Identification and Characterization of Epp, the Secreted Processing Protease for the \u3cem\u3eVibrio anguillarum\u3c/em\u3e EmpA Metalloprotease

The zinc metalloprotease EmpA is a virulence factor for the fish pathogen Vibrio anguillarum. Previous studies demonstrated that EmpA is secreted as a 46-kDa proenzyme that is activated extracellularly by the removal of an ∼10-kDa propeptide. We hypothesized that a specific protease is responsible for processing secreted pro-EmpA into mature EmpA. To identify the protease responsible for processing pro-EmpA, a minitransposon mutagenesis (using mini-Tn10Km) clone bank of V. anguillarum was screened for reduced protease activity due to insertions in undescribed genes. One mutant with reduced protease activity was identified. The region containing the mini-Tn10Km was cloned, sequenced, and found to contain epp, an open reading frame encoding a putative protease. Further characterization of epp was done using strain M101, created by single-crossover insertional mutagenesis. Protease activity was absent in M101 cultures even when empA protease activity was induced by salmon gastrointestinal mucus. When the epp mutation was complemented with a wild-type copy of epp (M102), protease activity was restored. Western blot analysis of sterile filtered culture supernatants from wild-type (M93Sm) cells, M101 cells, and M102 cells revealed that only pro-EmpA was present in M101supernatants; both pro-EmpA and mature EmpA were detected in M93Sm and M102 supernatants. When sterile filtered culture supernatants from the empA mutant strain (M99) and M101 were mixed, protease activity was restored. Western blot analysis revealed that pro-EmpA in M101 culture supernatant was processed to mature EmpA only after mixing with M99 culture supernatant. These data show that Epp is the EmpA-processing protease

The use of augmented reality technologies in the development of emotional intelligence of future specialists of socionomic professions under the conditions of adaptive learning.

In modern conditions, innovative augmented reality technologies are actively developing, which are widespread in many areas of human activity. Intro-duction of advanced developments in the process of professional training of future specialists of socionomic professions in the conditions of adaptive training, contributes to the implementation of the principles of a personalized approach and increase the overall level of competitiveness. The relevant sci-entific article is devoted to the theoretical and empirical analysis of the fea-tures of the implementation of augmented reality technologies in the con-struct of traditional psychological and pedagogical support aimed at the de-velopment of emotional intelligence of the future specialist. The interdisci-plinary approach was used while carrying out the researchwork at the ex-pense of the general fund of the state budget: "Adaptive system for individu-alization and personalization of professional training of future specialists in the conditions of blended learning". A comprehensive study of the imple-mentation of traditional psychological-pedagogical and innovative augment-ed reality technologies was conducted in the framework of scientific cooper-ation of STEAM-Laboratory, Laboratory of Psychophysiological Research and Laboratory of Psychology of Health in Bogdan Khmelnytsky Melitopol State Pedagogical University. The theoretical analysis considers the structural model of emotional intelligence of the future specialist of socionomic pro-fessions, which is represented by two structural components: intrapersonal construct of emotional intelligence (understanding and analysis of emotions; use of emotions to improve thinking and activity) and interpersonal construct of emotional intelligence (perception, evaluation and expression). emotions; conscious management of emotions). Each component mediates the inherent emotional intelligence of interpretive, regulatory, adaptive, stress-protective and activating functions. The algorithm of the empirical block of research is presented by two stages: ascertaining and forming research. According to the results of the statement, low indicators were found on most scales, reflecting the general level of emotional intelligence development of future specialists, actualizing the need to find and implement effective measures for the devel-opment of emotional intelligence components in modern higher education and taking into account information development and digitalization. As part of the formative stage of the research implementation, a comprehensive pro-gram "Development of emotional intelligence of future professionals" was tested, which integrated traditional psychological and pedagogical technolo-gies and innovative augmented reality technologies. This program is designed for 24 hours, 6 thematic classes of 4 hours. According to the results of a comprehensive ascertaining and shaping research, the effectiveness of the influence of augmented reality technologies on the general index of emo-tional intelligence is proved. The stepby- step model of integration of aug-mented reality components influencing the ability to analyze, understand and regulate emotional states into a complex program of emotional intelligence development is demonstrated. According to the results of the formative study, there is a dominance of high indicators of the following components: intrapersonal (50%), interpersonal (53.3%). Thus, we can say that in-trapersonal and interpersonal emotional intelligence together involve the ac-tualization of various cognitive processes and skills, and are related to each other. Empirical data were obtained as a result of conducting a psychodiag-nostic study on an innovative computer complex -psychotest

