10) from 30‐week‐old female Sprague‐Dawley rats were imaged with micro‐CT (3 μm nominal resolution) and sequential ground sections were then prepared. Bland–Altman plots were constructed to compare per cent porosity and mean canal diameter from micro‐CT (3D) versus histology (2D). The mean difference or bias (histology – micro‐CT; ±95% confidence interval) for per cent porosity was found to be −0.15% (±2.57%), which was not significantly different from zero (P= 0.720). Canal diameter had a bias (±95% confidence interval) of −5.73 μm (±4.02 μm) which was found to be significantly different from zero (P < 0.001). The results indicated that cortical porosity in rat bone can indeed be visualized by desktop micro‐CT. Quantitative assessment of per cent porosity provided unbiased results, whereas direct analysis of mean canal diameter was overestimated by micro‐CT. Thus, although higher resolution, such as that available from synchrotron micro‐CT, may ultimately be required for precise geometric measurements, desktop micro‐CT – which is far more accessible – is capable of yielding comparable measures of porosity and holds great promise for assessment of the 3D arrangement of cortical porosity in the rat.
Evaluation of a New Nutrition Education Curriculum and Factors Influencing Its Implementation

The study reported here evaluated a nutrition education curriculum through examining participant behavior change. A previously used curriculum (April September 2005 and 2006) was compared to a new curriculum (April September 2007) using preand post-program 24-hour dietary recalls and food behavior surveys. Participant, educator, and program variables were explored relative to behavior change. The new curriculum elicited behavior change similar to that elicited by the previous curriculum. Factors influencing behavior change included participant and educator race and educator experience. Participant and educator variables should be considered in future program/curriculum implementation.
Gut, bugs, and brain: Role of commensal bacteria in the control of central nervous system disease

The mammalian gastrointestinal track harbors a highly heterogeneous population of microbial organisms that are essential for the complete development of the immune system. The gut microbes or “microbiota,” coupled with host genetics, determine the development of both local microbial populations and the immune system to create a complex balance recently termed the “microbiome. ” Alterations of the gut microbiome may lead to dysregulation of immune responses both in the gut and in distal effector immune sites such as the central nervous system (CNS). Recent findings in experimental autoimmune encephalomyelitis, an animal model of human multiple sclerosis, suggest that altering certain bacterial populations present in the gut can lead to a proinflammatory condition that may result in the development of autoimmune diseases, in particular human multiple sclerosis. In contrast, other commensal bacteria and their antigenic products, when presented in the correct context, can protect against inflammation within the CNS. Ann Neurol 2011
Atomic force microscopy investigation of morphological changes in living keratinocytes treated with HgCl2 at not cytotoxic doses

Morphological changes of normal human keratinocyte cells have been monitored by means of atomic force microscopy after the exposure at a mercury solution containing HgCl2 at 10−7 M. The measurements have been carried out in contact mode in a thermostated liquid cell, to reproduce a cellular environment similar to the physiologic one. Remarkable alterations of the cellular morphology and volume have been revealed after few minutes from starting the exposure experiment, although the HgCl2 concentration is several orders of magnitudes lower than the cytotoxic value (10−4 M). The atomic force microscopy technique results to be a powerful mean to investigate modifications induced in the cell morphology by external chemical agents.
Can minocycline prevent the onset of Alzheimer's disease?

