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Microesferas de quitosana podem ser empregadas na área de biomaterias, em processos biotecnológicos e como adsorventes. Neste trabalho, foi empregada a técnica de atomização e coagulação para produção dessas microesferas, que permitiu o controle dos parâmetros de operação e por conseqüência a obtenção de microesferas de tamanho e faixas de tamanho específicos. Após a sua obtenção, as microesferas foram modificadas quimicamente com objetivo de estudar as resistências térmica, mecânica e química. Para isso foram empregadas três rotas distintas: a-) reticulação com glutaraldeído; b-) reticulação com epicloridrina e c-) acetilação. As microesferas preparadas apresentaram distribuição de tamanho da ordem de 140 µm com desvio padrão de 11,9 µm. Após as modificações químicas, as microesferas apresentaram temperatura de degradação térmica em torno de 300 ºC, aumento da estabilidade química à solução de HCl, e diminuição da resistência mecânica.Chitosan microspheres can be used as biomaterial, in biotechnology processes and as adsorbents. This work is concerned with the production of chitosan microspheres using spraying and coagulation processes, which allows us to control the operating parameters and to produce chitosan microspheres of several ranges and sizes. The microspheres were modified chemically in order to study their thermal, mechanical and chemical resistance. The methods used were: 1) crosslinking with glutaraldehyde; 2) crosslinking with epichlorohydrin; 3) acetylation. The microspheres obtained presented mean particle size of 140 µm and standard deviation of 11.9 µm. The modified microspheres showed thermal degradation around 300 ºC, an increase of chemical stability using HCl solution and a decrease of mechanical resistance.

Neste trabalho foi realizado um estudo de avaliação da degradação "in vitro" da blenda biorreabsorvível poli(beta-hidroxibutirato) (PHB) / poli(L-ácido lático) (PLLA). As blendas PHB/PLLA na forma de filmes, nas composições 0/100, 30/70, 50/50, 70/30 e 100/0 (% em massa), foram obtidas pelo método de evaporação do solvente. Os filmes obtidos foram imersos em solução tampão fosfato salino, PBS, (pH 7,4) por 12 meses em banho termostatizado à 37 °C. As amostras foram caracterizadas por MDSC, TG, DMA, SEM, WAXS e SAXS, antes e após o processo de degradação. Os resultados mostraram que a blenda PHB/PLLA é imiscível em todas as composições estudadas, apresentando morfologia com separação de fases. Observou-se que o PLLA perdeu propriedades mecânicas de tração mais rapidamente que o PHB, durante o processo de degradação, tornando-se mais quebradiço. Verificou-se também aumento no grau de cristalinidade e na espessura do período longo das cadeias poliméricas nas blendas em função da degradação, sugerindo aumento da lamela cristalina.In this work, "in vitro" degradation of poly(beta-hydroxybutyrate) (PHB) / poly(L-lactic acid) (PLLA) blends was evaluated. Films of PHB/PLLA blends, in the compositions 0/100, 30/70, 50/50, 70/30 and 100/0 (% weight), were fabricated by casting. The films obtained were immersed in a phosphate buffer solution, PBS (pH 7.4) for 12 months. The samples were characterized by MDSC, TG, DMA, SEM, WAXS and SAXS, before and after the degradation process. The results showed that the PHB/PLLA blends are immiscible and present phase separation as indicated by microscopy. The PLLA loses mechanical properties faster than PHB, during the degradation process. It was observed that the blends increased their crystallinity degree and long period thickness as a function of degradation time.

The influence of the calcification solution on the mineralization of chitosan membranes was analysed. Two factors were studied: pH and chemical composition (excess of calcium or phosphate ions) of solutions. Substrates (membranes) of chitosan underwent mineralization by soaking in calcification solutions for 7 days at 36.5 °C. As expected, SEM-EDS analyses demonstrated that higher pH and concentration of ions increases deposition, however, the morphology and composition of these deposits varies depending on which ion (phosphate or calcium) is in excess. Larger spherical deposits are observed when a calcium-rich solution is used instead of a phosphate-rich one. In the latter, the interconnectivity of the inorganic and organic (matrix) phases is very different, suggesting a different mechanism of deposition for each situation.

O presente trabalho trata da preparação e caracterização de membranas de quitosana porosas e densas, que possuem diversas aplicações tecnológicas. Para tanto, foram utilizadas as técnicas de microscopia (microscopia eletrônica de varredura - MEV - e microscopia de força atômica -MFA), titulação potenciométrica e espectroscopia no infravermelho (FTIR-ATR). A síntese das estruturas foi realizada através da secagem total ou parcial de uma solução acética de quitosana a 2,5% (w/w) seguida de coagulação em meio alcalino. Glutaraldeído foi utilizado para modificar quimicamente as membranas, tornando-as mais hidrofóbicas. A extensão deste processo pôde ser observada por FTIR-ATR, utilizando-se um plano fatorial onde foram variados o tempo de fixação e a concentração de glutaraldeído na solução de tratamento.

