RESUMEN
Burnout is an important public health issue at times of the COVID-19 pandemic. Current measures which focus on work-based burnout have limitations in length and/or relevance. When stepping into the post-pandemic as a new Norm Era, the burnout scale for the general population is urgently needed to fill the gap. This study aimed to develop a COVID-19 Burnout Views Scale (COVID-19 BVS) to measure burnout views of the general public in a Chinese context and examine its psychometric properties. A multiphase approach including literature review, expert consultation, and pilot testing was adopted in developing the scale. The scale was administered to a sample of 1,078 of the general public in Hong Kong with an average age of 34.45 years (SD = 12.47). Exploratory and Confirmatory Factor Analyses suggested a 5-item unidimensional model of COVID-19 BVS. The CFA results indicated that the COVID-19 BVS had a good model fit, as χ2 (10.054)/5 = 2.01, SRMR = 0.010, CFI = 0.998, RMSEA = 0.031. Five items were maintained in EFA with high internal consistency in terms of Cronbach's α of 0.845 and McDonald's ω coefficient of 0.87, and the corrected item-to-total correlations of 0.512 to 0.789 are way above the acceptable range. The KMO values of 0.841 and Bartlett's Test of Sphericity (p < 0.01) verified the normal distribution of the EFA and the adequacy of the EFA sampling. The analyses suggest that the COVID-19 BVS is a promising tool for assessing burnout views on the impacts of the epidemic on the Chinese general populations.
Asunto(s)
COVID-19 , Pandemias , Humanos , Adulto , COVID-19/epidemiología , Agotamiento Psicológico/epidemiología , Pueblo Asiatico , Hong Kong/epidemiologíaRESUMEN
Diverse processes in cancer are mediated by enzymes, which most proximally exert their function through their activity. High-fidelity methods to profile enzyme activity are therefore critical to understanding and targeting the pathological roles of enzymes in cancer. Here, we present an integrated set of methods for measuring specific protease activities across scales, and deploy these methods to study treatment response in an autochthonous model of Alk-mutant lung cancer. We leverage multiplexed nanosensors and machine learning to analyze in vivo protease activity dynamics in lung cancer, identifying significant dysregulation that includes enhanced cleavage of a peptide, S1, which rapidly returns to healthy levels with targeted therapy. Through direct on-tissue localization of protease activity, we pinpoint S1 cleavage to the tumor vasculature. To link protease activity to cellular function, we design a high-throughput method to isolate and characterize proteolytically active cells, uncovering a pro-angiogenic phenotype in S1-cleaving cells. These methods provide a framework for functional, multiscale characterization of protease dysregulation in cancer.
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Neoplasias Pulmonares , Péptido Hidrolasas , Endopeptidasas , Humanos , Neoplasias Pulmonares/genética , Péptido Hidrolasas/metabolismo , Proteolisis , Proteínas Tirosina Quinasas ReceptorasRESUMEN
BACKGROUND: Among individuals with nonvalvular atrial fibrillation (AF), the prevalence of obstructive sleep apnea (OSA) can be as high as 85%. Continuous positive airway pressure treatment for moderate or severe OSA might improve AF outcomes and quality of life, so early identification of OSA might be of value. However, screening questionnaires for OSA are suboptimal because they are weighted toward tiredness and loud snoring, which might be absent in AF patients. NoSAS (Neck, Obesity, Snoring, Age, Sex) is a new OSA questionnaire that excludes these parameters. Acoustic pharyngometry (AP) is a potential novel screening technique that measures pharyngeal cross-sectional area, which is reduced in patients with OSA. METHODS: We prospectively compared the accuracy of the NoSAS, the STOP-BANG questionnaire (Snoring, Tiredness, Observed apnea, blood Pressure, Body mass index, Age, Neck circumference and Gender), and AP with home sleep apnea testing (HSAT) in consecutive patients with nonvalvular AF. RESULTS: Of 188 participants, 86% had OSA and 49% had moderate or severe OSA. Mean Epworth Sleepiness Scale scores were low; 5.9 (SD, 3.9), indicating that most participants were not sleepy. Receiver operating characteristic curves for comparisons of screening tests with HSAT showed suboptimal accuracy. For moderate plus severe and severe only groups respectively, the area under the curve was 0.50 (95% confidence interval [CI], 0.42-0.58) and 0.42 (95% CI, 0.34-0.52) for AP, 0.65 (95% CI, 0.58-0.73) and 0.63 (95% CI, 0.52-0.74) for the STOP-BANG questionnaire, and 0.68 (95% CI, 0.60-0.75) and 0.69 (95% CI, 0.59-0.80) for the NoSAS. CONCLUSIONS: AP and NoSAS are not sufficiently accurate for screening AF patients for OSA. Because of the high rates of OSA in this cohort, the potential benefits of OSA treatment, and the suboptimal accuracy of current screening questionnaires, cardiologists should consider HSAT for AF patients.