I read with great interest the article by Chu and Pratico about the role of 5-lipoxygenase (5-LO) in the formation of b-amyloid in the brain. Previously, the same group reported that b-amyloid deposition in the brains of Tg2576 mice (a transgenic mouse model of Alzheimer’s disease [AD]) lacking 5-LO gene was reduced by 64% to 80% compared with Tg2576 controls. Subsequently, the present study showed that 5-LO regulated the formation of b-amyloid by activating the cAMP-response element binding protein (CREB), which in turn increased transcription of the c-secretase complex. Furthermore, 5-LO gene disruption or a specific 5-LO inhibitor results in a significant reduction of b-amyloid formation and CREB and c-secretase levels in the brain of Tg2576. These studies suggest that pharmacological inhibition of 5LO may be beneficial in the treatment and prevention of AD. Accumulating evidence suggests that the antibiotic drug minocycline would be a potential therapeutic drug for neurodegenerative and psychiatric diseases because this drug has neuroprotective, neurotrophic, and antineuroinflammatory actions in a number of animal models. Interestingly, minocycline protects against N-methyl-D-aspartate (NMDA)-induced cell death via inhibiting 5-LO activation, suggesting the role of 5-LO inhibition in the mechanism of its action. In the early stages of the AD-like b-amyloid pathology, minocycline treatment attenuated behavioral abnormalities, neuroinflammatory markers, and b-amyloid deposition in the transgenic mouse model of AD. Together, it is likely that a pharmacological approach targeting the early neuroinflammatory effects of b-amyloid might be a promising strategy to prevent or delay the onset of AD. Positron emission tomography studies using [C]PIB and [C](R)-PK11195 have demonstrated that b-amyloid deposition and microglial activation could be detected in the brain of patients with amnestic mild cognitive impairment (MCI). In addition, AD neuroimaging initiative research suggests that AD begins with b-amyloid accumulation in the brain, which ultimately leads to synaptic dysfunction, neurodegeneration, and cognitive or functional decline. Considering the inhibiting effects of minocycline on the b-amyloid formation and microglial activation in the brain, minocycline may prevent or delay the onset of AD from MCI. Taken together, minocycline, an inhibitor of both 5-LO and microglial activation, might be a therapeutic drug to prevent or delay the onset of AD. A further randomized, doubleblind, placebo-controlled study of minocycline in patients with MCI or normal subjects with early-stage b-amyloid deposition will be of great interest. Potential Conflicts of Interest
Role of sociodemographic factors in tuberculosis treatment outcome: A prospective study in Aligarh, Uttar Pradesh

Context: India has the highest tuberculosis (TB) burden in the world in terms of the absolute number of cases that occur each year. The directly observed treatment, short-course (DOTS) strategy is the most systematic and cost-effective approach to controlling this disease. Furthermore, understanding the specific reasons for treatment outcomes is important for the improvement of treatment systems. Aims: This study aimed to determine certain sociodemographic factors that contribute to TB treatment outcome. Settings and Design: A prospective study was conducted among 302 TB patients at four designated microscopy centers (DMCs) of Aligarh District, Uttar Pradesh from April 2012 to June 2013. Materials and Methods: Study subjects were followed up from the initiation to the end of treatment. Baseline data were collected using a pretested questionnaire. The subjects were interviewed at least 3 times each during their treatment. Outcomes were considered as per the Revised National Tuberculosis Control Programme (RNTCP) standard definition. "Cured" and "treatment completed" were considered good outcomes, while "default," "failure," and "death" were considered poor outcomes. Statistical Analysis Used: Data were analyzed in SPSS 17 using the chi-square test and univariate logistic regression to determine the association. The odds ratio (OR) and 95% confidence interval (CI) were calculated in logistic regression. Results: The majority of the subjects were male (72.2%) and in the age group of 15-30 years. Among the subjects, 53% resided in urban areas; 63.6% were literate; and 45% had a high standard of living index (SLI). Age group, literacy status, and SLI were found to be significantly associated with outcome (P < 0.05). On logistic regression, the 31-45-years age group, literacy, and high SLI were found to be significantly associated with good outcome.
Deficiency of Cardiomyocyte-specific MicroRNA-378 Contributes to the Development of Cardiac Fibrosis Involving a Transforming Growth Factor β (TGFβ1)-dependent Paracrine Mechanism*

