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Nicotinamide mononucleotide adenylyl transferase 2 (NMNAT2) is neuroprotective in numerous preclinical models of neurodegeneration. Here, we show that brain nmnat2 mRNA levels correlate positively with global cognitive function and negatively with AD pathology. In AD brains, NMNAT2 mRNA and protein levels are reduced. NMNAT2 shifts its solubility and colocalizes with aggregated Tau in AD brains, similar to chaperones, which aid in the clearance or refolding of misfolded proteins. Investigating the mechanism of this observation, we discover a novel chaperone function of NMNAT2, independent from its enzymatic activity. NMNAT2 complexes with heat shock protein 90 (HSP90) to refold aggregated protein substrates. NMNAT2's refoldase activity requires a unique C-terminal ATP site, activated in the presence of HSP90. Furthermore, deleting NMNAT2 function increases the vulnerability of cortical neurons to proteotoxic stress and excitotoxicity. Interestingly, NMNAT2 acts as a chaperone to reduce proteotoxic stress, while its enzymatic activity protects neurons from excitotoxicity. Taken together, our data indicate that NMNAT2 exerts its chaperone or enzymatic function in a context-dependent manner to maintain neuronal health.
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Doença de Alzheimer/metabolismo , Encéfalo/metabolismo , Proteínas de Choque Térmico HSP90/metabolismo , Chaperonas Moleculares/metabolismo , Nicotinamida-Nucleotídeo Adenililtransferase/metabolismo , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/genética , Doença de Alzheimer/fisiopatologia , Animais , Western Blotting , Encéfalo/patologia , Encéfalo/fisiopatologia , Células COS , Células Cultivadas , Chlorocebus aethiops , Cognição/fisiologia , Feminino , Proteínas de Choque Térmico HSP90/genética , Humanos , Masculino , Camundongos Transgênicos , Microscopia de Fluorescência , Pessoa de Meia-Idade , Chaperonas Moleculares/genética , Mutação , Neurônios/citologia , Neurônios/metabolismo , Nicotinamida-Nucleotídeo Adenililtransferase/genética , Ligação Proteica , Dobramento de Proteína , Estabilidade Proteica , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
The age-dependent progression of tau pathology is a major characteristic of tauopathies, including Alzheimer's disease (AD), and plays an important role in the behavioral phenotypes of AD, including memory deficits. Despite extensive molecular and cellular studies on tau pathology, it remains to be determined how it alters the neural circuit functions underlying learning and memory in vivo. In rTg4510 mice, a Tau-P301L tauopathy model, hippocampal place fields that support spatial memories are abnormal at old age (7-9 months) when tau tangles and neurodegeneration are extensive. However, it is unclear how the abnormality in the hippocampal circuit function arises and progresses with the age-dependent progression of tau pathology. Here we show that in young (2-4 months of age) rTg4510 mice, place fields of hippocampal CA1 cells are largely normal, with only subtle differences from those of age-matched wild-type control mice. Second, high-frequency ripple oscillations of local field potentials in the hippocampal CA1 area are significantly reduced in young rTg4510 mice, and even further deteriorated in old rTg4510 mice. The ripple reduction is associated with less bursty firing and altered synchrony of CA1 cells. Together, the data indicate that deficits in ripples and neuronal synchronization occur before overt deficits in place fields in these mice. The results reveal a tau-pathology-induced progression of hippocampal functional changes in vivo.
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Modelos Animais de Doenças , Progressão da Doença , Hipocampo/fisiopatologia , Neurônios , Tauopatias/fisiopatologia , Animais , Feminino , Hipocampo/patologia , Masculino , Camundongos , Camundongos da Linhagem 129 , Camundongos Transgênicos , Neurônios/patologia , Tauopatias/patologiaRESUMO
Honey is a popular natural food product with a very complex composition mainly consisting of both organic and inorganic constituents. The composition of honey is strongly influenced by both natural and anthropogenic factors, which vary based on its botanical and geographical origins. Although minerals and heavy metals are minor constituents of honey, they play vital role in determining its quality. There are several different analytical methods used to determine the chemical elements in honey. These methods are typically based on spectroscopy or spectrometry techniques (including atomic absorption spectrometry, atomic emission spectrometry, inductively coupled plasma mass spectrometry, and inductively coupled plasma optical emission spectrometry). This review compiles available scientific information on minerals and heavy metals in honey reported from all over the world. To date, 54 chemical elements in various types of honey have been identified and can be divided into 3 groups: major or macroelements (Na, K, Ca, Mg, P, S, Cl), minor or trace elements (Al, Cu, Pb, Zn, Mn, Cd, Tl, Co, Ni, Rb, Ba, Be, Bi, U, V, Fe, Pt, Pd, Te, Hf, Mo, Sn, Sb, La, I, Sm, Tb, Dy, Sd, Th, Pr, Nd, Tm, Yb, Lu, Gd, Ho, Er, Ce, Cr, As, B, Br, Cd, Hg, Se, Sr), and heavy metals (trace elements that have a specific gravity at least 5 times higher than that of water and inorganic sources). Chemical elements in honey samples throughout the world vary in terms of concentrations and are also influenced by environmental pollution.
