Изучение электрокинетических возможностей сепарации и концентрирования бактерий (S.aureus) в цельной крови четырехэлектродным биосенсорным чип-форматом с прозрачными микроэлектродами

The electrokinetic possibilities of separation and concentration of S.aureus bacteria in human whole blood (test mixture) were studied using a new design of a four-electrode biosensor chip format with transparent microelectrodes. In the measurements, a closed cell was used, into which the test mixture was poured. The electrokinetic movement of bacteria during concentration was observed when AC voltage was applied to the first and second annular external electrodes, and DC voltage was applied to the third and fourth semicircular central electrodes. The process of separation of erythrocytes and concentration of bacteria was recorded by an optical method. It is shown that the concentration of bacteria appears after 8 minutes and increases by 30 minutes in the region of the central electrodes under model conditions. The measurements were carried out on two biosensor chip formats having almost identical electrophysical parameters of capacitance and conductivity at frequencies from 100 Hz to 1 MHz. It is determined that the amplitude-frequency mode and biosensor chip format can be used to develop methodological support for the identification of bacterial species, but only by optical methods. It is shown that in order to increase the number of bacteria on both of the central electrode in the concentration mode, it is necessary to increase the speed of the electroosmotic flow at alternating current, by optimizing the composition of the medium and the parameters of the electrical regime in order to equalize the rates of separation and concentration processes.Изучены электрокинетические возможности сепарации и концентрирования бактерий S.aureus в цельной крови человека (исследуемая смесь) с помощью новой конструкции четырехэлектродного биосенсорного чип-формата с прозрачными микроэлектродами. В исследованиях использована закрытая ячейка, в которую заливали исследуемую смесь. Электрокинетическое перемещение бактерий при концентрировании наблюдалось при подаче на первый и второй кольцевые внешние электроды переменного напряжения, а на третий и четвертый полукруглые центральные электроды – постоянного напряжения. Процесс сепарации эритроцитов и концентрирования бактерий фиксировался оптическим методом для модельных условий и в цельной крови. Показано, что концентрирование бактерий проявляется через 8 минут и увеличивается к 30 минуте в области центральных электродов в модельных условиях. Измерения были проведены на биосенсорных чип-форматах, имеющих практически одинаковые электрофизические параметры емкости и проводимости на частотах от 100 Гц до 1 МГц. Установлено, что амплитудно-частотный режим и биосенсорный чип-формат могут быть использованы для разработки методического обеспечения идентификации вида бактерий, но только оптическими методами. Показано, что для увеличения количества бактерий на центральных электродах в режиме концентрирования необходимо повысить скорость электроосмотического потока на переменном токе путем оптимизации состава среды и параметров электрического режима, чтобы уравнять скорости процессов сепарации и концентрирования

A Genetic Signature of Spina Bifida Risk from Pathway-Informed Comprehensive Gene-Variant Analysis

Despite compelling epidemiological evidence that folic acid supplements reduce the frequency of neural tube defects (NTDs) in newborns, common variant association studies with folate metabolism genes have failed to explain the majority of NTD risk. The contribution of rare alleles as well as genetic interactions within the folate pathway have not been extensively studied in the context of NTDs. Thus, we sequenced the exons in 31 folate-related genes in a 480-member NTD case-control population to identify the full spectrum of allelic variation and determine whether rare alleles or obvious genetic interactions within this pathway affect NTD risk. We constructed a pathway model, predetermined independent of the data, which grouped genes into coherent sets reflecting the distinct metabolic compartments in the folate/one-carbon pathway (purine synthesis, pyrimidine synthesis, and homocysteine recycling to methionine). By integrating multiple variants based on these groupings, we uncovered two provocative, complex genetic risk signatures. Interestingly, these signatures differed by race/ethnicity: a Hispanic risk profile pointed to alterations in purine biosynthesis, whereas that in non-Hispanic whites implicated homocysteine metabolism. In contrast, parallel analyses that focused on individual alleles, or individual genes, as the units by which to assign risk revealed no compelling associations. These results suggest that the ability to layer pathway relationships onto clinical variant data can be uniquely informative for identifying genetic risk as well as for generating mechanistic hypotheses. Furthermore, the identification of ethnic-specific risk signatures for spina bifida resonated with epidemiological data suggesting that the underlying pathogenesis may differ between Hispanic and non-Hispanic groups