Surface functionalisation of nanodiamonds for human neural stem cell adhesion and proliferation.

Biological systems interact with nanostructured materials on a sub-cellular level. These interactions may govern cell behaviour and the precise control of a nanomaterial's structure and surface chemistry allow for a high degree of tunability to be achieved. Cells are surrounded by an extra-cellular matrix with nano-topographical properties. Diamond based materials, and specifically nanostructured diamond has attracted much attention due to its extreme electrical and mechanical properties, chemical inertness and biocompatibility. Here the interaction of nanodiamond monolayers with human Neural Stem Cells (hNSCs) has been investigated. The effect of altering surface functionalisation of nanodiamonds on hNSC adhesion and proliferation has shown that confluent cellular attachment occurs on oxygen terminated nanodiamonds (O-NDs), but not on hydrogen terminated nanodiamonds (H-NDs). Analysis of H and O-NDs by Atomic Force Microscopy, contact angle measurements and protein adsorption suggests that differences in topography, wettability, surface charge and protein adsorption of these surfaces may underlie the difference in cellular adhesion of hNSCs reported here

Biological membranes in EV biogenesis, stability, uptake, and cargo transfer: an ISEV position paper arising from the ISEV membranes and EVs workshop

Paracrine and endocrine roles have increasingly been ascribed to extracellular vesicles (EVs) generated by multicellular organisms. Central to the biogenesis, content, and function of EVs are their delimiting lipid bilayer membranes. To evaluate research progress on membranes and EVs, the International Society for Extracellular Vesicles (ISEV) conducted a workshop in March 2018 in Baltimore, Maryland, USA, bringing together key opinion leaders and hands-on researchers who were selected on the basis of submitted applications. The workshop was accompanied by two scientific surveys and covered four broad topics: EV biogenesis and release; EV uptake and fusion; technologies and strategies used to study EV membranes; and EV transfer and functional assays. In this ISEV position paper, we synthesize the results of the workshop and the related surveys to outline important outstanding questions about EV membranes and describe areas of consensus. The workshop discussions and survey responses reveal that while much progress has been made in the field, there are still several concepts that divide opinion. Good consensus exists in some areas, including particular aspects of EV biogenesis, uptake and downstream signalling. Areas with little to no consensus include EV storage and stability, as well as whether and how EVs fuse with target cells. Further research is needed in these key areas, as a better understanding of membrane biology will contribute substantially towards advancing the field of extracellular vesicles.Fil: Russell, Ashley E.. University Johns Hopkins; Estados UnidosFil: Sneider, Alexandra. University Johns Hopkins; Estados UnidosFil: Witwer, Kenneth W.. University Johns Hopkins; Estados UnidosFil: Bergese, Paolo. Università Degli Studi Di Brescia; ItaliaFil: Bhattacharyya, Suvendra N.. Indian Institute of Chemical Biology; IndiaFil: Cocks, Alexander. Cardiff University; Reino UnidoFil: Cocucci, Emanuele. Ohio State University; Estados UnidosFil: Erdbrügger, Uta. University of Virginia; Estados UnidosFil: Falcon Perez, Juan M.. Ikerbasque Basque Foundation for Science; EspañaFil: Freeman, David W.. National Institute On Aging National Institute for Helth ; Estados UnidosFil: Gallagher, Thomas M.. Loyola University Of Chicago; Estados UnidosFil: Hu, Shuaishuai. Technological University Dublin; IrlandaFil: Huang, Yiyao. University Johns Hopkins; Estados Unidos. Southern Medical University; ChinaFil: Jay, Steven M.. University of Maryland; Estados UnidosFil: Kano, Shin-ichi. The University of Alabama at Birmingham School of Medicine; Estados UnidosFil: Lavieu, Gregory. Institut Curie; FranciaFil: Leszczynska, Aleksandra. University of California at San Diego; Estados UnidosFil: Llorente, Alicia M.. Oslo University Hospital; NoruegaFil: Lu, Quan. Harvard University. Harvard School of Public Health; Estados UnidosFil: Mahairaki, Vasiliki. University Johns Hopkins; Estados UnidosFil: Muth, Dillon C.. University Johns Hopkins; Estados UnidosFil: Noren Hooten, Nicole. National Institute On Aging National Institute for Helth ; Estados UnidosFil: Ostrowski, Matias. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; ArgentinaFil: Prada, Ilaria. Consiglio Nazionale delle Ricerche; ItaliaFil: Sahoo, Susmita. Icahn School of Medicine at Mount Sinai ; Estados UnidosFil: Schøyen, Tine Hiorth. Uit The Arctic University Of Norway; Noruega. University Johns Hopkins; Estados UnidosFil: Sheng, Lifuy. University of Washington. School of Medicine; Estados UnidosFil: Tesch, Deanna. Shaw University; Estados UnidosFil: Van Niel, Guillaume. No especifíca;Fil: Vandenbroucke, Roosmarijn E.. University of Ghent; BélgicaFil: Verweij, Frederik J.. No especifíca;Fil: Villar, Ana V.. Universidad de Cantabria; EspañaFil: Wauben, Marca. University of Utrecht; Países BajosFil: Wehman, Ann M.. Universität Würzburg; AlemaniaFil: Ardavan, Arzhang. Peking University; ; ChinaFil: Carter, David Raul Francisco. Oxford Brookes University; Reino UnidoFil: Vader, Pieter. University Medical Center Utrecht; Países Bajo