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Active zones are specialized presynaptic structures critical for neurotransmission. We show that a neuronal maintenance factor, nicotinamide mononucleotide adenylyltransferase (NMNAT), is required for maintaining active zone structural integrity in Drosophila by interacting with the active zone protein, Bruchpilot (BRP), and shielding it from activity-induced ubiquitin-proteasome-mediated degradation. NMNAT localizes to the peri-active zone and interacts biochemically with BRP in an activity-dependent manner. Loss of NMNAT results in ubiquitination, mislocalization and aggregation of BRP, and subsequent active zone degeneration. We propose that, as a neuronal maintenance factor, NMNAT specifically maintains active zone structure by direct protein-protein interaction.
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Proteínas de Drosophila/genética , Drosophila/genética , Nicotinamida-Nucleotídeo Adenililtransferase/genética , Terminações Pré-Sinápticas/enzimologia , Animais , Drosophila/enzimologia , Proteínas de Drosophila/metabolismo , Regulação da Expressão Gênica , Nicotinamida-Nucleotídeo Adenililtransferase/antagonistas & inibidores , Nicotinamida-Nucleotídeo Adenililtransferase/metabolismo , Terminações Pré-Sinápticas/ultraestrutura , Complexo de Endopeptidases do Proteassoma , Ligação Proteica , Proteólise , RNA Interferente Pequeno/genética , Transdução de Sinais , Transmissão Sináptica/fisiologia , Ubiquitina/genética , Ubiquitina/metabolismo , UbiquitinaçãoRESUMO
Tauopathies, including Alzheimer's disease, are a group of neurodegenerative diseases characterized by abnormal tau hyperphosphorylation that leads to formation of neurofibrillary tangles. Drosophila models of tauopathy display prominent features of the human disease including compromised lifespan, impairments of learning, memory and locomotor functions and age-dependent neurodegeneration visible as vacuolization. Here, we use a Drosophila model of frontotemporal dementia with parkinsonism linked to chromosome 17 (FTDP-17), in order to study the neuroprotective capacity of a recently identified neuronal maintenance factor, nicotinamide mononucleotide (NAD) adenylyl transferase (NMNAT), a protein that has both NAD synthase and chaperone function. NMNAT is essential for maintaining neuronal integrity under normal conditions and has been shown to protect against several neurodegenerative conditions. However, its protective role in tauopathy has not been examined. Here, we show that overexpression of NMNAT significantly suppresses both behavioral and morphological deficits associated with tauopathy by means of reducing the levels of hyperphosphorylated tau oligomers. Importantly, the protective activity of NMNAT protein is independent of its NAD synthesis activity, indicating a role for direct protein-protein interaction. Next, we show that NMNAT interacts with phosphorylated tau in vivo and promotes the ubiquitination and clearance of toxic tau species. Consequently, apoptosis activation was significantly reduced in brains overexpressing NMNAT, and neurodegeneration was suppressed. Our report on the molecular basis of NMNAT-mediated neuroprotection in tauopathies opens future investigation of this factor in other protein foldopathies.
