RESUMO
Mammalian cells have evolved to function under Earth's gravity, but how they respond to microgravity remains largely unknown. Neural stem cells (NSCs) are essential for the maintenance of central nervous system (CNS) functions during development and the regeneration of all CNS cell populations. Here, we examined the behavior of space (SPC)-flown NSCs as they readapted to Earth's gravity. We found that most of these cells survived the space flight and self-renewed. Yet, some showed enhanced stress responses as well as autophagy-like behavior. To ascertain if the secretome from SPC-flown NSCs contained molecules inducing these responses, we incubated naïve, non-starved NSCs in a medium containing SPC-NSC secretome. We found a four-fold increase in stress responses. Proteomic analysis of the secretome revealed that the protein of the highest content produced by SPC-NSCs was secreted protein acidic and rich in cysteine (SPARC), which induces endoplasmic reticulum (ER) stress, resulting in the cell's demise. These results offer novel knowledge on the response of neural cells, particularly NSCs, subjected to space microgravity. Moreover, some secreted proteins have been identified as microgravity sensing, paving a new venue for future research aiming at targeting the SPARC metabolism. Although we did not establish a direct relationship between microgravity-induced stress and SPARC as a potential marker, these results represent the first step in the identification of gravity sensing molecules as targets to be modulated and to design effective countermeasures to mitigate intracranial hypertension in astronauts using structure-based protein design.
Assuntos
Células-Tronco Neurais , Voo Espacial , Animais , Humanos , Osteonectina , Proteômica , Neurônios , MamíferosRESUMO
Huntington's disease (HD), a genetic neurodegenerative disorder, primarily affects the striatum and cortex with progressive loss of medium-sized spiny neurons (MSNs) and pyramidal neurons, disrupting cortico-striatal circuitry. A promising regenerative therapeutic strategy of transplanting human neural stem cells (hNSCs) is challenged by the need for long-term functional integration. We previously described that, with short-term hNSC transplantation into the striatum of HD R6/2 mice, human cells differentiated into electrophysiologically active immature neurons, improving behavior and biochemical deficits. Here, we show that long-term (8 months) implantation of hNSCs into the striatum of HD zQ175 mice ameliorates behavioral deficits, increases brain-derived neurotrophic factor (BDNF) levels, and reduces mutant huntingtin (mHTT) accumulation. Patch clamp recordings, immunohistochemistry, single-nucleus RNA sequencing (RNA-seq), and electron microscopy demonstrate that hNSCs differentiate into diverse neuronal populations, including MSN- and interneuron-like cells, and form connections. Single-nucleus RNA-seq analysis also shows restoration of several mHTT-mediated transcriptional changes of endogenous striatal HD mouse cells. Remarkably, engrafted cells receive synaptic inputs, innervate host neurons, and improve membrane and synaptic properties. Overall, the findings support hNSC transplantation for further evaluation and clinical development for HD.
Assuntos
Doença de Huntington , Células-Tronco Neurais , Humanos , Camundongos , Animais , Doença de Huntington/genética , Doença de Huntington/terapia , Corpo Estriado , Neurônios , Fenótipo , Modelos Animais de Doenças , Camundongos Transgênicos , Proteína Huntingtina/genéticaRESUMO
Intracranial hypertension (ICP) and visual impairment intracranial pressure (VIIP) are some of the consequences of long-term space missions. Here we examined the behavior of oligodendrocyte progenitors (OLPs) after space flight using time-lapse microscopy. We show that most OLPs divided more than ground control (GC) counterparts did. Nonetheless, a subpopulation of OLPs flown to space presented a significant increase in autophagic cell death. Examination of the proteomic profile of the secretome of space flown OLPs (SPC-OLPs) revealed that the stress protein heat shock protein-90 beta "HSP-90ß" was the 5th most enriched (6.8 times) and the secreted protein acidic and rich in cysteine "SPARC" was the 7th most enriched (5.2 times), with respect to ground control cells. SPARC induces endoplasmic reticulum stress, which leads to autophagy. Given the roles and importance of these two proteins in mammalian cells' metabolism, their upregulation may hold the key to modulating cell proliferation and autophagy, in order to mitigate ICP and VIIP during and after space missions.
