Miracle-dz/UniFAR-FTUs huggingface dataset:
High Dose Vitamin D3 Improves Exercise Tolerance in Elderly Patients with Chronic Obstructive Pulmonary Disease, Chronic Obstructive Pulmonary Disease,

Background: In addition to airflow limitation, peripheral muscle dysfunction is a limiting factor in physical performance of patients with chronic obstructive pulmonary disease (COPD). Any measure to improve global muscle function in COPD patients such as vitamin D replacement seems to enhance exercise tolerance of these patients. Aim: The aim of present study was to study the effect of vitamin D replacement therapy on exercise tolerance of a group of stable COPD patients with vitamin D deficiency or insufficiency. Methods: It was an experimental, unblended open-label trial. A total number of 25 stable COPD patients were enrolled in the study in the period between March 2012 and OCT 2012 and received 50,000 IU vitamin D weekly for 4-8 weeks in addition to their standard medical therapy as before. They were also subdivided into two subgroups based on serum 25 hydroxy vitamin D,25(OH)D( <10 ng/ml and 10-30 ng/ml). Pulmonary function tests, six-minute walk distance test(6MWD) were measured at baseline and three months after the enrollment. Results: By the end of study, there was significant improvement in the distance walked during 6-minute walk test. This improvement was documented in both subgroups of patients. However, the perceived dyspnea during test did not differ from baseline value by the end of the study. Also, only patients with serum 25(OH) D, 10 -30 ng/ml showed the significant increase in their mean value of FEV1 by the end of the study. Conclusion: Vitamin D replacement therapy is an effective measure to improve exercise tolerance of stable COPD patients with vitamin D deficiency or insufficiency.
Cre recombinase induces DNA damage and tetraploidy in the absence of LoxP sites

The spatiotemporal manipulations of gene expression by the Cre recombinase (Cre) of bacteriophage P1 has become an essential asset to understanding mammalian genetics. Accumulating evidence suggests that Cre activity can, in addition to excising targeted loxP sites, induce cytotoxic effects, including abnormal cell cycle progression, genomic instability, and apoptosis, which can accelerate cancer progression. It is speculated that these defects are caused by Cre-induced DNA damage at off-target sites. Here we report the formation of tetraploid keratinocytes in the epidermis of keratin 5 and/or keratin 14 promoter-driven Cre (KRT5- and KRT14-Cre) expressing mouse skin. Biochemical analyses and flow cytometry demonstrated that Cre expression also induces DNA damage, genomic instability, and tetraploidy in HCT116 cells, and live-cell imaging revealed an extension of the G2 cell cycle phase followed by defective or skipping of mitosis as cause for the tetraploidy. Since tetraploidy eventually leads to aneuploidy, a hallmark of cancer, our findings highlight the importance of distinguishing non-specific cytopathic effects from specific Cre/loxP-driven genetic manipulations when using Cre-mediated gene deletions.
Pathological skeletal remains from ancient Egypt: the earliest case of diabetes mellitus?

The diagnosis of diabetes mellitus from skeletal remains is very difficult given the complexity of the disease and the fact that there are no pathological skeletal characteristics exclusively associated with diabetes mellitus. Skeletal identification of diabetes mellitus may only be possible through differential diagnosis, when several pathological changes are present. Skeletal and dental changes associated with diabetes mellitus include Charcot's joint (neuropathic arthropathy), osteoporosis, osteoarthritis, diffuse idiopathic skeletal hyperostosis (DISH, or Forestier's disease), adhesive capsulitis (frozen shoulder), dental caries, periodontal disease, and antemortem tooth loss. Skeletal remains of an adult male from the Egyptian archaeological site of Dayr al-Barsha, dated to the Middle Kingdom (ca. 2055–1650 BC), display a myriad of pathological conditions that, when considered together, likely indicate diabetes mellitus, specifically type 2 diabetes mellitus. This diagnosis represents the earliest, and possibly the only recorded archaeological skeletal evidence for this disease. Copyright © 2010 John Wiley & Sons.
Locus Coeruleus Activity Strengthens Prioritized Memories Under Arousal