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Modelos Animais de Doenças , Nicotinamida-Nucleotídeo Adenililtransferase/metabolismo , Tauopatias/metabolismo , Proteínas tau/antagonistas & inibidores , Animais , Drosophila , Fosforilação , Tauopatias/enzimologia , Tauopatias/patologia , Proteínas tau/metabolismoRESUMO
Tauopathies, characterized by neurofibrillary tangles (NFTs) of phosphorylated tau proteins, are a group of neurodegenerative diseases, including frontotemporal dementia and both sporadic and familial Alzheimer's disease. Forebrain-specific over-expression of human tau(P301L), a mutation associated with frontotemporal dementia with parkinsonism linked to chromosome 17, in rTg4510 mice results in the formation of NFTs, learning and memory impairment and massive neuronal death. Here, we show that the mRNA and protein levels of NMNAT2 (nicotinamide mononucleotide adenylyltransferase 2), a recently identified survival factor for maintaining neuronal health in peripheral nerves, are reduced in rTg4510 mice prior to the onset of neurodegeneration or cognitive deficits. Two functional cAMP-response elements (CREs) were identified in the nmnat2 promoter region. Both the total amount of phospho-CRE binding protein (CREB) and the pCREB bound to nmnat2 CRE sites in the cortex and the hippocampus of rTg4510 mice are significantly reduced, suggesting that NMNAT2 is a direct target of CREB under physiological conditions and that tau(P301L) overexpression down-regulates CREB-mediated transcription. We found that over-expressing NMNAT2 or its homolog NMNAT1, but not NMNAT3, in rTg4510 hippocampi from 6 weeks of age using recombinant adeno-associated viral vectors significantly reduced neurodegeneration caused by tau(P301L) over-expression at 5 months of age. In summary, our studies strongly support a protective role of NMNAT2 in the mammalian central nervous system. Decreased endogenous NMNAT2 function caused by reduced CREB signaling during pathological insults may be one of underlying mechanisms for neuronal death in tauopathies.
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Proteína de Ligação a CREB/genética , Regulação para Baixo , Nicotinamida-Nucleotídeo Adenililtransferase/genética , Tauopatias/genética , Transcrição Gênica , Animais , Sequência de Bases , Western Blotting , Primers do DNA , Modelos Animais de Doenças , Imunofluorescência , Hipocampo/patologia , Humanos , Camundongos , Camundongos Transgênicos , Regiões Promotoras Genéticas , Reação em Cadeia da Polimerase em Tempo Real , Tauopatias/patologiaRESUMO
BACKGROUND: Most patients with atherosclerotic cardiovascular disease fail to achieve guideline-directed low-density lipoprotein cholesterol (LDL-C) goals. Twice-yearly inclisiran lowers LDL-C by â¼50% when added to statins. OBJECTIVES: This study evaluated the effectiveness of an "inclisiran first" implementation strategy (adding inclisiran immediately upon failure to reach LDL-C <70 mg/dL despite receiving maximally tolerated statins) vs representative usual care in U.S. patients with atherosclerotic cardiovascular disease. METHODS: VICTORION-INITIATE, a prospective, pragmatically designed trial, randomized patients 1:1 to inclisiran (284 mg at days 0, 90, and 270) plus usual care (lipid management at treating physician's discretion) vs usual care alone. Primary endpoints were percentage change in LDL-C from baseline and statin discontinuation rates. RESULTS: We randomized 450 patients (30.9% women, 12.4% Black, 15.3% Hispanic); mean baseline LDL-C was 97.4 mg/dL. The "inclisiran first" strategy led to significantly greater reductions in LDL-C from baseline to day 330 vs usual care (60.0% vs 7.0%; P < 0.001). Statin discontinuation rates with "inclisiran first" (6.0%) were noninferior vs usual care (16.7%). More "inclisiran first" patients achieved LDL-C goals vs usual care (<70 mg/dL: 81.8% vs 22.2%; <55 mg/dL: 71.6% vs 8.9%; P < 0.001). Treatment-emergent adverse event (TEAE) and serious TEAE rates compared similarly between treatment strategies (62.8% vs 53.7% and 11.5% vs 13.4%, respectively). Injection-site TEAEs and TEAEs causing treatment withdrawal occurred more commonly with "inclisiran first" than usual care (10.3% vs 0.0% and 2.6% vs 0.0%, respectively). CONCLUSIONS: An "inclisiran first" implementation strategy led to greater LDL-C lowering compared with usual care without discouraging statin use or raising new safety concerns. (A Randomized, Multicenter, Open-label Trial Comparing the Effectiveness of an "Inclisiran First" Implementation Strategy to Usual Care on LDL Cholesterol [LDL-C] in Patients With Atherosclerotic Cardiovascular Disease and Elevated LDL-C [≥70 mg/dL] Despite Receiving Maximally Tolerated Statin Therapy [VICTORION-INITIATE]; NCT04929249).
