The application of the capacitive division technique to wide-field time-resolved fluorescence microscopy

Capacitive division and other charge-sharing techniques have become ubiquitous within modern technology. Almost all touchscreen devices depend on some form of charge sharing mechanism. The Capacitive-Division Imaging Readout, C-DIR, scheme developed for space/astronomy applications, is a proven concept which has benefited from widespread publication and several iterations of prototyping. In this study, we borrowed this idea and assessed its application in the field of life sciences, specifically, fluorescence lifetime imaging microscopy (FLIM). Firstly, the composite C-DIR camera system was developed using a prototype anode developed by Lapington et al in combination with advanced photomultiplier tube technology developed by Photek Limited, and ultra-fast NINO ASIC and high performance time-to-digital converter, HPTDC, readout electronics developed by CERN. Several issues like signal noise, timing jitter and image distortion required special attention to successfully tune the C-DIR system for obtaining FLIM measurements. The C-DIR was characterized in the context of current detector technologies used for time-resolved applications. The maximum achievable global event rate was determined to be a USB 2.0 hard limit of about 1MHz. The spatial resolution and timing performance were identified as 0.5 line-pairs/mm and 200ps FWHM, respectively, which was comparable to other wide-field fluorescence lifetime cameras. These results provided the basis for using the system in a real situation. Before this was possible, however, it was necessary to engineer a bespoke software platform for data acquisition which could cope with the data rates and reduce raw data emerging from the C-DIR system, producing a format compatible with widely used fitting software. The final stage of the project involved using the C-DIR for real science by reproducing real world experiments which allow for a fitness test of the system in the field. The first experiment involved a calcium calibration where the C-DIR system was calibrated using FLIM on a series of calcium buffers of known concentrations. This C-DIR was able accurately recover the lifetime values from the calcium buffers. The second shorter experiment involved using the calibrated system for the quantification of calcium within living tissue samples using fluorescence lifetime imaging. Results were consistent with those published in the literature which solidified the position of the C-DIR as a viable option for time-resolved fluorescence microscopy

Ultrasonography-First Imaging Algorithm for Acute Appendicitis Diagnosis

Acute appendicitis (AA) is the most common cause of acute abdominal pain requiring surgery in pediatric, adult, and pregnant patients. Several etiologies are believed to trigger luminal obstruction, which causes mucus and bacteria proliferation, resulting in inflammation and wall tension with subsequent necrosis and rupture of the appendix. Most AA patients present with the primary complaint of abdominal pain, which Murphy first described the characteristic diagnostic sequence seen in approximately 50% of patients as colicky centralized abdominal pain with subsequent vomiting with the migration of pain to the right lower quadrant (RLQ), specifically, the right iliac fossa. The typical AA patient will complain of colicky, periumbilical pain, which has progressively worsened over the past 24 hours that has become persistently sharp in the RLQ. Diagnostic imaging is essential in diagnosing AA, and it is critical for medical providers to quickly and accurately diagnose AA to reduce perforated appendix and negative appendectomy rates. Yet, there is not a universally accepted, widely utilized diagnostic imaging protocol for suspected AA patients. Ultrasonography (USG) and computerized axial tomography (CAT) scans are used most in diagnosing AA in all ages. Current evidence-based practice (EBP) literature shows that USG should be the first-line imaging modality followed by CAT scan as the second line in diagnosing AA in children and adults, to reduce ionizing radiation and cost burdens. The purpose of this Doctor of Nursing Practice (DNP) project was to develop, implement, and evaluate an educational intervention and an evidence-based USG-first algorithm for medical providers who treat patients with suspected AA. Pre- and post-test assessment questions were used to evaluate the implementation of knowledge and the AA clinical practice change over a six-to-eight-week period. Descriptive statistics were used to analyze demographic data, tabulating frequencies, and percentages. Pre- and post-assessment scores were analyzed via a paired t-test for matched samples. This project’s results displayed a statistically significant change in provider knowledge and may have improved clinical practice and patient outcomes. This project’s USG-first imaging algorithm for diagnosing acute appendicitis can likely be sustained in clinical nursing practice