Recent models posit that bursts of locus ceruleus (LC) activity amplify neural gain such that limited attention and encoding resources focus even more on prioritized mental representations under arousal. Here, we tested this hypothesis in human males and females using fMRI, neuromelanin MRI, and pupil dilation, a biomarker of arousal and LC activity. During scanning, participants performed a monetary incentive encoding task in which threat of punishment motivated them to prioritize encoding of scene images over superimposed objects. Threat of punishment elicited arousal and selectively enhanced memory for goal-relevant scenes. Furthermore, trial-level pupil dilations predicted better scene memory under threat, but were not related to object memory outcomes. fMRI analyses revealed that greater threat-evoked pupil dilations were positively associated with greater scene encoding activity in LC and parahippocampal cortex, a region specialized to process scene information. Across participants, this pattern of LC engagement for goal-relevant encoding was correlated with neuromelanin signal intensity, providing the first evidence that LC structure relates to its activation pattern during cognitive processing. Threat also reduced dynamic functional connectivity between high-priority (parahippocampal place area) and lower-priority (lateral occipital cortex) category-selective visual cortex in ways that predicted increased memory selectivity. Together, these findings support the idea that, under arousal, LC activity selectively strengthens prioritized memory representations by modulating local and functional network-level patterns of information processing. SIGNIFICANCE STATEMENT Adaptive behavior relies on the ability to select and store important information amid distraction. Prioritizing encoding of task-relevant inputs is especially critical in threatening or arousing situations, when forming these memories is essential for avoiding danger in the future. However, little is known about the arousal mechanisms that support such memory selectivity. Using fMRI, neuromelanin MRI, and pupil measures, we demonstrate that locus ceruleus (LC) activity amplifies neural gain such that limited encoding resources focus even more on prioritized mental representations under arousal. For the first time, we also show that LC structure relates to its involvement in threat-related encoding processes. These results shed new light on the brain mechanisms by which we process important information when it is most needed.
Invading cancer cells are predominantly in G0/G1 resulting in chemoresistance demonstrated by real-time FUCCI imaging

Invasive cancer cells are a critical target in order to prevent metastasis. In the present report, we demonstrate real-time visualization of cell cycle kinetics of invading cancer cells in 3-dimensional (3D) Gelfoam® histoculture, which is in vivo-like. A fluorescence ubiquitination cell cycle indicator (FUCCI) whereby G0/G1 cells express a red fluorescent protein and S/G2/M cells express a green fluorescent protein was used to determine the cell cycle position of invading and non-invading cells. With FUCCI 3D confocal imaging, we observed that cancer cells in G0/G1 phase in Gelfoam® histoculture migrated more rapidly and further than cancer cells in S/G2/M phases. Cancer cells ceased migrating when they entered S/G2/M phases and restarted migrating after cell division when the cells re-entered G0/G1. Migrating cancer cells also were resistant to cytotoxic chemotherapy, since they were preponderantly in G0/G1, where cytotoxic chemotherapy is not effective. The results of the present report suggest that novel therapy targeting G0/G1 cancer cells should be developed to prevent metastasis.
Loss of the Neuron-Specific F-Box Protein FBXO41 Models an Ataxia-Like Phenotype in Mice with Neuronal Migration Defects and Degeneration in the Cerebellum

The cerebellum is crucial for sensorimotor coordination. The cerebellar architecture not only requires proper development but also long-term integrity to ensure accurate functioning. Developmental defects such as impaired neuronal migration or neurodegeneration are thus detrimental to the cerebellum and can result in movement disorders including ataxias. In this study, we identify FBXO41 as a novel CNS-specific F-box protein that localizes to the centrosome and the cytoplasm of neurons and demonstrate that cytoplasmic FBXO41 promotes neuronal migration. Interestingly, deletion of the FBXO41 gene results in a severely ataxic gait in mice, which show delayed neuronal migration of granule neurons in the developing cerebellum in addition to deformities and degeneration of the mature cerebellum. We show that FBXO41 is a critical factor, not only for neuronal migration in the cerebellum, but also for its long-term integrity.
Neural Microstates Govern Perception of Auditory Input without Rhythmic Structure

