Trypanosoma cruzi infection changes the chromatin proteome profile of infected human cells.

Chagas disease is endemic in 22 Latin American countries, with approximately 8 million individuals infected worldwide and 10,000 deaths yearly. Trypanosoma cruzi presents an intracellular life cycle in mammalian hosts to sustain infection. Parasite infection activates host cell responses, promoting an unbalance in reactive oxygen species (ROS) in the intracellular environment inducing genomic DNA lesions in the host cell during infection. To further understand changes in host cell chromatin induced by parasite infection, we investigated alterations in chromatin caused by infection by performing quantitative proteomic analysis. DNA Damage Repair proteins, such as Poly-ADP-ribose Polymerase 1 (PARP-1) and X-Ray Repair Cross Complementing 6 (XRRC6), were recruited to the chromatin during infection. Also, changes in chromatin remodeling enzymes suggest that parasite infection may shape the epigenome of the host cells. Interestingly, the abundance of oxidative phosphorylation mitochondrial and vesicle-mediated transport proteins increased in the host chromatin at the final stages of infection. In addition, Apoptosis-inducing Factor (AIF) is translocated to the host cell nucleus upon infection, suggesting that cells enter parthanatos type of death. Altogether, this study reveals how parasites interfere with the host cells' responses at the chromatin level leading to significant crosstalk that support and disseminate infection. SIGNIFICANCE: The present study provides novel insights into the effects of Trypanosoma cruzi on the chromatin from the host cell. This manuscript investigated proteomic alterations in chromatin caused by parasite infection at early and late infection phases by performing a quantitative proteomic analysis. In this study, we revealed that parasites interfere with DNA metabolism in the early and late stages of infection. We identified that proteins related to DNA damage repair, oxidative phosphorylation, and vesicle-mediated transport have increased abundance at the host chromatin. Additionally, we have observed that Apoptosis-inducing Factor is translocated to the host cell nucleus upon infection, suggesting that the parasites could lead the cells to enter Parthanatos as a form of programmed cell death. The findings improve our understanding on how the parasites modulate the host cell chromatin to disseminate infection. In this study, we suggest a mechanistic parasite action towards host nucleus that could be used to indicate targets for future treatments.

Heart rate variability in patients with refractory epilepsy: The influence of generalized convulsive seizures.

OBJECTIVE: Patients with epilepsy, mainly drug-resistant, have reduced heart rate variability (HRV), linked to an increased risk of sudden death in various other diseases. In this context, it could play a role in SUDEP. Generalized convulsive seizures (GCS) are one of the most consensual risk factors for SUDEP. Our objective was to assess the influence of GCS in HRV parameters in patients with drug-resistant epilepsy. METHODS: We prospectively evaluated 121 patients with refractory epilepsy admitted to our Epilepsy Monitoring Unit. All patients underwent a 48-hour Holter recording. Only patients with GCS were included (n = 23), and we selected the first as the index seizure. We evaluated HRV (AVNN, SDNN, RMSSD, pNN50, LF, HF, and LF/HF) in 5-min epochs (diurnal and nocturnal baselines; preictal - 5 min before the seizure; ictal; postictal - 5 min after the seizure; and late postictal - >5 h after the seizure). These data were also compared with normative values from a healthy population (controlling for age and gender). RESULTS: We included 23 patients, with a median age of 36 (min-max, 16-55) years and 65% were female. Thirty percent had cardiovascular risk factors, but no previously known cardiac disease. HRV parameters AVNN, RMSSD, pNN50, and HF were significantly lower in the diurnal than in the nocturnal baseline, whereas the opposite occurred with LF/HF and HR. Diurnal baseline parameters were inferior to the normative population values (which includes only diurnal values). We found significant differences in HRV parameters between the analyzed periods, especially during the postictal period. All parameters but LF/HF suffered a reduction in that period. LF/HF increased in that period but did not reach statistical significance. Visually, there was a tendency for a global reduction in our patients' HRV parameters, namely AVNN, RMSSD, and pNN50, in each period, comparing with those from a normative healthy population. No significant differences were found in HRV between diurnal and nocturnal seizures, between temporal lobe and extra-temporal-lobe seizures, between seizures with and without postictal generalized EEG suppression, or between seizures of patients with and without cardiovascular risk factors. SIGNIFICANCE/CONCLUSION: Our work reinforces the evidence of autonomic cardiac dysfunction in patients with refractory epilepsy, at baseline and mainly in the postictal phase of a GCS. Those changes may have a role in some SUDEP cases. By identifying patients with worse autonomic cardiac function, HRV could fill the gap of a lacking SUDEP risk biomarker.