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Aterosclerose , LDL-Colesterol , Inibidores de Hidroximetilglutaril-CoA Redutases , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Aterosclerose/tratamento farmacológico , LDL-Colesterol/sangue , Inibidores de Hidroximetilglutaril-CoA Redutases/uso terapêutico , Oligonucleotídeos/uso terapêutico , Estudos Prospectivos , Resultado do TratamentoRESUMO
The aggresome pathway is activated when proteasomal clearance of misfolded proteins is hindered. Misfolded polyubiquitinated protein aggregates are recruited and transported to the aggresome via the microtubule network by a protein complex consisting of histone deacetylase 6 (HDAC6) and the dynein motor complex. The current model suggests that HDAC6 recognizes protein aggregates by binding directly to polyubiquitinated proteins. Here, we show that there are substantial amounts of unanchored ubiquitin in protein aggregates with solvent-accessible C termini. The ubiquitin-binding domain (ZnF-UBP) of HDAC6 binds exclusively to the unanchored C-terminal diglycine motif of ubiquitin instead of conjugated polyubiquitin. The unanchored ubiquitin C termini in the aggregates are generated in situ by aggregate-associated deubiquitinase ataxin-3. These results provide structural and mechanistic bases for the role of HDAC6 in aggresome formation and further suggest a novel ubiquitin-mediated signaling pathway, where the exposure of ubiquitin C termini within protein aggregates enables HDAC6 recognition and transport to the aggresome.
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Histona Desacetilases/metabolismo , Complexos Multiproteicos/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Proteínas Nucleares/metabolismo , Proteínas Repressoras/metabolismo , Transdução de Sinais/fisiologia , Ubiquitina/metabolismo , Proteínas Ubiquitinadas/metabolismo , Ataxina-3 , Linhagem Celular , Desacetilase 6 de Histona , Histona Desacetilases/genética , Humanos , Complexos Multiproteicos/genética , Proteínas do Tecido Nervoso/genética , Proteínas Nucleares/genética , Estrutura Terciária de Proteína , Transporte Proteico/fisiologia , Proteínas Repressoras/genética , Ubiquitina/genética , Proteínas Ubiquitinadas/genéticaRESUMO
Stress responses are cellular processes essential for maintenance of cellular integrity and defense against environmental and intracellular insults. Neurodegenerative conditions are linked with inadequate stress responses. Several stress-responsive genes encoding neuroprotective proteins have been identified, and among them, the heat shock proteins comprise an important group of molecular chaperones that have neuroprotective functions. However, evidence for other critical stress-responsive genes is lacking. Recent studies on the NAD synthesis enzyme nicotinamide mononucleotide adenylyltransferase (NMNAT) have uncovered a novel neuronal maintenance and protective function against activity-, injury-, or misfolded protein-induced degeneration in Drosophila and in mammalian neurons. Here, we show that NMNAT is also a novel stress response protein required for thermotolerance and mitigation of oxidative stress-induced shortened lifespan. NMNAT is transcriptionally regulated during various stress conditions including heat shock and hypoxia through heat shock factor (HSF) and hypoxia-inducible factor 1α in vivo. HSF binds to nmnat promoter and induces NMNAT expression under heat shock. In contrast, under hypoxia, HIF1α up-regulates NMNAT indirectly through the induction of HSF. Our studies provide an in vivo mechanism for transcriptional regulation of NMNAT under stress and establish an essential role for this neuroprotective factor in cellular stress response.