CONTEXTE: Chez les sujets présentant une fibrillation auriculaire (FA) non valvulaire, la prévalence de l'apnée obstructive du sommeil (AOS) peut atteindre 85 %. En cas d'AOS modérée ou sévère, un traitement par ventilation spontanée en pression positive continue peut améliorer les résultats liés à la FA et la qualité de vie du patient; un diagnostic précoce d'AOS pourrait donc être utile. Les questionnaires de dépistage de l'AOS ne sont toutefois pas optimaux parce qu'ils accordent une grande importance à la fatigue et aux ronflements sonores, des symptômes qui ne se manifestent pas nécessairement en cas de FA. Le questionnaire NoSAS (de l'anglais Neck, Obesity, Snoring, Age, Sex) est un nouvel outil d'évaluation de l'AOS qui ne tient pas compte de ces paramètres. La pharyngométrie acoustique (PA) pourrait aussi constituer une nouvelle technique de dépistage; elle mesure l'aire de section transversale du pharynx, qui est réduite chez les patients souffrant d'AOS. MÉTHODOLOGIE: Nous avons comparé de façon prospective la précision du score au questionnaire NoSAS, du score au questionnaire STOP-BANG (de l'anglais Snoring, Tiredness, Observed apnea, blood Pressure, Body mass index, Age, Neck circumference and Gender) et des résultats de la PA à celle du test d'apnée du sommeil à domicile (TASD) chez des patients consécutifs présentant une FA non valvulaire. RÉSULTATS: Sur les 188 participants, 86 % présentaient une AOS et 49 % souffraient d'AOS modérée ou sévère. Le score moyen sur l'échelle de somnolence d'Epworth était faible et se situait à 5,9 (écart-type : 3,9), ce qui indique que la plupart des participants ne ressentaient pas de somnolence. La comparaison entre les questionnaires de dépistage et le TASD effectuée au moyen des courbes caractéristiques de la performance des tests a révélé une précision sous-optimale. Dans les groupes souffrant d'AOS modérée ou sévère et d'AOS sévère seulement, les aires sous la courbe étaient respectivement de 0,50 (intervalle de confiance [IC] à 95 % : de 0,42 à 0,58) et de 0,42 (IC à 95 % : de 0,34 à 0,52) pour la PA, de 0,65 (IC à 95 % : de 0,58 à 0,73) et de 0,63 (IC à 95 % : de 0,52 à 0,74) pour le questionnaire STOP-BANG, et de 0,68 (IC à 95 % : de 0,60 à 0,75) et de 0,69 (IC à 95 % : de 0,59 à 0,80) pour le questionnaire NoSAS. CONCLUSIONS: La PA et le questionnaire NoSAS ne sont pas suffisamment précis pour dépister l'AOS chez les patients présentant une FA. Compte tenu de la forte prévalence de l'AOS dans cette cohorte, des bienfaits potentiels d'un traitement de l'AOS et de la précision sous-optimale des questionnaires de dépistage actuels, il conviendrait d'envisager un TASD chez les patients présentant une FA.
RESUMEN
Identifying molecular mediators of neural circuit development and/or function that contribute to circuit dysfunction when aberrantly reengaged in neurological disorders is of high importance. The role of the TWEAK/Fn14 pathway, which was recently reported to be a microglial/neuronal axis mediating synaptic refinement in experience-dependent visual development, has not been explored in synaptic function within the mature central nervous system. By combining electrophysiological and phosphoproteomic approaches, we show that TWEAK acutely dampens basal synaptic transmission and plasticity through neuronal Fn14 and impacts the phosphorylation state of pre- and postsynaptic proteins in adult mouse hippocampal slices. Importantly, this is relevant in two models featuring synaptic deficits. Blocking TWEAK/Fn14 signaling augments synaptic function in hippocampal slices from amyloid-beta-overexpressing mice. After stroke, genetic or pharmacological inhibition of TWEAK/Fn14 signaling augments basal synaptic transmission and normalizes plasticity. Our data support a glial/neuronal axis that critically modifies synaptic physiology and pathophysiology in different contexts in the mature brain and may be a therapeutic target for improving neurophysiological outcomes.