The secnidazole, a drug used in hepatic amebiasis, giardiasis and bacterial vaginosis, was enantioseparated on a semi-preparative scale by high performance liquid chromatography (HPLC), using chiral stationary phase (CSP) containing amylose tris (3, 5-dimethylphenyl) carbamate on silica gel (Chiralpak® AD). The experiments were carried out under normal phase conditions and the separation method exhibited very high selectivity and resolutions. The chromatographic parameters and the effects of column temperature on the retention time and selectivity were evaluated. Thermodynamic parameters were obtained from van't Hoff plots, and the elution order of the enantiomers was determined by circular dichorism. The enantiomers of secnidazole were separated under overloaded conditions with a good performance.

The influence of temperature on the equilibrium and mass-transfer parameters in the chromatographic separation of bupivacaine enantiomers was evaluated using a chiral stationary phase (CSP) based on diallyl-L-tartardiamide (Kromasil® CHI-TBB). The total and bed porosities as well as the linear equilibrium constant were obtained by pulse experiments using dilute solutions of inert compounds and bupivacaine enantiomers at different flow rates (1.0, 2.0, 3.0 and 4.0 m l /min) and temperatures (25, 35 and 45°C). The axial dispersion and overall mass-transfer coefficients were evaluated by van Deemter plot. A decrease in the linear equilibrium adsorption constants with increasing temperature was observed for both enantiomers. This chiral column showed good separation performance with a high order of magnitude for the overall mass-transfer coefficient. These values increased with increasing temperature. However, it was not possible to differentiate between these values at different temperatures due to poor standard deviations. Because of the high order of magnitude of the overall mass-transfer coefficient associated with a good resolution of bupivacaine enantiomers, it was concluded that convection effects dominated chiral separation in this column.

Isolation and purification of bioproducts from crude extracts can be obtained by affinity methods based on reversible binding of a specific molecule to ligand immobilized in a porous matrix. In the present work, nicrospheres based on chitosan matrix, which incorporated aminophenylboronic acid as a derivative, were prepared and characterized, aimed at developing a β-amylase adsorption process. Kinetic curves and adsorption isotheriru of the crude extracts as well as the breakthrough curves for a frontal chromatographic separation method of a commercial sample of β-amylase from soybean are presented. These results were compared to similar data obtained with a comercial microspheres gel based-on agarose. [ABSTRACT FROM AUTHOR]

The recovery and purification of the desired products from bioreactor outflow is greatly influenced by the nature of the products themselves, the size of the market, the need to achieve market and legally required specification standards; and the market value of the product. As the scale increases, economic methods of conducting product recovery and purification become not only important but essential for competitive process integration. It is therefore worthwhile to conceive a proper downstream scheme early in the process development, which requires laboratory development and process engineering efforts. Even small improvements can be very profitable in terms of the overall process. [ABSTRACT FROM AUTHOR]

Traumatic brain injury (TBI) is the leading cause of morbidity and mortality worldwide. Multiple injury models have been developed to study this neurological disorder. One such model is the lateral fluid percussion injury (LFPI) rodent model. The LFPI model can be generated with different surgical procedures that could affect the injury and be reflected in neurobehavioral dysfunction and acute electroencephalograph (EEG) changes. A craniectomy was performed either with a trephine hand drill or with a trephine electric drill that was centered over the left hemisphere of adult, male Sprague Dawley rats. Sham craniectomy groups were assessed by hand-drilled (Sham HMRI ) and electric-drilled (Sham EMRI ) to evaluate by magnetic resonance imaging (MRI). Then, TBI was induced in separate groups, (TBI H ) and (TBI E ), using a fluid-percussion device. Sham-injured rats (Sham H /Sham E ) underwent the same surgical procedures as the TBI rats. During the same surgery session, rats were implanted with screw and microwire electrodes positioned in the neocortex and hippocampus and the EEG activity was recorded 24 h for the first 7 days after TBI for assessing the acute EEG seizure and gamma event coupling. The electric drilling craniectomy induced greater tissue damage and sensorimotor deficits compared with the hand drill. Analysis of the EEG revealed acute seizures in at least one animal from each group after the procedure. Both TBI and Sham rats from the electric drill groups had a significant greater total number of seizures than the animals that were craniectomized manually ( p < 0.05). Similarly, EEG functional connectivity was lower in Sham E compared with Sham H rats. These results suggest that electrical versus hand-drilling craniectomies produce cortical injury in addition to the LFPI which increases the likelihood for acute post-traumatic seizures. Differences in the surgical approach could be one reason for the variability in the injury that makes it difficult to replicate results between preclinical TBI studies., (© The Author(s) 2024. Published by Mary Ann Liebert, Inc.)