Human perception fluctuates with the phase of neural oscillations in the presence of environmental rhythmic structure by which neural oscillations become entrained. However, in the absence of predictability afforded by rhythmic structure, we hypothesize that the neural dynamical states associated with optimal psychophysical performance are more complex than what has been described previously for rhythmic stimuli. The current electroencephalography study characterized the brain dynamics associated with optimal detection of gaps embedded in narrow-band acoustic noise stimuli lacking low-frequency rhythmic structure. Optimal gap detection was associated with three spectrotemporally distinct delta-governed neural microstates. Individual microstates were characterized by unique instantaneous combinations of neural phase in the delta, theta, and alpha frequency bands. Critically, gap detection was not predictable from local fluctuations in stimulus acoustics. The current results suggest that, in the absence of rhythmic structure to entrain neural oscillations, good performance hinges on complex neural states that vary from moment to moment. SIGNIFICANCE STATEMENT Our ability to hear faint sounds fluctuates together with slow brain activity that synchronizes with environmental rhythms. However, it is so far not known how brain activity at different time scales might interact to influence perception when there is no rhythm with which brain activity can synchronize. Here, we used electroencephalography to measure brain activity while participants listened for short silences that interrupted ongoing noise. We examined brain activity in three different frequency bands: delta, theta, and alpha. Participants' ability to detect gaps depended on different numbers of frequency bands—sometimes one, sometimes two, and sometimes three—at different times. Changes in the number of frequency bands that predict perception are a hallmark of a complex neural system.
Time-lapse, single cell based confocal imaging analysis of caspase activation and phosphatidylserine flipping during cellular apoptosis

Abstract Apoptosis is an important phenomenon for investigating the efficacy of anti-cancer drug candidates. The conventional assays for cellular apoptosis, such as enzyme-linked immunosorbent assay, absorbance monitoring for the activity of caspase, and flow cytometric assay, have focused only on biochemical events. We investigated the staurosporine (STS)-induced apoptosis of the murine macrophage RAW-264.7 cell using a cell based bioimaging technique. Using time-lapse confocal microscopy, we monitored caspase-3 activation during apoptosis by imaging the translocation of green fluorescent protein from the cytosol to the nuclei. Five hours after 1 μM STS treatment, caspase-3 was observed to be activated and membrane blebbing was observed simultaneously. Also, the loss of phosphatidylserine (PS) asymmetry in the phospholipid bilayer of plasma membrane during early apoptosis was monitored by imaging annexin-V labeled with fluorescein isocyanate binding to the externalized PS at various concentrations of STS. Moreover, disintegration of the plasma membrane during late apoptosis was confirmed using a nuclear dye, propidium iodide. The single cell based bioimaging data agreed well with those of the biochemical assays for caspase activation and morphological observation for membrane integrity.
NADPH Oxidase 1 Mediates α-Synucleinopathy in Parkinson's Disease

Accumulation of misfolded α-synuclein is the pathological hallmark of Parkinson's disease (PD). Nevertheless, little is known about the mechanism contributing to α-synuclein aggregation and its further toxicity to dopaminergic neurons. Since oxidative stress can increase the expression and aggregation levels of α-synuclein, NADPH oxidases (Noxs), which are responsible for reactive oxygen species generation, could be major players in α-synucleinopathy. Previously, we demonstrated that Nox1 is expressed in dopaminergic neurons of the PD animal models as well as postmortem brain tissue of PD patients, and is responsible for oxidative stress and subsequent neuronal degeneration. Here, using paraquat (PQ)-based in vitro and in vivo PD models, we show that Nox1 has a crucial role in modulating the behavior of α-synuclein expression and aggregation in dopaminergic neurons. We observed in differentiated human dopaminergic cells that Nox1 and α-synuclein expressions are increased under PQ exposure. Nox1 knockdown significantly reduced both α-synuclein expression and aggregation, supporting the role of Nox1 in this process. Furthermore, in rats exposed to PQ, the selective knockdown of Nox1 in the substantia nigra, using adeno-associated virus encoding Nox1-specific shRNA, largely attenuated the PQ-mediated increase of α-synuclein and ubiquitin expression levels as well as α-synuclein aggregates (proteinase K resistant) and A11 oligomers. Significant reductions in oxidative stress level and dopaminergic neuronal loss were also observed. Our data reveal a new mechanism by which α-synuclein becomes a neuropathologic protein through Nox1-mediated oxidative stress. This finding may be used to generate new therapeutic interventions that slower the rate of α-synuclein aggregation and the progression of PD pathogenesis.
Attention to Multiple Objects Facilitates Their Integration in Prefrontal and Parietal Cortex

