Human placental cytotrophoblasts attract monocytes and CD56(bright) natural killer cells via the actions of monocyte inflammatory protein 1alpha.

During human pregnancy, the specialized epithelial cells of the placenta (cytotrophoblasts) come into direct contact with immune cells in several locations. In the fetal compartment of the placenta, cytotrophoblast stem cells lie adjacent to macrophages (Hofbauer cells) that reside within the chorionic villus stroma. At sites of placental attachment to the mother, invasive cytotrophoblasts encounter specialized maternal natural killer (NK) cells (CD56(bright)), macrophages, and T cells that accumulate within the uterine wall during pregnancy. Here we tested the hypothesis that fetal cytotrophoblasts can direct the migration of these maternal immune cells. First, we assayed the chemotactic activity of cytotrophoblast conditioned medium samples, using human peripheral blood mononuclear cells as targets. The placental samples preferentially attracted NK cells (both CD56(dim) and CD56(bright)), monocytes, and T cells, suggesting that our hypothesis was correct. A screen to identify chemokine activity through the induction of a Ca(2)+ flux in cells transfected with individual chemokine receptors suggested that cytotrophoblasts secreted monocyte inflammatory protein (MIP)-1alpha. This was confirmed by localizing the corresponding mRNA and protein, both in vitro and in vivo. MIP-1alpha protein in conditioned medium was further characterized by immunoblotting and enzyme-linked immunosorbent assay. Immunodepletion of MIP-1alpha from cytotrophoblast conditioned medium showed that this chemokine was responsible for a significant portion of the induced monocyte and CD56(bright) NK cell chemotaxis. These data suggest the specific conclusion that cytotrophoblasts can attract monocytes and CD56(bright) NK cells by producing MIP-1alpha and the more general hypothesis that these cells may organize and act on leukocytes at the maternal-fetal interface.

Properties of laser-produced GaAs plasmas measured from highly resolved X-ray line shapes and ratios.

The properties of hot, dense plasmas generated by the irradiation of GaAs targets by the Titan laser at Lawrence Livermore National Laboratory were determined by the analysis of high resolution K shell spectra in the 9 keV to 11 keV range. The laser parameters, such as relatively long pulse duration and large focal spot, were chosen to produce a steady-state plasma with minimal edge gradients, and the time-integrated spectra were compared to non-LTE steady state spectrum simulations using the FLYCHK and NOMAD codes. The bulk plasma streaming velocity was measured from the energy shifts of the Ga He-like transitions and Li-like dielectronic satellites. The electron density and the electron energy distribution, both the thermal and the hot non-thermal components, were determined from the spectral line ratios. After accounting for the spectral line broadening contributions, the plasma turbulent motion was measured from the residual line widths. The ionization balance was determined from the ratios of the He-like through F-like spectral features. The detailed comparison of the experimental Ga spectrum and the spectrum simulated by the FLYCHK code indicates two significant discrepancies, the transition energy of a Li-like dielectronic satellite (designated t) and the calculated intensity of a He-like line (x), that should lead to improvements in the kinetics codes used to simulate the X-ray spectra from highly-charged ions.

Intracellular signal transduction: The role of endosomes.

Polypeptide hormones, growth factors, and other biologically significant molecules are specifically internalized by target cells. Exposure of cells to these ligands results in the formation of ligand-receptor complexes on the cell surface and subsequent internalization of these complexes into the endosomal apparatus (endosomes, or ENs). The study of ENs has identified several important functions for this unique cellular organelle. These include the dissociation of ligand from receptor and receptor recycling to the cell surface and the degradation of some internalized ligands, as well as the delivery of others to lysosomes. More recently, it has become apparent that ENs fulfill another critical role, that of signal transduction. In this article, we review the evidence substantiating this role for ENs and propose three models by which ENs participate in cell signaling.

Morphological and structural plasticity of grassland species in response to a gradient in saline-sodic soils.