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Proteínas de Ligação a DNA/metabolismo , Proteínas de Drosophila/metabolismo , Regulação Enzimológica da Expressão Gênica/fisiologia , Resposta ao Choque Térmico/fisiologia , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Longevidade/fisiologia , Nicotinamida-Nucleotídeo Adenililtransferase/biossíntese , Estresse Oxidativo/fisiologia , Fatores de Transcrição/metabolismo , Animais , Linhagem Celular , Proteínas de Ligação a DNA/genética , Proteínas de Drosophila/genética , Drosophila melanogaster , Fatores de Transcrição de Choque Térmico , Hipóxia/genética , Hipóxia/metabolismo , Nicotinamida-Nucleotídeo Adenililtransferase/genética , Fatores de Transcrição/genética , Transcrição Gênica/fisiologiaRESUMO
C9ORF72-hexanucleotide repeat expansions and ubiquilin-2 (UBQLN2) mutations are recently identified genetic markers in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). We investigate the relationship between C9ORF72 expansions and the clinical phenotype and neuropathology of ALS and FTLD. Genetic analysis and immunohistochemistry (IHC) were performed on autopsy-confirmed ALS (N = 75), FTLD-TDP (N = 30), AD (N = 14), and controls (N = 11). IHC for neurodegenerative disease pathology consisted of C9ORF72, UBQLN, p62, and TDP-43. A C9ORF72 expansion was identified in 19.4 % of ALS and 31 % of FTLD-TDP cases. ALS cases with C9ORF72 expansions frequently showed a bulbar onset of disease (57 %) and more rapid disease progression to death compared to non-expansion cases. Staining with C9ORF72 antibodies did not yield specific pathology. UBQLN pathology showed a highly distinct pattern in ALS and FTLD-TDP cases with the C9ORF72 expansion, with UBQLN-positive cytoplasmic inclusions in the cerebellar granular layer and extensive UBQLN-positive aggregates and dystrophic neurites in the hippocampal molecular layer and CA regions. These UBQLN pathologies were sufficiently unique to allow correct prediction of cases that were later confirmed to have C9ORF72 expansions by genetic analysis. UBQLN pathology partially co-localized with p62, and to a minor extent with TDP-43 positive dystrophic neurites and spinal cord skein-like inclusions. Our data indicate a pathophysiological link between C9ORF72 expansions and UBQLN proteins in ALS and FTLD-TDP that is associated with a highly characteristic pattern of UBQLN pathology. Our study indicates that this pathology is associated with alterations in clinical phenotype, and suggests that the presence of C9ORF72 repeat expansions may indicate a worse prognosis in ALS.
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Esclerose Lateral Amiotrófica/genética , Proteínas de Ciclo Celular/metabolismo , Degeneração Lobar Frontotemporal/genética , Proteínas/metabolismo , Ubiquitinas/metabolismo , Proteínas Adaptadoras de Transdução de Sinal , Idade de Início , Idoso , Idoso de 80 Anos ou mais , Esclerose Lateral Amiotrófica/metabolismo , Esclerose Lateral Amiotrófica/mortalidade , Esclerose Lateral Amiotrófica/patologia , Proteínas Relacionadas à Autofagia , Proteína C9orf72 , Proteínas de Ciclo Celular/genética , Expansão das Repetições de DNA , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Feminino , Degeneração Lobar Frontotemporal/mortalidade , Degeneração Lobar Frontotemporal/patologia , Humanos , Corpos de Inclusão/patologia , Masculino , Pessoa de Meia-Idade , Mutação/genética , Proteínas/genética , Ubiquitinas/genéticaRESUMO
BACKGROUND: Idiopathic Parkinson disease (PD) is a multi-system disorder with a multifactorial etiology and diverse clinical phenotype. Selective, regional dopaminergic neuronal degeneration in the substantia nigra and other CNS areas including the amygdala are observed in all patients. Apoptotic mechanisms resulting from oxidative stress and mitochondrial dysfunction have been implicated in the pathogenesis of the disease. Although the role of melatonin, a potent endogenous antioxidant, has been highlighted in PD there is no data on the expression of melatonin receptors in affected CNS regions. MATERIAL/METHODS: We conducted an RT-PCR-based study to determine the MT1 and MT2 receptors expression in whole brain post-mortem tissue from the amygdala and substantia nigra of well-characterized PD and control subjects. RESULTS: PD cases showed a statistically significant decrease of MT1 receptor expression in both substantia nigra (FC=5.11; p<0.05) and the amygdala (FC=3.11; p<0.001) versus normal controls. The expression of MT2 receptor expression was also decreased in both substantia nigra (FC=3.90; p<0.0001) and the amygdala (FC=1.91; p<0.001) versus normal controls. CONCLUSIONS: The results demonstrate a down-regulation of melatonin receptors in regions affected by PD, suggesting their possible involvement in the disease process. Future studies are needed to elucidate the role of melatonin and its receptors in the treatment/pathogenesis of PD.