Traumatic brain injury (TBI) is the leading cause of morbidity and mortality worldwide. Multiple injury models have been developed to study this neurological disorder. One such model is the lateral fluid-percussion injury (LFPI) rodent model. The LFPI model can be generated with different surgical procedures that could affect the injury and be reflected in neurobehavioral dysfunction and acute EEG changes. A craniectomy was performed either with a trephine hand drill or with a trephine electric drill that was centered over the left hemisphere of adult, male Sprague Dawley rats. Sham craniectomy groups were assessed by hand-drilled (ShamHMRI) and electric-drilled (ShamEMRI) to evaluate by MRI. Then, TBI was induced in separate groups (TBIH) and (TBIE) using a fluid-percussion device. Sham-injured rats (ShamH/ShamE) underwent the same surgical procedures as the TBI rats. During the same surgery session, rats were implanted with screw and microwire electrodes positioned in the neocortex and hippocampus and the EEG activity was recorded 24 hours for the first 7 days after TBI for assessing the acute EEG seizure and Gamma Event Coupling (GEC). The electric drilling craniectomy induced greater tissue damage and sensorimotor deficits compared to the hand drill. Analysis of the EEG revealed acute seizures in at least one animal from each group after the procedure. Both TBI and Sham rats from the electric drill groups had a significant greater total number of seizures than the animals that were craniectomized manually (p<0.05). Similarly, EEG functional connectivity was lower in ShamE compared to ShamH rats. These results suggest that electrical versus hand drilling craniectomies produce cortical injury in addition to the LFPI which increases the likelihood for acute post-traumatic seizures. Differences in the surgical approach could be one reason for the variability in the injury that makes it difficult to replicate results between preclinical TBI studies.

Functional connectivity (FC) after TBI is affected by an altered excitatory-inhibitory balance due to neuronal dysfunction, and the mechanistic changes observed could be reflected differently by contrasting methods. Local gamma event coupling FC (GEC-FC) is believed to represent multiunit fluctuations due to inhibitory dysfunction, and we hypothesized that FC derived from widespread, broadband amplitude signal (BBA-FC) would be different, reflecting broader mechanisms of functional disconnection. We tested this during sleep and active periods defined by high delta and theta EEG activity, respectively, at 1,7 and 28d after rat fluid-percussion-injury (FPI) or sham injury (n=6/group) using 10 indwelling, bilateral cortical and hippocampal electrodes. We also measured seizure and high-frequency oscillatory activity (HFOs) as markers of electrophysiological burden. BBA-FC analysis showed early hyperconnectivity constrained to ipsilateral sensory-cortex-to-CA1-hippocampus that transformed to mainly ipsilateral FC deficits by 28d compared to shams. These changes were conserved over active epochs, except at 28d when there were no differences to shams. In comparison, GEC-FC analysis showed large regions of hyperconnectivity early after injury within similar ipsilateral and intrahemispheric networks. GEC-FC weakened with time, but hyperconnectivity persisted at 28d compared to sham. Edge- and global connectivity measures revealed injury-related differences across time in GEC-FC as compared to BBA-FC, demonstrating greater sensitivity to FC changes post-injury. There was no significant association between sleep fragmentation, HFOs, or seizures with FC changes. The within-animal, spatial-temporal differences in BBA-FC and GEC-FC after injury may represent different mechanisms driving FC changes as a result of primary disconnection and interneuron loss., Significance Statement: The present study adds to the understanding of functional connectivity changes in preclinical models of traumatic brain injury. In previously reported literature, there is heterogeneity in the directionality of connectivity changes after injury, resulting from factors such as severity of injury, frequency band studied, and methodology used to calculate FC. This study aims to further clarify differential mechanisms that result in altered network topography after injury, by using Broadband Amplitude-Derived FC and Gamma Event Coupling-Derived FC in EEG. We found post-injury changes that differ in complexity and directionality between measures at and across timepoints. In conjunction with known results and future studies identifying different neural drivers underlying these changes, measures derived from this study could provide useful means from which to minimally-invasively study temporally-evolving pathology after TBI.