Assuntos
Células Precursoras de Oligodendrócitos , Voo Espacial , Animais , Osteonectina , Proteômica , Autofagia , Proliferação de Células , MamíferosRESUMO
Opioids are the most common medications for moderate to severe pain. Unfortunately, they also have addictive properties that have precipitated opioid misuse and the opioid epidemic. In the present study, we examined the effects of acute administration of oxycodone, a µ-opioid receptor (MOR) agonist, on Ca2+ transient activity of medium-sized spiny neurons (MSNs) in freely moving animals. Ca2+ imaging of MSNs in dopamine D1-Cre mice (expressing Cre predominantly in the direct pathway) or adenosine A2A-Cre mice (expressing Cre predominantly in the indirect pathway) was obtained with the aid of miniaturized microscopes (Miniscopes) and a genetically encoded Cre-dependent Ca2+ indicator (GCaMP6f). Systemic injections of oxycodone (3 mg/kg) increased locomotor activity yet, paradoxically, reduced concomitantly the number of active MSNs. The frequency of Ca2+ transients was significantly reduced in MSNs from A2A-Cre mice but not in those from D1-Cre mice. For comparative purposes, a separate group of mice was injected with a non-Cre dependent Ca2+ indicator in the cerebral cortex and the effects of the opioid also were tested. In contrast to MSNs, the frequency of Ca2+ transients in cortical pyramidal neurons was significantly increased by oxycodone administration. Additional electrophysiological studies in brain slices confirmed generalized inhibitory effects of oxycodone on MSNs, including membrane hyperpolarization, reduced excitability, and decreased frequency of spontaneous excitatory and inhibitory postsynaptic currents. These results demonstrate a dissociation between locomotion and striatal MSN activity after acute administration of oxycodone.
Assuntos
Cálcio , Oxicodona , Camundongos , Animais , Cálcio/metabolismo , Oxicodona/farmacologia , Oxicodona/metabolismo , Corpo Estriado/metabolismo , Neurônios/metabolismo , Dopamina/metabolismoRESUMO
Considering the imminence of long-term space travel, it is necessary to investigate the impact of space microgravity (SPC-µG) in order to determine if this environment has consequences on the astronauts' health, in particular, neural and cognitive functions. Neural stem cells (NSCs) are the basis for the regeneration of the central nervous system (CNS) cell populations and learning how weightlessness impacts NSCs in health and disease provides a critical tool for the potential mitigation of specific mechanisms leading to neurological disorders. In previous studies, we found that exposure to SPC-µG resulted in enhanced proliferation, a shortened cell cycle, and a larger cell diameter of NSCs compared to control cells. Here, we report the frequent occurrence of abnormal cell division (ACD) including incomplete cell division (ICD), where cytokinesis is not successfully completed, and multi-daughter cell division (MDCD) of NSCs following SPC-µG as well as secretome exposure compared to ground control (1G) NSCs. These findings provide new insights into the potential health implications of space travel and have far-reaching implications for understanding the mechanisms leading to the deleterious effects of long-term space travel as well as potential carcinogenic susceptibility. Knowledge of these mechanisms could help to develop preventive or corrective measures for successful long-term SPC-µG exposure.
Assuntos
Neoplasias Encefálicas , Células-Tronco Neurais , Ausência de Peso , Humanos , Ausência de Peso/efeitos adversos , Encéfalo/fisiologia , Neoplasias Encefálicas/etiologia , Autorrenovação CelularRESUMO
Huntington's disease (HD) is a genetic, inexorably fatal neurodegenerative disease. Patient average survivability is up to 20 years after the onset of symptoms. Those who suffer from the disease manifest motor, cognitive, and psychiatric impairments. There is indirect evidence suggesting that omega-3 polyunsaturated fatty acids (ω-3 PUFA) could have alleviating effects on most of HD symptoms. These include beneficial effects against cachexia and weight loss, decrease of cognitive impairment over time, and improvement of psychiatric symptoms such as depression and irritability. Furthermore, there is a positive correlation between consumption of ω-3 PUFAs in diets and prevalence of HD, as well as direct effects on the disease via release of serotonin. Unfortunately, to date, very few studies have examined the effects of ω-3 PUFAs in HD, both on the symptoms and on disease progression. This paper reviews evidence in the literature suggesting that ω-3 PUFAs can be used in neurodegenerative disorders. This information can be extrapolated to support further research of ω-3 PUFAs and their potential use for HD treatment.
