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This study evaluated an incentive-based negative reinforcement intervention to improve data recording of three educational care providers at a residential school for children and youth with intelle...

The Janus Kinase/Signal Transducers and Activators of Transcription (JAK/STAT) signaling pathway is utilized by numerous cytokines and interferons, and is essential for the development and function of both innate and adaptive immunity. Aberrant activation of the JAK/STAT pathway is evident in neuroinflammatory diseases such as Multiple Sclerosis and Parkinson's Disease. Innate immunity is the front line defender of the immune system and is composed of various cell types, including microglia, macrophages and neutrophils. Innate immune responses have both pathogenic and protective roles in neuroinflammation, depending on disease context and the microenvironment in the central nervous system. In this review, we discuss the role of innate immunity in the pathogenesis of neuroinflammatory diseases, how the JAK/STAT signaling pathway regulates the innate immune response, and finally, the potential for ameliorating neuroinflammation by utilization of JAK/STAT inhibitors.

We describe intervention with 2 adolescent male students who had autism spectrum disorder (ASD) and resisted haircutting performed by care providers at a residential school. The students were exposed to a graduated hierarchy of steps including the presence of hair clippers, and increased duration of hair clippers against their scalp and hair. Edible reinforcement was presented contingent on completion of a step without interfering behavior. Both students learned to tolerate all of the steps in the graduated hierarchy and a full haircut with maintenance at 2-, 4-, and 6-month follow-up. The study supports previous tolerance-training research with children and youth who have intellectual and developmental disabilities and resist personal care and hygiene routines.

Multiple sclerosis (MS) is associated with a dysregulated JAK/STAT signaling pathway. Suppressors of cytokine signaling (SOCS) negatively regulate the JAK/STAT pathway. MS patients with disease affecting the cerebellum display rapid progression and poor prognosis. Previous data from our laboratory demonstrates a severe, brain-targeted form of experimental autoimmune encephalomyelitis (EAE) in mice lacking Socs3 in myeloid cells (Socs3ΔLysM), with increased cerebellar infiltration of primed neutrophils. Here, we induced EAE in mice lacking Socs3 specifically in neutrophils (Socs3ΔLy6G; n=26) and Ly6G−/+ control mice (n=13), and examined disease progression, neutrophil populations and CD4+ T-cell polarization at disease peak. Ly6G−/+ mice exhibited clinical signs of classical EAE, whereas Socs3ΔLy6G mice exhibited clinical signs of brain-targeted EAE, similar to Socs3ΔLysM mice. An analysis of the cerebellar cell infiltrate in Socs3ΔLy6G mice demonstrated an increased cellularity (p=0.005), which was comprised of both an increase in the percentage (p=0.020) and total number (p

We assessed the utility of the Alere i Influenza A & B point of care influenza test (Ai-POCIT) with laboratory testing using RT-PCR. 270 adult hospital patients had both Ai-POCIT and laboratory influenza tests conducted on the same sample. Overall, 30% and 32% influenza tests were positive by Ai-POCIT and RT-PCR, respectively. The sensitivity of the Ai-POCIT for influenza A, influenza B and any influenza were 93%, 100%, and 95%, respectively. Specificity was 100% for both viruses, but an 11% test failure rate indicates the need for better training of users. We believe that the use of nasopharyngeal (NP) swabs resulted in the observed high performance of the Ai-POCIT in comparison to other published studies. Ai-POCIT was regarded as very useful by front line clinical staff for clinical decision making and acute bed management.