Objective . This study aims to develop and validate an end-to-end software platform, PyHFO, that streamlines the application of deep learning (DL) methodologies in detecting neurophysiological biomarkers for epileptogenic zones from EEG recordings. Approach . We introduced PyHFO, which enables time-efficient high-frequency oscillation (HFO) detection algorithms like short-term energy and Montreal Neurological Institute and Hospital detectors. It incorporates DL models for artifact and HFO with spike classification, designed to operate efficiently on standard computer hardware. Main results . The validation of PyHFO was conducted on three separate datasets: the first comprised solely of grid/strip electrodes, the second a combination of grid/strip and depth electrodes, and the third derived from rodent studies, which sampled the neocortex and hippocampus using depth electrodes. PyHFO demonstrated an ability to handle datasets efficiently, with optimization techniques enabling it to achieve speeds up to 50 times faster than traditional HFO detection applications. Users have the flexibility to employ our pre-trained DL model or use their EEG data for custom model training. Significance . PyHFO successfully bridges the computational challenge faced in applying DL techniques to EEG data analysis in epilepsy studies, presenting a feasible solution for both clinical and research settings. By offering a user-friendly and computationally efficient platform, PyHFO paves the way for broader adoption of advanced EEG data analysis tools in clinical practice and fosters potential for large-scale research collaborations., (Creative Commons Attribution license.)

Objective: This study was undertaken to assess reproducibility of the epilepsy outcome and phenotype in a lateral fluid percussion model of posttraumatic epilepsy (PTE) across three study sites., Methods: A total of 525 adult male Sprague Dawley rats were randomized to lateral fluid percussion-induced brain injury (FPI) or sham operation. Of these, 264 were assigned to magnetic resonance imaging (MRI cohort, 43 sham, 221 traumatic brain injury [TBI]) and 261 to electrophysiological follow-up (EEG cohort, 41 sham, 220 TBI). A major effort was made to harmonize the rats, materials, equipment, procedures, and monitoring systems. On the 7th post-TBI month, rats were video-EEG monitored for epilepsy diagnosis., Results: A total of 245 rats were video-EEG phenotyped for epilepsy on the 7th postinjury month (121 in MRI cohort, 124 in EEG cohort). In the whole cohort (n = 245), the prevalence of PTE in rats with TBI was 22%, being 27% in the MRI and 18% in the EEG cohort (p > .05). Prevalence of PTE did not differ between the three study sites (p > .05). The average seizure frequency was .317 ± .725 seizures/day at University of Eastern Finland (UEF; Finland), .085 ± .067 at Monash University (Monash; Australia), and .299 ± .266 at University of California, Los Angeles (UCLA; USA; p < .01 as compared to Monash). The average seizure duration did not differ between UEF (104 ± 48 s), Monash (90 ± 33 s), and UCLA (105 ± 473 s; p > .05). Of the 219 seizures, 53% occurred as part of a seizure cluster (≥3 seizures/24 h; p >.05 between the study sites). Of the 209 seizures, 56% occurred during lights-on period and 44% during lights-off period (p > .05 between the study sites)., Significance: The PTE phenotype induced by lateral FPI is reproducible in a multicenter design. Our study supports the feasibility of performing preclinical multicenter trials in PTE to increase statistical power and experimental rigor to produce clinically translatable data to combat epileptogenesis after TBI., (© 2023 The Authors. Epilepsia published by Wiley Periodicals LLC on behalf of International League Against Epilepsy.)

Objective: We used the lateral fluid percussion injury (LFPI) model of moderate-to-severe traumatic brain injury (TBI) to identify early plasma biomarkers predicting injury, early post-traumatic seizures or neuromotor functional recovery (neuroscores), considering the effect of levetiracetam, which is commonly given after severe TBI., Methods: Adult male Sprague-Dawley rats underwent left parietal LFPI, received levetiracetam (200 mg/kg bolus, 200 mg/kg/day subcutaneously for 7 days [7d]) or vehicle post-LFPI, and were continuously video-EEG recorded (n = 14/group). Sham (craniotomy only, n = 6), and naïve controls (n = 10) were also used. Neuroscores and plasma collection were done at 2d or 7d post-LFPI or equivalent timepoints in sham/naïve. Plasma protein biomarker levels were determined by reverse phase protein microarray and classified according to injury severity (LFPI vs. sham/control), levetiracetam treatment, early seizures, and 2d-to-7d neuroscore recovery, using machine learning., Results: Low 2d plasma levels of Thr 231 -phosphorylated tau protein (pTAU-Thr 231 ) and S100B combined (ROC AUC = 0.7790) predicted prior craniotomy surgery (diagnostic biomarker). Levetiracetam-treated LFPI rats were differentiated from vehicle treated by the 2d-HMGB1, 2d-pTAU-Thr 231 , and 2d-UCHL1 plasma levels combined (ROC AUC = 0.9394) (pharmacodynamic biomarker). Levetiracetam prevented the seizure effects on two biomarkers that predicted early seizures only among vehicle-treated LFPI rats: pTAU-Thr 231 (ROC AUC = 1) and UCHL1 (ROC AUC = 0.8333) (prognostic biomarker of early seizures among vehicle-treated LFPI rats). Levetiracetam-resistant early seizures were predicted by high 2d-IFNγ plasma levels (ROC AUC = 0.8750) (response biomarker). 2d-to-7d neuroscore recovery was best predicted by higher 2d-S100B, lower 2d-HMGB1, and 2d-to-7d increase in HMGB1 or decrease in TNF (P < 0.05) (prognostic biomarkers)., Significance: Antiseizure medications and early seizures need to be considered in the interpretation of early post-traumatic biomarkers., (© 2023 The Authors. Epilepsia Open published by Wiley Periodicals LLC on behalf of International League Against Epilepsy.)