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Degeneración Nerviosa/metabolismo , Transducción de Señal , Accidente Cerebrovascular/metabolismo , Sinapsis/metabolismo , Receptor de TWEAK/metabolismo , Animales , Citocina TWEAK/metabolismo , Modelos Animales de Enfermedad , Femenino , Hipocampo/fisiopatología , Masculino , Ratones Endogámicos C57BL , Ratones Transgénicos , Degeneración Nerviosa/fisiopatología , Plasticidad Neuronal/fisiología , Terminales Presinápticos/metabolismo , Accidente Cerebrovascular/fisiopatología , Transmisión Sináptica/fisiologíaRESUMEN
Recent years have seen the emergence of conditionally activated diagnostics and therapeutics that leverage protease-cleavable peptide linkers to enhance their specificity for cancer. However, due to a lack of methods to measure and localize protease activity directly within the tissue microenvironment, the design of protease-activated agents has been necessarily empirical, yielding suboptimal results when translated to patients. To address the need for spatially resolved protease activity profiling in cancer, we developed a new class of in situ probes that can be applied to fresh-frozen tissue sections in a manner analogous to immunofluorescence staining. These activatable zymography probes (AZP) detected dysregulated protease activity in human prostate cancer biopsy samples, enabling disease classification. AZPs were leveraged within a generalizable framework to design conditional cancer diagnostics and therapeutics and showcased in the Hi-Myc mouse model of prostate cancer, which models features of early pathogenesis. Multiplexed screening against barcoded substrates yielded a peptide, S16, that was robustly and specifically cleaved by tumor-associated metalloproteinases in the Hi-Myc model. In situ labeling with an AZP incorporating S16 revealed a potential role of metalloproteinase dysregulation in proliferative, premalignant Hi-Myc prostatic glands. Systemic administration of an in vivo imaging probe incorporating S16 perfectly classified diseased and healthy prostates, supporting the relevance of ex vivo activity assays to in vivo translation. We envision AZPs will enable new insights into the biology of protease dysregulation in cancer and accelerate the development of conditional diagnostics and therapeutics for multiple cancer types. SIGNIFICANCE: Visualization of protease activity within the native tissue context using AZPs provides new biological insights into protease dysregulation in cancer and guides the design of conditional diagnostics and therapeutics.
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Modelos Animales de Enfermedad , Sondas Moleculares/química , Péptido Hidrolasas/análisis , Péptido Hidrolasas/metabolismo , Neoplasias de la Próstata/patología , Proteínas Proto-Oncogénicas c-myc/genética , Animales , Humanos , Masculino , Ratones , Imagen Molecular , Neoplasias de la Próstata/enzimología , ProteolisisRESUMEN
The molecular basis of the earliest neuronal changes that lead to Alzheimer's disease (AD) is unclear. Here, we analyze neural cells derived from sporadic AD (SAD), APOE4 gene-edited and control induced pluripotent stem cells (iPSCs). We observe major differences in iPSC-derived neural progenitor (NP) cells and neurons in gene networks related to neuronal differentiation, neurogenesis, and synaptic transmission. The iPSC-derived neural cells from SAD patients exhibit accelerated neural differentiation and reduced progenitor cell renewal. Moreover, a similar phenotype appears in NP cells and cerebral organoids derived from APOE4 iPSCs. Impaired function of the transcriptional repressor REST is strongly implicated in the altered transcriptome and differentiation state. SAD and APOE4 expression result in reduced REST nuclear translocation and chromatin binding, and disruption of the nuclear lamina. Thus, dysregulation of neural gene networks may set in motion the pathologic cascade that leads to AD.
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Enfermedad de Alzheimer/genética , Enfermedad de Alzheimer/patología , Redes Reguladoras de Genes , Células Madre Pluripotentes Inducidas/metabolismo , Neuronas/metabolismo , Proteínas Represoras/metabolismo , Anciano , Anciano de 80 o más Años , Péptidos beta-Amiloides/metabolismo , Apolipoproteínas E/metabolismo , Diferenciación Celular/genética , Reprogramación Celular/genética , Fibroblastos/patología , Regulación de la Expresión Génica , Humanos , Persona de Mediana Edad , Células-Madre Neurales/metabolismo , Neurogénesis/genética , Lámina Nuclear/metabolismoRESUMEN
Although amyotrophic lateral sclerosis (ALS), also known as Lou Gehrig's disease, was first described in 1874, a flurry of genetic discoveries in the last 10 years has markedly increased our understanding of this disease. These findings have not only enhanced our knowledge of mechanisms leading to ALS, but also have revealed that ALS shares many genetic causes with another neurodegenerative disease, frontotemporal lobar dementia (FTLD). In this review, we survey how recent genetic studies have bridged our mechanistic understanding of these two related diseases and how the genetics behind ALS and FTLD point to complex disorders, implicating non-neuronal cell types in disease pathophysiology. The involvement of non-neuronal cell types is consistent with a non-cell autonomous component in these diseases. This is further supported by studies that identified a critical role of immune-associated genes within ALS/FTLD and other neurodegenerative disorders. The molecular functions of these genes support an emerging concept that various non-autonomous functions are involved in neurodegeneration. Further insights into such a mechanism(s) will ultimately lead to a better understanding of potential routes of therapeutic intervention. Facts ALS and FTLD are severe neurodegenerative disorders on the same disease spectrum. Multiple cellular processes including dysregulation of RNA homeostasis, imbalance of proteostasis, contribute to ALS/FTLD pathogenesis. Aberrant function in non-neuronal cell types, including microglia, contributes to ALS/FTLD. Strong neuroimmune and neuroinflammatory components are associated with ALS/FTLD patients. Open Questions Why can patients with similar mutations have different disease manifestations, i.e., why do C9ORF72 mutations lead to motor neuron loss in some patients while others exhibit loss of neurons in the frontotemporal lobe? Do ALS causal mutations result in microglial dysfunction and contribute to ALS/FTLD pathology? How do microglia normally act to mitigate neurodegeneration in ALS/FTLD? To what extent do cellular signaling pathways mediate non-cell autonomous communications between distinct central nervous system (CNS) cell types during disease? Is it possible to therapeutically target specific cell types in the CNS?