Background: MicroRNA-378, a cardiomyocyte-specific miRNA, is down-regulated during heart failure. Results: miR-378 inhibition induced TGFβ1 expression, which correlated with the activity of c-Fos, c-Jun, and Ras. Conditioned media of miR-378-depleted myocytes induced fibroblast activation by utilizing TGFβ1-dependent paracrine mechanisms. Conclusion: miR-378 is a negative regulator of TGFβ1 and cardiac fibrosis. Significance: miR-378 offers therapeutic potential for the management of heart failure. Understanding the regulation of cardiac fibrosis is critical for controlling adverse cardiac remodeling during heart failure. Previously we identified miR-378 as a cardiomyocyte-abundant miRNA down-regulated in several experimental models of cardiac hypertrophy and in patients with heart failure. To understand the consequence of miR-378 down-regulation during cardiac remodeling, our current study employed a locked nucleic acid-modified antimiR to target miR-378 in vivo. Results showed development of cardiomyocyte hypertrophy and fibrosis in mouse hearts. Mechanistically, miR-378 depletion was found to induce TGFβ1 expression in mouse hearts and in cultured cardiomyocytes. Among various secreted cytokines in the conditioned-media of miR-378-depleted cardiomyocytes, only TGFβ1 levels were found to be increased. The increase was prevented by miR-378 expression. Treatment of cardiac fibroblasts with the conditioned media of miR-378-depleted myocytes activated pSMAD2/3 and induced fibrotic gene expression. This effect was counteracted by including a TGFβ1-neutralizing antibody in the conditioned-medium. In cardiomyocytes, adenoviruses expressing dominant negative N-Ras or c-Jun prevented antimiR-mediated induction of TGFβ1 mRNA, documenting the importance of Ras and AP-1 signaling in this response. Our study demonstrates that reduction of miR-378 during pathological conditions contributes to cardiac remodeling by promoting paracrine release of profibrotic cytokine, TGFβ1 from cardiomyocytes. Our data imply that the presence in cardiomyocyte of miR-378 plays a critical role in the protection of neighboring fibroblasts from activation by pro-fibrotic stimuli.
A randomized, pilot trial of etanercept in dermatomyositis

The aims of this pilot study were to assess (1) the safety and tolerability of etanercept in dermatomyositis (DM); (2) the feasibility and safety of a forced prednisone taper; and (3) outcome measures, including those recommended by the International Myositis Assessment Clinical Study (IMACS) group.
Arachidonic Acid Induces Direct Interaction of the p67phox-Rac Complex with the Phagocyte Oxidase Nox2, Leading to Superoxide Production*

Background: The mechanism whereby arachidonic acid (AA) activates the phagocyte oxidase Nox2 is not well understood, except for the AA-induced conformational change of p47phox, a partner of the Nox2 activator p67phox. Results: AA also triggers Rac-GTP formation and Nox2 interaction with the p67phox·Rac-GTP complex. Conclusion: AA regulates Nox2 assembly at multiple steps. Significance: p67phox-Nox2 interaction is a novel regulatory step. The phagocyte NADPH oxidase Nox2, heterodimerized with p22phox in the membrane, is dormant in resting cells but becomes activated upon cell stimulation to produce superoxide, a precursor of microbicidal oxidants. Nox2 activation requires two switches to be turned on simultaneously: a conformational change of the cytosolic protein p47phox and GDP/GTP exchange on the small GTPase Rac. These proteins, in an active form, bind to their respective targets, p22phox and p67phox, leading to productive oxidase assembly at the membrane. Although arachidonic acid (AA) efficiently activates Nox2 both in vivo and in vitro, the mechanism has not been fully understood, except that AA induces p47phox conformational change. Here we show that AA elicits GDP-to-GTP exchange on Rac at the cellular level, consistent with its role as a potent Nox2 activator. However, even when constitutively active forms of p47phox and Rac1 are both expressed in HeLa cells, superoxide production by Nox2 is scarcely induced in the absence of AA. These active proteins also fail to effectively activate Nox2 in a cell-free reconstituted system without AA. Without affecting Rac-GTP binding to p67phox, AA induces the direct interaction of Rac-GTP-bound p67phox with the C-terminal cytosolic region of Nox2. p67phox-Rac-Nox2 assembly and superoxide production are both abrogated by alanine substitution for Tyr-198, Leu-199, and Val-204 in the p67phox activation domain that localizes the C-terminal to the Rac-binding domain. Thus the “third” switch (AA-inducible interaction of p67phox·Rac-GTP with Nox2) is required to be turned on at the same time for Nox2 activation.
Yeast Importin-α (Srp1) Performs Distinct Roles in the Import of Nuclear Proteins and in Targeting Proteasomes to the Nucleus*