Objective: We examined whether posttraumatic epilepsy (PTE) is associated with measurable perturbations in gut microbiome., Methods: Adult Sprague Dawley rats were subjected to lateral fluid percussion injury (LFPI). PTE was examined 7 months after LFPI, during 4-week continuous video-electroencephalographic monitoring. 16S ribosomal RNA gene sequencing was performed in fecal samples collected before LFPI/sham-LFPI and 1 week, 1 month, and 7 months thereafter. Longitudinal analyses of alpha diversity, beta diversity, and differential microbial abundance were performed. Short-chain fatty acids (SCFAs) were measured in fecal samples collected before LFPI by liquid chromatography with tandem mass spectrometry., Results: Alpha diversity changed over time in both LFPI and sham-LFPI subjects; no association was observed between alpha diversity and LFPI, the severity of post-LFPI neuromotor impairments, and PTE. LFPI produced significant changes in beta diversity and selective changes in microbial abundances associated with the severity of neuromotor impairments. No association between LFPI-dependent microbial perturbations and PTE was detected. PTE was associated with beta diversity irrespective of timepoint vis-à-vis LFPI, including at baseline. Preexistent fecal microbial abundances of four amplicon sequence variants belonging to the Lachnospiraceae family (three enriched and one depleted) predicted the risk of PTE, with area under the curve (AUC) of .73. Global SCFA content was associated with the increased risk of PTE, with AUC of .722, and with 2-methylbutyric (depleted), valeric (depleted), isobutyric (enriched), and isovaleric (enriched) acids being the most important factors (AUC = .717). When the analyses of baseline microbial and SCFA compositions were combined, AUC to predict PTE increased to .78., Significance: Whereas LFPI produces no perturbations in the gut microbiome that are associated with PTE, the risk of PTE can be stratified based on preexistent microbial abundances and SCFA content., (© 2022 International League Against Epilepsy.)

Establishing a cause-and-effect relationship for an adverse event is one of the key steps in preventing them and involves multiple people, resources, and steps, thus requiring a root cause analysis. Here, we describe a root cause analysis performed in the nuclear medicine department for an event involving Na 131 I contamination. Oral administration of Na 131 I in a capsule minimizes the risk of contamination and spills. However, the patient must be able to swallow a capsule. Na 131 I in capsule form is currently in widespread use for treatment of hyperthyroidism and thyroid cancer. Na 131 I in liquid form is rarely available immediately on demand and must be ordered at least 24-48 h in advance of the planned administration. The events leading to the incident, immediate remedial steps taken, and subsequent root cause analysis are described. The corrective actions taken after the root cause analysis, as well as the subsequent effectiveness of these actions, are mentioned. There may be one or multiple causes for an adverse event. It is important to identify the root cause. Corrective actions derived from the root cause can help prevent similar adverse events in the future. Therapeutic procedures in nuclear medicine involve unsealed radioactive sources, further adding a separate layer of immediate steps and reporting to the root cause analysis itself., (© 2021 by the Society of Nuclear Medicine and Molecular Imaging.)