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Esclerosis Amiotrófica Lateral , Proteína C9orf72 , Demencia Frontotemporal , Neuronas Motoras , Mutación , Esclerosis Amiotrófica Lateral/genética , Esclerosis Amiotrófica Lateral/metabolismo , Esclerosis Amiotrófica Lateral/patología , Animales , Proteína C9orf72/genética , Proteína C9orf72/metabolismo , Demencia Frontotemporal/genética , Demencia Frontotemporal/metabolismo , Demencia Frontotemporal/patología , Humanos , Neuronas Motoras/metabolismo , Neuronas Motoras/patologíaRESUMEN
In mammals, the environment plays a critical role in promoting the final steps in neuronal development during the early postnatal period. While epigenetic factors are thought to contribute to this process, the underlying molecular mechanisms remain poorly understood. Here, we show that in the brain during early life, the DNA methyltransferase DNMT3A transiently binds across transcribed regions of lowly expressed genes, and its binding specifies the pattern of DNA methylation at CA sequences (mCA) within these genes. We find that DNMT3A occupancy and mCA deposition within the transcribed regions of genes is negatively regulated by gene transcription and may be modified by early-life experience. Once deposited, mCA is bound by the methyl-DNA-binding protein MECP2 and functions in a rheostat-like manner to fine-tune the cell-type-specific transcription of genes that are critical for brain function.
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ADN (Citosina-5-)-Metiltransferasas/genética , ADN (Citosina-5-)-Metiltransferasas/metabolismo , Metilación de ADN , Epigénesis Genética , Neuronas/metabolismo , Animales , Encéfalo/citología , Encéfalo/metabolismo , ADN Metiltransferasa 3A , Regulación del Desarrollo de la Expresión Génica , Humanos , Proteína 2 de Unión a Metil-CpG , Ratones , Transcripción Genética , Activación TranscripcionalRESUMEN
Diverse molecular mechanisms regulate synaptic composition and function in the mammalian nervous system. The multifunctional protein arginine methyltransferase 8 (PRMT8) possesses both methyltransferase and phospholipase activities. Here we examine the role of this neuron-specific protein in hippocampal plasticity and cognitive function. PRMT8 protein localizes to synaptic sites, and conditional whole-brain Prmt8 deletion results in altered levels of multiple synaptic proteins in the hippocampus, using both male and female mice. Interestingly, these altered protein levels are due to post-transcriptional mechanisms as the corresponding mRNA levels are unaffected. Strikingly, electrophysiological recordings from hippocampal slices of mice lacking PRMT8 reveal multiple defects in excitatory synaptic function and plasticity. Furthermore, behavioral analyses show that PRMT8 conditional knock-out mice exhibit impaired hippocampal-dependent fear learning. Together, these findings establish PRMT8 as an important component of the molecular machinery required for hippocampal neuronal function.SIGNIFICANCE STATEMENT Numerous molecular processes are critically required for normal brain function. Here we use mice lacking protein arginine methyltransferase 8 (PRMT8) in the brain to examine how loss of this protein affects the structure and function of neurons in the hippocampus. We find that PRMT8 localizes to the sites of communication between neurons. Hippocampal neurons from mice lacking PRMT8 have no detectable structural differences compared with controls; however, multiple aspects of their function are altered. Consistently, we find that mice lacking PRMT8 also exhibit reduced hippocampus-dependent memory. Together, our findings establish important roles for PRMT8 in regulating neuron function and cognition in the mammalian brain.
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Hipocampo/fisiopatología , Trastornos de la Memoria/fisiopatología , Trastornos Mentales/fisiopatología , Proteína-Arginina N-Metiltransferasas/metabolismo , Sinapsis/metabolismo , Transmisión Sináptica , Animales , Femenino , Hipocampo/patología , Masculino , Trastornos de la Memoria/complicaciones , Trastornos de la Memoria/patología , Trastornos Mentales/complicaciones , Trastornos Mentales/patología , Ratones , Ratones Noqueados , Plasticidad Neuronal , Proteína-Arginina N-Metiltransferasas/genética , Sinapsis/patologíaRESUMEN
Osteoporotic patients, incapacitated due to vertebral compression fractures (VCF), suffer grave financial and clinical burden. Current clinical treatments focus on symptoms' management but do not combat the issue at the source. In this pilot study, allogeneic, porcine mesenchymal stem cells, overexpressing the BMP6 gene (MSC-BMP6), were suspended in fibrin gel and implanted into a vertebral defect to investigate their effect on bone regeneration in a clinically relevant, large animal pig model. To check the effect of the BMP6-modified cells on bone regeneration, a fibrin gel only construct was used for comparison. Bone healing was evaluated in vivo at 6 and 12 weeks and ex vivo at 6 months. In vivo CT showed bone regeneration within 6 weeks of implantation in the MSC-BMP6 group while only minor bone formation was seen in the defect site of the control group. After 6 months, ex vivo analysis demonstrated enhanced bone regeneration in the BMP6-MSC group, as compared to control. This preclinical study presents an innovative, potentially minimally invasive, technique that can be used to induce bone regeneration using allogeneic gene modified MSCs and therefore revolutionize current treatment of challenging conditions, such as osteoporosis-related VCFs.
