Characterization of a distinct population of circulating human non-adherent endothelial forming cells and their recruitment via intercellular adhesion molecule-3

Circulating vascular progenitor cells contribute to the pathological vasculogenesis of cancer whilst on the other hand offer much promise in therapeutic revascularization in post-occlusion intervention in cardiovascular disease. However, their characterization has been hampered by the many variables to produce them as well as their described phenotypic and functional heterogeneity. Herein we have isolated, enriched for and then characterized a human umbilical cord blood derived CD133+ population of non-adherent endothelial forming cells (naEFCs) which expressed the hematopoietic progenitor cell markers (CD133, CD34, CD117, CD90 and CD38) together with mature endothelial cell markers (VEGFR2, CD144 and CD31). These cells also expressed low levels of CD45 but did not express the lymphoid markers (CD3, CD4, CD8)or myeloid markers (CD11b and CD14) which distinguishes them from ‘early’ endothelial progenitor cells (EPCs). Functional studies demonstrated that these naEFCs (i) bound Ulex europaeus lectin, (ii)demonstrated acetylated-low density lipoprotein uptake, (iii) increased vascular cell adhesion molecule (VCAM-1) surface expression in response to tumor necrosis factor and (iv) in co-culture with mature endothelial cells increased the number of tubes, tubule branching and loops in a 3- dimensional in vitro matrix. More importantly, naEFCs placed in vivo generated new lumen containing vasculature lined by CD144 expressing human endothelial cells (ECs). Extensive genomic and proteomic analyses of the naEFCs showed that intercellular adhesion molecule (ICAM)-3 is expressed on their cell surface but not on mature endothelial cells. Furthermore, functional analysis demonstrated that ICAM-3 mediated the rolling and adhesive events of the naEFCs under shear stress. We suggest that the distinct population of naEFCs identified and characterized here represents a new valuable therapeutic target to control aberrant vasculogenesis.Sarah L. Appleby, Michaelia P. Cockshell, Jyotsna B. Pippal, Emma J. Thompson, Jeffrey M. Barrett, Katie Tooley, Shaundeep Sen, Wai Yan Sun, Randall Grose, Ian Nicholson, Vitalina Levina, Ira Cooke, Gert Talbo, Angel F. Lopez and Claudine S. Bonde

Examining the Influence of the Human Gut Microbiota on Cognition and Stress: A Systematic Review of the Literature

The gut microbiota is seen as an emerging biotechnology that can be manipulated to enhance or preserve cognition and physiological outputs of anxiety and depression in clinical conditions. However, the existence of such interactions in healthy young individuals in both non-stressful and stressful environments is unclear. The aim of this systematic review was to examine the relationship between the human gut microbiota, including modulators of the microbiota on cognition, brain function and/or stress, anxiety and depression. A total of n = 25 eligible research articles from a possible 3853 published between October 2018 and August 2021 were identified and included. Two study design methods for synthesis were identified: cross-sectional or pre/post intervention. Few cross-sectional design studies that linked microbiota to cognition, brain activity/structure or mental wellbeing endpoints existed (n = 6); however, correlations between microbiota diversity and composition and areas of the brain related to cognitive functions (memory and visual processing) were observed. Intervention studies targeting the gut microbiota to improve cognition, brain structure/function or emotional well-being (n = 19) generally resulted in improved brain activity and/or cognition (6/8), and improvements in depression and anxiety scores (5/8). Despite inherit limitations in studies reviewed, available evidence suggests that gut microbiota is linked to brain connectivity and cognitive performance and that modulation of gut microbiota could be a promising strategy for enhancing cognition and emotional well-being in stressed and non-stressed situations

A Systematic Review of the Effect of Dietary Supplements on Cognitive Performance in Healthy Young Adults and Military Personnel

Intake of dietary supplements has increased, despite evidence that some of these have adverse side effects and uncertainty about their effectiveness. This systematic review examined the evidence for the cognitive benefits of a wide range of dietary supplements in healthy young adult samples; the aim was to identify if any might be useful for optimising cognitive performance during deployment in military personnel. Searches were conducted in 9 databases and 13 grey literature repositories for relevant studies published between January 2000 and June 2017. Eligible studies recruited healthy young adults (18–35 years), administered a legal dietary supplement, included a comparison control group, and assessed cognitive outcome(s). Thirty-seven of 394 identified studies met inclusion criteria and were included for synthesis. Most research was deemed of low quality (72.97%; SIGN50 guidelines), highlighting the need for sound empirical research in this area. Nonetheless, we suggest that tyrosine or caffeine could be used in healthy young adults in a military context to enhance cognitive performance when personnel are sleep-deprived. Caffeine also has the potential benefit of improving vigilance and attention during sustained operations offering little opportunity for sleep. Inconsistent findings and methodological limitations preclude firm recommendations about the use of other specific dietary supplements

Optimization of the non-invasive 13C-sucrose breath test in a rat model of methotrexate-induced mucositis