Assuntos
Disfunção Cognitiva , Ácidos Graxos Ômega-3 , Doença de Huntington , Doenças Neurodegenerativas , Progressão da Doença , Ácidos Graxos Ômega-3/uso terapêutico , Humanos , Doença de Huntington/tratamento farmacológicoRESUMO
Patients with a history of cancer are increasingly common in the dental office. Treating cancer patients requires a multidisciplinary team, which should include the dentist, in order to control the complications that occur in the oral cavity and also to recover the patient undergoing treatment in any of its types: surgical, medical, radiotherapeutic, or its possible combinations. Dental implants can be a safe and predictable treatment option for prosthetic rehabilitation. The aim of this paper is to describe in retrospect the success rate of osseointegrated implants in oncology and non-oncology patients placed by the Master of Dentistry in Oncology and Immunocompromised Patients, as well as the Master of Medicine, Surgery and Oral Implantology of the University of Barcelona Dental Hospital, between July 2011 and March 2016. 466 patients were reviewed, with a total of 1405 implants placed, considering the oncological history of the patients and the implant success rate. The total success rate in the concerned period was 96.65%. When comparing cancer patients with healthy ones, the success rate has been 93.02% in the first case, and 97.16% in the latter. According to the literature review, our results encourage implant placement in cancer patients, it is important to recognize that this is an analysis of a complex care pathway with a large number of confounding variables. However, the findings should not be considered as generalizable.
Assuntos
Implantes Dentários , Neoplasias , Implantação Dentária Endóssea , Prótese Dentária Fixada por Implante , Falha de Restauração Dentária , Humanos , Estudos Retrospectivos , Resultado do TratamentoRESUMO
Abstract Patients with a history of cancer are increasingly common in the dental office. Treating cancer patients requires a multidisciplinary team, which should include the dentist, in order to control the complications that occur in the oral cavity and also to recover the patient undergoing treatment in any of its types: surgical, medical, radiotherapeutic, or its possible combinations. Dental implants can be a safe and predictable treatment option for prosthetic rehabilitation. The aim of this paper is to describe in retrospect the success rate of osseointegrated implants in oncology and non-oncology patients placed by the Master of Dentistry in Oncology and Immunocompromised Patients, as well as the Master of Medicine, Surgery and Oral Implantology of the University of Barcelona Dental Hospital, between July 2011 and March 2016. 466 patients were reviewed, with a total of 1405 implants placed, considering the oncological history of the patients and the implant success rate. The total success rate in the concerned period was 96.65%. When comparing cancer patients with healthy ones, the success rate has been 93.02% in the first case, and 97.16% in the latter. According to the literature review, our results encourage implant placement in cancer patients, it is important to recognize that this is an analysis of a complex care pathway with a large number of confounding variables. However, the findings should not be considered as generalizable.
Resumo Pacientes com histórico de câncer são cada vez mais comuns no consultório odontológico. O tratamento de pacientes com câncer requer uma equipe multidisciplinar, que deve incluir o dentista, a fim de controlar as complicações que ocorrem na cavidade oral e também para tratar o paciente com qualquer uma das modalidades de tratamento: cirúrgica, médica, radioterápica ou suas possíveis combinações. Os implantes dentários podem ser uma opção de tratamento segura e previsível para reabilitação protética. O objetivo deste artigo é propor um estudo retrospectivo sobre a taxa de sucesso de implantes osseointegrados em pacientes oncológicos e não oncológicos atendidos no Mestrado em Odontologia em Pacientes Oncológicos e Imunodeprimidos, bem como no Mestrado em Medicina, Cirurgia e Implantodontia Oral do Hospital Odontológico da Universidade de Barcelona, entre julho de 2011 e março de 2016. Foram revisados 466 pacientes, com um total de 1405 implantes instalados, considerando o histórico oncológico dos pacientes e a taxa de sucesso do implante. Resultados: A taxa de sucesso total no período em questão foi de 96,65%. Na comparação entre pacientes com câncer e saudáveis, a taxa de sucesso foi de 93,02% no primeiro caso e 97,16% no segundo. Conclusão: De acordo com a revisão da literatura, nossos resultados encorajam a colocação de implantes em pacientes com câncer, é importante reconhecer que esta é uma análise complexa que requer cuidado devido ao grande número de variáveis. No entanto, os resultados não devem ser considerados de forma generalizada.