RESUMEN
Cyclin-dependent kinase 5 (Cdk5) is a serine/threonine kinase implicated in synaptic plasticity, behavior, and cognition, yet its synaptic function remains poorly understood. Here, we report that physiological Cdk5 signaling in rat hippocampal CA1 neurons regulates homeostatic synaptic transmission using an unexpectedly rapid mechanism that is different from all known slow homeostatic regulators, such as beta amyloid (Aß) and activity-regulated cytoskeleton-associated protein (Arc, aka Arg3.1). Interestingly, overproduction of the potent Cdk5 activator p25 reduces synapse density, and dynamically regulates synaptic size by suppressing or enhancing Aß/Arc production. Moreover, chronic overproduction of p25, seen in Alzheimer's patients, induces initially concurrent reduction in synapse density and increase in synaptic size characteristic of the early Alzheimer-like pathology, and later persistent synapse elimination in intact brains. These results identify Cdk5 as the regulator of a novel rapid form of homeostasis at central synapses and p25 as the first molecule capable of initiating the early Alzheimer's synaptic pathology.
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Región CA1 Hipocampal/enzimología , Región CA1 Hipocampal/patología , Quinasa 5 Dependiente de la Ciclina/metabolismo , Homeostasis/fisiología , Sinapsis/enzimología , Sinapsis/patología , Enfermedad de Alzheimer/metabolismo , Enfermedad de Alzheimer/patología , Péptidos beta-Amiloides/metabolismo , Animales , Células Cultivadas , Quinasa 5 Dependiente de la Ciclina/genética , Proteínas del Citoesqueleto/metabolismo , Modelos Animales de Enfermedad , Femenino , Masculino , Microscopía Electrónica , Proteínas del Tejido Nervioso/metabolismo , Plasticidad Neuronal/fisiología , Técnicas de Placa-Clamp , Fosfotransferasas/genética , Fosfotransferasas/metabolismo , Ratas , Ratas Transgénicas , Técnicas de Cultivo de TejidosRESUMEN
Osteoporosis affects more than 200 million people worldwide leading to more than 2 million fractures in the United States alone. Unfortunately, surgical treatment is limited in patients with low bone mass. Parathyroid hormone (PTH) was shown to induce fracture repair in animals by activating mesenchymal stem cells (MSCs). However, it would be less effective in patients with fewer and/or dysfunctional MSCs due to aging and comorbidities. To address this, we evaluated the efficacy of combination i.v. MSC and PTH therapy versus monotherapy and untreated controls, in a rat model of osteoporotic vertebral bone defects. The results demonstrated that combination therapy significantly increased new bone formation versus monotherapies and no treatment by 2 weeks (P < 0.05). Mechanistically, we found that PTH significantly enhanced MSC migration to the lumbar region, where the MSCs differentiated into bone-forming cells. Finally, we used allogeneic porcine MSCs and observed similar findings in a clinically relevant minipig model of vertebral defects. Collectively, these results demonstrate that in addition to its anabolic effects, PTH functions as an adjuvant to i.v. MSC therapy by enhancing migration to heal bone loss. This systemic approach could be attractive for various fragility fractures, especially using allogeneic cells that do not require invasive tissue harvest.