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Regulação para Baixo , Doença de Parkinson/genética , Doença de Parkinson/metabolismo , Receptor MT1 de Melatonina/genética , Receptor MT1 de Melatonina/metabolismo , Receptor MT2 de Melatonina/genética , Receptor MT2 de Melatonina/metabolismo , Idoso , Idoso de 80 Anos ou mais , Tonsila do Cerebelo/metabolismo , Tonsila do Cerebelo/patologia , Estudos de Casos e Controles , Estudos de Coortes , Feminino , Humanos , Imuno-Histoquímica , Masculino , Doença de Parkinson/patologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Substância Negra/metabolismo , Substância Negra/patologiaRESUMO
Human neurodegenerative diseases arise from a wide array of genetic and environmental factors. Despite the diversity in etiology, many of these diseases are considered "conformational" in nature, characterized by the accumulation of pathological, misfolded proteins. These misfolded proteins can induce cellular stress by overloading the proteolytic machinery, ultimately resulting in the accumulation and deposition of aggregated protein species that are cytotoxic. Misfolded proteins may also form aberrant, non-physiological protein-protein interactions leading to the sequestration of other normal proteins essential for cellular functions. The progression of such disease may therefore be viewed as a failure of normal protein homeostasis, a process that involves a network of molecules regulating the synthesis, folding, translocation and clearance of proteins. Molecular chaperones are highly conserved proteins involved in the folding of nascent proteins, and the repair of proteins that have lost their typical conformations. These functions have therefore made molecular chaperones an active area of investigation within the field of conformational diseases. This review will discuss the role of molecular chaperones in neurodegenerative diseases, highlighting their functional classification, regulation, and therapeutic potential for such diseases.
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Chaperonas Moleculares/química , Doenças Neurodegenerativas/fisiopatologia , Conformação Proteica , Dobramento de Proteína , Resposta ao Choque Térmico/fisiologia , Humanos , Chaperonas Moleculares/classificação , Chaperonas Moleculares/genética , Chaperonas Moleculares/metabolismo , Mutação , Processamento de Proteína Pós-TraducionalRESUMO
The title compound, C(9)H(11)NO(3), was prepared by an Aldol reaction of 2,3-dihydro-1H-pyrrolizin-1-one with formaldehyde. The asymmetric unit contains six mol-ecules. The pyrrolizine ring system in each mol-ecule is planar, the maximum atomic deviation being 0.066â (2)â Å. In the crystal structure, mol-ecules are liked together by an extensive O-Hâ¯O hydrogen-bonding network.
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The data presented in this article are related to the research article entitled "NiCoPt/Graphene-dot Nanosponge as a Highly Stable Electrocatalyst for Efficient Hydrogen Evolution Reaction in Acidic Electrolyte (N.-A. Nguyen et al., 2020) [1]. This article reports a simple method to synthesize NiCoPt/Graphene-dot as an electrocatalyst with low Pt loading but high hydrogen evolution reaction (HER) performance. The morphology of NiCoPt/Graphene-dot was analyzed by scanning electron microscopy (SEM) and high-resolution transmission electron microscopy (HRTEM) techniques. The structural and chemical properties of NiCoPt/Graphene-dot were investigated by using X-ray Powder Diffraction (XRD) and X-ray photoelectron spectroscopy (XPS) techniques.
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OBJECTIVE: The objective of this study was to develop a low-cost kit for the detection of subclinical mastitis (SCM) and to check its validity, reproducibility, and efficacy at the field level. MATERIALS AND METHODS: A total of 550 quarter milk samples from crossbred dairy cows were collected, of which 400 milk samples were used to validate the newly developed BLRI mastitis test (BMT) kit to justify its efficacy as an individual test kit in detecting SCM based on somatic cell count (SCC) by direct microscopic count (DMC). The efficacy of the newly developed BMT was compared with the California Mastitis Test (CMT) kit. Another 150 milk samples were subjected to SCC determined by DMC and DCC (De Laval cell counter®) categorized by CMT and BMT scores. RESULTS: A SCM test kit, namely, BMT kit was successfully developed in this study. The percentage accuracy of CMT and BMT were 76.75% and 75.75%; sensitivity 69.36% and 67.56%; specificity 85.95% and 85.85%; positive predictive value 86.03% and 85.71%; negative predictive value 69.23% and 68%, respectively. A p value of 0.001 was found for both CMT and BMT. However, CMT and BMT had no significant difference in sensitivity (p = 0.778). Average SCCs (cells/ml) determined by DCC and DMC, respectively, were mostly corresponded to the SCC ranges of both CMT and BMT scores. CONCLUSION: The newly developed BMT kit is an independent, cheap, farmer-friendly, first country made, and reliable SCM diagnostic test kit that can be used at field condition.
