Cellular Environment Directs Differentiation of Human Umbilical Cord Blood�CDerived Neural Stem Cells In Vitro

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.     

Characterization of juvenile hormone esterase from genetically-determined wing morphs of the cricket, Gryllus rubens

Long-winged (LW) vs short-winged (SW) genetic stocks of the cricket Gryllus rubens differ in plasma juvenile hormone esterase (JHE) activity during the last stadium. These activity differences may be important in morph determination. In the present study, plasma JHEs from the LW vs SW stocks were characterized with respect to a variety of kinetic and physical characteristics. Gel permeation chromatography of LW or SW plasma each resulted in a single JHE peak of high molecular weight (190 kDa). This molecular weight is about twice as high as that of JHEs from most other insects. The apparent Michaelis constant for JH III ranged from 47 to 81 nM. Like JHEs from other insects, the enzyme from G. rubens was inhibited strongly by trifluoropropanone transition-state analogs and weakly by the general esterase inhibitors, eserine and DFP. JHEs from LW and SW plasma exhibited no significant differences in K_M, inhibition by trifluoropropanone or general esterase inhibitors, thermal denaturation profiles or pH profiles. The absence of K_M differences between LW and SW JHEs indicate that the 2–4 fold higher enzyme activity in LW plasma, previous documented in assays employing saturating substrate concentration, will exist under physiological substrate concentrations. Two isoforms (pI = 5.1, 4.2−4.1) were identified in SW plasma but only the more acidic form was observed in LW plasma. This is the first documentation of genetically-determined differences in JHE isozymes in any insect species. However, the functional significance of these isoform differences, if any, remains to be established. These results provide no evidence that the plasma JHE activity differences between LW and SW stocks results from allozymes or isozymes with altered kinetic or stability characteristics.     

Functional incorporation of the pore forming segment of AChR M2 into tethered bilayer lipid membranes

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.     

Molecules and morphology suggest cryptic species diversity and an overall complex taxonomy of fish scale geckos, genus Geckolepis

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.     

Interspecific variation in floral fragrances within the genus Narcissus (Amaryllidaceae)

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.