Advances in Engineering and Application of Optogenetic Indicators for Neuroscience

Our ability to investigate the brain is limited by available technologies that can record biological processes in vivo with suitable spatiotemporal resolution. Advances in optogenetics now enable optical recording and perturbation of central physiological processes within the intact brains of model organisms. By monitoring key signaling molecules noninvasively, we can better appreciate how information is processed and integrated within intact circuits. In this review, we describe recent efforts engineering genetically-encoded fluorescence indicators to monitor neuronal activity. We summarize recent advances of sensors for calcium, potassium, voltage, and select neurotransmitters, focusing on their molecular design, properties, and current limitations. We also highlight impressive applications of these sensors in neuroscience research. We adopt the view that advances in sensor engineering will yield enduring insights on systems neuroscience. Neuroscientists are eager to adopt suitable tools for imaging neural activity in vivo, making this a golden age for engineering optogenetic indicators. Keywords: optogenetic tools; neuroscience; calcium sensor; voltage sensor; neurotransmitter

Integration of Disability Studies and Culinary Instruction into Nutrition Curriculum Development

The Academy of Nutrition and Dietetics is currently promoting the advancement of cultural humility as a means for nutritional professionals to better appreciate the lived experience and identities of others as well as the power dynamics that create health inequalities. At the same time, the Academy has struggled to advance services to underserved populations such as those with Intellectual and Developmental Disabilities (IDD) and autism spectrum disorder. Here the authors argue that both goals can be achieved by involving nutrition students in the teaching of food preparation skills to individuals with IDD. Dietetic students emerge with an enhanced understanding of marginalized populations and individuals with IDD develop important life skills contributing to improved dietary quality. Two programs based upon the Active Engagement protocol are described herein, both of which advance food skills and cultural humility amongst participants

Conversion of red fluorescent protein into a bright blue probe

We used a red chromophore formation pathway, in which the anionic red chromophore is formed from the neutral blue intermediate, to suggest a rational design strategy to develop blue fluorescent proteins with a tyrosine-based chromophore. The strategy was applied to red fluorescent proteins of the different genetic backgrounds, such as TagRFP, mCherry, HcRed1, M355NA, and mKeima, which all were converted into blue probes. Further improvement of the blue variant of TagRFP by random mutagenesis resulted in an enhanced monomeric protein, mTagBFP, characterized by the substantially higher brightness, the faster chromophore maturation, and the higher pH stability than blue fluorescent proteins with a histidine in the chromophore. The detailed biochemical and photochemical analysis indicates that mTagBFP is the true monomeric protein tag for multicolor and lifetime imaging, as well as the outstanding donor for green fluorescent proteins in Forster resonance energy transfer applications

МЕТОД ФОРМУВАННЯ АСОЦІАТИВНИХ ПРАВИЛ З БАЗИ ДАНИХ SIEM – СИСТЕМИ НА ОСНОВІ ТЕОРІЇ НЕЧІТКИХ МНОЖИН ТА ЛІНГВІСТИЧНИХ ТЕРМІВ