Introduction: Impaired lower extremity perfusion is a hallmark of peripheral vascular disease (PVD) and is particularly problematic in diabetic patients, who suffer from high rates of PVD, ulceration, and lower extremity amputation. The ability to non-invasively detect deficits in microvascular perfusion within vascular territories, or angiosomes, of the feet may provide information related to tissue viability, assist with classification of diabetic wounds, and guide future therapeutic interventions. In this study, we sought to quantify volumetric microvascular perfusion within specific angiosomes containing non-healing foot ulcers in diabetic patients with critical limb ischemia. Methods: Twenty-eight diabetic patients (mean age, 68 ± 13 yrs) with non-healing ulcers underwent SPECT/CT imaging of the feet following a resting injection of 99m Tc-tetrofosmin (dose, 550.6 ± 37 Mbq). CT images were segmented into five angiosomes and used for quantifying relative radiotracer uptake (Fig. 1A), expressed as standardized uptake values (SUVs). SUVs were assessed for each patient in the angiosome containing the non-healing ulcers, which were rated using the Wagner Grade Classification. Results: SPECT/CT imaging allowed for visualization of resting perfusion deficits in ulcer sites (Fig. 1B) and quantitative image analysis revealed variability in angiosome perfusion based on Wagner Grade of ulcers. Angiosomes containing Wagner Grade IV ulcers exhibited significantly lower SUVs when compared to SUVs of angiosomes containing Wagner Grade I, II, and III ulcers (Fig. 1C). Conclusions: SPECT/CT imaging provides a useful non-invasive tool for evaluating a wide clinical spectrum of ulcers within associated angiosomes of the foot and can be performed without requiring pharmacological or exercise stress. Future application of SPECT/CT imaging may provide additional value for detection and targeting of ischemic tissue for therapeutic interventions.

The initiation and elongation stages of translation are directed by codon–anticodon interactions. In contrast, a release factor protein mediates stop codon recognition prior to polypeptide chain release. Previous studies have identified specific regions of eukaryotic release factor one (eRF1) that are important for decoding each stop codon. The cavity model for eukaryotic stop codon recognition suggests that three binding pockets/cavities located on the surface of eRF1's domain one are key elements in stop codon recognition. Thus, the model predicts that amino acid changes in or near these cavities should influence termination in a stop codon-dependent manner. Previous studies have suggested that the TASNIKS and YCF motifs within eRF1 domain one play important roles in stop codon recognition. These motifs are highly conserved in standard code organisms that use UAA, UAG, and UGA as stop codons, but are more divergent in variant code organisms that have reassigned a subset of stop codons to sense codons. In the current study, we separately introduced TASNIKS and YCF motifs from six variant code organisms into eRF1 of Saccharomyces cerevisiae to determine their effect on stop codon recognition in vivo. We also examined the consequences of additional changes at residues located between the TASNIKS and YCF motifs. Overall, our results indicate that changes near cavities two and three frequently mediated significant effects on stop codon selectivity. In particular, changes in the YCF motif, rather than the TASNIKS motif, correlated most consistently with variant code stop codon selectivity.

Parkinson's Disease (PD) is an age-related, chronic neurodegenerative disorder. At present, there are no disease-modifying therapies to prevent PD progression. Activated microglia and neuroinflammation are associated with the pathogenesis and progression of PD. Accumulation of α-synuclein (α-SYN) in the brain is a core feature of PD and leads to microglial activation, inflammatory cytokine/chemokine production, and ultimately to neurodegeneration. Given the importance of the JAK/STAT pathway in activating microglia and inducing cytokine/chemokine expression, we investigated the therapeutic potential of inhibiting the JAK/STAT pathway using the JAK1/2 inhibitor, AZD1480.In vitro, α-SYN exposure activated the JAK/STAT pathway in microglia and macrophages, and treatment with AZD1480 inhibited α-SYN-induced major histocompatibility complex Class II and inflammatory gene expression in microglia and macrophages by reducing STAT1 and STAT3 activation. Forin vivostudies, we used a rat model of PD induced by viral overexpression of α-SYN. AZD1480 treatment inhibited α-SYN-induced neuroinflammation by suppressing microglial activation, macrophage and CD4+T-cell infiltration and production of proinflammatory cytokines/chemokines. Numerous genes involved in cell–cell signaling, nervous system development and function, inflammatory diseases/processes, and neurological diseases are enhanced in the substantia nigra of rats with α-SYN overexpression, and inhibited upon treatment with AZD1480. Importantly, inhibition of the JAK/STAT pathway prevented the degeneration of dopaminergic neuronsin vivo. These results indicate that inhibiting the JAK/STAT pathway can prevent neuroinflammation and neurodegeneration by suppressing activation of innate and adaptive immune responses to α-SYN. Furthermore, this suggests the feasibility of targeting the JAK/STAT pathway as a neuroprotective therapy for neurodegenerative diseases.SIGNIFICANCE STATEMENTα-SYN plays a central role in the pathophysiology of PD through initiation of neuroinflammatory responses. Using an α-SYN overexpression PD model, we demonstrate a beneficial therapeutic effect of AZD1480, a specific inhibitor of JAK1/2, in suppressing neuroinflammation and neurodegeneration. Our findings document that inhibition of the JAK/STAT pathway influences both innate and adaptive immune responses by suppressing α-SYN-induced microglia and macrophage activation and CD4+T-cell recruitment into the CNS, ultimately suppressing neurodegeneration. These findings are the first documentation that suppression of the JAK/STAT pathway disrupts the circuitry of neuroinflammation and neurodegeneration, thus attenuating PD pathogenesis. JAK inhibitors may be a viable therapeutic option for the treatment of PD patients.