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Fish assemblage structure in Panchet, a large tropical reservoir along river Damodar, major tributary of river Ganga in India, was studied along the spatial gradient of the impoundment. Fish samples were collected bimonthly from October 2014 to September 2016. Fish community structure in terms of species composition, relative abundance, and trophic and conservation status was recorded. Spatial fish diversity was analyzed from riverine, transitional, and lacustrine zones. Sixty-two fish species were recorded wherein the family Cyprinidae dominated in number of species (26) followed by Bagridae (5). Shannon diversity index, evenness index, and species richness did not reveal significant variation (p > 0.05) across different zones. Relative abundance of individuals and species richness were maximum in the transitional zone and minimum in the lacustrine zone. Multi-dimensional scaling of fish assemblage revealed similar pattern in the riverine and transitional zones. Trophic guild of fishes indicated dominance of carnivorous species followed by planktivores. Conservation status (IUCN 2017.3) showed 4 fish species in Near Threatened category. The study suggests conservation of fish habitats to maintain diversity and sustained production. The baseline information generated in the study will be beneficial for monitoring the alteration in fish assemblage, conservation of fish diversity, and management planning.
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Peixes/classificação , Lagos , Rios , Animais , Biodiversidade , Ecossistema , Pesqueiros , Peixes/crescimento & desenvolvimento , Cadeia Alimentar , Índia , Densidade Demográfica , Dinâmica Populacional , Estações do AnoRESUMO
MicroRNAs (miRs) are 18~23 nucleotides long non-coding RNAs that regulate gene expression. To explore whether miR alterations in tauopathy contribute to pathological conditions, we first determined which hippocampal miRs are altered at the presymptomatic and symptomatic stages of tauopathy using rTg4510 mice (Tau mice), a well-characterized tauopathy model. miR-RNA pairing analysis using QIAGEN Ingenuity Pathway Analysis (IPA) revealed 401 genes that can be regulated by 71 miRs altered in Tau hippocampi at the presymptomatic stage. Among several miRs confirmed with real-time qPCR, miR142 (-3p and -5p) in Tau hippocampi were significantly upregulated by two-weeks of age and onward. Transcriptome studies by RNAseq and IPA revealed several overlapping biological and disease associated pathways affected by either Tau or miR142 overexpression, including Signal Transducer and Activator of Transcription 3 (Stat3) and Tumor Necrosis Factor Receptor 2 (Tnfr2) signaling pathways. Similar to what was observed in Tau brains, overexpressing miR142 in wildtype cortical neurons augments mRNA levels of Glial Fibrillary Acidic Protein (Gfap) and Colony Stimulating Factor 1 (Csf1), accompanied by a significant increase in microglia and reactive astrocyte numbers. Taken together, our study suggests that miR alterations by Tau overexpression may contribute to the neuroinflammation observed in Tau brains.
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MicroRNAs/metabolismo , Tauopatias/genética , Proteínas tau/metabolismo , Envelhecimento/genética , Animais , Feminino , Redes Reguladoras de Genes , Hipocampo/metabolismo , Humanos , Camundongos Endogâmicos ICR , Camundongos Transgênicos , MicroRNAs/genética , Microglia/metabolismo , Neurônios/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transcrição Gênica , Transcriptoma/genética , Regulação para Cima/genéticaRESUMO
Nicotinamide mononucleotide adenylyl transferase 2 (NMNAT2) is a key neuronal maintenance factor and provides potent neuroprotection in numerous preclinical models of neurological disorders. NMNAT2 is significantly reduced in Alzheimer's, Huntington's, Parkinson's diseases. Here we developed a Meso Scale Discovery (MSD)-based screening platform to quantify endogenous NMNAT2 in cortical neurons. The high sensitivity and large dynamic range of this NMNAT2-MSD platform allowed us to screen the Sigma LOPAC library consisting of 1280 compounds. This library had a 2.89% hit rate, with 24 NMNAT2 positive and 13 negative modulators identified. Western analysis was conducted to validate and determine the dose-dependency of identified modulators. Caffeine, one identified NMNAT2 positive-modulator, when systemically administered restored NMNAT2 expression in rTg4510 tauopathy mice to normal levels. We confirmed in a cell culture model that four selected positive-modulators exerted NMNAT2-specific neuroprotection against vincristine-induced cell death while four selected NMNAT2 negative modulators reduced neuronal viability in an NMNAT2-dependent manner. Many of the identified NMNAT2 positive modulators are predicted to increase cAMP concentration, suggesting that neuronal NMNAT2 levels are tightly regulated by cAMP signaling. Taken together, our findings indicate that the NMNAT2-MSD platform provides a sensitive phenotypic screen to detect NMNAT2 in neurons.