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Regeneración Ósea/efectos de los fármacos , Trasplante de Células Madre Mesenquimatosas/métodos , Células Madre Mesenquimatosas/efectos de los fármacos , Osteoporosis/terapia , Hormona Paratiroidea/farmacología , Fracturas de la Columna Vertebral/terapia , Animales , Diferenciación Celular/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Terapia Combinada , Modelos Animales de Enfermedad , Femenino , Humanos , Células Madre Mesenquimatosas/citología , Osteoporosis/complicaciones , Ratas , Fracturas de la Columna Vertebral/etiología , PorcinosRESUMEN
Mechanisms underlying motor neuron degeneration in spinal muscular atrophy (SMA), the leading inherited cause of infant mortality, remain largely unknown. Many studies have established the importance of hyperphosphorylation of the microtubule-associated protein tau in various neurodegenerative disorders, including Alzheimer's and Parkinson's diseases. However, tau phosphorylation in SMA pathogenesis has yet to be investigated. Here we show that tau phosphorylation on serine 202 (S202) and threonine 205 (T205) is increased significantly in SMA motor neurons using two SMA mouse models and human SMA patient spinal cord samples. Interestingly, phosphorylated tau does not form aggregates in motor neurons or neuromuscular junctions (NMJs), even at late stages of SMA disease, distinguishing it from other tauopathies. Hyperphosphorylation of tau on S202 and T205 is mediated by cyclin-dependent kinase 5 (Cdk5) in SMA disease condition, because tau phosphorylation at these sites is significantly reduced in Cdk5 knock-out mice; genetic knock-out of Cdk5 activating subunit p35 in an SMA mouse model also leads to reduced tau phosphorylation on S202 and T205 in the SMA;p35(-/-) compound mutant mice. In addition, expression of the phosphorylation-deficient tauS202A,T205A mutant alleviates motor neuron defects in a zebrafish SMA model in vivo and mouse motor neuron degeneration in culture, whereas expression of phosphorylation-mimetic tauS202E,T205E promotes motor neuron defects. More importantly, genetic knock-out of tau in SMA mice rescues synapse stripping on motor neurons, NMJ denervation, and motor neuron degeneration in vivo. Altogether, our findings suggest a novel mechanism for SMA pathogenesis in which hyperphosphorylation of non-aggregating tau by Cdk5 contributes to motor neuron degeneration.
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Quinasa 5 Dependiente de la Ciclina/metabolismo , Neuronas Motoras/patología , Atrofia Muscular Espinal , Degeneración Nerviosa/etiología , Médula Espinal/patología , Proteínas tau/metabolismo , Animales , Células Cultivadas , Modelos Animales de Enfermedad , Femenino , Regulación de la Expresión Génica/genética , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Proteínas de Homeodominio/metabolismo , Humanos , Inmunoprecipitación , Lactante , Recién Nacido , Masculino , Ratones , Ratones Transgénicos , Neuronas Motoras/metabolismo , Músculo Esquelético/patología , Atrofia Muscular Espinal/complicaciones , Atrofia Muscular Espinal/genética , Atrofia Muscular Espinal/patología , Proteínas del Tejido Nervioso/metabolismo , Unión Neuromuscular/metabolismo , Unión Neuromuscular/patología , Proteínas Nucleares/metabolismo , Oligodesoxirribonucleótidos Antisentido/farmacología , Fosforilación , Proteínas Represoras/metabolismo , Proteína 1 para la Supervivencia de la Neurona Motora/genética , Proteína 1 para la Supervivencia de la Neurona Motora/metabolismo , Pez Cebra , Proteínas tau/deficiencia , Proteínas tau/genéticaRESUMEN
Perturbations in fast-spiking parvalbumin (PV) interneurons are hypothesized to be a major component of various neuropsychiatric disorders; however, the mechanisms regulating PV interneurons remain mostly unknown. Recently, cyclin-dependent kinase 5 (Cdk5) has been shown to function as a major regulator of synaptic plasticity. Here, we demonstrate that genetic ablation of Cdk5 in PV interneurons in mouse brain leads to an increase in GABAergic neurotransmission and impaired synaptic plasticity. PVCre;fCdk5 mice display a range of behavioral abnormalities, including decreased anxiety and memory impairment. Our results reveal a central role of Cdk5 expressed in PV interneurons in gating inhibitory neurotransmission and underscore the importance of such regulation during behavioral tasks. Our findings suggest that Cdk5 can be considered a promising therapeutic target in a variety of conditions attributed to inhibitory interneuronal dysfunction, such as epilepsy, anxiety disorders, and schizophrenia.
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Ansiedad/psicología , Quinasa 5 Dependiente de la Ciclina/genética , Inhibición Psicológica , Interneuronas/metabolismo , Trastornos de la Memoria/psicología , Parvalbúminas/metabolismo , Animales , Ansiedad/genética , Conducta Animal/fisiología , Interneuronas/enzimología , Aprendizaje por Laberinto/fisiología , Trastornos de la Memoria/genética , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Actividad Motora/genética , Actividad Motora/fisiología , Transmisión Sináptica/genética , Transmisión Sináptica/fisiología , Vesículas Sinápticas/ultraestructura , Ácido gamma-Aminobutírico/metabolismoRESUMEN
The use of multicomponent scaffolds for cell implantation has necessitated sophisticated techniques for tracking of cell survival in vivo. Bioluminescent imaging (BLI) has emerged as a noninvasive tool for evaluating the therapeutic potential of cell-based tissue engineering strategies. However, the ability to use BLI measurements to longitudinally assess large 3D cellular constructs in vivo and the effects of potential confounding factors are poorly understood. In this study, luciferase-expressing human mesenchymal stem cells (hMSCs) were delivered subcutaneously within agarose and RGD-functionalized alginate hydrogel vehicles to investigate the impact of construct composition and tissue formation on BLI signal. Results showed that alginate constructs exhibited twofold greater BLI counts than agarose constructs at comparable hMSC doses. However, each hydrogel type produced a linear correlation between BLI counts and live cell number, indicating that within a given material, relative differences in cell number could be accurately assessed at early time points. The survival efficiency of delivered hMSCs was highest for the lower cell doses embedded within alginate matrix. BLI signal remained predictive of live cell number through 1 week in vivo, although the strength of correlation decreased over time. Irrespective of hydrogel type or initial hMSC seeding dose, all constructs demonstrated a degree of vascularization and development of a fibrotic capsule after 1 week. Formation of tissue within and adjacent to the constructs was accompanied by an attenuation of BLI signal during the initial period of the image acquisition time-frame. In alginate constructs only, greater vessel volume led to a delayed rise in BLI signal following luciferin delivery. This study identified vascular and fibrotic tissue ingrowth as potential confounding variables for longitudinal BLI studies. Further investigation into the complexities of noninvasive BLI data acquisition from multicomponent constructs, following implantation and subsequent tissue formation, is warranted.
