Regeneration of blood vessels within diabetic wounds after treatment with mesenchymal stem cells

Diabetes is a chronic disease that affects more than 30 million Americans. This disorder leads to a variety of acute and chronic complications, including diabetic ulcers (chronic wounds). Chronic wounds often persist due to poor regeneration of the blood supply which is essential to bring nutrients for healing. Particularly, diabetic individuals are prone to damage in their peripheral tissues which leads to a high prevalence of ulcers in their extremities, often leading to limb amputations. The aim of this study is to improve healing outcomes for diabetics through the use of mesenchymal stem cells (MSCs) to stimulate healing, in which vasculogenesis is an important aspect. Catecholamines such as epinephrine (adrenaline) are prevalent in diabetic foot ulcer tissue and have been shown to inhibit wound healing. In this study, healing rates of type II diabetic mice wounds were evaluated when human MSCs were delivered within a collagen scaffold (IntegraTM) and treated with Timolol, a beta blocker that inhibits the effects of epinephrine. We examined wounded mice after 7 days that had received either no MSCs (control), MSCs, or MSCs treated with timolol for blood vessel development using immunohistochemical staining and confocal fluorescence microscopy. Blood vessel biomarkers GSL-I Isolectin B4 and CD31 were used to stain the wound tissue and fluorescent imaging data was quantified using software. Our results indicate that wound tissue treated with MSCs and timolol had the highest blood vessel regeneration and it was statistically significant when compared to control levels. Additionally, a Fluorescent in situ Hybridization (FISH) protocol to identify human chromosomes was successfully implemented using positive and negative control slides so that human MSCs can be identified when delivered to mouse wound tissue. Future experiments will examine how long the MSCs persist and whether they migrate outside the wound tissue bed

Outcome of protease inhibitor substitution with nevirapine in HIV-1 infected children

<p>Abstract</p> <p>Background</p> <p>Protease inhibitors (PIs) have been associated with metabolic complications. There is a trend to switch to simpler therapy to improve these disturbances. We report a case-series describing the effects in metabolic abnormalities in seven HIV-infected children, previously treated with protease inhibitor (PI) after switching to nevirapine.</p> <p>Methods</p> <p>Seven children with stable PI-containing regimen and a long lasting HIV-1 RNA < 50 copies/ml were switched to nevirapine. All patients were naïve to non nucleoside reverse transcriptase inhibitor. PIs were switched to nevirapine. Preentry nucleoside reverse transcriptase inhibitors were maintained. The substitution of PIs with nevirapine was made when the patient showed hyperlipidemia or lipodystrophy or the physician and/or the patient's willingness to simplify. Clinical, laboratory data and anthropometric parameters were assessed every 3 months. Dual-energy X-Ray absorptiometry scans (DXA) was performed at baseline and at 12 months.</p> <p>Results</p> <p>Seven HIV-infected children were enrolled. Median age: 130 months (99,177). Median baseline CD4%: 32%. All had HIV-1 RNA < 50 copies/ml. Median length of preentry PI-therapy was 47 months (28, 91). Median age at the beginning of nevirapine was 120 months (99,177). Median decrease in cholesterol in 7.2 mmol/L was observed (P = 0.09), from baseline to 12 months. HDL-cholesterol increased in 5.1 mmol/L (P = 0.03) throughout the study period. No significant changes were observed in DXA with regard to body fat, but changes in total body bone mineral content and lean body content were significant. CD4% remained stable. All patients but one maintained viral load < 50 copies/ml at 12 months. The patient with virologic failure referred bad adherence. Children referred to take medication more easily.</p> <p>Conclusion</p> <p>PI substitution with nevirapine improved lipid profile in our patients, although this strategy did not show significant changes in body fat or lipodystrophy.</p

Changes in phytoplankton composition in a Mediterranean coastal lagoon in the Cullera Estany (Comunitat Valenciana, Spain)