Purpose In order to determine the sensitivity and specificity of the test and to optimize experimental conditions utilizing the SBT in a rat model of chemotherapy-induced small intestinal damage. Methods Initially, a 13C-sucrose dose-response study was performed in rats to determine an optimal sucrose concentration for the SBT; then applied to assess chemotherapy-induced intestinal damage. A further study was conducted to establish a SBT time-course of methotrexate-induced small intestinal damage and repair. Animals were killed at 96 or 144 h. Results A sucrose concentration of 0.25 g/ml was optimal (20% CV) for reproducibility and detection of intestinal damage. Maximal damage occurred at 72 h, small intestinal repair was initiated by 96 h and continued at 144 h post-MTX, as determined by the SBT and confirmed by biochemical analyses. Levels of sensitivity and specificity for the SBT were 98 and 94%, respectively. Conclusions The SBT is a reliable non-invasive marker of small intestinal health and damage with a high degree of sensitivity and specificity.

Resistance of Grape Rootstocks to Crown Gall

Crown gall can develop on grapevines wounded by freezing temperatures, mechanical damage or grafting. Increased demand for grape varieties grafted to phylloxera resistant rootstocks has led to increased incidence of crown gall at graft unions. Therefore a search for plant material that is resistant to crown gall has been undertaken. Previous studies on resistance have produced confusing and in some cases contradictory results, probably due to the difference in methods and agrobacterial strains used in each investigation (5, 11)

Crown Gall of Grape Rootstocks

Crown gall can develop on grapevines wounded by freezing temperatures, mechanical damage or grafting. Pathogenic agrobacteria residing within the vascular system are able to initiate tumor growth in damaged plant cells. Sampling from 21 sites in Oregon vineyards and nurseries yielded diverse populations of pathogenic agrobacteria. Most of the pathogenic strains isolated were biotype 3 (Agrobacterium vitis), but there were some biotype 2 strains isolated from five sites that were highly virulent. Several methods have been proposed for minimizing crown gall in grapevines. These include: 1) heat treatments to kill bacteria within plants, 2) planting grafted grapevines growing on rootstocks that are tolerant to pathogenic agrobacteria and 3) planting clean grapevines into disease-free soil. Research at Oregon State University over the past few years has resulted in progress in these three areas. 1) Heat treatments. agrobacteria subjected to 54°C (129°F) for 30 minutes killed 97 different strains isolated from grape. These same heat treatments were used on dormant cuttings of rootstocks and scion wood infiltrated with antibiotic resistant strains of pathogenic agrobacteria. No pathogenic agrobacteria were detected from sap extractions of heat-treated cuttings. The plants showed no adverse growth effects from the heat treatments when compared to non-heated controls, but one grower reported that the heat-treated rootstocks did not graft well. 2) Crown gall tolerant rootstocks. Two hybrid rootstocks (V. riparia X V. rupestris) 101-24 Millardet et de Grasset and 3309 Couderc were the most tolerant to pathogenic agrobacteria grape isolates, while 420 A Milardet et de Grasset (V riparia X V. berlandieri) was the most susceptible to crown gall of six rootstocks tested. 3) Clean grapevines in clean soil. Shoot tip culture and propagation is one way to acquire bacteria free plants (2) and soil solarization has been shown to reduce the populations of pathogenic agrobacteria in planting sites (4)

Hematopoietic properties of naEFCs.

<p>naEFCs were seeded in MethoCult and growth factors GM-CSF, IL-3, SCF and EPO for 14 days prior to colony counting and staining with May Grunwald/Giemsa to assess cellular morphology. naEFCs formed blast-forming unit-erythroid (BFU-E), colony-forming units (CFU)-GEMM, -GM, -G and -M colonies in methylcellulose. Colony formation was photographed and quantified after 14 days and compared between naEFCs and freshly isolated CD133⁺ and CD133⁻ cells (mean ± sem, n = 3).</p

Gene expression analysis of naEFCs versus HUVEC.

<p>In (A), a heat map illustrating the hierarchical clustering of Log2 relative gene expression in 3 separate HUVEC and naEFC samples. In (B), scatter data showing the average gene expression data in naEFCs and HUVEC. The dots represent the gene expression of UCB CD133+ 4 day cultured naEFCs versus HUVEC. The diagonal lines indicate the cut off value of 1.5 fold activation and genes coloured on the basis of expression level (yellow, evenly expressed genes; blue, naEFC upregulated genes; red, naEFC downregulated genes). In (C), ICAM-3 mRNA levels in naEFCs and HUVEC as determined by qPCR with relative gene expression normalised to CycA. Data are expressed as relative fold change (mean ± sem) normalised to HUVEC, n = 3,*<i>p</i><0.05 versus HUVEC. In (D–F), flow cytometric analysis of ICAM-3 on (D) naEFCs, (E) HUVEC and (F) freshly isolated peripheral blood CD133⁺CD117⁺ gated cells. Light dotted line represents the unstained control and the dark line represents cells stained for ICAM-3. One representative experiment is shown n≥3.</p

MALDI-tof/tof-MS of N-glysocylated naEFC membrane proteins using hydrazide-bead capture.

<p>The consensus motif for N-linked glycosylation is highlighted in bold.</p