The JAK/STAT pathway is critical for development, regulation, and termination of immune responses, and dysregulation of the JAK/STAT pathway, that is, hyperactivation, has pathological implications in autoimmune and neuroinflammatory diseases. Suppressor of cytokine signaling 3 (SOCS3) regulates STAT3 activation in response to cytokines that play important roles in the pathogenesis of neuroinflammatory diseases, including IL-6 and IL-23. We previously demonstrated that myeloid lineage–specific deletion of SOCS3 resulted in a severe, nonresolving atypical form of experimental autoimmune encephalomyelitis (EAE), characterized by lesions, inflammatory infiltrates, elevated STAT activation, and elevated cytokine and chemokine expression in the cerebellum. Clinically, these mice exhibit ataxia and tremors. In this study, we provide a detailed analysis of this model, demonstrating that the atypical EAE observed in LysMCre-SOCS3fl/fl mice is characterized by extensive neutrophil infiltration into the cerebellum and brainstem, increased inducible NO synthase levels in the cerebellum and brainstem, and prominent axonal damage. Importantly, infiltrating SOCS3-deficient neutrophils produce high levels of CXCL2, CCL2, CXCL10, NO, TNF-α, and IL-1β. Kinetic studies demonstrate that neutrophil infiltration into the cerebellum and brainstem of LysMCre-SOCS3fl/fl mice closely correlates with atypical EAE clinical symptoms. Ab-mediated depletion of neutrophils converts the atypical phenotype to the classical EAE phenotype and, in some cases, a mixed atypical/classical phenotype. Blocking CXCR2 signaling ameliorates atypical EAE development by reducing neutrophil infiltration into the cerebellum/brainstem. Thus, neutrophils lacking SOCS3 display elevated STAT3 activation and expression of proinflammatory mediators and play a critical role in the development of atypical EAE.

The vacuole is the major site of intracellular Ca2+ storage in yeast and functions to maintain cytosolic Ca2+ levels within a narrow physiological range. In this study, we examined how cellular Ca2+ homeostasis is maintained in a vps33Delta vacuolar biogenesis mutant. We found that growth of the vps33Delta strain was sensitive to high or low extracellular Ca2+. This strain could not properly regulate cytosolic Ca2+ levels and was able to retain only a small fraction of its total cellular Ca2+ in a nonexchangeable intracellular pool. Surprisingly, the vps33Delta strain contained more total cellular Ca2+ than the wild type strain. Because most cellular Ca2+ is normally found within the vacuole, this suggested that other intracellular compartments compensated for the reduced capacity to store Ca2+ within the vacuole of this strain. To test this hypothesis, we examined the contribution of the Golgi-localized Ca2+ ATPase Pmr1p in the maintenance of cellular Ca2+ homeostasis. We found that a vps33Delta/pmr1Delta strain was hypersensitive to high extracellular Ca2+. In addition, certain combinations of mutations effecting both vacuolar and Golgi Ca2+ transport resulted in synthetic lethality. These results indicate that the Golgi apparatus plays a significant role in maintaining Ca2+ homeostasis when vacuolar biogenesis is compromised.

