Expression of neuronal markers suggests heterogeneity of chick sympathoadrenal cells prior to invasion of the adrenal anlagen

We have analyzed the distribution of neural crest-derived precursors and the expression of catecholaminergic and neuronal markers in developing adrenal tissue of chick embryos. Undifferentiated neural crest cells are found in presumptive adrenal regions from embryonic day 3 (E3) onward. An increasing proportion of cells expressing tyrosine hydroxylase (TH) mRNA indicates catecholaminergic differentiation of precursors not only in primary sympathetic ganglia, but also in presumptive adrenal regions. Whereas precursors and differentiating cells show mesenchymal distribution until E5, discrete adrenal anlagen form during E6. Even during E5, catecholaminergic cells with low or undetectable neurofilament M (NF-M) mRNA expression prevail in positions at which adrenal anlagen become distinct during E6. The predominance of TH-positive and NF-M-negative cells is maintained throughout embryogenesis in adrenal tissue. RNA encoding SCG10, a pan-neuronal marker like NF-M, is strongly expressed throughout adrenal anlagen during E6 but is found at reduced levels in chromaffin cells compared with neuronal cells at E15. Two additional neuronal markers, synaptotagmin 1 and neurexin 1, are expressed at low to undetectable levels in developing chromaffin cells throughout embryogenesis. The developmental regulation of neuronal markers shows at least three different patterns among the four mRNAs analyzed. Importantly, there is no generalized downregulation of neuronal markers in developing adrenal anlagen. Thus, our observations question the classical concept of chromaffin differentiation from a common sympathoadrenal progenitor expressing neuronal properties and suggest alternative models with changing instructive signals or separate progenitor populations for sympathetic neuronal and chromaffin endocrine cells.Chaya Kalcheim and Klaus Unsicker are supported by the Deutsche Forschungsgemeinschaft (SFB 488)     

Ultrastructure of the spermatozoon of <Emphasis Type="Italic">Talpa romana</Emphasis> (Mammalia,Lipotyphla)

In this study, we used SEM and TEM to investigate the ultrastructure of spermatozoa from the cauda epididymis of Talpa romana. For comparison, we also analysed spermatozoa from the cauda epididymis of T. europaea captured in the same area. The male gamete of T. romana has a flattened head with an elliptic profile, consisting of a large acrosome and a nuclear region separated by a thin subacrosomal space. At the tip of the nucleus, the subacrosomal space ends in a finger-shaped projection. The tail includes a connecting piece, middle piece, principal piece and end piece. The male gametes of T. romana are substantially similar to those of T. europaea. A comparison with other species of insectivores permits extension of the similarity of sperm features to Scalopus aquaticus and Condylura cristata. Many spermatozoa from the cauda epididymis of T. romana and T. europaea have the tail bent at the annulus, and this is always associated with remnants of cytoplasmic droplets. This morphology is considered to be a common phenomenon.     

Multiple imputation procedures allow the rescue of missing data: an application to determine serum tumor necrosis factor (TNF) concentration values during the treatment of rheumatoid arthritis patients with anti-TNF therapy

Longitudinal studies aimed at evaluating patients clinical response to specific therapeutic treatments are frequently summarized in incomplete datasets due to missing data. Multivariate statistical procedures use only complete cases, deleting any case with missing data. MI and MIANALYZE procedures of the SAS software perform multiple imputations based on the Markov Chain Monte Carlo method to replace each missing value with a plausible value and to evaluate the efficiency of such missing data treatment. The objective of this work was to compare the evaluation of differences in the increase of serum TNF concentrations depending on the -308 TNF promoter genotype of rheumatoid arthritis (RA) patients receiving anti-TNF therapy with and without multiple imputations of missing data based on mixed models for repeated measures. Our results indicate that the relative efficiency of our multiple imputation model is greater than 98% and that the related inference was significant (p-value < 0.001). We established that under both approaches serum TNF levels in RA patients bearing the G/A -308 TNF promoter genotype displayed a significantly (p-value < 0.0001) increased ability to produce TNF over time than the G/G patient group, as they received successively doses of anti-TNF therapy.     

ISCOM-based vaccines: the second decade

The immunostimulating complex or 'iscom' was first described 20 years ago as an antigen delivery system with powerful immunostimulating activity. Iscoms are cage-like structures, typically 40 nm in diameter, that are comprised of antigen, cholesterol, phospholipid and saponin. ISCOM-based vaccines have been shown to promote both antibody and cellular immune responses in a variety of experimental animal models. This review focuses on the evaluation of ISCOM-based vaccines in animals over the past 10 years, as well as examining the progress that has been achieved in the development of human vaccines based on ISCOM adjuvant technology.     