HIDE AND SEEK: MOLECULAR BARCODING CLARIFIES THE DISTRIBUTION OF TWO CRYPTIC DUCKWEED SPECIES ACROSS ALBERTA

Regional and global biodiversity may be underestimated due to the presence of cryptic species: species that are morphologically similar, but genetically distinct. Here we focus on two cryptic duckweed species, Lemna minor and L. turionifera, which have overlapping geographic ranges and are easily mistaken for one another. We developed species-specific primers based on DNA barcoding sequences to facilitate the rapid identification of these two monomorphic duckweeds, allowing us to investigate their presence and distribution in Alberta, Canada. While current reports indicate the presence of L. turionifera (and the morphologically distinct L. trisulca) in Alberta, our data indicate that L. minor is also present, predominantly in the southern part of the province. Thus, this paper (1) contributes to the accuracy and completeness of a regional flora, and (2) provides useful and flexible tools for the rapid molecular identification of cryptic Lemna species, species of wide interest in such diverse fields as biotechnology, toxicology, bioremediation, and ecology.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author

Identification and Characterization of Epp, the Secreted Processing Protease for the Vibrio anguillarum EmpA Metalloprotease ▿

The zinc metalloprotease EmpA is a virulence factor for the fish pathogen Vibrio anguillarum. Previous studies demonstrated that EmpA is secreted as a 46-kDa proenzyme that is activated extracellularly by the removal of an ∼10-kDa propeptide. We hypothesized that a specific protease is responsible for processing secreted pro-EmpA into mature EmpA. To identify the protease responsible for processing pro-EmpA, a minitransposon mutagenesis (using mini-Tn10Km) clone bank of V. anguillarum was screened for reduced protease activity due to insertions in undescribed genes. One mutant with reduced protease activity was identified. The region containing the mini-Tn10Km was cloned, sequenced, and found to contain epp, an open reading frame encoding a putative protease. Further characterization of epp was done using strain M101, created by single-crossover insertional mutagenesis. Protease activity was absent in M101 cultures even when empA protease activity was induced by salmon gastrointestinal mucus. When the epp mutation was complemented with a wild-type copy of epp (M102), protease activity was restored. Western blot analysis of sterile filtered culture supernatants from wild-type (M93Sm) cells, M101 cells, and M102 cells revealed that only pro-EmpA was present in M101supernatants; both pro-EmpA and mature EmpA were detected in M93Sm and M102 supernatants. When sterile filtered culture supernatants from the empA mutant strain (M99) and M101 were mixed, protease activity was restored. Western blot analysis revealed that pro-EmpA in M101 culture supernatant was processed to mature EmpA only after mixing with M99 culture supernatant. These data show that Epp is the EmpA-processing protease

Quantitative Electroencephalography After Pediatric Anterior Circulation Stroke

OBJECTIVE: Regional differences were investigated in quantitative EEG (QEEG) characteristics and associations of QEEG to hemodynamics after pediatric acute stroke. METHODS: Quantitative EEG was analyzed, including power in delta, theta, alpha, and beta bands, alpha-delta power ratio, total power, and spectral edge frequency from 11 children with unilateral, anterior circulation strokes during the first 24 hours of continuous EEG recording. Differences between injured and uninjured hemispheres were assessed using multivariate dynamic structural equations modeling. Dynamic structural equations modeling was applied to six children with hemorrhagic stroke undergoing arterial blood pressure, heart rate, and cerebral oximetry monitoring to investigate associations between hemodynamics with QEEG adjacent to anterior circulation regions. RESULTS: All patients with acute ischemic stroke (n = 5) had lower alpha and beta power and spectral edge frequency on injured compared with uninjured regions. This was not consistent after hemorrhagic stroke (n = 6). All hemorrhagic stroke patients demonstrated negative association of total power with arterial blood pressure within injured regions. No consistency was observed for direction or strength of association in other QEEG measures to arterial blood pressure nor were such consistent relationships observed for any QEEG measure studied in relation to heart rate or cerebral oximetry. CONCLUSIONS: After pediatric anterior circulation acute ischemic stroke, reduced spectral edge frequency and alpha and beta power can be observed on injured as compared with noninjured regions. After pediatric anterior circulation hemorrhagic stroke, total power can be negatively associated with arterial blood pressure within injured regions. Larger studies are needed to understand conditions in which QEEG patterns manifest and relate to hemodynamics and brain penumbra

Quantitative Electroencephalography After Pediatric Anterior Circulation Stroke

OBJECTIVE: Regional differences were investigated in quantitative EEG (QEEG) characteristics and associations of QEEG to hemodynamics after pediatric acute stroke. METHODS: Quantitative EEG was analyzed, including power in delta, theta, alpha, and beta bands, alpha-delta power ratio, total power, and spectral edge frequency from 11 children with unilateral, anterior circulation strokes during the first 24 hours of continuous EEG recording. Differences between injured and uninjured hemispheres were assessed using multivariate dynamic structural equations modeling. Dynamic structural equations modeling was applied to six children with hemorrhagic stroke undergoing arterial blood pressure, heart rate, and cerebral oximetry monitoring to investigate associations between hemodynamics with QEEG adjacent to anterior circulation regions. RESULTS: All patients with acute ischemic stroke (n = 5) had lower alpha and beta power and spectral edge frequency on injured compared with uninjured regions. This was not consistent after hemorrhagic stroke (n = 6). All hemorrhagic stroke patients demonstrated negative association of total power with arterial blood pressure within injured regions. No consistency was observed for direction or strength of association in other QEEG measures to arterial blood pressure nor were such consistent relationships observed for any QEEG measure studied in relation to heart rate or cerebral oximetry. CONCLUSIONS: After pediatric anterior circulation acute ischemic stroke, reduced spectral edge frequency and alpha and beta power can be observed on injured as compared with noninjured regions. After pediatric anterior circulation hemorrhagic stroke, total power can be negatively associated with arterial blood pressure within injured regions. Larger studies are needed to understand conditions in which QEEG patterns manifest and relate to hemodynamics and brain penumbra

