Mesenchymal Stem Cell Spheroids Retain Osteogenic Phenotype Through α2β1 Signaling.

Unlabelled: The induction of mesenchymal stem cells (MSCs) toward the osteoblastic lineage using osteogenic supplements prior to implantation is one approach under examination to enhance their bone-forming potential. MSCs rapidly lose their induced phenotype upon removal of the soluble stimuli; however, their bone-forming potential can be sustained when provided with continued instruction via extracellular matrix (ECM) cues. In comparison with dissociated cells, MSC spheroids exhibit improved survival and secretion of trophic factors while maintaining their osteogenic potential. We hypothesized that entrapment of MSC spheroids formed from osteogenically induced cells would exhibit better preservation of their bone-forming potential than would dissociated cells from monolayer culture. Spheroids exhibited comparable osteogenic potential and increased proangiogenic potential with or without osteogenic preconditioning versus monolayer-cultured MSCs. Spheroids were then entrapped in collagen hydrogels, and the osteogenic stimulus was removed. In comparison with entrapped dissociated MSCs, spheroids exhibited significantly increased markers of osteogenic differentiation. The capacity of MSC spheroids to retain their osteogenic phenotype upon withdrawal of inductive cues was mediated by α2β1 integrin binding to cell-secreted ECM. These results demonstrate the capacity of spheroidal culture to sustain the mineral-producing phenotype of MSCs, thus enhancing their contribution toward bone formation and repair.SignificanceDespite the promise of mesenchymal stem cells (MSCs) for cell-based therapies for tissue repair and regeneration, there is little evidence that transplanted MSCs directly contribute to new bone formation, suggesting that induced cells rapidly lose their osteogenic phenotype or undergo apoptosis. In comparison with dissociated cells, MSC spheroids exhibit increased trophic factor secretion and improved cell survival. The loss of phenotype represents a significant clinical challenge for cell therapies, yet there is no evidence for whether MSC spheroids retain their osteogenic phenotype upon entrapment in a clinically relevant biomaterial. These findings demonstrate that MSC spheroids retain their osteogenic phenotype better than do dissociated MSCs, and this is due to integrin engagement with the cell-secreted extracellular matrix. These data provide evidence for a novel approach for potentiating the use of MSCs in bone repair

Similar variation in carbon storage between deciduous and evergreen treeline species across elevational gradients

Background and Aims The most plausible explanation for treeline formation so far is provided by the growth limitation hypothesis (GLH), which proposes that carbon sinks are more restricted by low temperatures than by carbon sources. Evidence supporting the GLH has been strong in evergreen, but less and weaker in deciduous treeline species. Here a test is made of the GLH in deciduous-evergreen mixed species forests across elevational gradients, with the hypothesis that deciduous treeline species show a different carbon storage trend from that shown by evergreen species across elevations. Methods Tree growth and concentrations of non-structural carbohydrates (NSCs) in foliage, branch sapwood and stem sapwood tissues were measured at four elevations in six deciduous-evergreen treeline ecotones (including treeline) in the southern Andes of Chile (40°S, Nothofagus pumilio and Nothofagus betuloides; 46°S, Nothofagus pumilio and Pinus sylvestris) and in the Swiss Alps (46°N, Larix decidua and Pinus cembra). Key Results Tree growth (basal area increment) decreased with elevation for all species. Regardless of foliar habit, NSCs did not deplete across elevations, indicating no shortage of carbon storage in any of the investigated tissues. Rather, NSCs increased significantly with elevation in leaves (P < 0·001) and branch sapwood (P = 0·012) tissues. Deciduous species showed significantly higher NSCs than evergreens for all tissues; on average, the former had 11 % (leaves), 158 % (branch) and 103 % (sapwood) significantly (P < 0·001) higher NSCs than the latter. Finally, deciduous species had higher NSC (particularly starch) increases with elevation than evergreens for stem sapwood, but the opposite was true for leaves and branch sapwood. Conclusions Considering the observed decrease in tree growth and increase in NSCs with elevation, it is concluded that both deciduous and evergreen treeline species are sink limited when faced with decreasing temperatures. Despite the overall higher requirements of deciduous tree species for carbon storage, no indication was found of carbon limitation in deciduous species in the alpine treeline ecoton

Single-provenance mature conifers show higher non-structural carbohydrate storage and reduced growth in a drier location