Assuntos
Córtex Cerebral/citologia , Ensaios de Triagem em Larga Escala/métodos , Neurônios/efeitos dos fármacos , Nicotinamida-Nucleotídeo Adenililtransferase/metabolismo , Bibliotecas de Moléculas Pequenas/farmacologia , Animais , Cafeína/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Estimulantes do Sistema Nervoso Central/farmacologia , Córtex Cerebral/metabolismo , Expressão Gênica/efeitos dos fármacos , Camundongos Knockout , Camundongos Transgênicos , Neurônios/metabolismo , Nicotinamida-Nucleotídeo Adenililtransferase/genética , Reprodutibilidade dos Testes , Tauopatias/genética , Tauopatias/metabolismoRESUMO
Acute liver failure (ALF) is an acute medical emergency which carries high mortality without liver transplantation. Various hepatotropic viruses, drug induced liver injury, auto immune hepatitis, and metabolic liver diseases are the commonly implicated etiologic agents. Liver involvement in dengue hemorrhagic fever (DHF) is quite common, but acute liver failure is its rare complication. Neurological complications are also commonly seen in DHF. Ateenage girl presented with high grade fever and subconjunctival hemorrhage, and later developed jaundice due to acute liver failure. Liver transplantation could not be offered due to fungemia. During hospital stay, she had seizures and intracranial hemorrhage culminating in brain death. ALF with neurological involvement is a rare but very important and fatal complication of DHF; and it should be considered as a cause of acute liver failure, especially in endemic areas.
Assuntos
Doenças da Túnica Conjuntiva/complicações , Hepatite/diagnóstico por imagem , Icterícia/complicações , Falência Hepática Aguda/etiologia , Dengue Grave/diagnóstico , Adolescente , Antivirais/uso terapêutico , Evolução Fatal , Feminino , Hepatite/tratamento farmacológico , Humanos , Dengue Grave/complicaçõesRESUMO
Nicotinamide mononucleotide adenylyl transferases (NMNATs) are essential neuronal maintenance factors postulated to preserve neuronal function and protect against axonal degeneration in various neurodegenerative disease states. We used in vitro and in vivo approaches to assess the impact of NMNAT2 reduction on cellular and physiological functions induced by treatment with a vinca alkaloid (vincristine) and a taxane-based (paclitaxel) chemotherapeutic agent. NMNAT2 null (NMNAT2-/-) mutant mice die at birth and cannot be used to probe functions of NMNAT2 in adult animals. Nonetheless, primary cortical cultures derived from NMNAT2-/- embryos showed reduced cell viability in response to either vincristine or paclitaxel treatment whereas those derived from NMNAT2 heterozygous (NMNAT2+/-) mice were preferentially sensitive to vincristine-induced degeneration. Adult NMNAT2+/- mice, which survive to adulthood, exhibited a 50% reduction of NMNAT2 protein levels in dorsal root ganglia relative to wildtype (WT) mice with no change in levels of other NMNAT isoforms (NMNAT1 or NMNAT3), NMNAT enzyme activity (i.e. NAD/NADH levels) or microtubule associated protein-2 (MAP2) or neurofilament protein levels. We therefore compared the impact of NMNAT2 knockdown on the development and maintenance of chemotherapy-induced peripheral neuropathy induced by vincristine and paclitaxel treatment using NMNAT2+/- and WT mice. NMNAT2+/- did not differ from WT mice in either the development or maintenance of either mechanical or cold allodynia induced by either vincristine or paclitaxel treatment. Intradermal injection of capsaicin, the pungent ingredient in hot chili peppers, produced equivalent hypersensitivity in NMNAT2+/- and WT mice receiving vehicle in lieu of paclitaxel. Capsaicin-evoked hypersensitivity was enhanced by prior paclitaxel treatment but did not differ in either NMNAT2+/- or WT mice. Thus, capsaicin failed to unmask differences in nociceptive behaviors in either paclitaxel-treated or paclitaxel-untreated NMNAT2+/- and WT mice. Moreover, no differences in motor behavior were detected between genotypes in the rotarod test. Our studies do not preclude the possibility that complete knockout of NMNAT2 in a conditional knockout animal could unmask a role for NMNAT2 in protection against detrimental effects of chemotherapeutic treatment.