Assuntos
Humanos , Implantes Dentários , Neoplasias , Estudos Retrospectivos , Resultado do Tratamento , Prótese Dentária Fixada por Implante , Falha de Restauração Dentária , Implantação Dentária EndósseaRESUMO
In the present study, we characterized the effects of bath application of the proconvulsant drug 4-aminopyridine (4-AP) alone or in combination with GABAA and/or GABAB receptor antagonists, in cortical dysplasia (CD type I and CD type IIa/b), tuberous sclerosis complex (TSC), and non-CD cortical tissue samples from pediatric epilepsy surgery patients. Whole-cell patch clamp recordings in current and voltage clamp modes were obtained from cortical pyramidal neurons (CPNs), interneurons, and balloon/giant cells. In pyramidal neurons, bath application of 4-AP produced an increase in spontaneous synaptic activity as well as rhythmic membrane oscillations. In current clamp mode, these oscillations were generally depolarizing or biphasic and were accompanied by increased membrane conductance. In interneurons, membrane oscillations were consistently depolarizing and accompanied by bursts of action potentials. In a subset of balloon/giant cells from CD type IIb and TSC cases, respectively, 4-AP induced very low-amplitude, slow membrane oscillations that echoed the rhythmic oscillations from pyramidal neurons and interneurons. Bicuculline reduced the amplitude of membrane oscillations induced by 4-AP, indicating that they were mediated principally by GABAA receptors. 4-AP alone or in combination with bicuculline increased cortical excitability but did not induce seizure-like discharges. Ictal activity was observed in pyramidal neurons and interneurons from CD and TSC cases only when phaclofen, a GABAB receptor antagonist, was added to the 4-AP and bicuculline solution. These results emphasize the critical and permissive role of GABAB receptors in the transition to an ictal state in pediatric CD tissue and highlight the importance of these receptors as a potential therapeutic target in pediatric epilepsy.
RESUMO
Pathological high-frequency oscillations (HFOs), specifically fast ripples (FRs, >250â¯Hz), are pathognomonic of an active epileptogenic zone. However, the origin of FRs remains unknown. Here we explored the correlation between FRs recorded with intraoperative pre-resection electrocorticography (ECoG) and spontaneous synaptic activity recorded ex vivo from cortical tissue samples resected for the treatment of pharmacoresistant epilepsy. The cohort included 47 children (ages 0.22-9.99â¯yr) with focal cortical dysplasias (CD types I and II), tuberous sclerosis complex (TSC) and non-CD pathologies. Whole-cell patch clamp recordings were obtained from pyramidal neurons and interneurons in cortical regions that were positive or negative for pathological HFOs, defined as FR band oscillations (250-500â¯Hz) at ECoG. The frequency of spontaneous excitatory and inhibitory postsynaptic currents (sEPSCs and IPSCs, respectively) was compared between HFO+ and HFO- regions. Regardless of pathological substrate, regions positive for FRs displayed significantly increased frequencies of sIPSCs compared with regions negative for FRs. In contrast, the frequency of sEPSCs was similar in both regions. In about one third of cases (nâ¯=â¯17), pacemaker GABA synaptic activity (PGA) was observed. In the vast majority (nâ¯=â¯15), PGA occurred in HFO+ areas. Further, fast-spiking interneurons displayed signs of hyperexcitability exclusively in HFO+ areas. These results indicate that, in pediatric epilepsy patients, increased GABA synaptic activity is associated with interictal FRs in the epileptogenic zone and suggest an active role of GABAergic interneurons in the generation of pathological HFOs. Increased GABA synaptic activity could serve to dampen excessive excitability of cortical pyramidal neurons in the epileptogenic zone, but it could also promote neuronal network synchrony.
Assuntos
Epilepsia/fisiopatologia , Neurônios GABAérgicos/patologia , Interneurônios/patologia , Ondas Encefálicas/fisiologia , Criança , Pré-Escolar , Eletrocorticografia , Epilepsia/cirurgia , Feminino , Humanos , Lactente , Masculino , Sinapses/patologia , Ácido gama-Aminobutírico/metabolismoRESUMO
Huntington's disease (HD), an inherited neurodegenerative disorder that principally affects striatum and cerebral cortex, is generally thought to have an adult onset. However, a small percentage of cases develop symptoms before 20 years of age. This juvenile variant suggests that brain development may be altered in HD. Indeed, recent evidence supports an important role of normal huntingtin during embryonic brain development and mutations in this protein cause cortical abnormalities. Functional studies also demonstrated that the cerebral cortex becomes hyperexcitable with disease progression. In this review, we examine clinical and experimental evidence that cortical development is altered in HD. We also provide preliminary evidence that cortical pyramidal neurons from R6/2 mice, a model of juvenile HD, are hyperexcitable and display dysmorphic processes as early as postnatal day 7. Further, some symptomatic mice present with anatomical abnormalities reminiscent of human focal cortical dysplasia, which could explain the occurrence of epileptic seizures in this genetic mouse model and in children with juvenile HD. Finally, we discuss recent treatments aimed at correcting abnormal brain development.