PI3K-AKT, JAK2-STAT3 pathways and cell-cell contact regulate maspin subcellular localization.

BACKGROUND: Maspin (SERPINB5) is a potential tumor suppressor gene with pleiotropic biological activities, including regulation of cell proliferation, death, adhesion, migration and gene expression. Several studies indicate that nuclear localization is essential for maspin tumor suppression activity. We have previously shown that the EGFR activation leads to maspin nuclear localization in MCF-10A cells. The present study investigated which EGFR downstream signaling molecules are involved in maspin nuclear localization and explored a possible role of cell-cell contact in this process. METHODS: MCF-10A cells were treated with pharmacological inhibitors against EGFR downstream pathways followed by EGF treatment. Maspin subcellular localization was determined by immunofluorescence. Proteomic and interactome analyses were conducted to identify maspin-binding proteins in EGF-treated cells only. To investigate the role of cell-cell contact these cells were either treated with chelating agents or plated on different cell densities. Maspin and E-cadherin subcellular localization was determined by immunofluorescence. RESULTS: We found that PI3K-Akt and JAK2-STAT3, but not MAP kinase pathway, regulate EGF-induced maspin nuclear accumulation in MCF-10A cells. We observed that maspin is predominantly nuclear in sparse cell culture, but it is redistributed to the cytoplasm in confluent cells even in the presence of EGF. Proteomic and interactome results suggest a role of maspin on post-transcriptional and translation regulation, protein folding and cell-cell adhesion. CONCLUSIONS: Maspin nuclear accumulation is determined by an interplay between EGFR (via PI3K-Akt and JAK2-STAT3 pathways) and cell-cell contact. Video Abstract.

Ocular involvement in systemic lupus erythematosus patients: a paradigm shift based on the experience of a tertiary referral center.

OBJECTIVE: To evaluate ocular involvement in a cohort of systemic lupus erythematosus (SLE) patients of a tertiary referral center and to compare the results with the existing literature. METHODS: Patients underwent a complete ophthalmological evaluation, including visual acuity, slit-lamp examination, fluorescein staining, Schirmer-I test, Goldmann applanation tonometry, fundoscopy, 10-2 automated threshold visual fields, fundus autofluorescence and spectral-domain optical coherence tomography to screen for hydroxychloroquine (HCQ) macular toxicity. RESULTS: A total of 161 patients (16 men and 145 women) were enrolled in this study. The mean age was 47.6 years and the mean disease duration was 11.5 years. Fifty patients (31.1%) had at least one ocular manifestation of SLE. The most frequent manifestation was dry eye syndrome (12.4%), immediately followed by cataracts (11.2%) and HCQ macular toxicity (11.2%). Among patients with HCQ maculopathy, two presented with an atypical spectral-domain optical coherence tomography pattern. Five patients (3.1%) presented with glaucoma, two patients (1.2%) presented with SLE retinopathy while only one presented with lupus choroidopathy (0.6%). CONCLUSIONS: Compared with previous studies, we conclude there has been a significant reduction in disease-related ocular complications, particularly those associated with poor systemic disease control. On the other hand, drug and age-related complications are assuming a prominent role in the ophthalmic care of these patients.

