Recreating the perivascular niche ex vivo using a microfluidic approach

Stem cell niches are composed of numerous microenvironmental features, including soluble and insoluble factors, cues from other cells, and the extracellular matrix (ECM), which collectively serve to maintain stem cell quiescence and promote their ability to support tissue homeostasis. A hallmark of many adult stem cell niches is their proximity to the vasculature in vivo, a feature common to neural stem cells, mesenchymal stem cells (MSCs) from bone marrow and adipose tissue, hematopoietic stem cells, and many tumor stem cells. In this study, we describe a novel 3D microfluidic device (MFD) as a model system in which to study the molecular regulation of perivascular stem cell niches. Endothelial cells (ECs) suspended within 3D fibrin gels patterned in the device adjacent to stromal cells (either fibroblasts or bone marrow‐derived MSCs) executed a morphogenetic process akin to vasculogenesis, forming a primitive vascular plexus and maturing into a robust capillary network with hollow well‐defined lumens. Both MSCs and fibroblasts formed pericytic associations with the ECs but promoted capillary morphogenesis with distinct kinetics. Biochemical assays within the niche revealed that the perivascular association of MSCs required interaction between their α6β1 integrin receptor and EC‐deposited laminin. These studies demonstrate the potential of this physiologically relevant ex vivo model system to study how proximity to blood vessels may influence stem cell multipotency. Biotechnol. Bioeng. 2010;107: 1020–1028. © 2010 Wiley Periodicals, Inc.     

Development of 4-cell mouse embryos after re-vitrification

This paper reports studies on the effects of re-vitrification by the CPS (Closed Pulled Straw) method on the development of 4-cell stage mouse embryos. The procedure involved culturing 2-cell mouse embryos in G-1 medium until the 4-cell stage followed by the division of the normal 4-cell stage embryos into a control group (non-vitrified) and two experimental subgroups (vitrified and re-vitrified). Embryos in the vitrified subgroup were cryopreserved by the CPS vitrification method. In the second experimental subgroup (re-vitrified), embryos that were already vitrified were warmed and cryopreserved again by the same method. There was no significant reduction in the rate of blastocyst formation after vitrification and re-vitrification. However, re-vitrification reduced the total cell number, ICM (inner cell mass) percent and blastocyst diameter (P<0.05). These results showed that vitrification and re-vitrification by the CPS method did not negatively affect the development of vitrified-warmed 4-cell mouse embryos, whereas re-vitrification significantly reduced both the cell number and diameter of blastocysts.     

Chromoblastomycosis due to Fonsecaea pedrosoi and F. monophora in Cuba

We report two cases of chromoblastomycosis due to Fonsecaea pedrosoi and F. monophora in otherwise healthy Cuban males. Direct microscopic examination of biopsies revealed muriform cells, the hallmark of chromoblastomycosis. The suspected agents were recovered in culture, identified on the basis of morphological criteria and confirmed by sequencing of the internal transcribed spacer regions of rDNA. Final treatment consisted of surgical excision. The patients were successfully cured since there was no relapse after a follow-up of more than a year. In vitro antifungal susceptibility testing of both isolates showed that itraconazole and posaconazole had potent activity. High MICs of amphotericin B (2 μg/ml), fluconazole (>64 μg/ml), anidulafungin (8 μg/ml) and caspofungin (8 μg/ml) were found.     

Towards a better understanding of agronomic and soil basis for possible charcoal root rot control and production improvement in bean

Abstract

Macrophomina phaseolina causes charcoal root rot (CRR) in numerous crops worldwide. However, structural progression of CRR epidemics in association with bean agro-ecosystems is little understood. Thus, progression of CRR epidemics in interaction with 13 agro-ecological variables was characterised in 48 commercial bean fields. Frequency of pathogen isolated from root was assessed at vegetative (V3), flowering-podding (R6-7) and pod maturity (R9) growth stages. Two principal factors accounting for 89% of total data variance characterised CRR epidemics. Rhizobial nodulation and cultivation–depth, clay–pH, date–preceding crop, urea–bean class and urea–herbicide interactions accounted for 50% CRR epidemic dynamics. Sixty-five percent of production variance was explained by soil clay, bean class, colonisation, planting date and depth, cultivation method, preceding crop, nodulation, herbicide and urea application. The present findings enabled us to explain a noticeable part of variations in productivity among the bean CRR pathosystems with the help of certain agricultural and environmental events for sustainable agriculture purposes.     