Host compatibility of the cloud forest mistletoe Psittacanthus schiedeanus (Loranthaceae) in Central Veracruz, Mexico

The consequences of the seed deposition of the parasitic mistletoe Psittacanthus schiedeanus were evaluated in a 32-mo study. We conducted a field seed inoculation experiment to determine variation in seed adhesion, seed germination, seedling establishment, and plant survival to reproduction among five host species and to evaluate whether these post-dispersal processes explain mistletoe prevalence and specificity at the regional scale. Seeds without an exocarp were inoculated onto branches of individuals of the five most common host species identified in nature in central Veracruz, México. Seed fate was monitored for 2 yr, at weekly intervals for the first 2 mo and at 2-mo intervals thereafter. The height and diameter of experimental host branches and canopy cover above them were measured to see if these factors affected mistletoe establishment. Significant differences in seed attachment and seed germination were found among host species. Fewer seeds remained attached on experimental branches of Quercus germana than those of Liquidambar styraciflua, Acacia pennatula, and Platanus mexicana. Although significant differences in seed germination were observed among species (significantly greater on A. pennatula), >70% of mistletoe seeds germinated within the first 5 wk on all host species. Towards the end of the inoculation experiment, more mistletoe seedlings survived, grew, and then flowered on Liquidambar styraciflua than on A. pennatula, P. mexicana, Q. germana, or Q. leiophylla. Host branch initial height and diameter did not affect seedling survival, but seedlings survived better on trees where the canopy was more open. Our results suggest that Liquidambar styraciflua is the most compatible host species with P. schiedeanus in central Veracruz. Not surprisingly, Liquidambar is by far the most common host tree for P. schiedeanus in this area as well. We suggest that the observed local specialization is a result of seed dispersal as consequence of bird foraging and territorial behavior, host abundance, and host species compatibility.     

Altered Expression of Natural Cytotoxicity Receptors and NKG2D on Peripheral Blood NK Cell Subsets in Breast Cancer Patients

Human natural killer (NK) cells are considered professional cytotoxic cells that are integrated into the effector branch of innate immunity during antiviral and antitumoral responses. The purpose of this study was to examine the peripheral distribution and expression of NK cell activation receptors from the fresh peripheral blood mononuclear cells of 30 breast cancer patients prior to any form of treatment (including surgery, chemotherapy, and radiotherapy), 10 benign breast pathology patients, and 24 control individuals. CD3⁻CD56^dimCD16^bright NK cells (CD56^dim NK) and CD3⁻CD56^brightCD16^dim/− NK cells (CD56^bright NK) were identified using flow cytometry. The circulating counts of CD56^dim and CD56^bright NK cells were not significantly different between the groups evaluated, nor were the counts of other leukocyte subsets between the breast cancer patients and benign breast pathology patients. However, in CD56^dim NK cells, NKp44 expression was higher in breast cancer patients (P = .0302), whereas NKp30 (P = .0005), NKp46 (P = .0298), and NKG2D (P = .0005) expression was lower with respect to healthy donors. In CD56^bright NK cells, NKp30 (P = .0007), NKp46 (P = .0012), and NKG2D (P = .0069) expression was lower in breast cancer patients compared with control group. Only NKG2D in CD56^bright NK cells (P = .0208) and CD56^dim NK cells (P = .0439) showed difference between benign breast pathology and breast cancer patients. Collectively, the current study showed phenotypic alterations in activation receptors on CD56^dim and CD56^bright NK cells, suggesting that breast cancer patients have decreased NK cell cytotoxicity.     

Limitations to the Use of Species-Distribution Models for Environmental-Impact Assessments in the Amazon

Species-distribution models (SDM) are tools with potential to inform environmental-impact studies (EIA). However, they are not always appropriate and may result in improper and expensive mitigation and compensation if their limitations are not understood by decision makers. Here, we examine the use of SDM for frogs that were used in impact assessment using data obtained from the EIA of a hydroelectric project located in the Amazon Basin in Brazil. The results show that lack of knowledge of species distributions limits the appropriate use of SDM in the Amazon region for most target species. Because most of these targets are newly described and their distributions poorly known, data about their distributions are insufficient to be effectively used in SDM. Surveys that are mandatory for the EIA are often conducted only near the area under assessment, and so models must extrapolate well beyond the sampled area to inform decisions made at much larger spatial scales, such as defining areas to be used to offset the negative effects of the projects. Using distributions of better-known species in simulations, we show that geographical-extrapolations based on limited information of species ranges often lead to spurious results. We conclude that the use of SDM as evidence to support project-licensing decisions in the Amazon requires much greater area sampling for impact studies, or, alternatively, integrated and comparative survey strategies, to improve biodiversity sampling. When more detailed distribution information is unavailable, SDM will produce results that generate uncertain and untestable decisions regarding impact assessment. In many cases, SDM is unlikely to be better than the use of expert opinion.     

Biosynthesis of glycoproteins in Candida albicans: Biochemical characterization of dolichol phosphate glucose synthase

A mixed membrane fraction isolated from C. albicans yeast cells catalyzed the transfer of glucose from UDP-Glc into three classes of endogenous acceptors: glucolipid, glycoprotein and lipid-linked oligosaccharides. About 80 of the total radioactivity transferred into these products corresponded to the glucolipid which was identified as dolichol phosphate glucose by several criteria. The remainder was detected in about equal proportions in the other two fractions. Conditions that stimulated or inhibited glucolipid synthesis did not affect the extent of glycoprotein labeling. The synthesis of dolichol phosphate glucose exhibited a K_mof 104 M UDP-Glc and was stimulated by Mg²⁺but not by Mn²⁺or Ca²⁺. The latter cations were, however, better stimulators of glycoprotein labeling than Mg²⁺. Most nucleotides strongly inhibited the synthesis of dolichol phosphate glucose, UMP being a competitive inhibitor with a K_iof 100 M. The dolichol phosphate glucose synthase reaction was reversed about 57 by 0.62 mM UDP but not by UMP.