Quantitative and qualitative analysis of ictal vocalization in focal epilepsy syndromes.

PURPOSE: To investigate the frequency, localizing significance, and intensity characteristics of ictal vocalization in different focal epilepsy syndromes. METHODS: Up to four consecutive focal seizures were evaluated in 277 patients with lesional focal epilepsy, excluding isolated auras and subclinical EEG seizure patterns. Vocalization was considered to be present if observed in at least one of the analyzed seizures and not being of speech quality. Intensity features of ictal vocalization were analyzed in a subsample of 17 patients with temporal and 19 with extratemporal epilepsy syndrome. RESULTS: Ictal vocalization was observed in 37% of the patients (102/277) with similar frequency amongst different focal epilepsy syndromes. Localizing significance was found for its co-occurrence with ictal automatisms, which identified patients with temporal seizure onset with a sensitivity of 92% and specificity of 70%. Quantitative analysis of vocalization intensity allowed to distinguish seizures of frontal from temporal lobe origin based on the intensity range (p = 0.0003), intensity variation (p < 0.0001), as well as the intensity increase rate at the beginning of the vocalization (p = 0.003), which were significantly higher in frontal lobe seizures. No significant difference was found for mean intensity and mean vocalization duration. CONCLUSIONS: Although ictal vocalization is similarly common in different focal epilepsies, it shows localizing significance when taken into account the co-occurring seizure semiology. It especially increases the localizing value of automatisms, predicting a temporal seizure onset with a sensitivity of 92% and specificity of 70%. Quantitative parameters of the intensity dynamic objectively distinguished frontal lobe seizures, establishing an observer independent tool for semiological seizure evaluation.

Alzheimer's disease: A review of its visual system neuropathology. Optical coherence tomography-a potential role as a study tool in vivo.

Alzheimer's disease (AD) is a prevalent, long-term progressive degenerative disorder with great social impact. It is currently thought that, in addition to neurodegeneration, vascular changes also play a role in the pathophysiology of the disease. Visual symptoms are frequent and are an early clinical manifestation; a number of psychophysiologic changes occur in visual function, including visual field defects, abnormal contrast sensitivity, abnormalities in color vision, depth perception deficits, and motion detection abnormalities. These visual changes were initially believed to be solely due to neurodegeneration in the posterior visual pathway. However, evidence from pathology studies in both animal models of AD and humans has demonstrated that neurodegeneration also takes place in the anterior visual pathway, with involvement of the retinal ganglion cells' (RGCs) dendrites, somata, and axons in the optic nerve. These studies additionally showed that patients with AD have changes in retinal and choroidal microvasculature. Pathology findings have been corroborated in in-vivo assessment of the retina and optic nerve head (ONH), as well as the retinal and choroidal vasculature. Optical coherence tomography (OCT) in particular has shown great utility in the assessment of these changes, and it may become a useful tool for early detection and monitoring disease progression in AD. The authors make a review of the current understanding of retinal and choroidal pathological changes in patients with AD, with particular focus on in-vivo evidence of retinal and choroidal neurodegenerative and microvascular changes using OCT technology.

SenseMyHeart: A cloud service and API for wearable heart monitors.

In the era of ubiquitous computing, the growing adoption of wearable systems and body sensor networks is trailing the path for new research and software for cardiovascular intensity, energy expenditure and stress and fatigue detection through cardiovascular monitoring. Several systems have received clinical-certification and provide huge amounts of reliable heart-related data in a continuous basis. PhysioNet provides equally reliable open-source software tools for ECG processing and analysis that can be combined with these devices. However, this software remains difficult to use in a mobile environment and for researchers unfamiliar with Linux-based systems. In the present paper we present an approach that aims at tackling these limitations by developing a cloud service that provides an API for a PhysioNet-based pipeline for ECG processing and Heart Rate Variability measurement. We describe the proposed solution, along with its advantages and tradeoffs. We also present some client tools (windows and Android) and several projects where the developed cloud service has been used successfully as a standard for Heart Rate and Heart Rate Variability studies in different scenarios.