The Cullera Estany is a coastal lagoon located in a highly intensified agriculture and tourist area in Valencia. This coastal lagoon has connections with the sea that produce marine intrusion and generate a freshwater interface. Four sampling campaigns were carried out during 2010 in order to analyse the phytoplankton composition and its relation to nutrient content through a Redundancy Analysis. Temperature, dissolved oxygen, nitrite and salinity are the main factors controlling the dynamics of phytoplankton community. During July and October, there is water column stratification; meanwhile in March, there is a well-mixed water column. In addition, in May and July campaigns, hypoxia/anoxia conditions are detected at the bottom. The most abundant phytoplankton groups are Diatoms and Cryptophyceae. Diatoms and Cyanophyceae respond positively to temperature while Cryptophyceae, Prasinophyceae and Dinophyceae respond to high salinity and dissolved oxygen values. Furthermore, picoplankton is correlated inversely with nutrient concentrations.This research work has been supported by the Generalitat Valenciana.Pachés Giner, MAV.; Romero Gil, I.; Martínez Guijarro, MR.; Martí Insa, CM.; Ferrer Polo, J. (2014). Changes in phytoplankton composition in a Mediterranean coastal lagoon in the Cullera Estany (Comunitat Valenciana, Spain). Water and Environment Journal. 28(1):135-144. doi:10.1111/wej.12020S135144281Agencia Estatal de Meteorologia. AEMET 2011 http://www.aemet.es/es/portadaTer Braak, C. J. F. (1986). Canonical Correspondence Analysis: A New Eigenvector Technique for Multivariate Direct Gradient Analysis. Ecology, 67(5), 1167-1179. doi:10.2307/1938672Brogueira, M. J., Oliveira, M. do R., & Cabeçadas, G. (2007). Phytoplankton community structure defined by key environmental variables in Tagus estuary, Portugal. Marine Environmental Research, 64(5), 616-628. doi:10.1016/j.marenvres.2007.06.007Caroppo, C. (2000). The contribution of picophytoplankton to community structure in a Mediterranean brackish environment. Journal of Plankton Research, 22(2), 381-397. doi:10.1093/plankt/22.2.381Cloern, J. (2001). Our evolving conceptual model of the coastal eutrophication problem. Marine Ecology Progress Series, 210, 223-253. doi:10.3354/meps210223Domingues, R. B., Barbosa, A., & Galvão, H. (2008). Constraints on the use of phytoplankton as a biological quality element within the Water Framework Directive in Portuguese waters. Marine Pollution Bulletin, 56(8), 1389-1395. doi:10.1016/j.marpolbul.2008.05.006Falco , S. 2003 Comportamiento de los nutrientes en un estuario estratificado: Caso del Delta del EbroReview Lecture - Picoplankton. (1986). Proceedings of the Royal Society of London. Series B. Biological Sciences, 228(1250), 1-30. doi:10.1098/rspb.1986.0037Havskum, H., Schlüter, L., Scharek, R., Berdalet, E., & Jacquet, S. (2004). Routine quantification of phytoplankton groups—microscopy or pigment analyses? Marine Ecology Progress Series, 273, 31-42. doi:10.3354/meps273031Johnson, P. W., & Sieburth, J. M. (1979). Chroococcoid cyanobacteria in the sea: A ubiquitous and diverse phototrophic biomass1. Limnology and Oceanography, 24(5), 928-935. doi:10.4319/lo.1979.24.5.0928Kirkwood , D. Aminot , A. Pertilla , M. 1991 Report on the Results of the Fourth Intercomparison Exercise for Nutrients in Sea Water ICES Cooperative Research Report 174Lidón, A., Ramos, C., & Rodrigo, A. (1999). Comparison of drainage estimation methods in irrigated citrus orchards. Irrigation Science, 19(1), 25-36. doi:10.1007/s002710050068Martí , C.M. 2010 Caracterización ecológica y establecimiento de los criterios para determinar el potencial ecológico en las salinas de la Comunidad ValencianaMösso, C., Sierra, J. P., Rodilla, M., Romero, I., Falco, S., González del Río, J., & Sánchez-Arcilla, A. (2007). High Vertical Resolution Sampling in Density Interfaces of Estuaries and River Plumes. Estuaries and Coasts, 31(2), 258-268. doi:10.1007/s12237-007-9009-4Novarino, G. (2003). A companion to the identification of cryptomonad flagellates (Cryptophyceae = Cryptomonadea). Hydrobiologia, 502(1-3), 225-270. doi:10.1023/b:hydr.0000004284.12535.25Pachés, M., Romero, I., Hermosilla, Z., & Martinez-Guijarro, R. (2012). PHYMED: An ecological classification system for the Water Framework Directive based on phytoplankton community composition. Ecological Indicators, 19, 15-23. doi:10.1016/j.ecolind.2011.07.003Pérez-Ruzafa, A., Gilabert, J., Gutiérrez, J. M., Fernández, A. I., Marcos, C., & Sabah, S. (2002). Hydrobiologia, 475/476, 359-369. doi:10.1023/a:1020343510060Puigserver, M., Ramon, G., Moyà, G., & Martínez-Taberner, A. (2002). Hydrobiologia, 475/476, 493-504. doi:10.1023/a:1020368215511Ramos, C., Agut, A., & Lidón, A. . (2002). Nitrate leaching in important crops of the Valencian Community region (Spain). Environmental Pollution, 118(2), 215-223. doi:10.1016/s0269-7491(01)00314-1Ramos, M. C. (2001). Rainfall distribution patterns and their change over time in a Mediterranean area. Theoretical and Applied Climatology, 69(3-4), 163-170. doi:10.1007/s007040170022Reynolds, C. S. (2006). The Ecology of Phytoplankton. doi:10.1017/cbo9780511542145Rojo , C. Miracle , R. 1984 Fluctuación estacional de las poblaciones fitoplanctónicas del Estany de Cullera (Valencia) Anales de biología, 2 (Sección especial, 2) Universidad de Murcia (Spain) 161 168Suikkanen, S., Laamanen, M., & Huttunen, M. (2007). Long-term changes in summer phytoplankton communities of the open northern Baltic Sea. Estuarine, Coastal and Shelf Science, 71(3-4), 580-592. doi:10.1016/j.ecss.2006.09.004Vargo , G.A. 1978 Using a Fluorescence Microscope. Phytoplankton Manual Monographs on Oceanography Methodology UNESCO 108 112Vicente , E. Miracle , M.R. 1988 Estructura y función de los procariotas en dos ecosistemas lagunares costeros: L'albufera de Valencia y l'Estany de Cullera Actas del Congreso de Biologia Ambiental. II Congreso Mundial Vasco 79 108 ISBN 84-7585-146-

