RESUMO
The majority of biopsies in both basic research and translational cancer studies are preserved in the format of archived formalin-fixed paraffin-embedded (FFPE) samples. Profiling histone modifications in archived FFPE tissues is critically important to understand gene regulation in human disease. The required input for current genome-wide histone modification profiling studies from FFPE samples is either 10-20 tissue sections or whole tissue blocks, which prevents better resolved analyses. But it is desirable to consume a minimal amount of FFPE tissue sections in the analysis as clinical tissues of interest are limited. Here, we present FFPE tissue with antibody-guided chromatin tagmentation with sequencing (FACT-seq), the first highly sensitive method to efficiently profile histone modifications in FFPE tissues by combining a novel fusion protein of hyperactive Tn5 transposase and protein A (T7-pA-Tn5) transposition and T7 in vitro transcription. FACT-seq generates high-quality chromatin profiles from different histone modifications with low number of FFPE nuclei. We proved a very small piece of FFPE tissue section containing â¼4000 nuclei is sufficient to decode H3K27ac modifications with FACT-seq. H3K27ac FACT-seq revealed disease-specific super enhancers in the archived FFPE human colorectal and human glioblastoma cancer tissue. In summary, FACT-seq allows decoding the histone modifications in archival FFPE tissues with high sensitivity and help researchers to better understand epigenetic regulation in cancer and human disease.
Assuntos
Cromatina/metabolismo , Epigênese Genética , Histonas/análise , Animais , Linhagem Celular , Humanos , Camundongos , Processamento de Proteína Pós-Traducional , Proteína Estafilocócica A/metabolismo , Transposases/metabolismoRESUMO
PURPOSE: The diagnosis of polycystic ovary syndrome (PCOS) remains a challenge to clinicians due to heterogeneous clinical presentation and diagnostic criteria. This study investigated the utilization of Anti-Müllerian hormone (AMH) alone or replacing polycystic ovarian morphology (PCOM) in the PCOS diagnostic criteria. METHODS: A total of 401 women were categorised as PCOS (n:154), nonPCOS with polycystic ovarian morphology (PCOM) (n:105), and nonPCOS with normal ovarian morphology (NOM) (n:142). First, the diagnostic performance of AMH for PCOS diagnosis in Rotterdam, Androgen Excess Society, and National Institutes of Health (NIH) criteria was analyzed. Second, AMH was used instead of PCOM in Rotterdam criteria and we searched diagnostic performance for PCOS phenotypes. RESULTS: AMH levels were positively correlated with LH, testosterone, hirsutism score, menstrual cycle length, and antral follicle count (p < 0.05). AMH alone had specificity and sensitivity for PCOS diagnosis were 84.9% and 72.4% in Rotterdam (AUC: 0.866); 84.4% and 72% in Androgen Excess Society (AUC: 0.857); 83.3% and 66.4% in National Institute of Health criteria (AUC: 0.825). AMH alone had satisfactory diagnostic potential for phenotype A, but not other phenotypes. The replacement of PCOM with AMH in Rotterdam criteria had a high diagnostic potential for PCOS (AUC: 0.934, sensitivity:97.4%, specificity: 90.67%). Phenotype A and phenotype D were diagnosed with 100% sensitivity and 94.5% specificity. Phenotype C was recognised with 96.15% sensitivity and 94.5% specificity. CONCLUSION: AMH may be used with high diagnostic accuracy instead of PCOM in the Rotterdam PCOS criteria.
Assuntos
Síndrome do Ovário Policístico , Humanos , Feminino , Síndrome do Ovário Policístico/diagnóstico , Hormônio Antimülleriano , Androgênios , FenótipoRESUMO
PURPOSE: To investigate and provide objective documentation of the possible differences in the axonal reinnervation process of facial muscles after hypoglossal-facial nerve anastomosis. Then, to search for the presence of the trigemino-hypoglossal reflex and determine whether it indicates better peripheral recovery. METHODS: Electrophysiological examination performed on 20 patients who had undergone VII-XII anastomosis, with follow-up periods of more than 2 years. RESULTS: The mean follow-up time after surgery was 4.1 ± 1.3 years (range 2-8 years). The degrees of axonal reinnervation for the orbicularis oculi (OOc) and orbicularis oris (OOr) were 46.91 ± 19.77 and 32.65 ± 14.85, respectively. And the difference between these muscles was statistically significant (p = 0.018) in favor of the OOc. In addition, R1 blink reflexes that were not followed by R2 components were observed in 30% of the patients. However, these 6 patients with short-latency potential did not differ from the others in terms of latency, the amplitude of compound muscle action potential (CMAP), and degree of axonal reinnervation (p > 0.05) at both muscles (OOc and OOr). CONCLUSION: The recoveries of the lower face and upper face are different after VII-XII anastomosis, and in our patients the OOc healed better. In addition, R1 blink reflexes that were not followed by R2 components were observed in 30% of the patients. However, the patients with these blink reflexes did not have better peripheral healing in their neuromuscular units, which suggests that the blink reflex is not an indicator for peripheral recovery.