Gene expression patterns associated with blood-feeding in the malaria mosquito Anopheles gambiae

BACKGROUND: Blood feeding, or hematophagy, is a behavior exhibited by female mosquitoes required both for reproduction and for transmission of pathogens. We determined the expression patterns of 3,068 ESTs, representing ~2,000 unique gene transcripts using cDNA microarrays in adult female Anopheles gambiae at selected times during the first two days following blood ingestion, at 5 and 30 min during a 40 minute blood meal and at 0, 1, 3, 5, 12, 16, 24 and 48 hours after completion of the blood meal and compared their expression to transcript levels in mosquitoes with access only to a sugar solution. RESULTS: In blood-fed mosquitoes, 413 unique transcripts, approximately 25% of the total, were expressed at least two-fold above or below their levels in the sugar-fed mosquitoes, at one or more time points. These differentially expressed gene products were clustered using k-means clustering into Early Genes, Middle Genes, and Late Genes, containing 144, 130, and 139 unique transcripts, respectively. Several genes from each group were analyzed by quantitative real-time PCR in order to validate the microarray results. CONCLUSION: The expression patterns and annotation of the genes in these three groups (Early, Middle, and Late genes) are discussed in the context of female mosquitoes' physiological responses to blood feeding, including blood digestion, peritrophic matrix formation, egg development, and immunity

Integrins of Anopheles gambiae and a putative role of a new beta integrin, BINT2, in phagocytosis of E. coli

Contains fulltext : 35755.pdf (publisher's version ) (Closed access)Mosquitoes use effective immune responses, including phagocytosis, to fight microbial infection. Here we show that in an Anopheles gambiae immune responsive cell line, RGD recognizing receptors play an important role in the phagocytic response, suggesting overlap between molecular components implicated in adhesion and phagocytosis. Integrins are a major class of adhesive receptors that recognize ligands containing an RGD motif. We have cloned a gene encoding a new beta integrin, BINT2, and demonstrated its involvement in Escherichia coli engulfment. Based on molecular modeling, we propose a structural reason for the role of BINT2, but not BINT1, on phagocytosis of Gram-negative bacteria. Using bioinformatic tools, we have identified and compared the complete A. gambiae integrin repertoire as a prelude to a future systematic functional study

Close association of invading Plasmodium berghei and beta integrin in the Anopheles gambiae midgut.

We have used confocal microscopy and an antibody against Anopheles gambiae beta integrin to study this protein's distribution in the mosquito midgut and its relationship to invading Plasmodium berghei parasites. An extensive reorganization of integrin is seen to take place in the midgut epithelial cells following the uptake of either non-infected or parasite-infected blood meal, probably reflecting the reshaping of the gut due to the presence of the food bolus and the peritrophic membrane that surrounds it. Furthermore, malaria parasites are coated with beta integrin immediately upon entry into the epithelium, independent of whether they develop intra- or extracellularly. Although this coat is shed a few days after the invasion, beta integrin remains concentrated in the cells surrounding the maturing oocyst for several days. Finally, the antibody detects a structural change in the midgut epithelial cells in the immediate vicinity of the invading ookinete, which is consistent with Plasmodium-induced apoptosis followed by wound healing. This intimate association suggests a specific role of beta integrin in the invasion process

Brain Tissue-Derived Extracellular Vesicles in Alzheimer's Disease Display Altered Key Protein Levels Including Cell Type-Specific Markers

Background: Brain tissue-derived extracellular vesicles (bdEVs) play neurodegenerative and protective roles, including in Alzheimer's disease (AD). Extracellular vesicles (EVs) may also leave the brain to betray the state of the CNS in the periphery. Only a few studies have profiled the proteome of bdEVs and source brain tissue. Additionally, studies focusing on bdEV cell type-specific surface markers are rare. Objective: We aimed to reveal the pathological mechanisms inside the brain by profiling the tissue and bdEV proteomes in AD patients. In addition, to indicate targets for capturing and molecular profiling of bdEVs in the periphery, CNS cell-specific markers were profiled on the intact bdEV surface. Methods: bdEVs were separated and followed by EV counting and sizing. Brain tissue and bdEVs from age-matched AD patients and controls were then proteomically profiled. Total tau (t-tau), phosphorylated tau (p-tau), and antioxidant peroxiredoxins (PRDX) 1 and 6 were measured by immunoassay in an independent bdEV separation. Neuron, microglia, astrocyte, and endothelia markers were detected on intact EVs by multiplexed ELISA. Results: Overall, concentration of recovered bdEVs was not affected by AD. Proteome differences between AD and control were more pronounced for bdEVs than for brain tissue. Levels of t-tau, p-tau, PRDX1, and PRDX6 were significantly elevated in AD bdEVs compared with controls. Release of certain cell-specific bdEV markers was increased in AD. Conclusion: Several bdEV proteins are involved in AD mechanisms and may be used for disease monitoring. The identified CNS cell markers may be useful tools for peripheral bdEV capture