Agriculture impairs stream ecosystem functioning in a tropical catchment

The expansion of agriculture is particularly worrying in tropical regions of the world, where native forests are being replaced by crops at alarming rates, with severe consequences for biodiversity and ecosystems. However, there is little information about the potential effects of agriculture on the functioning of tropical streams, which is essential if we are to assess the condition and ecological integrity of these ecosystems. We conducted a litter decomposition experiment in streams within a tropical catchment, which were subjected to different degrees of agricultural influence: low (protected area, PA), medium (buffer area, BA) and high (agricultural area, AA). We quantified decomposition rates of litter enclosed within coarse-mesh and fine-mesh bags, which allowed the distinction of microbial and detritivore-mediated decomposition pathways. We used litter of three riparian species representing a gradient in litter quality (Alnus acuminate > Ficus irtsipida > Querns burnelioides), and examined detritivore assemblages through the contents of litterbags and benthic samples. We found that the increasing agricultural influence promoted microbial decomposition, probably due to nutrient-mediated stimulation; and inhibited detritivore-mediated and total decomposition because of reduced detritivore numbers, most likely caused by pesticides and sedimentation. Effects were evident for Alnus and Ficus, but not for Querns, which was barely decomposed across the gradient. Our study provides key evidence about the impact of agriculture on topical stream ecosystem functioning, which is associated to changes in stream assemblages and may have far-reaching repercussions for global biochemical cycles. (C) 2020 The Authors. Published by Elsevier B.V.Thisworkwas supported by the National Secretariat for Science, Technology and Innovation (SENACYT; project APY-GC-2018B-052; contract no. 259-2018) and the Ministry of Economy and Finance (MEF; project 019910.001). AC was supported by a fellowship from SENACYT (contract no. 001-2015) and by the National Research System of Panama (SNI; PhD category; contract no. 186-2018). GC was supported by a fellowship from IFARHU-SENACYT (contract no. 270-2018-1011)

Global population genetic structure and demographic trajectories of the black soldier fly, Hermetia illucens