Since growth is more sensitive to drought than photosynthesis, trees inhabiting dry regions are expected to exhibit higher carbohydrate storage and less growth than their conspecifics from more humid regions. However, the same pattern can be the result of different genotypes inhabiting contrasting humidity conditions. To test if reduced growth and high carbohydrate storage are environmentally driven by drought, we examined the growth and non-structural carbohydrate (NSC) concentrations in single-provenance stands of mature trees of Pinus contorta Douglas and Pinus ponderosa Douglas ex C. Lawson planted at contrasting humidity conditions (900 versus 300 mm of annual precipitation) in Patagonia, Chile. Individual tree growth was measured for each species and at each location as mean basal area increment of the last 10 years (BAI10), annual shoot elongation for the period 2011-14, and needle length for 2013 and 2014 cohorts. Additionally, needle, branch, stem sapwood and roots were collected from each sampled tree to determine soluble sugars, starch and total NSC concentrations. The two species showed lower mean BAI10 and 2013 needle length in the dry site; P. ponderosa also had lower annual shoot extension for 2011 and 2014, and lower 2014 needle length, in the dry than in the mesic site. By contrast, NSC concentrations of all woody tissues for both species were either similar or higher in the dry site when compared with the mesic site. Patterns of starch and sugars were substantially different: starch concentrations were similar between sites except for roots of P. ponderosa, which were higher in the dry site, while sugar concentrations of all woody tissues in both species were higher in the dry site. Overall, our study provides evidence that reduced growth along with carbon (C) accumulation is an environmentally driven response to drought. Furthermore, the significant accumulation of low-molecular weight sugars in the dry site is compatible with a prioritized C allocation for osmoregulation. However, since this accumulation did not come at the expense of reduced starch, it is unlikely that growth was limited by C supply in the dry site

Stickleback Gasterosteus aculeatus (Gasterosteidae) – a new species for ichthyofauna of Western Podolian Prydnistrovya

Описано новий для водойм Захiдноподiльського Приднiстров’я вид – Gasterosteus aculeatus Linnaeus, 1758 (Gasterosteidae). Вивчено змiни у пластичних і меристичних ознаках, рацiонi живлення, вiдтвореннi порівняно з іншими водоймами України. Показано, що умови існування туводної форми колючки у водних об’єктах регіону є сприятливими. Зроблено висновок про високу ймовірність подальшого збільшення чисельності колючки та розширення її ареалу. Описан новый для водоемов Заподноподольского Приднестровья вид - GasterosteusaculeatusLinnaeus, 1758 (Gasterosteidae). Изучены изменения в пластических и меристичних признакам, рационе питания, воспроизведении по сравнению с другими водоемами Украины. Показано, что условия существования туводних формы колючки в водных объектах региона благоприятны. Сделан вывод о высокой вероятности дальнейшего увеличения численности колючки и расширение ее ареала.There was described a new species for fish fauna of Western Podolian Prydnistrovya – stickleback Gasterosteus aculeatus Linnaeus, 1758 (Gasterosteidae). The changes of plastic and meristic characters, diet and propagation in comparison with other water bodies of Ukraine were studied. It was shown that present conditions of nonmigratory form of stickleback in waters of the region are favourable. There was made a conclusion on high possibility of further increase of the stickleback number and enlargement of its distribution range.

The virtual cathode chamber

We describe the operating principle and the first experimental results obtained with gas micro-strip detectors realized with anodes only on the active side, the multiplying field being provided from the back-plane and drift electrodes. For high rate operation, the detector has to be implemented on electron conducting supports, with resistivity around 10$^{11}\Omega$ cm. By construction, the ³Virtual Cathode Chamber² is not subjected to the possibility of discharges between anodes and cathodes, thus avoiding one of the most dangerous problems met with standard micro-strip chambers

A practical attack on the fixed RC4 in the wep mode

Abstract. In this paper we revisit a known but ignored weakness of the RC4 keystream generator, where secret state info leaks to the generated keystream, and show that this leakage, also known as Jenkins’ correlation or the RC4 glimpse, can be used to attack RC4 in several modes. Our main result is a practical key recovery attack on RC4 when an IV modifier is concatenated to the beginning of a secret root key to generate a session key. As opposed to the WEP attack from [FMS01] the new attack is applicable even in the case where the first 256 bytes of the keystream are thrown and its complexity grows only linearly with the length of the key. In an exemplifying parameter setting the attack recoversa16-bytekeyin2 48 steps using 2 17 short keystreams generated from different chosen IVs. A second attacked mode is when the IV succeeds the secret root key. We mount a key recovery attack that recovers the secret root key by analyzing a single word from 2 22 keystreams generated from different IVs, improving the attack from [FMS01] on this mode. A third result is an attack on RC4 that is applicable when the attacker can inject faults to the execution of RC4. The attacker derives the internal state and the secret key by analyzing 2 14 faulted keystreams generated from this key