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751.
Viable suspended hepatocytes were prepared from surgical biopsy specimens of pig and human liver by digestion with collagenase. Initial perfusion of the tissue through cannulated blood vessels with 0.5 mM EGTA followed by 0.2% collagenase gave the best results. 20−870 × 106 cells of which 60–95 % excluded trypan blue were obtained from 5–30 g pig liver pieces, while results with human liver specimens were usually less satisfactory. In some experiments, however, viable cells, as judged by vital stain exclusion and ability to synthesize lipids were obtained in sufficient yield. In the pig hepatocytes glycerolipid synthesis from [3H] glycerol and oxidation and esterification of [14C] oleic acid had the same characteristics as those observed earlier in rat hepatocytes.  相似文献   
752.
The purpose of this study was to evaluate Cellvibrio japonicus as a potential host strain for one‐step bioconversion of hemicellulose polymers to value‐added products. C. japonicus could be cultivated on all main lignocellulose monosaccharides as well as xylan polymers as a sole carbon source. This is particularly interesting as most industrially relevant bacteria are neither able to depolymerize wood polymers nor metabolize most hemicellulose monosaccharides. As a result, lignocellulose raw materials typically have to be degraded employing additional processes while the complete conversion of all lignocellulose sugars remains a challenge. Exemplary for a value‐added product, a one‐step conversion of xylan polymers to mono‐rhamnolipid biosurfactants with C. japonicus after transformation with the plasmid pSynPro8oT carrying the genes rhlAB was demonstrated. As achieved product yields in this one‐step bioconversion process are comparably low, many challenges remain to be overcome for application on an industrial scale. Nonetheless, this study provides a first step in the search for establishing a future host strain for bioeconomy, which will ideally be used for bioconversion of lignocellulose polymers with as little exhaustive pretreatment as possible.  相似文献   
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Cord blood–derived neural stem cells (NSCs) are proposed as an alternative cell source to repair brain damage upon transplantation. However, there is a lack of data showing how these cells are driven to generate desired phenotypes by recipient nervous tissue. Previous research indicates that local environment provides signals driving the fate of stem cells. To investigate the impact of these local cues interaction, the authors used a model of cord blood–derived NSCs co-cultured with different rat brain–specific primary cultures, creating the neural-like microenvironment conditions in vitro. Neuronal and astro-, oligo-, and microglia cell cultures were obtained by the previously described methods. The CMFDA-labeled neural stem cells originated from, non-transformed human umbilical cord blood cell line (HUCB-NSCs) established in a laboratory. The authors show that the close vicinity of astrocytes and oligodendrocytes promotes neuronal differentiation of HUCB-NSCs, whereas postmitotic neurons induce oligodendrogliogenesis of these cells. In turn, microglia or endothelial cells do not favor any phenotypes of their neural commitment. Studies have confirmed that HUCB-NSCs can read cues from the neurogenic microenvironment, attaining features of neurons, astrocytes, or oligodendrocytes. The specific responses of neurally committed cord blood–derived cells, reported in this work, are very much similar to those described previously for NSCs derived from other “more typical” sources. This further proves their genuine neural nature. Apart from having a better insight into the neurogenesis in the adult brain, these findings might be important when predicting cord blood cell derivative behavior after their transplantation for neurological disorders.  相似文献   
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Tethered bilayer lipid membranes (tBLMs) are robust and flexible model platforms for the investigation of various membrane related processes. They are especially suited to study the incorporation and function of ion channel proteins, where a high background resistance of the membrane is essential. Synthetic M2 peptides, analogues of the transmembrane fragment of the acetylcholine receptor, could be incorporated into two different membrane architectures. The functional reconstitution and the formation of a conducting pore are shown by electrochemical impedance spectroscopy (EIS). The pore is selective for small monovalent cations, while bulky ions cannot pass. This is a significant step towards a novel biosensing approach. We envision a device, where a stable and insulating membrane would be attached to an electronic read-out unit and embedded proteins would serve as actual sensing units.  相似文献   
758.
The current classification of the Malagasy-Comoroan geckos of the genus Geckolepis recognizes three valid species and is based on morphological differences only. Species allocation of individuals is difficult, mainly because of the insufficiently known degree of variation and the frequent partial loss of the integument in preserved specimens. Here we study Geckolepis specimens from almost their entire known range. We combine molecular and morphological data to estimate species richness in the genus, and to assess the taxonomic validity of the morphological differences proposed. Analyses of mitochondrial (12S, ND4) and nuclear (RAG1, CMOS) gene sequences support three major clades in addition to the morphologically distinct G. polylepis (included for 12S only), and the presence of various divergent lineages within these clades, some occurring in sympatry. Among lineages, the external morphology seems to be comparatively conservative, and differences are faint in many cases. For each major lineage, we summarize the most diagnostic morphological characters that distinguish it from other lineages. The combined molecular and morphological data strongly indicate the presence of highly divergent lineages, three of which are tentatively referred to the names Geckolepis maculata, G. polylepis and G. typica, whereas three other major lineages are considered Confirmed Candidate Species. Among the remaining lineages, differentiation is shallower, and these may qualify for Deep Conspecific Lineages. In conclusion, our analysis provides evidence for an underestimation of species richness in the genus and a complex evolutionary history, not reflected by current Geckolepis species-level taxonomy.  相似文献   
759.
To improve our understanding of the floral biology, pollination, and systematics of the genus Narcissus, a comparative study was made of flower volatiles from nine species native to southern Spain using headspace collection and GC-MS analysis. The species fell into three fragrance types based on the identity of their major volatiles. In all but one species the fragrances consisted mainly of monoterpene isoprenoids mixed with benzenoids: in six species trans-ß-ocimene occurred in high proportions, in two others it was lacking; the last species had a fragrance dominated by fatty acid derivatives, mixed with terpenoids. Two of the species showed marked intraspecific variation in many of their volatiles. When the volatile data matrix of all species was subjected to cluster analyses and the resulting phenetic trees compared with currently recognized taxonomic groups, there was no congruence at the subgeneric level. However, there was considerable agreement at the sectional level, although in most sections we studied only a single species. This apparent agreement was stronger when the volatiles were analyzed according to shared biosynthetic pathways rather than treated individually, pointing to the higher value of using biosynthetic pathways for uncovering and confirming phenetic, and probable evolutionary, relationships among species. In terms of possible selective pressures from pollinators in shaping fragrance chemistry, available information on the pollination of our species suggested an association between fragrance and types of pollinators. Two pollinator-fragrance groups were apparent: (1) species pollinated by insects that include butterflies and moths displayed fragrances containing volatiles typical of moth-pollinated flowers, most particularly indole combined with high amounts of esters, and (2) species visited exclusively by insects other than butterflies and moths, especially by bees and flies, had fragrances lacking this combination of volatiles. Narcissus assoanus was unusual among our species in having both fragrance chemotypes. Future pollination studies of Narcissus in the field are needed to test the reliability and predictability of the proposed fragrance-pollinator associations.  相似文献   
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