Assuntos
Córtex Cerebral/crescimento & desenvolvimento , Córtex Cerebral/fisiopatologia , Excitabilidade Cortical , Doença de Huntington/fisiopatologia , Neurônios/fisiologia , Animais , Córtex Cerebral/patologia , Modelos Animais de Doenças , Humanos , Proteína Huntingtina/genética , Doença de Huntington/patologia , Camundongos Transgênicos , Neurônios/patologiaRESUMO
Neural stem/progenitor cell (NS/PC)-based therapies have shown exciting potential for regeneration of the central nervous system (CNS) and NS/PC cultures represent an important resource for disease modeling and drug screening. However, significant challenges limiting clinical translation remain, such as generating large numbers of cells required for model cultures or transplantation, maintaining physiologically representative phenotypes ex vivo and directing NS/PC differentiation into specific fates. Here, we report that culture of human NS/PCs in 3D, hyaluronic acid (HA)-rich biomaterial microenvironments increased differentiation toward oligodendrocytes and neurons over 2D cultures on laminin-coated glass. Moreover, NS/PCs in 3D culture exhibited a significant reduction in differentiation into reactive astrocytes. Many NS/PC-derived neurons in 3D, HA-based hydrogels expressed synaptophysin, indicating synapse formation, and displayed electrophysiological characteristics of immature neurons. While inclusion of integrin-binding, RGD peptides into hydrogels resulted in a modest increase in numbers of viable NS/PCs, no combination of laminin-derived, adhesive peptides affected differentiation outcomes. Notably, 3D cultures of differentiating NS/PCs were maintained for at least 70 days in medium with minimal growth factor supplementation. In sum, results demonstrate the use of 3D, HA-based biomaterials for long-term expansion and differentiation of NS/PCs toward oligodendroglial and neuronal fates, while inhibiting astroglial fates. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 704-718, 2019.
Assuntos
Técnicas de Cultura de Células , Ácido Hialurônico/química , Hidrogéis/química , Células-Tronco Neurais/metabolismo , Oligopeptídeos/química , Linhagem Celular , Humanos , Células-Tronco Neurais/citologiaRESUMO
The present study was designed to examine the potential cellular antiseizure mechanisms of everolimus, a mechanistic target of rapamycin (mTOR) pathway blocker, in pediatric epilepsy cases. Cortical tissue samples obtained from pediatric patients (n = 11, ages 0.67-6.75 years) undergoing surgical resections for the treatment of their pharmacoresistant epilepsy were examined electrophysiologically in ex vivo slices. The cohort included mTOR-mediated pathologies (tuberous sclerosis complex [TSC] and severe cortical dysplasia [CD]) as well as non-mTOR-mediated pathologies (tumor and perinatal infarct). Bath application of everolimus (2 µm) had practically no effect on spontaneous inhibitory postsynaptic activity. In contrast, long-term application of everolimus reduced spontaneous excitatory postsynaptic activity, burst discharges induced by blockade of γ-aminobutyric acid A (GABAA) receptors, and epileptiform activity generated by 4-aminopyridine, a K+ channel blocker. The antiseizure effects were more pronounced in TSC and CD cases, whereas in non-mTOR-mediated pathologies, the effects were subtle at best. These results support further clinical trials of everolimus in mTOR pathway-mediated pathologies and emphasize that the effects require sustained exposure over time.
RESUMO
The principal symptoms of Huntington's disease (HD), chorea, cognitive deficits, and psychiatric symptoms are associated with the massive loss of striatal and cortical projection neurons. As current drug therapies only partially alleviate symptoms, finding alternative treatments has become peremptory. Cell replacement using stem cells is a rapidly expanding field that offers such an alternative. In this review, we examine recent studies that use mesenchymal cells, as well as pluripotent, cell-derived products in animal models of HD. Additionally, we provide further electrophysiological characterization of a human neural stem cell line, ESI-017, which has already demonstrated disease-modifying properties in two mouse models of HD. Overall, the field of regenerative medicine represents a viable and promising avenue for the treatment of neurodegenerative disorders including HD.