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Hidrogel de Polietilenoglicol-Dimetacrilato , Mediciones Luminiscentes/métodos , Células Madre Mesenquimatosas/citología , Andamios del Tejido/química , Adulto , Animales , Supervivencia Celular/efectos de los fármacos , Femenino , Proteínas Fluorescentes Verdes/metabolismo , Humanos , Hidrogel de Polietilenoglicol-Dimetacrilato/farmacología , Luciferasas/metabolismo , Masculino , Células Madre Mesenquimatosas/efectos de los fármacos , Neovascularización Fisiológica/efectos de los fármacos , Implantación de Prótesis , Ratas Desnudas , Adulto JovenRESUMEN
Allografts may be useful in craniofacial bone repair, although they often fail to integrate with the host bone. We hypothesized that intermittent administration of parathyroid hormone (PTH) would enhance mesenchymal stem cell recruitment and differentiation, resulting in allograft osseointegration in cranial membranous bones. Calvarial bone defects were created in transgenic mice, in which luciferase is expressed under the control of the osteocalcin promoter. The mice were given implants of allografts with or without daily PTH treatment. Bioluminescence imaging (BLI) was performed to monitor host osteprogenitor differentiation at the implantation site. Bone formation was evaluated with the aid of fluorescence imaging (FLI) and microcomputed tomography (µCT) as well as histological analyses. Reverse transcription polymerase chain reaction (RT-PCR) was performed to evaluate the expression of key osteogenic and angiogenic genes. Osteoprogenitor differentiation, as detected by BLI, in mice treated with an allograft implant and PTH was over 2-fold higher than those in mice treated with an allograft implant without PTH. FLI also demonstrated that the bone mineralization process in PTH-treated allografts was significantly higher than that in untreated allografts. The µCT scans revealed a significant increase in bone formation in allograft + PTH treated mice comparing to allograft + PBS treated mice. The osteogenic genes osteocalcin (Oc/Bglap) and integrin binding sialoprotein (Ibsp) were upregulated in the allograft + PTH treated animals. In summary, PTH treatment enhances osteoprogenitor differentiation and augments bone formation around structural allografts. The precise mechanism is not clear, but we show that infiltration pattern of mast cells, associated with the formation of fibrotic tissue, in the defect site is significantly affected by the PTH treatment.
Asunto(s)
Huesos/efectos de los fármacos , Huesos/fisiología , Osteogénesis/efectos de los fármacos , Osteogénesis/fisiología , Hormona Paratiroidea/farmacología , Aloinjertos/efectos de los fármacos , Aloinjertos/fisiología , Animales , Trasplante Óseo/métodos , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/genética , Diferenciación Celular/fisiología , Expresión Génica/efectos de los fármacos , Expresión Génica/genética , Expresión Génica/fisiología , Mastocitos/efectos de los fármacos , Mastocitos/fisiología , Células Madre Mesenquimatosas/efectos de los fármacos , Células Madre Mesenquimatosas/fisiología , Ratones , Ratones Transgénicos , Neovascularización Fisiológica/efectos de los fármacos , Neovascularización Fisiológica/genética , Neovascularización Fisiológica/fisiología , Osteocalcina/genética , Osteogénesis/genética , Regiones Promotoras Genéticas/genética , Sialoglicoproteínas/genética , Trasplante Homólogo/métodosRESUMEN
Defects in DNA repair have been extensively linked to neurodegenerative diseases, but the exact mechanisms remain poorly understood. We found that FUS, an RNA/DNA-binding protein that has been linked to amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration, is important for the DNA damage response (DDR). The function of FUS in DDR involved a direct interaction with histone deacetylase 1 (HDAC1), and the recruitment of FUS to double-stranded break sites was important for proper DDR signaling. Notably, FUS proteins carrying familial ALS mutations were defective in DDR and DNA repair and showed a diminished interaction with HDAC1. Moreover, we observed increased DNA damage in human ALS patients harboring FUS mutations. Our findings suggest that an impaired DDR and DNA repair may contribute to the pathogenesis of neurodegenerative diseases linked to FUS mutations.