Assuntos
Nervo Facial , Paralisia Facial , Anastomose Cirúrgica , Músculos Faciais/cirurgia , Nervo Facial/cirurgia , Paralisia Facial/cirurgia , Humanos , Nervo Hipoglosso/cirurgia , ReflexoRESUMO
Resective epilepsy surgery has been accepted as an effective treatment for patients with medically intractable temporal lobe epilepsy (TLE) to control the seizures and to limit cognitive dysfunction. Complete resection of the epileptic zone, and therefore the success of the surgery, depends on the identification of the seizure focus. Reliable lateralizing semiologic signs, together with other presurgical assessments, are of great importance for an accurate identification of the seizure focus. In this respect, this study evaluated the frequency of semiologic signs in medically intractable temporal lobe epilepsy (TLE) together with the lateralizing values and variations according to the age and gender groups. Two hundred seventy-three seizures of 55 patients of the Adult Epilepsy Monitoring Unit of Gazi University Faculty of Medicine with the diagnosis of medically intractable TLE, whose epileptic foci were detected through noninvasive presurgical procedures and seizures were controlled successfully after anterior temporal lobectomy (ATL), were analyzed retrospectively. Seizure semiologies of the patients were evaluated in terms of lateralizing values, and it was inquired whether age/gender causes any variation. Versive head rotation, unilateral dystonic limb posturing, asymmetric tonic limb posturing, and the combination of unilateral hand automatisms and dystonic posturing were determined as the semiologic signs with the highest lateralizing values (90-100%). While hand automatisms were observed frequently in the group with early seizure-onset age (onset age ≤ 2), asymmetric tonic limb posturing was detected as more frequent in the group with later seizure-onset age (onset age > 2; p < .005). In addition to this, semiologic signs were noted to be different between male and female groups; psychic and autonomic auras and ictal emotional signs were associated with women (p < .005).
Assuntos
Epilepsia do Lobo Temporal/fisiopatologia , Lateralidade Funcional , Adolescente , Adulto , Idade de Início , Lobectomia Temporal Anterior , Sistema Nervoso Autônomo/fisiopatologia , Progressão da Doença , Epilepsia Resistente a Medicamentos/fisiopatologia , Epilepsia Resistente a Medicamentos/psicologia , Epilepsia Resistente a Medicamentos/cirurgia , Eletroencefalografia , Emoções , Epilepsia Generalizada/fisiopatologia , Epilepsia do Lobo Temporal/psicologia , Epilepsia do Lobo Temporal/cirurgia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Procedimentos Neurocirúrgicos , Postura , Estudos Retrospectivos , Caracteres Sexuais , Adulto JovemRESUMO
Despite the explanations of many lateralization findings, body turning in focal epilepsy has been rarely investigated. One of the aims of this study was to evaluate the role of ictal body turning in the lateralization of focal epilepsies. The records of 263 patients with focal epilepsy (temporal lobe epilepsy (TLE), n=178; extratemporal lobe epilepsy (ETLE), n=85) who underwent prolonged video-EEG monitoring during presurgical epilepsy evaluation were reviewed. Preoperative findings (TLE, n=16; ETLE, n=6) and postoperative outcomes (TLE, n=7) of patients with focal epilepsy with ictal body turning were assessed. For the evaluation of ictal body turning, two definitions were proposed. Nonversive body turning (NVBT) was used to denote at least a 90° nonforced (without tonic or clonic component) rotation of the upper (shoulder) and lower (hip) parts of the body around the body axis for a minimum of 3s. Versive body turning (VBT) was used to denote at least a 90° forced (with tonic or clonic component) rotation of the upper (shoulder) and lower (hip) parts of the body around the body axis for a minimum of 3s. Nonversive body turning was observed in 6% (n=11) of patients with TLE and 2% (n=2) of patients with ETLE. For VBT, these ratios were 5% (n=8) and 7% (n=6) for patients with TLE and ETLE, respectively. Nonversive body turning was frequently oriented to the same side as the epileptogenic zone (EZ) in TLE and ETLE seizures (76% and 80%, respectively). If the amount of NVBT was greater than 180°, then it was 80% to the same side in TLE seizures. Versive body turning was observed in 86% of the TLE seizures, and 55% of the ETLE seizures were found to be contralateral to the EZ. When present with head turning, NVBT ipsilateral to the EZ and VBT contralateral to the EZ were more valuable for lateralization. In TLE seizures, a significant correlation was found between the head turning and body turning onsets and durations. Our study demonstrated that ictal body turning is a rarely observed but reliable lateralization finding in TLE and ETLE seizures, which also probably has the same pathophysiological mechanism as head turning in TLE seizures.