Background: Srp1 imports proteins containing nuclear localization signals (NLS) into the nucleus. Results: Srp1 can also target proteasomes to the nucleus. Conclusion: Srp1 binds Sts1 to specifically target proteasomes to the nucleus. Significance: The nuclear targeting of proteasomes by Srp1 is distinct from its well established role in NLS-mediated import. Srp1 (importin-α) can translocate proteins that contain a nuclear localization signal (NLS) into the nucleus. The loss of Srp1 is lethal, although several temperature-sensitive mutants have been described. Among these mutants, srp1-31 displays the characteristic nuclear import defect of importin-α mutants, whereas srp1-49 shows a defect in protein degradation. We characterized these and additional srp1 mutants to determine whether distinct mechanisms were required for intracellular proteolysis and the import of NLS-containing proteins. We determined that srp1 mutants that failed to import NLS-containing proteins (srp1-31 and srp1-55) successfully localized proteasomes to the nucleus. In contrast, srp1 mutants that did not target proteasomes to the nucleus (srp1-49 and srp1-E402Q) were able to import NLS-containing proteins. The proteasome targeting defect of specific srp1 mutants caused stabilization of nuclear substrates and overall accumulation of multiubiquitylated proteins. Co-expression of a member of each class of srp1 mutants corrected both the proteasome localization defect and the import of NLS-containing proteins. These findings indicate that the targeting of proteasomes to the nucleus occurs by a mechanism distinct from the Srp1-mediated import of nuclear proteins.
The role of the P2Y12 receptor in the vessel wall response to injury and inflammation

Platelets play a fundamental role in thrombosis, but they are increasingly regarded as discrete immune cells due to their pro-inflammatory actions on leukocytes and the endothelium. The P2Y12 receptor is responsible for amplifying and sustaining platelet activation. As a result, P2Y12 inhibitors, such as ticagrelor and clopidogrel, form an essential component of acute coronary syndrome treatment in preventing thrombotic events. Furthermore, P2Y12 receptors have also been identified on vascular smooth muscle cells where they mediate cell contraction and mitogenic responses. This thesis investigates the role of P2Y12 in mediating the vessel wall response to injury and to inflammation, specifically in atherogenesis, and considers the use of P2Y12 inhibitors in modulating these effects. I have shown, using a murine ferric chloride injury model, that platelet P2Y12 is crucial in mediating thrombus and neointima formation following vessel injury, and that P2Y12 inhibition at the time of injury was sufficient to prevent their formation. To investigate the role of P2Y12 in atherogenesis, a novel ApoE/P2Y12 double knockout mouse strain was generated and fed a high-fat western diet for 12 weeks, before assessing atherosclerotic burden. Compared to ApoE-/- controls, P2Y12-deficency led to a moderate reduction in lesion area, particularly in areas of the vasculature prone to turbulent blood flow, such as the aortic arch. These vessels exhibited characteristics of established disease, with advanced lesions, and so atheroma was also assessed after 4 weeks on a western diet. I demonstrated, using bone marrow transplantations, that platelet P2Y12-deficiency profoundly reduced platelet reactivity compared to ApoE-/- controls, but had no effect on atheroma; whereas vessel wall P2Y12-deficiency significantly attenuated lesion area and led to a less mature lesion phenotype. Treating mice with ticagrelor or clopidogrel for 4 weeks achieved effective platelet P2Y12 inhibition, compared to mice treated with mannitol as a control, but did not block the vessel wall P2Y12 effect on atherogenesis. These data describe a previously unknown role for vessel wall P2Y12 in potentiating early atherogenesis. Despite its major contribution to platelet reactivity, platelet P2Y12 had no effect at this early stage, and yet is vital in later disease in atherothrombosis and modulating neointima formation following vessel injury. This work emphasises the complexity of cellular roles within atherogenesis, and how the impact of these cell populations may change as disease evolves. Further work is required to clarify the extent to which vessel wall and platelet P2Y12 influence atherogenesis throughout disease progression, in order to provide a clearer understanding of potential pharmacological targets in the prevention and management of atherosclerosis.
Arrested preoligodendrocyte maturation contributes to myelination failure in premature infants

The major form of magnetic resonance imaging–defined white matter injury (WMI) comprises diffuse lesions where the burden of small necrotic foci (microscopic necrosis) is poorly defined. We hypothesized that myelination failure associated with diffuse WMI involves an aberrant injury response linked to arrested preoligodendrocyte (preOL) maturation in reactive astrocyte‐rich lesions.