Assuntos
Doença de Huntington/fisiopatologia , Doença de Huntington/terapia , Transplante de Células-Tronco , Animais , Modelos Animais de Doenças , Humanos , RoedoresRESUMO
Huntington's disease (HD) is an inherited neurodegenerative disorder with no disease-modifying treatment. Expansion of the glutamine-encoding repeat in the Huntingtin (HTT) gene causes broad effects that are a challenge for single treatment strategies. Strategies based on human stem cells offer a promising option. We evaluated efficacy of transplanting a good manufacturing practice (GMP)-grade human embryonic stem cell-derived neural stem cell (hNSC) line into striatum of HD modeled mice. In HD fragment model R6/2 mice, transplants improve motor deficits, rescue synaptic alterations, and are contacted by nerve terminals from mouse cells. Furthermore, implanted hNSCs are electrophysiologically active. hNSCs also improved motor and late-stage cognitive impairment in a second HD model, Q140 knockin mice. Disease-modifying activity is suggested by the reduction of aberrant accumulation of mutant HTT protein and expression of brain-derived neurotrophic factor (BDNF) in both models. These findings hold promise for future development of stem cell-based therapies.
Assuntos
Cognição , Doença de Huntington/terapia , Atividade Motora , Células-Tronco Neurais/transplante , Recuperação de Função Fisiológica , Animais , Linhagem Celular , Modelos Animais de Doenças , Xenoenxertos , Células-Tronco Embrionárias Humanas/metabolismo , Células-Tronco Embrionárias Humanas/patologia , Humanos , Doença de Huntington/metabolismo , Doença de Huntington/patologia , Doença de Huntington/fisiopatologia , Camundongos , Células-Tronco Neurais/metabolismo , Células-Tronco Neurais/patologiaRESUMO
Huntington's disease (HD) is a fatal genetic disorder characterized by cell death of medium-sized spiny neurons (MSNs) in the striatum, traditionally attributed to excessive glutamate inputs and/or receptor sensitivity. While changes in corticostriatal projections have typically been studied in mouse models of HD, morphological and functional alterations in thalamostriatal projections have received less attention. In this study, an adeno-associated virus expressing channelrhodopsin-2 under the calcium/calmodulin-dependent protein kinase IIα promoter was injected into the sensorimotor cortex or the thalamic centromedian-parafascicular nuclear complex in the R6/2 mouse model of HD, to permit selective activation of corticostriatal or thalamostriatal projections, respectively. In symptomatic R6/2 mice, peak amplitudes and areas of corticostriatal glutamate AMPA and NMDA receptor-mediated responses were reduced. In contrast, although peak amplitudes of AMPA and NMDA receptor-mediated thalamostriatal responses also were reduced, the areas remained unchanged due to an increase in response decay times. Blockade of glutamate reuptake further increased response areas and slowed rise and decay times of NMDA responses. These effects appeared more pronounced at thalamostriatal synapses of R6/2 mice, suggesting increased activation of extrasynaptic NMDA receptors. In addition, the probability of glutamate release was higher at thalamostriatal than corticostriatal synapses, particularly in R6/2 mice. Morphological studies indicated that the density of all excitatory synaptic contacts onto MSNs was reduced, which matches the basic electrophysiological findings of reduced amplitudes. There was a consistent reduction in the area of spines but little change in presynaptic terminal size, indicating that the postsynaptic spine may be more significantly affected than presynaptic terminals. These results highlight the significant and differential contribution of the thalamostriatal projection to glutamate excitotoxicity in HD.