Background The black soldier fly (Hermetia illucens) is the most promising insect candidate for nutrient-recycling through bioconversion of organic waste into biomass, thereby improving sustainability of protein supplies for animal feed and facilitating transition to a circular economy. Contrary to conventional livestock, genetic resources of farmed insects remain poorly characterised. We present the first comprehensive population genetic characterisation of H. illucens. Based on 15 novel microsatellite markers, we genotyped and analysed 2862 individuals from 150 wild and captive populations originating from 57 countries on seven subcontinents. Results We identified 16 well-distinguished genetic clusters indicating substantial global population structure. The data revealed genetic hotspots in central South America and successive northwards range expansions within the indigenous ranges of the Americas. Colonisations and naturalisations of largely unique genetic profiles occurred on all non-native continents, either preceded by demographically independent founder events from various single sources or involving admixture scenarios. A decisive primarily admixed Polynesian bridgehead population serially colonised the entire Australasian region and its secondarily admixed descendants successively mediated invasions into Africa and Europe. Conversely, captive populations from several continents traced back to a single North American origin and exhibit considerably reduced genetic diversity, although some farmed strains carry distinct genetic signatures. We highlight genetic footprints characteristic of progressing domestication due to increasing socio-economic importance of H. illucens, and ongoing introgression between domesticated strains globally traded for large-scale farming and wild populations in some regions. Conclusions We document the dynamic population genetic history of a cosmopolitan dipteran of South American origin shaped by striking geographic patterns. These reflect both ancient dispersal routes, and stochastic and heterogeneous anthropogenic introductions during the last century leading to pronounced diversification of worldwide structure of H. illucens. Upon the recent advent of its agronomic commercialisation, however, current human-mediated translocations of the black soldier fly largely involve genetically highly uniform domesticated strains, which meanwhile threaten the genetic integrity of differentiated unique local resources through introgression. Our in-depth reconstruction of the contemporary and historical demographic trajectories of H. illucens emphasises benchmarking potential for applied future research on this emerging model of the prospering insect-livestock sector.Peer reviewe

The interplay between the Rab27A effectors Slp4-a and MyRIP controls hormone-evoked Weibel-Palade body exocytosis.

Weibel-Palade body (WPB) exocytosis underlies hormone-evoked VWF secretion from endothelial cells (ECs). We identify new endogenous components of the WPB: Rab3B, Rab3D, and the Rab27A/Rab3 effector Slp4-a (granuphilin), and determine their role in WPB exocytosis. We show that Rab3B, Rab3D, and Rab27A contribute to Slp4-a localization to WPBs. siRNA knockdown of Slp4-a, MyRIP, Rab3B, Rab3D, Rab27A, or Rab3B/Rab27A, or overexpression of EGFP-Slp4-a or EGFP-MyRIP showed that Slp4-a is a positive and MyRIP a negative regulator of WPB exocytosis and that Rab27A alone mediates these effects. We found that ECs maintain a constant amount of cellular Rab27A irrespective of the WPB pool size and that Rab27A (and Rab3s) cycle between WPBs and a cytosolic pool. The dynamic redistribution of Rab proteins markedly decreased the Rab27A concentration on individual WPBs with increasing WPB number per cell. Despite this, the probability of WPB release was independent of WPB pool size showing that WPB exocytosis is not determined simply by the absolute amount of Rab27A and its effectors on WPBs. Instead, we propose that the probability of release is determined by the fractional occupancy of WPB-Rab27A by Slp4-a and MyRIP, with the balance favoring exocytosis

Height and timing of growth spurt during puberty in young people living with vertically acquired HIV in Europe and Thailand.

OBJECTIVE: The aim of this study was to describe growth during puberty in young people with vertically acquired HIV. DESIGN: Pooled data from 12 paediatric HIV cohorts in Europe and Thailand. METHODS: One thousand and ninety-four children initiating a nonnucleoside reverse transcriptase inhibitor or boosted protease inhibitor based regimen aged 1-10 years were included. Super Imposition by Translation And Rotation (SITAR) models described growth from age 8 years using three parameters (average height, timing and shape of the growth spurt), dependent on age and height-for-age z-score (HAZ) (WHO references) at antiretroviral therapy (ART) initiation. Multivariate regression explored characteristics associated with these three parameters. RESULTS: At ART initiation, median age and HAZ was 6.4 [interquartile range (IQR): 2.8, 9.0] years and -1.2 (IQR: -2.3 to -0.2), respectively. Median follow-up was 9.1 (IQR: 6.9, 11.4) years. In girls, older age and lower HAZ at ART initiation were independently associated with a growth spurt which occurred 0.41 (95% confidence interval 0.20-0.62) years later in children starting ART age 6 to 10 years compared with 1 to 2 years and 1.50 (1.21-1.78) years later in those starting with HAZ less than -3 compared with HAZ at least -1. Later growth spurts in girls resulted in continued height growth into later adolescence. In boys starting ART with HAZ less than -1, growth spurts were later in children starting ART in the oldest age group, but for HAZ at least -1, there was no association with age. Girls and boys who initiated ART with HAZ at least -1 maintained a similar height to the WHO reference mean. CONCLUSION: Stunting at ART initiation was associated with later growth spurts in girls. Children with HAZ at least -1 at ART initiation grew in height at the level expected in HIV negative children of a comparable age