Rationale: The Epilepsy Bioinformatics Study for Antiepileptogenic Therapy (EpiBioS4Rx) Centre without walls is an NIH funded multicenter consortium. One of EpiBioS4Rx projects is a preclinical post-traumatic epileptogenesis biomarker study that involves three study sites: The University of Eastern Finland, Monash University (Melbourne) and the University of California Los Angeles. Our objective is to create a platform for evaluating biomarkers and testing new antiepileptogenic treatments for post-traumatic epilepsy (PTE) using the lateral fluid percussion injury (FPI) model in rats. As only 30-50% of rats with severe lateral FPI develop PTE by 6 months post-injury, prolonged video-EEG monitoring is crucial to identify animals with PTE. Our objective is to harmonize the surgical and data collection procedures, equipment, and data analysis for chronic EEG recording in order to phenotype PTE in this rat model across the three study sites., Methods: Traumatic brain injury (TBI) was induced using lateral FPI in adult male Sprague-Dawley rats aged 11-12 weeks. Animals were divided into two cohorts: a) the long-term video-EEG follow-up cohort (Specific Aim 1), which was implanted with EEG electrodes within 24 h after the injury; and b) the magnetic resonance imaging (MRI) follow-up cohort (Specific Aim 2), at 5 months after lateral FPI. Four cortical epidural screw electrodes (2 ipsilateral, 2 contralateral) and three intracerebral bipolar electrodes were implanted (septal CA1 and the dentate gyrus, layers II and VI of the perilesional cortex both anterior and posterior to the injury site). During the 7th post-TBI month, animals underwent 4 weeks of continuous video-EEG recordings to diagnose of PTE., Results: All centers harmonized the induction of TBI and surgical procedures for the implantation of EEG recordings, utilizing 4 or more EEG recording channels to cover areas ipsilateral and contralateral to the brain injury, perilesional cortex and the hippocampus and dentate gyrus. Ground and reference screw electrodes were implanted. At all sites the minimum sampling rate was 512 Hz, utilizing a finite impulse response (FIR) and impedance below 10 KΩ through the entire recording. As part of the quality control criteria we avoided electrical noise, and monitoring changes in impedance over time and the appearance of noise on the recordings. To reduce electrical noise, we regularly checked the integrity of the cables, stability of the EEG recording cap and the appropriate connection of the electrodes with the cables. Following the pipeline presented in this article and after applying the quality control criteria to our EEG recordings all of the sites were successful to phenotype seizure in chronic EEG recordings of animals after TBI., Discussion: Despite differences in video-EEG acquisition equipment used, the three centers were able to consistently phenotype seizures in the lateral fluid-percussion model applying the pipeline presented here. The harmonization of methodology will help to improve the rigor of preclinical research, improving reproducibility of pre-clinical research in the search of biomarkers and therapies to prevent antiepileptogenesis., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Studies of chronic epilepsy show pathological high frequency oscillations (HFOs) are associated with brain areas capable of generating epileptic seizures. Only a few of these studies have focused on HFOs during the development of epilepsy, but results suggest pathological HFOs could be a biomarker of epileptogenesis. The Epilepsy Bioinformatics Study for Antiepileptogenic Therapy" (EpiBioS4Rx) is a multi-center project designed to identify biomarkers of epileptogenesis after a traumatic brain injury (TBI) and evaluate treatments that could modify or prevent the development of post-traumatic epilepsy. One goal of the EpiBioS4Rx project is to assess whether HFOs could be a biomarker of post-traumatic epileptogenesis. The current study describes the work towards this goal, including the development of common surgical procedures and EEG protocols, an interim analysis of the EEG for HFOs, and identifying issues that need to be addressed for a robust biomarker analysis. At three participating sites - University of Eastern Finland (UEF), Monash University in Melbourne (Melbourne) and University of California, Los Angeles (UCLA) - TBI was induced in adult male Sprague-Dawley rats by lateral fluid-percussion injury. After injury and in sham-operated controls, rats were implanted with screw and microwire electrodes positioned in neocortex and hippocampus to record EEG. A separate group of rats had serial magnetic resonance imaging after injury and then implanted with electrodes at 6 months. Recordings 28 days post-injury were available from UEF and UCLA, but not Melbourne due to technical issues with their EEG files. Analysis of recordings from 4 rats - UEF and UCLA each had one TBI and one sham-operated control - showed EEG contained evidence of HFOs. Computer-automated algorithms detected a total of 1,819 putative HFOs and of these only 40 events (2%) were detected by all three sites. Manual review of all events verified 130 events as HFO and the remainder as false positives. Review of the 40 events detected by all three sites was associated with 88% agreement. This initial report from the EpiBioS4Rx Consortium demonstrates the standardization of EEG electrode placements, recording protocol and long-term EEG monitoring, and differences in detection algorithm HFO results between sites. Additional work on detection strategy, detection algorithm performance, and training in HFO review will be performed to establish a robust, preclinical evaluation of HFOs as a biomarker of post-traumatic epileptogenesis., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Multi-center preclinical studies can facilitate the discovery of biomarkers of antiepileptogenesis and thus facilitate the diagnosis and treatment development of patients at risk of developing post-traumatic epilepsy. However, these studies are often limited by the difficulty in harmonizing experimental protocols between laboratories. Here, we assess whether the production of traumatic brain injury (TBI) using the lateral fluid-percussion injury (FPI) in adult male Sprague-Dawley rats (12 weeks at the time of injury) was harmonized between three laboratories - located in the University of Eastern Finland (UEF), Monash University in Melbourne, Australia (Melbourne) and The University