Assuntos
Córtex Cerebral/fisiopatologia , Corpo Estriado/fisiopatologia , Doença de Huntington/fisiopatologia , Tálamo/fisiopatologia , Animais , Córtex Cerebral/patologia , Corpo Estriado/patologia , Modelos Animais de Doenças , Feminino , Ácido Glutâmico/metabolismo , Doença de Huntington/patologia , Imuno-Histoquímica , Masculino , Camundongos Transgênicos , Microscopia Eletrônica , Vias Neurais/patologia , Vias Neurais/fisiopatologia , Neurônios/patologia , Neurônios/fisiologia , Optogenética , Técnicas de Patch-Clamp , Receptores de AMPA/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Sinapses/patologia , Sinapses/fisiologia , Tálamo/patologia , Técnicas de Cultura de TecidosRESUMO
We tested the hypothesis that exposure of glut3+/- mice to a ketogenic diet ameliorates autism-like features, which include aberrant behavior and electrographic seizures. We first investigated the life course sex-specific changes in basal plasma-cerebrospinal fluid (CSF)-brain metabolic profile, brain glucose transport/uptake, glucose and monocarboxylate transporter proteins, and adenosine triphosphate (ATP) in the presence or absence of systemic insulin administration. Glut3+/- male but not female mice (5 months of age) displayed reduced CSF glucose/lactate concentrations with no change in brain Glut1, Mct2, glucose uptake or ATP. Exogenous insulin-induced hypoglycemia increased brain glucose uptake in glut3+/- males alone. Higher plasma-CSF ketones (ß-hydroxybutyrate) and lower brain Glut3 in females vs males proved protective in the former while enhancing vulnerability in the latter. As a consequence, increased synaptic proteins (neuroligin4 and SAPAP1) with spontaneous excitatory postsynaptic activity subsequently reduced hippocampal glucose content and increased brain amyloid ß1-40 deposition in an age-dependent manner in glut3+/- males but not females (4 to 24 months of age). We then explored the protective effect of a ketogenic diet on ultrasonic vocalization, sociability, spatial learning and memory, and electroencephalogram seizures in male mice (7 days to 6 to 8 months of age) alone. A ketogenic diet partially restored sociability without affecting perturbed vocalization, spatial learning and memory, and reduced seizure events. We conclude that (1) sex-specific and age-dependent perturbations underlie the phenotype of glut3+/- mice, and (2) a ketogenic diet ameliorates seizures caused by increased cortical excitation and improves sociability, but fails to rescue vocalization and cognitive deficits in glut3+/- male mice.
Assuntos
Comportamento Animal/fisiologia , Encéfalo/metabolismo , Dieta Cetogênica , Transportador de Glucose Tipo 3/metabolismo , Convulsões/dietoterapia , Comportamento Social , Animais , Encéfalo/fisiopatologia , Moléculas de Adesão Celular Neuronais/genética , Moléculas de Adesão Celular Neuronais/metabolismo , Eletroencefalografia , Feminino , Glucose/metabolismo , Transportador de Glucose Tipo 3/genética , Masculino , Memória/fisiologia , Camundongos , Camundongos Knockout , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , Convulsões/metabolismo , Convulsões/fisiopatologia , Fatores Sexuais , Aprendizagem Espacial/fisiologia , Vocalização Animal/fisiologiaRESUMO
Antecedentes: Chimó es el nombre en Venezuela de una sustancia viscosa de color negruzco, cuyo componente principal deriva de la hoja de tabaco. Su consumo se ha asociado a alteraciones sistémicas y patologías bucales. Objetivo: Identificar in vitro la erosión dental asociada a exposición al chimó. Métodos: Se seleccionaron 30 dientes, 10 control negativo, 10 control positivo y 10 grupo experimental que se expusieron a soluciones de saliva artificial, Coca-Cola® light y chimó respectivamente, durante 20 semanas. Se analizó clínicamente la superficie y el análisis ultraestructural se efectuó con microscopio electrónico de barrido. Resultados: Clínicamente, se evidenció cambio de color en el grupo control negativo; cambio de color y superficial y pérdida de brillo en el grupo control positivo; y cambio de color y superficial en el grupo experimental (p = 0,000). El análisis ultraestructural indica que el grupo control negativo no presentó alteraciones morfológicas en la superficie del esmalte. En la escala de valores de grabado ácido utilizada, el grupo control positivo fue tipo 4 y el grupo experimental mostró estructura adamantina erosionada con imágenes similares a los patrones de grabado tipo 3-4. En cuanto a la clasificación según su gravedad, el grupo experimental correspondió al grado 1, con pérdida de esmalte sin involucrar dentina. Conclusión: En las condiciones experimentales de este estudio, el chimó produjo pérdida de la superficie del esmalte tanto clínica como ultraestructuralmente, con patrones de erosión tipos 3 y 4 de la escala de valores del grabado ácido y grado 1 según su gravedad.