Deletion of C/EBP homologous protein (Chop) in C57Bl/6 mice dissociates obesity from insulin resistance.

AIMS/HYPOTHESIS: Endoplasmic reticulum (ER) stress has been implicated in the development of type 2 diabetes, via effects on obesity, insulin resistance and pancreatic beta cell health. C/EBP homologous protein (CHOP) is induced by ER stress and has a central role in apoptotic execution pathways triggered by ER stress. The aim of this study was to characterise the role of CHOP in obesity and insulin resistance. METHODS: Metabolic studies were performed in Chop ( -/- ) and wild-type C57Bl/6 mice, and included euglycaemic-hyperinsulinaemic clamps and indirect calorimetry. The inflammatory state of liver and adipose tissue was determined by quantitative RT-PCR, immunohistology and macrophage cultures. Viability and absence of ER stress in islets of Langerhans was determined by electron microscopy, islet culture and quantitative RT-PCR. RESULTS: Systemic deletion of Chop induced abdominal obesity and hepatic steatosis. Despite marked obesity, Chop ( -/- ) mice had preserved normal glucose tolerance and insulin sensitivity. This discrepancy was accompanied by lower levels of pro-inflammatory cytokines and less infiltration of immune cells into fat and liver. CONCLUSIONS/INTERPRETATION: These observations suggest that insulin resistance is not induced by fat accumulation per se, but rather by the inflammation induced by ectopic fat. CHOP may play a key role in the crosstalk between excessive fat deposition and induction of inflammation-mediated insulin resistance.

Dysprosody during epileptic seizures lateralizes to the nondominant hemisphere.

OBJECTIVE: In human speech, the changes in intonation, rhythm, or stress reflect emotions or intentions and are called prosody. Dysprosody is the impairment of prosody and has been described in stroke and neurodegenerative disorders. Reports in epilepsy patients are limited to case reports. METHODS: We assessed prosody qualitatively and quantitatively in 967 focal epilepsy patients. The qualitative assessment was performed by 2 native German speakers, and the quantitative frequency analysis used linguistic software tools. For the quantitative analysis, the formant F0 (a frequency peak, which is an approximation of pitch) and the further spectral frequency peaks of our patients' voices were analyzed. RESULTS: We found 26 patients with ictal dysprosody through qualitative analysis (2.7% of all focal epilepsies). The qualitative changes affected mostly the pitch and the loss of melody. The seizure patterns at the time of ictal dysprosody were always in the nondominant hemisphere (100%) and were mostly right temporal (n = 22; 84.6%). Quantitative analysis of 15 audio samples (11 patients) showed a change in the frequency of formant F0 of several patients and a reduction of frequency variation during ictal speech, expressed as the SD of formant F0 (ictal 14.1 vs interictal 27.2). CONCLUSIONS: Ictal dysprosody localizes seizure onset or propagation to the nondominant temporal lobe. This information can be used in the evaluation of patients considered for resective epilepsy surgery.

Associating ECG features with firefighter's activities.

In this paper we associate features obtained from ECG signals with the expected levels of stress of real firefighters in action when facing specific events such as fires or car accidents. Five firefighters were monitored using wearable technology collecting ECG signals. Heart rate and heart rate variability features were analyzed in consecutive 5-min intervals during several types of events. A questionnaire was used to rank these types of events according to stress and fatigue and a measure of association was applied to compare this ranking to the ECG features. Results indicate associations between this ranking and both heart rate and heart rate variability features extracted in the time domain. Finally, an example of differences in inter personal responses to stressful events is shown and discussed, motivating future challenges within this research field.

Compressed domain topographic classification for capsule endoscopy.