Selective attention is known to interact with perceptual organization. In visual scenes, individual objects that are distinct and discriminable may occur on their own, or in groups such as a stack of books. The main objective of this study is to probe the neural interaction that occurs between individual objects when attention is directed toward one or more objects. Here we record steady-state visual evoked potentials via electrocorticography to directly assess the responses to individual stimuli and to their interaction. When human participants attend to two adjacent stimuli, prefrontal and parietal cortex shows a selective enhancement of only the neural interaction between stimuli, but not the responses to individual stimuli. When only one stimulus is attended, the neural response to that stimulus is selectively enhanced in prefrontal and parietal cortex. In contrast, early visual areas generally manifest responses to individual stimuli and to their interaction regardless of attentional task, although a subset of the responses is modulated similarly to prefrontal and parietal cortex. Thus, the neural representation of the visual scene as one progresses up the cortical hierarchy becomes more highly task-specific and represents either individual stimuli or their interaction, depending on the behavioral goal. Attention to multiple objects facilitates an integration of objects akin to perceptual grouping. SIGNIFICANCE STATEMENT Individual objects in a visual scene are seen as distinct entities or as parts of a whole. Here we examine how attention to multiple objects affects their neural representation. Previous studies measured single-cell or fMRI responses and obtained only aggregate measures that combined the activity to individual stimuli as well as their potential interaction. Here, we directly measure electrocorticographic steady-state responses corresponding to individual objects and to their interaction using a frequency-tagging technique. Attention to two stimuli increases the interaction component that is a hallmark for perceptual integration of stimuli. Furthermore, this stimulus-specific interaction is represented in prefrontal and parietal cortex in a task-dependent manner.
Hypoxic Preconditioning Attenuated Simulated Ischemia/Reperfusion Injury in Cultured Bone Marrow Stromal Cells

Aim: Bone marrow stromal cells (BMSCs) are a promising candidate for cell therapy in ischemic stroke. However, the majority of BMSCs are readily lost after transplantation because of apoptosis due to ischemia-reperfusion (I/R) injury. In the present study, we aimed to evaluate the effects of hypoxia preconditioning (HP) on simulated I/R injury in cultured BMSCs. Methods: Four generations of BMSCs were used as subjects. BMSCs were divided into 6 groups, including normal group, I/R group(I/R followed 0 h HP), 2 h-HP group(I/R followed 2 h HP ), 4 h-HP group(I/R followed 4 h HP), 6 h-HP group (I/R followed 6 h HP) and 8 h-HP group(I/R followed 8 h HP) 2 h-HP group(I/R followed 2 h HP ), 4 h-HP group(I/R followed 4 h HP), 6 h-HP group (I/R followed 6 h HP) and 8 h-HP group(I/R followed 8 h HP). BMSCs were subjected to HP by exposing the cells to hypoxia (2%O2). After 12 h of reoxygenation, BMSCs were suffered from 3 h ischemia (<0.5% O2 and serum deprivation) and subsequent reperfusion (I/R). Cell viability, hypoxia-inducible factor (HIF) 1-α, caspase-3 and apoptosis were tested using MTT, ELISA and immunofluorescence staining respectively. Results: There were no changes in cell viability after 2 h to 8 h HP in cultured BMSCs as compared to the normal group. But HP upregulated the content of HIF-1α in BMSCs and protected the BMSCs from later I/R injury, as evidenced by increased cell viability and decreased Caspase-3 release and apoptosis, especially in 8 h-HP group. Conclusion: HP attenuated the apoptosis induced by I/R injury via co-regulating the expression of HIF-1α and Caspase-3 in BMSCs, and thereby might play a beneficial effect on cell therapy
Robust Pacemaking in Substantia Nigra Dopaminergic Neurons