The abundance and distribution of species can be ascribed to both environmental heterogeneity and stress tolerance, with the latter measure sometimes associated with phenotypic plasticity. Although phenotypic plasticity varies predictably in response to common forms of stress, we lack a mechanistic understanding of the response of species to high saline-sodic soils. We compared the phenotypic plasticity of three pairs of high and low saline-sodic tolerant congeners from the families Poaceae (Leymus chinensis versus L. secalinus), Fabaceae (Lespedeza davurica versus L. bicolor) and Asteraceae (Artemisia mongolica versus A. sieversiana) in a controlled pot experiment in the Songnen grassland, China. The low tolerant species, L. secalinus and A. sieversiana exhibited higher plasticity in response to soil salinity and sodicity than their paired congeners. Highly tolerant species, L. chinensis and A. mongolica, had higher values for several important morphological traits, such as shoot length and total biomass under the high saline-sodic soil treatment than their paired congeners. In contrast, congeners from the family Fabaceae, L. davurica and L. bicolor, did not exhibit significantly different plasticity in response to soil salinity and sodicity. All species held a constant reproductive effort in response to saline-sodic soil stress. The different responses between low and high tolerant species offer an explanation for the distribution patterns of these species in the Songnen grassland. Highly tolerant species showed less morphological plasticity over a range of saline-sodic conditions than their paired congeners, which may manifest as an inability to compete with co-occurring species in locations where saline-sodic soils are absent.

A role for tyrosine phosphorylation in both activation and inhibition of the insulin receptor tyrosine kinase in vivo.

Upon insulin binding, a conformational change in the insulin receptor (IR) leads to IR beta-subunit autophosphorylation, an increase in IR beta-subunit exogenous tyrosine kinase activity, and the rapid endocytosis of the ligand-receptor complex into endosomes. Previous work has shown that upon internalization, rat hepatic endosomal IRs manifest increased autophosphorylating and exogenous tyrosine kinase activity compared to IRs located at the plasma membrane. As this period of enhanced activity is associated with reduced endosomal IR beta-subunit phosphotyrosine content, it has been proposed that partial dephosphorylation of the internalized IR beta-subunit by an endosomally located phosphotyrosine phosphatase(s) [PTPase(s)] mediates this effect. To test whether endosomal PTPase activity was required for internalization-dependent augmentation of IR tyrosine kinase activity, the present study used the peroxovanadium PTPase inhibitor, bisperoxo(1,10-phenanthroline)oxovanadate anion [bpV(phen)], to block IR dephosphorylation within this subcellular compartment. Rats were pretreated with bpV(phen) before receiving insulin (1.5 micrograms/100 g BW). bpV(phen) inhibited the dephosphorylation of 32P-labeled hepatic endosomal IR by approximately 97% at 15 min post-bpV(phen) injection and prevented a decrease in IR beta-subunit phosphotyrosine content after IR internalization. Fifteen-minute bpV(phen) pretreatment produced a significant reduction (75%; P < 0.001) in maximal insulin-stimulated endosomal IR exogenous kinase activity and decreased IR autophosphorylating activity by 4.3-fold in this subcellular fraction. In conclusion, these findings suggest that an hepatic endosomal PTPase(s) regulates internalization-dependent increases in IR exogenous tyrosine kinase activity.

Desiccation-induced loss of seed viability is associated with a 10-fold increase in CO2 evolution in seeds of the rare tropical rainforest tree Idiospermum australiense.

• Here the relationship was investigated between metabolic activity, state of hydration and seed viability in the desiccation-intolerant (recalcitrant) seeds of Idiospermum australiense, a rare and primitive angiosperm tree restricted to wet tropical forest. • Seed CO2 evolution rate, R, was monitored in fully hydrated (control) seeds and seeds that were allowed to desiccate under ambient conditions over a period of c. 90 d. • During desiccation R increased dramatically toward a peak at a seed relative water content of 39 ± 3% (relative to maximum water content, which corresponded to 0.45 ± 0.03 g water g-1 d. wt) followed by a decline toward zero with total desiccation. This peak constituted a 10-fold increase in mean R, relative to the control. Exposing seeds to O2 -free air at this peak induced a further large, but transient, increase in CO2 evolution, indicating that the peak developed in the presence of oxidative phosphorylation, rather than due to the absence of it. • The magnitude and mode of the observed increase in CO2 evolution in response to desiccation is unlike any reported so far and thus adds new information about metabolic changes that may occur as the water content of desiccation-intolerant seeds declines.

Ragaglitazar: the pharmacokinetics, pharmacodynamics, and tolerability of a novel dual PPAR alpha and gamma agonist in healthy subjects and patients with type 2 diabetes.