Report: Forum on Ministerial Formation if Black Church Traditions

Church leadership is always developed contextually, never in a vacuum. Clergy leaders are shaped theologically, rhetorically, and culturally in distinct communities of faith. These communities of faith may not use the language of formation, but they are forming women and men for ministry. The task of field education is to make possible the dance between the formation students receive before they attend seminary and the formation they receive upon matriculation. Each student must begin a dance that will last a lifetime. The black church’s traditions of formation, when not excluded from the dance floor, enrich the church’s leaders across the divides of culture, gender, and theology

A wide field fluorescence lifetime imaging system using a light sheet microscope

Fluorescence lifetime imaging microscopy (FLIM) has allowed scientists to discern information about the chemical properties of biological processes and has become a vital tool in the life sciences and medical research communities. Measuring the spatial lifetime distribution of the fluorophores as well as the intensity distribution enables users to discern vital information about the chemical environment. It however, remains challenging and often involves slow scanning. We present a new microscope system based on light sheet illumination that uses a micro channel plate (MCP) device called a Capacitive Division Imaging Readout (CDIR) which has been developed by Photek Ltd. The device uses an array of capacitors to move the charge site from the MCP to four pre-amplifiers and time-over-threshold discriminators. This camera has the ability to image photons as well as measure the arrival time, enabling high speed FLIM imaging of biological samples

Endothelial cells and angiogenesis in the horse in health and disease—A review

The cardiovascular system is the first functional organ in the embryo, and its blood vessels form a widespread conductive network within the organism. Blood vessels develop de novo, by the differentiation of endothelial progenitor cells (vasculogenesis) or by angiogenesis, which is the formation of new blood vessels from existing ones. This review presents an overview of the current knowledge on physiological and pathological angiogenesis in the horse including studies on equine endothelial cells. Principal study fields in equine angiogenesis research were identified: equine endothelial progenitor cells; equine endothelial cells and angiogenesis (heterogeneity, markers and assessment); endothelial regulatory molecules in equine angiogenesis; angiogenesis research in equine reproduction (ovary, uterus, placenta and conceptus, testis); angiogenesis research in pathological conditions (tumours, ocular pathologies, equine wound healing, musculoskeletal system and laminitis). The review also includes a table that summarizes in vitro studies on equine endothelial cells, either describing the isolation procedure or using previously isolated endothelial cells. A particular challenge of the review was that results published are fragmentary and sometimes even contradictory, raising more questions than they answer. In conclusion, angiogenesis is a major factor in several diseases frequently occurring in horses, but relatively few studies focus on angiogenesis in the horse. The challenge for the future is therefore to continue exploring new therapeutic angiogenesis strategies for horses to fill in the missing pieces of the puzzle

Evasion of anti-growth signaling: a key step in tumorigenesis and potential target for treatment and prophylaxis by natural compounds

The evasion of anti-growth signaling is an important characteristic of cancer cells. In order to continue to proliferate, cancer cells must somehow uncouple themselves from the many signals that exist to slow down cell growth. Here, we define the anti-growth signaling process, and review several important pathways involved in growth signaling: p53, phosphatase and tensin homolog (PTEN), retinoblastoma protein (Rb), Hippo, growth differentiation factor 15 (GDF15), AT-rich interactive domain 1A (ARID1A), Notch, insulin-like growth factor (IGF), and Krüppel-like factor 5 (KLF5) pathways. Aberrations in these processes in cancer cells involve mutations and thus the suppression of genes that prevent growth, as well as mutation and activation of genes involved in driving cell growth. Using these pathways as examples, we prioritize molecular targets that might be leveraged to promote anti-growth signaling in cancer cells. Interestingly, naturally-occurring phytochemicals found in human diets (either singly or as mixtures) may promote anti-growth signaling, and do so without the potentially adverse effects associated with synthetic chemicals. We review examples of naturally-occurring phytochemicals that may be applied to prevent cancer by antagonizing growth signaling, and propose one phytochemical for each pathway. These are: epigallocatechin-3-gallate (EGCG) for the Rb pathway, luteolin for p53, curcumin for PTEN, porphyrins for Hippo, genistein for GDF15, resveratrol for ARID1A, withaferin A for Notch and diguelin for the IGF1-receptor pathway. The coordination of anti-growth signaling and natural compound studies will provide insight into the future application of these compounds in the clinical setting

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Brassboard TREESS kit

Issued as R&D status reports no. 1-11, and Performance and cost reports no. 1-11, Project no. A-326

Reflectivity arch measurement system

Issued as Final report, Project no. A-3519Final report has title: Reflectivity arch measurement syste