Introduction of Beauveria bassiana as a biological control agent against the Ommatissus lybicus in Kerman province

Dubas bug, Ommatissus lybicus is one of the major sucking pests of date palm. Data palm is the most important economic crop in Iran. In this research, we isolated the pathogenic strain KB 201, KB 206, KB 211, KB 215, KB 220 and KB 512, of entomopathogenic fungi Beauveria bassiana from soil and insects, which produced aerial and submerged conidia and blastospores in the laboratory conditions. We investigated the best conditions for the production and utilisation of the spore suspension to spray the nymph of O. lybicus, which is one of the important pests in Kerman province (Iran). A total of 180 isolates that naturally infected by O. lybicus were collected from date palm in Kerman province during spring and summer seasons 2010–2011. The pathogen city test was carried out with direct spray. To bioassay the isolates, three concentrations of the spore suspension were prepared as follows: 1?×?10⁶, 1?×?10⁷ and 1?×?10⁸?conidia/ml. The pests were sprayed by aerial conidial suspension, which was prepared by 0.01% Tween 80 in distilled water, and the controls were sprayed by 0.01% Tween 80in distilled water. After spraying the pests, the plates were incubated at 25?±?1?°C and 80% of relative humidity. Then, the treated pests were monitored every day for the fungal growth and mortality.     

Odonata fauna of Khuzestan province (Iran) with a note on an Oriental species Trithemis aurora (Burmeister, 1839) as a new record for the south-west of Iran

This paper presents a survey of spider species that have not been previously recorded for Israel. Twenty species, twelve genera and two families (Mysmenidae and Phyxelididae) are recorded for the first time in Israel. Nine species, Agroeca parva Bosmans, 2011, Aulonia kratochvili Dunin et al., 1986, Ero flammeola Simon, 1881, Hogna ferox (Lucas, 1838), Maculoncus parvipalpus Wunderlich, 1995, Neon rayi (Simon, 1875), Pardosa aenigmatica Tongiorgi, 1966 and Phyxelida anatolica Griswold, 1990, are illustrated. Tarentula jaffa Strand, 1913, syn. n. is synonymised with Hogna ferox (Lucas, 1838), and Hahnia carmelita Levy, 2007, syn. n. is synonymised with Hahnia nava (Westring, 1851). A possible synonymy of the widespread Prodidomus rufus Hentz, 1847 with P. hispanicus Dalmas, 1919 known from the Iberian Peninsula is discussed. An original combination is restored for Trochosa urbana O.P.-Cambridge, 1876.     

Reduced efficacy of multiple doses of CpG-matured dendritic cell tumor vaccine in an experimental model

CpG motifs have been advanced as agents that stimulate the maturation of DCs for immunotherapy. The present study tested the hypothesis that multiple doses of CpG-matured DC vaccine would be efficacious for complete eradication of experimentally-induced tumor. Accordingly, WEHI164 cells were implanted subcutaneously in the flank of BALB/c mice. During DC culture, tumor lysate was added to immature DCs followed by addition of CpG or non-CpG control 4–6 h later. A total of three doses of CpG or non-CpG control-matured DCs were injected around tumors. The results showed that multiple doses of CpG-matured DCs led to considerable decrease in cytotoxicity of lymphocytes and significantly increased tumor growth rate compared to a single dose. Further, mice which received three doses of the vaccine also displayed significant FoxP3 in tumor tissue. In conclusion, multiple doses of CpG-matured DCs exhibited decreased anti-tumor immunity in association with increased expression of FoxP3.     

Insulin-like growth factor 1 (IGF-I) improves hepatic differentiation of human bone marrow-derived mesenchymal stem cells

The ability of MSCs (mesenchymal stem cells) to differentiate between other cell types makes these cells an attractive therapeutic tool for cell transplantation. This project was designed to improve transdifferentiation of human MSCs into liver cells using IGF-I (insulin-like growth factor 1) which, despite its important role in liver development, has not been used for in vitro hepatic differentiation. In the present study, the MSCs derived from healthy human bone marrow samples were cultured and characterized by immunophenotyping and differentiation potential into osteoblast and adipocytes. Transdifferentiation into hepatocyte-like cells was performed in the presence/absence of IGF-I in combination with predefined hepatic differentiation cocktail. To evaluate transdifferentiation, morphological features, immuno-cytochemical staining of specific biological markers and hepatic functions were assessed. Morphological assessment and evaluation of glycogen content, albumin and AFP (α-feto protein) expression as well as albumin and urea secretion revealed statistically significant difference between experimental groups compared with the control. Morphology and function (albumin secretion) of IGF-I-treated cells were significantly better than IGF-I-free experimental group. To the best of our knowledge, our study is the first to demonstrate that the combination of IGF-I with the predefined hepatic differentiation cocktail will significantly improve the morphological features of the differentiated cells and albumin secretion.