Ragaglitazar is a novel dual peroxisome proliferator-activated receptor (PPAR) alpha and gamma agonist intended to restore insulin sensitivity and correct diabetic dyslipidemia. These studies assessed single-dose pharmacokinetics and tolerability of ragaglitazar in healthy subjects, as well as multiple-dose pharmacokinetics, pharmacodynamics, and tolerability of ragaglitazar in healthy subjects and in patients with type 2 diabetes. Healthy subjects received a single oral dose (1-120 mg), and healthy subjects and type 2 diabetic patients received a loading dose and thereafter once-daily doses (0.5-16 mg) of ragaglitazar for 6 and 20 days, respectively. Ragaglitazar was rapidly absorbed (tmax: 1.5-1.7 h), with mean AUC0-24 h and Cmax proportional to dose after single and multiple dosing; t1/2 was 80 hours following a single dose and 104 hours in healthy subjects and 122 hours in patients after multiple dosing. Administration of 4 mg ragaglitazar to patients (n = 4) for 21 days resulted in mean decreases from baseline in fasting levels of plasma glucose (18%), C-peptide (18%), fructosamine (6%), triglycerides (36%), free fatty acids (49%), total cholesterol (11%), low-density lipoprotein (LDL) cholesterol (21%), and very low-density lipoprotein (VLDL) cholesterol (15%), as well as an increase in high-density lipoprotein (HDL) cholesterol (33%). Overall, ragaglitazar was well tolerated; with multiple dosing, there was a higher incidence of adverse events for patients that, at the highest dose level (16 mg), included peripheral edema and anemia.

Mechanisms of action of cyclic nucleotides on a bursting pacemaker and silent neuron in Aplysia.

Cyclic nucleotides are believed to mediate a long-lasting synaptic hyperpolarization in the bursting pacemaker neuron, R15, and are capable of inducing bursting pacemaker activity in the usually silent metacerebral giant cell. Steady state voltage clamp techniques were used to examine the alterations of membrane characteristics produced in these different cell types by cyclic nucleotides. In both cells, IBMX, a phosphodiesterase inhibitor, increased two components: (1) voltage-dependent sodium current and (2) slope conductance believed to reflect potassium flux. The effects of 8-benzylthio-cAMP were identical to those of IBMX in the metacerebral cell. In R15, 8-benzylthio-cAMP affected only the slope conductance. These results are discussed in terms of cyclic nucleotide of bursting pacemaker activity.

Synthesis and fast axonal transport of proteins in the isolated Aplysia nervous system.

Fast axonal transport and neuronal protein synthesis was studied in the isolated nervous system of Aplysia californica. The abdominal ganglion with attached pleural-abdominal connectives (PAC) was removed and the ganglion pulse-labelled with [35S]methionine for 30 min in vitro. The axon containing connectives were ligated 24-28 mm from the ganglia and the system was perfused with chase media for 6-72 h to allow labelled rapidly transported proteins to accumulate at the ligature. One-dimensional polyacrylamide gel electrophoresis (PAGE) and fluorography was used to analyze the distribution of rapidly transported proteins along the right PAC. By 12 h, a significant accumulation of labelled proteins at the ligature was present but the build up was not complete until 48 h when almost no trailing of rapidly transported proteins was observed. Quantitation of the transport profiles of several rapidly transported proteins suggested a discontinuous release of proteins from the cell body. Analysis using two-dimensional PAGE revealed 10 major groups of rapidly transported proteins. These proteins were all identified among the total complement of newly synthesized proteins in cell R2. Not all rapidly transported proteins are cleared from the cell body at the same rate. Several of the major groups were no longer present in the neuron cell body 24 h after labelling, indicating that these species are selectively exported; others were still present after 3 days, suggesting that these proteins with a longer residence time have functions in both somatic and axonal regions of the neuron.

Assessing mercury health effects in gold workers near El Callao, Venezuela.

Mercury exposure and health status were examined in 40 gold workers in the area surrounding El Callao, Venezuela. Concentrations of mercury in workplace air were measured on 3 successive days, and spot urine and hair samples were also taken for analysis. Subjects underwent a physical examination and completed a questionnaire regarding employment history, work activities involving mercury exposure, use of protective clothing and equipment, and frequency of 37 symptoms associated with mercury toxicity. A complete set of health data was collected for 29 of the subjects. Use of protective equipment was limited, and 17.9%, 24.1%, and 48.3% of subjects had mercury concentrations in air, hair, and urine, respectively, above contemporary occupational exposure guidelines. Physical examination found the workers to be generally healthy and without overt symptoms of mercury toxicity. The frequency of psychoneurological, gastrointestinal, cardio-respiratory, and dermal symptoms was unrelated to any of the measures of mercury exposure. Two subjects had modestly elevated urinary levels of N-acetyl beta-D-glucosaminidase. Despite substantial occupational exposure to mercury among a number of the subjects, few adverse health effects were observed that were plausibly related to mercury.