Asunto(s)
Daño del ADN/fisiología , Histona Desacetilasa 1/metabolismo , Neuronas/metabolismo , Proteína FUS de Unión a ARN/metabolismo , Esclerosis Amiotrófica Lateral/genética , Esclerosis Amiotrófica Lateral/metabolismo , Esclerosis Amiotrófica Lateral/patología , Animales , Células Cultivadas , Células HEK293 , Histona Desacetilasa 1/genética , Humanos , Ratones , Neuronas/patología , Unión Proteica/fisiología , Proteína FUS de Unión a ARN/genéticaRESUMEN
INTRODUCTION: Depression is predicted to become the world's second leading cause of disability by 2020 according to the World Health Organization. Cognitive behavioral intervention (CBI), recognized as a viable and effective treatment for depression, is becoming more widely used among Chinese clients. However, information about the application of this Western approach in the Chinese population is very limited. METHODS: This paper discusses adaptations of CBI protocols for Chinese patients, considering the major Chinese cultural characteristics of predestination, losing face, avoiding conflict, and Yin-Yang balance (PLAY) for persons with depression. RESULTS: Illustrated is the application of the PLAY protocol in the actual case of a 35-year-old woman with depression. Implications for integrating Chinese cultural characteristics with CBI are discussed. DISCUSSION: There is evidence for adaptations of CBI for enhancing its effectiveness among Chinese people within their cultural context. Since there are limited studies on cultural-sensitive CBI for Chinese people, the conclusions drawn from this study are only preliminary. Further studies that verify the findings reported in this paper are necessary.
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Terapia Cognitivo-Conductual/métodos , Cultura , Depresión/terapia , Adulto , Depresión/etnología , Depresión/psicología , Femenino , Humanos , Resultado del TratamientoRESUMEN
Cyclin-dependent kinase 5 (Cdk5) is associated with synaptic plasticity and cognitive function. Previous reports have demonstrated that Cdk5 is necessary for memory formation, although others have reported Cdk5 conditional knockout mouse models exhibiting enhanced learning and memory. Furthermore, how Cdk5 acts in specific cell populations to affect behavior and cognitive outcomes remains unclear. Here we conduct a behavioral characterization of a forebrain-specific Cdk5 conditional knockout mouse model under the αCaMKII promoter, in which Cdk5 is ablated in excitatory pyramidal neurons of the forebrain. The Cdk5 conditional knockouts exhibit hyperactivity in the open field, reduced anxiety, and reduced behavioral despair. Moreover, the Cdk5 conditional knockouts also display impaired spatial learning in the Morris water maze and are severely impaired in contextual fear memory, which correspond to deficits in synaptic transmission. Remarkably, the hyperactivity of the Cdk5 conditional knockouts can be ameliorated by the administration of lithium chloride, an inhibitor of GSK3ß signaling. Collectively, our data reveal that Cdk5 ablation from forebrain excitatory neurons results in deleterious effects on emotional and cognitive behavior and highlight a key role for Cdk5 in regulating the GSK3ß signaling pathway.
Asunto(s)
Cognición , Quinasa 5 Dependiente de la Ciclina/metabolismo , Hipercinesia/metabolismo , Prosencéfalo/metabolismo , Células Piramidales/metabolismo , Animales , Quinasa 5 Dependiente de la Ciclina/genética , Técnicas de Inactivación de Genes , Ratones , Ratones Endogámicos C57BL , Ratones NoqueadosRESUMEN
BACKGROUND: Brown adipose tissue plays a pivotal role in mammal metabolism and thermogenesis. It has a great therapeutic potential in several metabolic disorders such as obesity and diabetes. Mesenchymal stem cells (MSCs) are suitable candidates for brown adipose tissue formation de novo. Pparγ2 and C/ebpα are nucleic receptors known to mediate adipogenic differentiation. We hypothesized that overexpression of the Pparγ2 and C/ebpα genes in MSCs would lead to the formation of adipose tissue. MATERIALS & METHODS: MSCs bearing the Luc reporter gene were transfected to overexpress Pparγ2 and C/ebpα. Differentiation of nucleofected cells was evaluated in vitro and in vivo following ectopic implantation of the cells in C3H/HeN mice. RESULTS: After implantation, the engineered cells survived for 5 weeks and brown adipose-like tissue was observed in histological samples. Immunostaining and bioluminescent imaging showed new adipocytes expressing Luc and the brown adipose tissue marker, UCP1, in vitro and in vivo. CONCLUSION: We show that gene delivery of transcription factors into MSCs generates brown adipose tissue in vitro and in vivo.