Th1 and Th17 hypercytokinemia as early host response signature in severe pandemic influenza

Abstract Introduction Human host immune response following infection with the new variant of A/H1N1 pandemic influenza virus (nvH1N1) is poorly understood. We utilize here systemic cytokine and antibody levels in evaluating differences in early immune response in both mild and severe patients infected with nvH1N1. Methods We profiled 29 cytokines and chemokines and evaluated the haemagglutination inhibition activity as quantitative and qualitative measurements of host immune responses in serum obtained during the first five days after symptoms onset, in two cohorts of nvH1N1 infected patients. Severe patients required hospitalization (n = 20), due to respiratory insufficiency (10 of them were admitted to the intensive care unit), while mild patients had exclusively flu-like symptoms (n = 15). A group of healthy donors was included as control (n = 15). Differences in levels of mediators between groups were assessed by using the non parametric U-Mann Whitney test. Association between variables was determined by calculating the Spearman correlation coefficient. Viral load was performed in serum by using real-time PCR targeting the neuraminidase gene. Results Increased levels of innate-immunity mediators (IP-10, MCP-1, MIP-1β), and the absence of anti-nvH1N1 antibodies, characterized the early response to nvH1N1 infection in both hospitalized and mild patients. High systemic levels of type-II interferon (IFN-γ) and also of a group of mediators involved in the development of T-helper 17 (IL-8, IL-9, IL-17, IL-6) and T-helper 1 (TNF-α, IL-15, IL-12p70) responses were exclusively found in hospitalized patients. IL-15, IL-12p70, IL-6 constituted a hallmark of critical illness in our study. A significant inverse association was found between IL-6, IL-8 and PaO2 in critical patients. Conclusions While infection with the nvH1N1 induces a typical innate response in both mild and severe patients, severe disease with respiratory involvement is characterized by early secretion of Th17 and Th1 cytokines usually associated with cell mediated immunity but also commonly linked to the pathogenesis of autoimmune/inflammatory diseases. The exact role of Th1 and Th17 mediators in the evolution of nvH1N1 mild and severe disease merits further investigation as to the detrimental or beneficial role these cytokines play in severe illness

STXBP1 promotes Weibel-Palade body exocytosis through its interaction with the Rab27A effector Slp4-a.

Vascular endothelial cells contain unique rod-shaped secretory organelles, called Weibel-Palade bodies (WPBs), which contain the hemostatic protein von Willebrand factor (VWF) and a cocktail of angiogenic and inflammatory mediators. We have shown that the Rab27A effector synaptotagmin-like protein 4-a (Slp4-a) plays a critical role in regulating hormone-evoked WPB exocytosis. Using a nonbiased proteomic screen for targets for Slp4-a, we now identify syntaxin-binding protein 1 (STXBP1) and syntaxin-2 and -3 as endogenous Slp4-a binding partners in endothelial cells. Coimmunoprecipitations showed that STXBP1 interacts with syntaxin-2 and -3, but not with syntaxin-4. Small interfering RNA-mediated silencing of STXBP1 expression impaired histamine- and forskolin-induced VWF secretion. To further substantiate the role of STXBP1, we isolated blood outgrowth endothelial cells (BOECs) from an early infantile epileptic encephalopathy type 4 (EIEE4) patient carrying a de novo mutation in STXBP1. STXBP1-haploinsufficient EIEE4 BOECs contained similar numbers of morphologically normal WPBs compared with control BOECs of healthy donors; however, EIEE4 BOECs displayed significantly impaired histamine- and forskolin-stimulated VWF secretion. Based on these findings, we propose that the Rab27A-Slp4-a complex on WPB promotes exocytosis through an interaction with STXBP1, thereby controlling the release of vaso-active substances in the vasculature