Early hormonal therapy stabilizes hemodynamics during donor procurement.

BACKGROUND: Hemodynamic instability has been implicated in the loss of otherwise transplantable organs. We examined the hypothesis that administration of hormonal therapy early during donor management would stabilize hemodynamics and increase the number of organs procured. METHODS: We retrospectively analyzed 133 consecutive donor records from a single organ procurement organization. Controls (C) received no early hormonal therapy. A steroid group (S) received methylprednisolone only and a combination hormonal therapy group (CH) received thyroxine, methylprednisolone, dextrose, and insulin at the start of donor management (t(0h)). Adrenergic support was adjusted to maintain mean arterial blood pressure (MAP) at > or =60 mm Hg. Doses of adrenergic agents were assessed at t(0h), 4 hours (t(4h)), and just prior to procurement (t(proc)). RESULTS: Baseline characteristics were similar in all groups. Dosages of adrenergic agents decreased over time in all groups. A significant decrease in adrenergic requirements was seen in the CH group compared with the C group at t(4h) and t(proc). A trend toward decreased adrenergic requirements was noted in S compared with C at t(4h) and t(proc). Slightly more total organs were procured from S and CH compared with C. CONCLUSIONS: Significantly less adrenergic support was required with early use of CH. A similar (although nonsignificant) reduction was seen with S. The benefit(s) of CH vs corticosteroids alone remains uncertain and requires further study.

Initial effects of the toxic waste spill (Aznalcóllar mine accident) on the aquatic macrofauna of the Guadalquivir Estuary.

The initial effects of the toxic waste spill in April 1998 at the Aznalcóllar mine (SW Spain) on the nektonic community of the Guadalquivir Estuary were examined at three sampling sites using univariate and multivariate techniques. Since studied communities showed a considerable seasonal trend, only seasonally homogenous periods were compared to analyse effects of the spill: May-August 1997 (before spill) and May-August 1998 (after spill). Results of both techniques (two-way nested ANOVA and ANOSIM tests, P > 0.05) indicated that there was no significant difference between the nektonic community of the estuary before and after the spill (monthly number of species, abundance, biomass and similarity among samples). Conversely, an unusually high density was observed at the outer sampling site immediately after the spill for species typical of more stagnant estuarine habitats. This feature seems to indicate that the fauna in the estuarine area through which the untreated water penetrated into the main course may have been disturbed. Results also suggest that this initial sudden input of fresh water to the estuary could have enhanced the effects of an increased river flow (a drop in the salinity). Nevertheless, longer temporal series of data, especially for permanent estuarine inhabitants, are recommended before conclusions can be drawn on the effects of the toxic waste spill on estuarine communities.

Addressing developmental needs of pregnant adolescents.

To appropriately meet the needs of pregnant adolescents, health care providers must evaluate the cognitive and psychosocial development of these clients. Differences in early, middle, and late adolescents who are pregnant are described. The impact of adolescent stages on accomplishing the developmental tasks of pregnancy are discussed. Nursing implications for assessment, communication techniques, teaching strategies, and family interventions are presented. A case study is included.

PIP: Each year in the US more than 1 million women younger than age 20 years become pregnant, of whom approximately 50% give birth. In order to interact effectively with pregnant adolescents, one needs to understand where they are along their course of cognitive and psychosocial development. Adolescents move through the developmental tasks of adolescence and pregnancy at different rates. Identification of at which developmental stage a given adolescent is should therefore be determined by nurses and other health care providers before interventions are planned. There can be significant differences in the maturity levels and abilities of early, middle, and late adolescents. Differences in early, middle, and late pregnant adolescents are described. The impact of adolescent stages on accomplishing the developmental tasks of pregnancy are discussed, followed by consideration of nursing implications for assessment, communication techniques, teaching strategies, and family interventions. A case study is presented.

Hemorrhage after bone marrow harvest: a case presentation.

The purpose of this article is to describe the usual procedure and postoperative recovery after an allogeneic bone marrow harvest and to present a case study of an unusual complication of hemorrhage. The case study describes a donor who experienced hemorrhage with severe pain, muscle spasms, and prolonged limitations in range of motion and ambulation. Oncology nurses should inform donors to promptly report persistent pain, spasms, and muscle weakness. Should hemorrhage occur, blood loss should be evaluated, bedrest should be maintained, and cold packs should be applied to the area. Although excessive bleeding is a rare occurrence, nurses should be alert for this complication to prevent pain and activity impairment.