Dopaminergic neurons of the substantia nigra pars compacta are autonomous pacemakers. This activity is responsible for the sustained release of dopamine necessary for the proper functioning of target structures, such as the striatum. Somatodendritic L-type Ca2+ channels have long been viewed as important, if not necessary, for this activity. The studies reported here challenge this viewpoint. Using a combination of optical and electrophysiological approaches in brain slices, it was found that antagonism of L-type Ca2+ channel effectively stopped dendritic Ca2+ oscillations but left autonomous pacemaking unchanged. Moreover, damping intracellular Ca2+ oscillations with exogenous buffer had little effect on pacemaking rate. Although not necessary for pacemaking, L-type channels helped support pacemaking when challenged with cationic channel blockers. Simulations suggested that the insensitivity to antagonism of L-type channels reflected the multichannel nature of the pacemaking process. The robustness of pacemaking underscores its biological importance and provides a framework for understanding how therapeutics targeting L-type Ca2+ channels might protect dopaminergic neurons in Parkinson's disease without compromising their function.
Mechanisms of Regulation of Oligodendrocyte Development by p38 Mitogen-Activated Protein Kinase

Many extracellular and intrinsic factors regulate oligodendrocyte development, but their signaling pathways remain poorly understood. Although the p38 mitogen-activated protein kinase (MAPK)-dependent pathway is implicated in oligodendrocyte progenitor cell (OPC) lineage progression, its molecular targets involved in myelinogenesis are mostly unidentified. We have analyzed mechanisms by which p38MAPK regulates oligodendrocyte development and demonstrate that p38MAPK inhibition prevents OPC lineage progression and inhibits MBP (myelin basic protein) promoter activity and Sox10 function. In white-matter tissue, differential levels of MAPK phosphorylation are observed in oligodendrocyte lineage cells. Phosphorylated p38MAPK was found in CC1- and CNP-expressing differentiated oligodendrocytes of the adult brain and was temporally associated with a decline in the levels of phosphorylated extracellular signal-regulated kinase (ERK) in cells of this lineage. PDGF stimulates the phosphorylation of ERK, p38MAPK, and c-Jun N-terminal kinase (JNK), and p38MAPK inhibition was associated with increased ERK, JNK, and c-Jun phosphorylation. In the presence of PDGF, simultaneous inhibition of p38MAPK and either MAPK kinase (MEK) or JNK significantly alleviates the repression of myelin gene expression and lineage progression induced by p38MAPK inhibition alone. Dominant-negative c-Jun reverses the inhibition of myelin promoter activity by active MEK1 or dominant-negative p38MAPKα mutants, and phosphorylated c-Jun was detected at the MBP promoter after p38MAPK inhibition, indicating c-Jun as a negative mediator of p38MAPK action. Our findings indicate that p38MAPK activity in the brain supports myelin gene expression through distinct mechanisms via positive and negative regulatory targets. We show that oligodendrocyte differentiation involves p38-mediated Sox10 regulation and cross talk with parallel ERK and JNK pathways to repress c-Jun activity.
Improved gold chloride staining method for anatomical analysis of sensory nerve endings in the shoulder capsule and labrum as examples of loose and dense fibrous tissues

Abstract Consistency in gold chloride staining is essential for anatomical analysis of sensory nerve endings. The gold chloride stain for this purpose has been modified by many investigators, but often yields inconsistent staining, which makes it difficult to differentiate structures and to determine nerve ending distribution in large tissue samples. We introduce additional steps and major changes to the modified Gairns’ protocol. We controlled the temperature and mixing rate during tissue staining to achieve consistent staining and complete solution penetration. We subjected samples to sucrose dehydration to improve cutting efficiency. We then exposed samples to a solution containing lemon juice, formic acid and paraformaldehyde to produce optimal tissue transparency with minimal tissue deformity. We extended the time for gold chloride impregnation 1.5 fold. Gold chloride was reduced in the labrum using 25% formic acid in water for 18 h and in the capsule using 25% formic acid in citrate phosphate buffer for 2 h. Citrate binds gold nanoparticles, which minimizes aggregation in the tissue. We stored samples in fresh ultrapure water at 4° C to slow reduction and to maintain color contrast in the tissue. Tissue samples were embedded in Tissue Tek and sectioned at 80 and 100 μm instead of using glycerin and teasing the tissue apart as in Gairns’ modified gold chloride method. We attached sections directly to gelatin subbed slides after sectioning with a cryostat. The slides then were processed and coverslipped with Permount. Staining consistency was demonstrated throughout the tissue sections and neural structures were clearly identifiable.