Background: In Venezuela, chimó is a blackish goo derived from tobacco leaf. Its consumption has been associated with systemic and oral diseases. Purpose: To identify in vitro dental erosion associated to chimó exposure. Methods: The sample consisted of 30 teeth that were assigned 10 to the negative control group, another 10 to the positive control group, and 10 to the experimental group, which were exposed respectively to artificial saliva, Coca-Cola® light, and chimó for 20 weeks. Tooth surfaces were analyzed clinically and structurally, the latter through scanning electron microscope. Results: Clinical observations showed surface color change in the negative control group; color change and surface gloss loss in the positive control group; color and surface changes in the experimental group (p = 0.000). Ultrastructural analysis showed no enamel surface alterations in the negative control group. Acid etching scale values were 4 for the positive control group and 3-4 for the experimental group. The latter had signs of enamel erosion. In terms of severity of damage, the experimental group was type 1, that is, enamel loss without involving dentin. Conclusion: Under these experimental conditions, chimó caused clinical and ultrastructural enamel surface loss with type 3-4 erosion patterns, and type 1 severity.
Assuntos
Humanos , Esmalte Dentário , Nicotiana , Erosão DentáriaRESUMO
INTRODUCTION: Smoking is one of the main public health problems in developed countries. Despite extensive evidence on the effects of smoking on both oral and general health, the rate of smoking cessation is not promising. MATERIAL AND METHODS: To review the evidence on knowledge and programs for smoking cessation developed by dentists, a literature review was carried out on programs for smoking cessation from the dentist's perspective, as well as a review of behavioral guidelines that have been recently proposed for these interventions. We used the keywords "Tobacco", "Smoking Prevention", "Public Health" AND "Dentistry", to identify controlled studies, systematic reviews and meta-analyses published between 1999 and 2014, in Google Scholar, SCOPUS and PubMed. RESULTS: Out of 177 studies found, 35 were considered, and these were divided into 2 groups of 20 and 15 articles respectively, according to type of study and inclusion criteria. CONCLUSIONS: There is considerable scientific evidence describing the programs for smoking cessation used in dentistry, which support their effectiveness. Overall, these are brief behavioral interventions complemented by pharmacological treatment, with the participation of the entire dental team. KEY WORDS: Dentistry, nicotine, smoking cessation, tobacco.
RESUMO
Our inability to adequately treat many patients with refractory epilepsy caused by focal cortical dysplasia (FCD), surgical inaccessibility and failures are significant clinical drawbacks. The targeting of physiologic features of epileptogenesis in FCD and colocalizing functionality has enhanced completeness of surgical resection, the main determinant of outcome. Electroencephalography (EEG)-functional magnetic resonance imaging (fMRI) and magnetoencephalography are helpful in guiding electrode implantation and surgical treatment, and high-frequency oscillations help defining the extent of the epileptogenic dysplasia. Ultra high-field MRI has a role in understanding the laminar organization of the cortex, and fluorodeoxyglucose-positron emission tomography (FDG-PET) is highly sensitive for detecting FCD in MRI-negative cases. Multimodal imaging is clinically valuable, either by improving the rate of postoperative seizure freedom or by reducing postoperative deficits. However, there is no level 1 evidence that it improves outcomes. Proof for a specific effect of antiepileptic drugs (AEDs) in FCD is lacking. Pathogenic mutations recently described in mammalian target of rapamycin (mTOR) genes in FCD have yielded important insights into novel treatment options with mTOR inhibitors, which might represent an example of personalized treatment of epilepsy based on the known mechanisms of disease. The ketogenic diet (KD) has been demonstrated to be particularly effective in children with epilepsy caused by structural abnormalities, especially FCD. It attenuates epigenetic chromatin modifications, a master regulator for gene expression and functional adaptation of the cell, thereby modifying disease progression. This could imply lasting benefit of dietary manipulation. Neurostimulation techniques have produced variable clinical outcomes in FCD. In widespread dysplasias, vagus nerve stimulation (VNS) has achieved responder rates >50%; however, the efficacy of noninvasive cranial nerve stimulation modalities such as transcutaneous VNS (tVNS) and noninvasive (nVNS) requires further study. Although review of current strategies underscores the serious shortcomings of treatment-resistant cases, initial evidence from novel approaches suggests that future success is possible.