of California, Los Angeles, USA (UCLA). These laboratories are part of the international multicenter-based project, the Epilepsy Bioinformatics Study for Antiepileptogenesis Therapy (EpiBioS4Rx). Lateral FPI was induced in adult male Sprague-Dawley rats. The success of methodological harmonization was assessed by performing inter-site comparison of injury parameters including duration of anesthesia during surgery, impact pressure, post-impact transient apnea, post-impact seizure-like behavior, acute mortality (<72 h post-injury), time to self-right after the impact, and severity of the injury (assessed with the neuroscore). The data was collected using Common Data Elements and Case Report Forms. The acute mortality was 15% (UEF), 50% (Melbourne) and 57% (UCLA) (p < 0.001). The sites differed in the duration of anesthesia, the shortest being at UEF < Melbourne < UCLA (p < 0.001). The impact pressure used also differed between the sites, the highest being in UEF > Melbourne > UCLA (p < 0.001). The impact pressure associated with the severity of the functional deficits (low neuroscore) (P < 0.05) only at UEF, but not at any of the other sites. Additionally, the sites differed in the duration of post-impact transient apnea (p < 0.001) and time to self-right (P < 0.001), the highest values in both parameters was registered in Melbourne. Post-impact seizure-like behavior was observed in 51% (UEF), 25% (Melbourne) and 2% (UCLA) of rats (p < 0.001). Despite the differences in means when all sites were compared there was significant overlap in injury parameters between the sites. The data reflects the technical difficulties in the production of lateral FPI across multiple sites. On the other hand, the data can be used to model the heterogeneity in human cohorts with closed-head injury. Our animal cohort will provide a good starting point to investigate the factors associated with epileptogenesis after lateral FPI., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Post-traumatic epilepsy is the architype of acquired epilepsies, wherein a brain insult initiates an epileptogenic process culminating in an unprovoked seizure after weeks, months or years. Identifying biomarkers of such process is a prerequisite for developing and implementing targeted therapies aimed at preventing the development of epilepsy. Currently, there are no validated electrophysiological biomarkers of post-traumatic epileptogenesis. Experimental EEG studies using the lateral fluid percussion injury model have identified three candidate biomarkers of post-traumatic epileptogenesis: pathological high-frequency oscillations (HFOs, 80-300 Hz); repetitive HFOs and spikes (rHFOSs); and reduction in sleep spindle duration and dominant frequency at the transition from stage III to rapid eye movement sleep. EEG studies in humans have yielded conflicting data; recent evidence suggests that epileptiform abnormalities detected acutely after traumatic brain injury carry a significantly increased risk of subsequent epilepsy. Well-designed studies are required to validate these promising findings, and ultimately establish whether there are post-traumatic electrophysiological features which can guide the development of 'antiepileptogenic' therapies., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The Epilepsy Bioinformatics Study for Antiepileptogenic Therapy (EpiBioS4Rx) is an international, multicenter, multidisciplinary study aimed at preventing epileptogenesis (EpiBioS4Rx: https://epibios.loni.usc.edu/). One of the study's major objectives is the discovery of diagnostic, prognostic, and predictive plasma protein and microRNA (miRNA) biomarkers that are sensitive, specific, and translatable to the human condition. Epilepsy due to structural brain abnormalities, secondary to neurological insults such as traumatic brain injury (TBI), currently represents ∼50% of all epilepsy cases. In the preclinical EpiBioS4Rx study, TBI was induced in adult male Sprague Dawley rats using a standardized protocol for lateral fluid-percussion injury. Whole blood was collected from the tail vein at baseline and 2, 9 and 30 days post-injury and processed for plasma separation. Biomaterial properties, sample preparation and integrity, and choice of analysis platform can significantly impact measured marker levels and, in turn, interpretation with respect to injury and/or other variables. We present here the results of procedural harmonization for the first 320 rats included in the EpiBioS4Rx study study, from three international research centers, and preliminary proteomic and miRNA analyses. We also discuss experimental considerations for establishing rigorous quality controls with the goal of harmonizing operating procedures across study sites, and delivering high-quality specimens for preclinical biomarker discovery in a rat model of post-traumatic epilepsy (PTE)., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Background: The effective non-invasive identification of coronary artery disease (CAD) and its proper referral for invasive treatment are still unresolved issues. We evaluated our quantification of myocardium at risk (MAR) from our second generation 3D MPI/CTA fusion framework for the detection and localization of obstructive coronary disease., Methods: Studies from 48 patients who had rest/stress MPI, CTA, and ICA were analyzed from 3 different institutions. From the CTA, a 3D biventricular surface of the myocardium with superimposed coronaries was extracted and fused to the perfusion distribution. Significant lesions were identified from CTA readings and positioned on the fused display. Three estimates of MAR were computed on the 3D LV surface on the basis of the MPI alone (MAR p ), the CTA alone (MAR a ), and the fused information (MAR f ). The extents of areas at risk were used to generate ROC curves using ICA anatomical findings as reference standard., Results: Areas under the ROC curve (AUC) for CAD detection using MAR f was 0.88 (CI = 0.75-0.95) and for MAR p and MAR a were, respectively 0.82 (CI = 0.69-0.92) and 0.75 (CI = 0.60-0.86) using the ≥70% stenosis criterion. AUCs for CAD localization (all vessels) using MAR f showed significantly higher performance than either MAR a or MAR p or both., Conclusions: Using ICA as the reference standard, MAR as the quantitative parameter, and AUC to measure diagnostic performance, MPI-CTA fusion imaging provided incremental diagnostic information compared to MPI or CTA alone for the diagnosis and localization of CAD.