Assuntos
Encéfalo/fisiopatologia , Epilepsia do Lobo Temporal/fisiopatologia , Lateralidade Funcional/fisiologia , Rotação , Convulsões/fisiopatologia , Adolescente , Adulto , Encéfalo/cirurgia , Eletroencefalografia , Epilepsias Parciais/fisiopatologia , Epilepsias Parciais/cirurgia , Epilepsia do Lobo Temporal/cirurgia , Feminino , Humanos , Masculino , Convulsões/cirurgia , Gravação em Vídeo , Adulto JovemRESUMO
When two sufficiently different stimuli are presented to each eye, perception alternates between them. This binocular rivalry is conceived as a competition for representation in the single stream of visual consciousness. The magnocellular (M) and parvocellular (P) pathways, originating in the retina, encode disparate information, but their potentially different contributions to binocular rivalry have not been determined. Here, we used functional magnetic resonance imaging to measure the human lateral geniculate nucleus (LGN), where the M and P neurons are segregated into layers receiving input from a single eye. We had three participants (one male, two females) and used achromatic stimuli to avoid contributions from color opponent neurons that may have confounded previous studies. We observed activity in the eye-specific regions of LGN correlated with perception, with similar magnitudes during rivalry or physical stimuli alternations, also similar in the M and P regions. These results suggest that LGN activity reflects our perceptions during binocular rivalry and is not simply an artifact of color opponency. Further, perception appears to be a global phenomenon in the LGN, not just limited to a single information channel.
Assuntos
Corpos Geniculados , Visão Binocular , Feminino , Humanos , Masculino , Visão Binocular/fisiologia , Corpos Geniculados/fisiologia , Retina , Neurônios , Estado de Consciência , Percepção Visual/fisiologia , Estimulação Luminosa/métodos , Vias Visuais/fisiologiaRESUMO
OBJECTIVES: Cardiac disturbances and rhythm abnormalities which potentially lead sudden unexpected death in epilepsy, have been extensively studied in focal epilepsies. However, studies including generalized epilepsies are scarce and it is not clear whether electrocardiogram parameters reflecting vulnerability to ventricular arrhythmias differ between these groups. METHODS: Medical records of patients who were followed in epilepsy department of a tertiary center between October 2015 and September 2016 were retrospectively reviewed. 66 generalized and 64 focal epilepsy patients with eligible electrophysiological data were analyzed. QTc interval, QTcd and other electrocardiographic indices were compared between patients with focal vs generalized epilepsy. Another analysis was performed in order to disclose any difference between patients with epilepsy (n:130) and psychogenic non-epileptic seizures. A two-tailed p valueâ¯<â¯0.05 was considered significant. RESULTS: There was no difference in terms of QTc and QTcd between patients with focal and generalized epilepsy [median: 406â¯ms vs 404â¯ms, pâ¯=â¯0.119; and median: 46â¯ms vs 44â¯ms, pâ¯=â¯0.497, respectively]. However patients with epilepsy were found to have longer QTc and QTcd when compared to ones with psychogenic non-epileptic seizures (pâ¯=â¯0.035 and pâ¯<â¯0.001, respectively). CONCLUSION: Current findings demonstrate that patients with epilepsy have longer QTc and QTcd than patients with pure psychogenic non-epileptic seizures. Since there was no difference between patients with focal and generalized epilepsy; QTc interval, QTcd and potential susceptibility to cardiac arrhythmias as a result, could be a consequence of epilepsy itself regardless of origin.
Assuntos
Epilepsias Parciais , Epilepsia Generalizada , Epilepsia , Arritmias Cardíacas/etiologia , Eletrocardiografia , Epilepsia/complicações , Humanos , Estudos Retrospectivos , ConvulsõesRESUMO
There is ample support for developmental regulation of glioblastoma stem cells. To examine how cell lineage controls glioblastoma stem cell function, we present a cross-species epigenome analysis of mouse and human glioblastoma stem cells. We analyze and compare the chromatin-accessibility landscape of nine mouse glioblastoma stem cell cultures of three defined origins and 60 patient-derived glioblastoma stem cell cultures by assay for transposase-accessible chromatin using sequencing. This separates the mouse cultures according to cell of origin and identifies three human glioblastoma stem cell clusters that show overlapping characteristics with each of the mouse groups, and a distribution along an axis of proneural to mesenchymal phenotypes. The epigenetic-based human glioblastoma stem cell clusters display distinct functional properties and can separate patient survival. Cross-species analyses reveals conserved epigenetic regulation of mouse and human glioblastoma stem cells. We conclude that epigenetic control of glioblastoma stem cells primarily is dictated by developmental origin which impacts clinically relevant glioblastoma stem cell properties and patient survival.