Resting-state functional MRI connectivity impact on epilepsy surgery plan and surgical candidacy: prospective clinical work

OBJECTIVE: The authors\u27 goal was to prospectively quantify the impact of resting-state functional MRI (rs-fMRI) on pediatric epilepsy surgery planning. METHODS: Fifty-one consecutive patients (3 months to 20 years old) with intractable epilepsy underwent rs-fMRI for presurgical evaluation. The team reviewed the following available diagnostic data: video-electroencephalography (n = 51), structural MRI (n = 51), FDG-PET (n = 42), magnetoencephalography (n = 5), and neuropsychological testing (n = 51) results to formulate an initial surgery plan blinded to the rs-fMRI findings. Subsequent to this discussion, the connectivity results were revealed and final recommendations were established. Changes between pre- and post-rs-fMRI treatment plans were determined, and changes in surgery recommendation were compared using McNemar\u27s test. RESULTS: Resting-state fMRI was successfully performed in 50 (98%) of 51 cases and changed the seizure onset zone localization in 44 (88%) of 50 patients. The connectivity results prompted 6 additional studies, eliminated the ordering of 11 further diagnostic studies, and changed the intracranial monitoring plan in 10 cases. The connectivity results significantly altered surgery planning with the addition of 13 surgeries, but it did not eliminate planned surgeries (p = 0.003). Among the 38 epilepsy surgeries performed, the final surgical approach changed due to rs-fMRI findings in 22 cases (58%), including 8 (28%) of 29 in which extraoperative direct electrical stimulation mapping was averted. CONCLUSIONS: This study demonstrates the impact of rs-fMRI connectivity results on the decision-making for pediatric epilepsy surgery by providing new information about the location of eloquent cortex and the seizure onset zone. Additionally, connectivity results may increase the proportion of patients considered eligible for surgery while optimizing the need for further testing

Source-sink connectivity: A novel interictal EEG marker for seizure localization

Over 15 million epilepsy patients worldwide have drug-resistant epilepsy (DRE). Successful surgery is a standard of care treatment for DRE but can only be achieved through complete resection or disconnection of the epileptogenic zone (EZ), the brain region(s) where seizures originate. Surgical success rates vary between 20-80% because no clinically validated biological markers of the EZ exist. Localizing the EZ is a costly and time-consuming process beginning with non-invasive neuroimaging and often followed by days to weeks of intracranial EEG (iEEG) monitoring. Clinicians visually inspect iEEG data to identify abnormal activity (e.g., low-voltage high frequency activity) on individual channels occurring immediately before seizures or spikes that occur on interictal iEEG (i.e., between seizures). In the end, the clinical standard mainly relies on a small proportion of the iEEG data captured to assist in EZ localization (minutes of seizure data versus days of recordings), missing opportunities to leverage these largely ignored interictal data to better diagnose and treat patients. Intracranial EEG offers a unique opportunity to observe epileptic cortical network dynamics but waiting for seizures increases patient risks associated with invasive monitoring. In this study, we aim to leverage interictal iEEG data by developing a new network-based interictal iEEG marker of the EZ. We hypothesize that when a patient is not clinically seizing, it is because the EZ is inhibited by other regions. We developed an algorithm that identifies two groups of nodes from the interictal iEEG network: those that are continuously inhibiting a set of neighboring nodes ("sources") and the inhibited nodes themselves ("sinks"). Specifically, patient-specific dynamical network models (DNMs) were estimated from minutes of iEEG and their connectivity properties revealed top sources and sinks in the network, with each node being quantified by source-sink metrics (SSMs). We validated the SSMs in a retrospective analysis of 65 patients by using the SSMs of the annotated EZ to predict surgical outcomes. The SSMs predicted outcomes with an accuracy of 79% compared to an accuracy of 43% for clinicians' predictions (surgical success rate of this dataset). In failed outcomes, we identified regions of the brain with high SSMs that were untreated. When compared to high frequency oscillations, the most commonly proposed interictal iEEG feature for EZ localization, SSMs outperformed in predictive power (by a factor of 1.2) suggesting SSMs may be an interictal iEEG fingerprint of the EZ. Competing Interest Statement The authors have declared no competing interest