Background: Disruptions to the microcirculation may contribute to foot ulceration in patients with diabetes mellitus and critical limb ischemia (CLI). Single photon emission computed tomography (SPECT)/CT imaging assesses microvascular perfusion in the foot, but has not been evaluated in foot

Straight-line motion, albeit simple, manifest itself in numerous applications, from running steam engines and oil wells to manufacturing parts with straight edges and sides. The drive to maximize production creates a need for continuously running assembly-line manufacturing comprised of precise, individually optimized components. While there are many so-called straight-line generating mechanisms, few actually produce a true straight-line, most generate only approximate straight-line. Featured an eight-link rhomboidal system with length constraints, the Peaucellier mechanism is one that actually produces a true straight line intrinsically. This paper presents a study on the dimension synthesis of the Peaucellier mechanism, namely by identifying the correct ratio of linkage lengths to produce the longest straight line stroke. In addition to designing for stroke, another objective of interest is to attain a desired velocity profile along the path. Kinematic analysis of the velocity profile on the mechanism will render the creation of input angular velocity standards based on desired stroke speed. Given the stroke and velocity specifications, specific steps to size the dimensions of the mechanism developed as result of this study will be presented.Copyright © 2011 by ASME

Objective: Increasing evidence indicates that the thalamus may be a location of early neurodegeneration in multiple sclerosis (MS). Our objective was to identify the presence of gray matter volume loss and thinning in patients with radiologically isolated syndrome (RIS). Methods: Sixty-three participants were included in this case-control study. Twenty-one patients with RIS were age- and sex-matched to 42 healthy controls in a 1:2 ratio. All participants underwent brain MRIs on a single 3T scanner. After lesion segmentation and inpainting, 1 mm 3 -isometric T1-weighted images were submitted to FreeSurfer (v5.2). Normalized cortical and deep gray matter volumes were compared between patients with RIS and controls using t tests, and thalamic volumes were correlated with white matter lesion volumes using Pearson correlation. Exploratory cortical thickness maps were created. Results: Although traditional normalized total gray and white matter volumes were not statistically different between patients with RIS and controls, normalized left (0.0046 ± 0.0005 vs 0.0049 ± 0.0004, p = 0.006), right (0.0045 ± 0.0005 vs 0.0048 ± 0.0004, p = 0.008), and mean (0.0045 ± 0.0005 vs 0.0049 ± 0.0004, p = 0.004) thalamic volumes were significantly lower in patients with RIS (n = 21, mean age 41.9 ± 12.7 years) than in controls (n = 42, mean age 41.4 ± 11.2 years). Thalamic volumes correlated modestly with white matter lesion volumes (range: r = −0.35 to −0.47). Conclusion: Our data provide novel evidence of thalamic atrophy in RIS and are consistent with previous reports in early MS stages. Thalamic volume loss is present early in CNS demyelinating disease and should be further investigated as a metric associated with neurodegeneration.

We conducted a retrospective cohort study of children with catheter-associated bloodstream infections (BSIs) due toEscherichia coliand/orKlebsiella. Risk factors for poor outcome (ie, death or recurrence of infection) were receipt of mechanical ventilation (adjusted odds ratio [aOR], 4.6 [95% confidence interval {CI}, 1.39-16.30]) and receipt of total parenteral nutrition (aOR, 3.5 [95% CI, 1.1-10.8]). A significant proportion of children with catheter-associated BSI were treated successfully without catheter removal.

Parkinson’s Disease (PD) is an age-related, chronic neurodegenerative disorder. At present, there are no therapies to prevent PD progression. Activated microglia and subsequent neuroinflammation associate with the pathogenesis and progression of PD. Accumulation of α-synuclein (α-syn) in the brain is a core feature of PD and leads to microglial activation, inflammatory cytokines/chemokines production and neurodegeneration. Given the importance of the JAK/STAT pathway in activating microglia and inducing cytokine/chemokine expression, we investigated the therapeutic potential of inhibiting the JAK/STAT pathway using the JAK1/2 inhibitor, AZD1480. We utilized an in vivo rat model of PD induced by viral overexpression of α-syn. We find that AZD1480 treatment inhibits α-syn-induced neuroinflammation by inhibiting microglia activation and CD3+ T-cell infiltration, and neurodegeneration by preventing the degeneration of dopaminergic neurons in vivo. In vitro, AZD1480 inhibits α-syn- and IFN-γ-induced MHC Class II, CD40, iNOS, TNF-α and IL-6 expression in microglia and macrophages by reducing STAT1 and STAT3 activation. These results indicate that inhibiting the JAK/STAT pathway can prevent neuroinflammation and neurodegeneration by suppressing activation of innate and adaptive immune responses to α-syn and inflammatory cytokines in this PD model. Furthermore, this suggests the feasibility of targeting the JAK/STAT pathway as neuroprotective therapy for neurodegenerative diseases.