Assuntos
Glioblastoma , Linhagem da Célula/genética , Cromatina/genética , Epigênese Genética , Glioblastoma/genética , Humanos , Células-Tronco NeoplásicasRESUMO
Real-world scenes elicit anticipatory representation in long-term memory (LTM) and working memory (WM) resulting in boundary extension (BE). Would the same results hold for nonscene patterns of objects? In Experiment 1A (LTM-paradigm), 15 regular or 15 random object-patterns were sequentially presented (10 s each); a boundary-rating-task followed. Both pattern-types elicited BE. Surprisingly, regularity (greater pattern predictability) did not enhance BE. Instead, random patterns more consistently yielded BE. Perhaps random object-groupings enhance BE (e.g., more scene-like). Objects within regular patterns were colored to create object-groupings; this increased BE compared to regular patterns without color groups, but only in 1 of the 2 tests (Experiment 1B, LTM-paradigm). In Experiment 2 (WM-paradigm), a briefly presented pattern, regular or random, was followed by a 2-s masked interval and a boundary-rating-task on each trial. Here, results reversed: patterns now elicited boundary restriction (whether or not a concurrent task had been presented to prevent verbalization). Under time pressure, without real-world layout and meaning, pattern elements may not have cohered into a unified expanse, failing to elicit BE and resulting in loss of peripheral content. We propose 2 classes of visual content that may determine anticipatory representation: content that embellishes space versus content that defines space. (PsycInfo Database Record (c) 2020 APA, all rights reserved).
Assuntos
Antecipação Psicológica/fisiologia , Percepção de Cores/fisiologia , Reconhecimento Visual de Modelos/fisiologia , Desempenho Psicomotor/fisiologia , Percepção Espacial/fisiologia , Memória Espacial/fisiologia , Adolescente , Adulto , Feminino , Humanos , Masculino , Adulto JovemRESUMO
The visual system can efficiently summarize various lower-level and higher-level features of ensembles. However, no research to date has directly investigated how different features interact with each other within a single summary and whether people can efficiently report an integrated summary of two feature dimensions. In the first two experiments, we specifically investigated whether individuals can integrate spatial and size information to report a bound spatial summary, the center of mass (CoM), as efficiently as the centroid, which is devoid of size information. Both experiments revealed that viewers were equally accurate in extracting the centroid and the CoM, with the size distribution inadvertently affecting the centroid estimates. In the final experiment, we investigated whether encouraging observers to attend to individual item size would cause the centroid estimates to be biased toward the CoM. When item size was task-irrelevant, as in the centroid task, observers were able to selectively focus on spatial location, eliminating any impact from the size distribution. Taken together, these findings demonstrate that viewers are capable of extracting integrated summaries, possibly through a mechanism that allows them to represent the spatial distribution of sizes. We discuss possible mechanisms that may support the extraction of integrated summaries, and highlight the need to consider multilevel mechanisms extending beyond simple feature- and object-based mechanisms.
Assuntos
Percepção de Tamanho , Percepção Espacial , Adulto , Feminino , Humanos , Masculino , Estimulação Luminosa , Adulto JovemRESUMO
People represent summary statistics of visual scenes, but it is not fully clear whether such summary statistics are extracted automatically. To determine whether different levels of summary representation (i.e., at the perceptual-group or the entire-display level) may be formed differently, in two experiments we investigated how people extracted summary statistics for displays consisting of spatially segregated groups. Participants were asked to report the mean sizes of either entire sets or perceptual groups in precue and postcue conditions. There was no precueing advantage in the mean size estimations of entire sets. However, when these precues identified target perceptual groups, participants reported the perceptual-group means more accurately than when postcues were used. In the postcue condition, participants were biased toward the entire-set mean even when they were probed to report the perceptual-group mean. There was also greater bias toward the entire-set mean for more erroneous perceptual-group summaries. These findings suggest that ensemble representations are extracted more efficiently for the whole than for the perceptual parts and that ensemble perception is not a uniform process across perceptual groups and entire sets.