The nature of calibrators in immunoassays: are they commutable with test samples? Must they be?

Immunoassay systems require calibration protocols that are normally more sophisticated than many analytical techniques in routine clinical use. Calibrators used in such assays may differ significantly from the analyte in clinical specimens. Differences in the properties of calibrators, or reference materials, from those of clinical specimens may include: species origin of the calibrator for an analyte; integrity of the molecular species; matrix of the calibration solution; addition of preservative agents. Owing to the large number of potential differences in the properties of calibrators and those of serum specimens that may affect immunoassay results, the concept of commutability that we originated and first applied to enzyme activity measurements can readily be applied to immunoassay determinations. We specifically examined the properties of calibration materials in nine commercial immunoassay tests for human thyrotropin. Significant non-commutability of materials was demonstrated. The measured results obtained with authentic patient sera differed by a factor of approximately two fold between the techniques exhibiting the lowest and greatest numeric results. Considerably larger intermethod biases were found for calibration materials. Multivariate analysis revealed that the patient sera formed a highly focussed pattern. The calibration materials for one instrument system also focussed in this group. Other calibrators formed three foci indicating similar patterns of commutability within each of the three groups. Clustering was independent of the amount of thyrotropin in the patient specimens, but appeared to be concentration-dependent for at least some of the calibrators. Thus the availability of a common calibration material appears feasible, but not presently available in many commercial products. A processed human serum, a candidate material for use in our proficiency testing program, was projected in the same cluster as authentic patient sera indicating that this material has intermethod properties identical to patient sera (i.e. fully commutable).

Multivariate methods and artificial neural networks in the assessment of the response of infaunal assemblages to sediment metal contamination and organic enrichment.

A 4-year annual sediment survey was conducted in an organically enriched tidal channel to compare the performance of univariate community descriptors, traditional multivariate techniques (TM) and artificial neural networks (AANs), in the assessment of infaunal responses to moderate levels of sediment metal contamination. Both TM approaches and the SOM ANN revealed spatiotemporal patterns of environmental and biological variables, suggesting a causal relationship between them and further highlighting subsets of taxa and sediment variables as potential main drivers of those patterns. Namely, high values of non-natural metals and organic content prompted high abundances of opportunists, while high values of natural metals yielded typical tolerant assemblages of organically enriched areas. The two approaches yielded identical final results but ANNs showed the following advantages over TM: ability to generalise results, powerful visualization tools and the ability to account simultaneously for sediment and faunal variables in the same analysis. Therefore, the SOM ANN, combined with the K-means clustering algorithm, is suggested as a promising tool for the assessment of the ecological quality of estuarine infaunal communities, although further work is needed to ensure the accuracy of the method.

Nekton response to freshwater inputs in a temperate European estuary with regulated riverine inflow.

The aim of this 12-year study was to assess the nekton (fish, decapod crustaceans) response to freshwater inputs (rainfall, dam discharges) in a temperate estuary with regulated riverine inflow. Although interannual variability in river discharges to the Guadalquivir estuary has been extremely high since the construction of a dam in 1930, a significant decreasing trend in the dam's discharges has been observed in the last 80 years. During this study, an alternation of wet, standard and dry years occurred in the estuarine area but no significant long-term trend was observed. River discharge, in turn, showed a considerable interannual variability and a significantly decreasing long-term trend. Freshwater inputs had an immediate effect on estuarine salinity and turbidity, and consequently on prey availability (mysids). Although 124 nektonic species were collected, only 47 of them (adding up to 99.7% of total abundance) were regularly present in the estuary: 32 marine migrants, 13 estuarine species and 2 diadromous species. Well-defined temporal changes in species composition and abundance yielded clear seasonal patterns in the estuarine nektonic community. Considerable intermonth and interannual changes were occasionally observed relating to freshwater inputs, mainly in winter/autumn of wet years. Thus, within each two-month period, some significant interannual differences in the nektonic community were also observed, with marine migrants tending to be more abundant in dry years. However, changes in the studied nektonic community did not show long-term trends. In conclusion, natural and human-controlled freshwater inputs currently play a significant role in determining the physicochemical conditions and the biota of the Guadalquivir estuary. However, although freshwater input seemed to transitorily affect the estuarine nekton, either directly (flushing out) or indirectly (through changes in salinity, turbidity and prey availability), a quick reestablishment of the estuarine nekton (strong resilience) was observed following freshwater inputs together with the recovery of environmental conditions within the estuary.