Molecular dynamics simulations have been performed on the interface between linear saturated hydrocarbons and water in the presence of an asphaltene molecule by measuring the properties such as mean square displacement, radial distribution function, density profile using ave/spatial command, and interfacial tension (IFT) by OPLS and TIP3P FF (force fields). The box of simulation contained one particle of asphaltene, 100 linear saturated hydrocarbons molecules, and 300 water molecules in mixture with interfacial appropriate positioning. The main results show that a small amount of asphaltene in the interface does not significantly alter the data of IFT and that the aliphatic and aromatic groups have preferred orientation., Competing Interests: The authors declare no competing financial interest.

Single photon emission computed tomography (SPECT) myocardial perfusion imaging has attained widespread clinical acceptance as a standard of care for patients with known or suspected coronary artery disease. A significant contribution to this success has been the use of computer techniques to provide objective quantitative assessment in interpreting these studies. We have implemented the Emory Cardiac Toolbox (ECTb) as a pipeline to distribute the software tools that we and others have researched, developed, and validated to be clinically useful so that diagnosticians everywhere can benefit from our work. Our experience has demonstrated that integration of all software tools in a common platform is the optimal approach to promote both accuracy and efficiency. Important attributes of the ECTb approach are (1) our extensive number of normal perfusion databases for SPECT and positron emission tomography (PET) studies, each created with at least 150 patients; (2) our use of Fourier analysis of regional thickening to ensure proper temporal resolution and to allow accurate measurement of left ventricular function and dyssynchrony; (3) our development of PET tools to quantify myocardial hibernation and viability; (4) our development of 3-dimensional displays and the use of these displays as a platform for image fusion of perfusion and computed tomography angiography; and (5) the use of expert systems for decision support. ECTb is an important tool for extracting quantitative parameters from all types of cardiac radionuclide distributions. ECTb should continue to play an important role in establishing cardiac SPECT and PET for flow, function, metabolism, and innervation clinical applications.

Background: Transient ischemic dilation (TID) has been established as an important independent marker of severe and extensive coronary artery disease (CAD) in myocardial perfusion imaging (MPI). The accuracy of the TID index is dependent on a well-determined threshold (normal limits) between normal and abnormal values for each study protocol. To date, the effects of neither gender nor attenuation correction (AC) on TID normal limits have been established. Thus, the objectives of this study were to determine if AC processing changes the normal value of the TID index and if there were gender-related differences in the TID index of normal patients who had undergone rest/exercise-stress technetium-99m sestamibi MPI., Methods and Results: Seventy-five patients (33 women, 42 men; mean age, 57.7 +/- 11.7 y and 55.9 +/- 10.0 y, respectively) with less than a 5% likelihood of CAD, who had undergone low-dose rest/high-dose exercise-stress Tc-99m sestamibi MPI, were studied. All studies were acquired using simultaneous emission/transmission scans and were corrected for attenuation, scatter, and resolution effects using the ExSPECT II method. Both the AC and non-AC studies were analyzed using the Emory Cardiac Toolbox (ECTb; Syntermed, Inc, Atlanta, Ga) quantitative software. The TID index was calculated automatically as the ratio of stress mean left ventricular volumes to rest mean left ventricular volumes by ECTb. Patients were grouped by gender and the TID indices from AC and non-AC studies were compared. Linear regressions of the TID index and body mass index were analyzed to exclude differences in body size between male and female patients as a confounding factor in gender-related differences in TID. The TID index upper normal limits were calculated as the mean value plus 2 standard deviations (SDs). AC processing did not change the TID index significantly whether the genders were combined or separated (AC TID = 0.97 +/- 0.14 vs non-AC TID = 0.98 +/- 0.12 for all patients). Female patients showed higher mean TID indices than male patients in both AC (1.01 +/- 0.15 vs 0.95 +/- 0.12) and non-AC studies (1.00 +/- 0.15 vs. 0.97 +/- 0.10), but this difference was statistically significant only in AC studies (p = .03). TID indices remained constant across the range of body mass index studied. The TID index upper normal limit was 1.31 for female and 1.18 for male patients., Conclusion: TID normal values for rest/exercise-stress Tc-99m sestamibi MPI are gender-dependent and not affected by AC processing. Thus, diagnosticians should take into account these gender-related differences, as compared with the traditional value generated from mostly male populations, to ensure both men and women have the same overall accuracy of using the TID index in the diagnosis and prognosis of CAD.