In this paper we compare the classification accuracy of using compressed domain color (CDC) descriptors versus traditional full decoded images, for the purposes of topographic classification of wireless capsule endoscopy images. Results using a dataset of 26469 images, divided into stomach, small intestine and large intestine show a difference in classification accuracy below 1%. We also show that errors are mostly located near zone transitions (the pylorus and the ileocecal valve) and motivate the need for other visual descriptors (e.g. shape, motion) for addressing these specific areas. We conclude we can use the advantages of CDC in this type of classification with minor accuracy sacrifice.

Bioinformatics construction of the human cell surfaceome.

Cell surface proteins are excellent targets for diagnostic and therapeutic interventions. By using bioinformatics tools, we generated a catalog of 3,702 transmembrane proteins located at the surface of human cells (human cell surfaceome). We explored the genetic diversity of the human cell surfaceome at different levels, including the distribution of polymorphisms, conservation among eukaryotic species, and patterns of gene expression. By integrating expression information from a variety of sources, we were able to identify surfaceome genes with a restricted expression in normal tissues and/or differential expression in tumors, important characteristics for putative tumor targets. A high-throughput and efficient quantitative real-time PCR approach was used to validate 593 surfaceome genes selected on the basis of their expression pattern in normal and tumor samples. A number of candidates were identified as potential diagnostic and therapeutic targets for colorectal tumors and glioblastoma. Several candidate genes were also identified as coding for cell surface cancer/testis antigens. The human cell surfaceome will serve as a reference for further studies aimed at characterizing tumor targets at the surface of human cells.

A novel dry active electrode for EEG recording.

The design and testing of a "dry" active electrode for electroencephalographic recording is described. A comparative study between the EEG signals recorded in human volunteers simultaneously with the classical Ag-AgCl and "dry" active electrodes was carried out and the reported preliminary results are consistent with a better performance of these devices over the conventional Ag-AgCl electrodes.

A new and fast nonlinear method for association analysis of biosignals.

In this paper, we present some original theoretical aspects of a fast nonlinear association measure based on the work of Cramér. The features of this new measure--the V measure--when applied to biosignals are also shown using simulated time series. A comparative study with other well-known association measures available in the literature of biosignals is presented. V was found to be twice as fast and more robust to nonlinearities than the classical cross-correlation ratio (r2) and more than 100 times faster than the nonlinear regression coefficient (h2), presenting similar behavior in the presence of nonlinear simulated situations. This new measure is very fast and versatile. It is appropriate to deal with nonlinear relations presenting usually a sharp peak in the association function enabling a high degree of selectivity for maxima detection. It seems to constitute an improvement over linear methods of association which is faster and more robust to the existing nonlinearities. It can be used as an alternative to more complex nonlinear association measures when computational speed is an important feature.

SIGIF: a digital signal interchange format with application in neurophysiology.

In this paper, the authors describe a biomedical digital-signal interchange format. The format supports both raw and processed data, multiple segments, several signal structures and representations, and an open architecture. Its versatility and adaptability allows the software to take advantage of any particular features of the acquisition hardware. The format has been used and improved in routine work during a five-year period involving the cooperation between two hospitals and one engineering research center. In order to support the format, an object oriented C language library has been developed and is also shortly described.

Scalp EEG recording: interictal/ictal location and spreading of epileptiform events.

The relevance of scalp EEG recording on the selection of patients for epilepsy surgery usually is based on the concordance between the location of the epileptogenic focus and the presumed ictal origin of clinical seizures visually identified or video recorded. Bilateralisation and/or spreading of the epileptiform events are among the causes of the relative lack of agreement between scalp topography and origin of EEG potentials. Computer methods mainly if they are adaptive and use multistrategic approaches improve the accuracy on the detection of the epileptogenic focus, extracting relevant information on the location and spreading of epileptiform events.

The role of computer based techniques in patient monitoring: technical note.

In this paper the requirements of Neurophysiology and Neurotraumatology monitoring are analyzed. As a result a set of designated systems were developed by the authors a short description of which is given in the paper. Finally the future perspectives and problems to be faced are briefly described.