The article presents a method of forming fuzzy associative rules with weighted attributes from the database (DB) of the SIEM to supplement its knowledge base (KB) in order to more effectively detect cyber incidents that occur during the operation of special information and communication systems (SICS). The problems that reduce the effectiveness of the application of existing methods for solving the problem of forming associative rules based on the analysis of information located in the database of cyber protection systems are considered. An analysis of publications devoted to methods in which attempts were made to eliminate these problems was made. The basic idea of eliminating the shortcomings inherent in known methods is formulated, which consists in finding a compromise between reducing the time of the computing algorithm that implements the method in practice and reducing information losses as a result of its operation. An improved method of finding associative rules from SIEM databases is proposed, which is based on the theory of fuzzy sets and linguistic terms. The problem of finding fuzzy associative rules with weighted attributes is formulated. The mathematical apparatus that forms the basis of the implementation of the method is given. An algorithm for finding frequent sets of elements, including the values of the signs of cyber incidents and the classes to which they belong, is proposed, which implements the first stage of the proposed method. The peculiarities of the structure of the test data sets used for training and testing of cyber protection systems were analyzed, and based on its results, a conclusion was drawn about the possibility of improving the considered algorithm. A graphic illustration of the idea of improving the algorithm for finding frequent sets of elements is given and the essence of its improvement is described. An improved algorithm for finding frequent sets of elements of the considered method is proposed and its main advantages are given.У статті представлено метод формування нечітких асоціативних правил із зваженими атрибутами з бази даних (БД) SIEM – системи для поповнення її бази знань (БЗ) з метою більш ефективного виявлення нею кіберінцидентів, які виникають в ході функціонування спеціальних інформаційно – комунікаційних систем (СІКС). Розглянуто проблеми, які знижують ефективність застосування існуючих методів для вирішення задачі формування асоціативних правил на основі аналізу інформації, що знаходиться у БД систем кіберзахисту. Проведено аналіз публікацій, присвячених методам, в яких здійснено спроби усунення наведених проблем. Сформульовано основну ідею усунення недоліків, що є властивими відомим методам, яка полягає в знаходженні компромісу між зменшенням часу роботи обчислювального алгоритму, який реалізує на практиці метод та зменшенням інформаційних втрат в результаті його роботи. Запропоновано удосконалений метод пошуку асоціативних правил з БД SIEM – систем, в основу якого покладено теорію нечітких множин та лінгвістичних термів. Сформульовано задачу пошуку нечітких асоціативних правил із зваженими атрибутами. Наведено математичний апарат, який покладено в основу реалізації метода. Запропоновано алгоритм пошуку частих наборів елементів, що включають значення ознак кіберінцидентів та класів, до яких вони відносяться та який реалізує перший етап запропонованого методу. Проаналізовано особливості структури тестових наборів даних, які використовуються для навчання та тестування систем кіберзахисту та на основі його результатів зроблено висновок про можливість удосконалення розглянутого алгоритму. Наведено графічну ілюстрацію ідеї удосконалення алгоритму пошуку частих наборів елементів та описано суть його удосконалення. Запропоновано удосконалений алгоритм пошуку частих наборів елементів розглянутого методу та наведено його основні переваги

Anterior Lumbar Interbody Implants: Importance of the Interdevice Distance

Object. The implantation of interbody fusion cages allows for the restoration of disc height and the enlargement of the neuroforaminal space. The purpose of this study was to compare the extent of subsidence occurring after conventional cage placement compared to a novel wider cage placement technique. Methods. This study is a retrospective evaluation of radiographs of patients who underwent stand-alone single level anterior lumbar interbody fusion with lordotic titanium cages and rhBMP-2. Fifty-three patients were evaluated: 39 patients had wide cage placement (6 mm interdevice distance) and 14 had narrow cage placement (2 mm interdevice distance). Anterior and posterior intervertebral disc space heights were measured post-operatively and at follow-up imaging. Results. The decrease in anterior intervertebral disc space height was 2.05 mm versus 3.92 mm (P < .005) and 1.08 mm versus 3.06 mm in posterior disc space height for the wide cage placement and the narrow cage placement respectively. The proportion of patients with subsidence greater than 2 mm was 41.0% in the wide cage patients and 85.7% for the narrow cage patients (P < .005). Conclusions. The wider cage placement significantly reduced the amount of subsidence while allowing for a greater exposed surface area for interbody fusion

Determination of the Beta Ray Energy Spectrum from the Absorption Curves of Beta Rays

<p>(a) The decrease in food-discovery time between the first tests on two successive days; (b) the positive correlation between colony size and the number of workers searching (mean values for the first tests on the two successive days are presented). The asterisk indicates a significant difference.</p

A new design for a green calcium indicator with a smaller size and a reduced number of calcium-binding sites

Genetically encoded calcium indicators (GECIs) are mainly represented by two- or one-fluorophore-based sensors. One type of two-fluorophore-based sensor, carrying Opsanus troponin C (TnC) as the Ca2+-binding moiety, has two binding sites for calcium ions, providing a linear response to calcium ions. One-fluorophore-based sensors have four Ca2+-binding sites but are better suited for in vivo experiments. Herein, we describe a novel design for a one-fluorophore-based GECI with two Ca2+-binding sites. The engineered sensor, called NTnC, uses TnC as the Ca2+-binding moiety, inserted in the mNeonGreen fluorescent protein. Monomeric NTnC has higher brightness and pH-stability in vitro compared with the standard GECI GCaMP6s. In addition, NTnC shows an inverted fluorescence response to Ca2+. Using NTnC, we have visualized Ca2+ dynamics during spontaneous activity of neuronal cultures as confirmed by control NTnC and its mutant, in which the affinity to Ca2+ is eliminated. Using whole-cell patch clamp, we have demonstrated that NTnC dynamics in neurons are similar to those of GCaMP6s and allow robust detection of single action potentials. Finally, we have used NTnC to visualize Ca2+ neuronal activity in vivo in the V1 cortical area in awake and freely moving mice using two-photon microscopy or an nVista miniaturized microscope