High frequency properties of a planar ion trap fabricated on a chip.

We report on the measurement of the high frequency properties of a planar Penning ion trap fabricated on a chip. Two types of chips have been measured: the first manufactured by photolithographic metal deposition on a p-doped silicon substrate and the second made with printed circuit board technology on an alumina substrate. The input capacitances and the admittances between the different trap's electrodes play a critical role in the electronic detection of the trapped particles. The measured input capacitances of the photolithographic chip amount to 65-76 pF, while the values for the printed circuit board chips are in the range of 3-5 pF. The latter are small enough for detecting non-destructively a single trapped electron or ion with a specifically tuned LC resonator. We have also measured a mutual capacitance of ∼85 fF between two of the trap's electrodes in the printed circuit board chip. This enables the detection of single, or very few, trapped particles in a broader range of charge-to-mass ratios with a simple resistor on the chip. We provide analytic calculations of the capacitances and discuss their origin and possible further reduction.

Loss of PRP4K drives anoikis resistance in part by dysregulation of epidermal growth factor receptor endosomal trafficking.

Anoikis acts as a critical barrier to metastasis by inducing cell death upon cancer cell detachment from the extracellular matrix (ECM), thereby preventing tumor cell dissemination to secondary sites. The induction of anoikis requires the lysosomal-mediated downregulation of epidermal growth factor receptors (EGFRs) leading to termination of pro-survival signaling. In this study, we demonstrate that depletion of pre-mRNA splicing factor 4 kinase (PRP4K; also known as PRPF4B) causes dysregulation of EGFR trafficking and anoikis resistance. We also report a novel cytoplasmic localization of PRP4K at the late endosome, and demonstrate both nuclear and cytoplasmic localization in breast, lung and ovarian cancer tissue. Mechanistically, depletion of PRP4K leads to reduced EGFR degradation following cell detachment from the ECM and correlates with increased TrkB, vimentin and Zeb1 expression. As a result, PRP4K loss promotes sustained growth factor signaling and increased cellular resistance to anoikis in vitro and in a novel zebrafish xenotransplantation model of anoikis sensitivity, as well as increased metastasis in a mouse model of ovarian cancer. Thus, PRP4K may serve as a potential biomarker of anoikis sensitivity in ovarian and other epithelial cancers.

Application of computational models to estimate organ radiation dose in rainbow trout from uptake of molybdenum-99 with comparison to iodine-131.

This study compares three anatomical phantoms for rainbow trout (Oncorhynchus mykiss) for the purpose of estimating organ radiation dose and dose rates from molybdenum-99 ((99)Mo) uptake in the liver and GI tract. Model comparison and refinement is important to the process of determining accurate doses and dose rates to the whole body and the various organs. Accurate and consistent dosimetry is crucial to the determination of appropriate dose-effect relationships for use in environmental risk assessment. The computational phantoms considered are (1) a geometrically defined model employing anatomically relevant organ size and location, (2) voxel reconstruction of internal anatomy obtained from CT imaging, and (3) a new model utilizing NURBS surfaces to refine the model in (2). Dose Conversion Factors (DCFs) for whole body as well as selected organs of O. mykiss were computed using Monte Carlo modeling and combined with empirical models for predicting activity concentration to estimate dose rates and ultimately determine cumulative radiation dose (µGy) to selected organs after several half-lives of (99)Mo. The computational models provided similar results, especially for organs that were both the source and target of radiation (less than 30% difference between all models). Values in the empirical model as well as the 14 day cumulative organ doses determined from (99)Mo uptake are compared to similar models developed previously for (131)I. Finally, consideration is given to treating the GI tract as a solid organ compared to partitioning it into gut contents and GI wall, which resulted in an order of magnitude difference in estimated dose for most organs.

Development and comparison of computational models for estimation of absorbed organ radiation dose in rainbow trout (Oncorhynchus mykiss) from uptake of iodine-131.

This study develops and compares different, increasingly detailed anatomical phantoms for rainbow trout (Oncorhynchus mykiss) for the purpose of estimating organ absorbed radiation dose and dose rates from (131)I uptake in multiple organs. The models considered are: a simplistic geometry considering a single organ, a more specific geometry employing additional organs with anatomically relevant size and location, and voxel reconstruction of internal anatomy obtained from CT imaging (referred to as CSUTROUT). Dose Conversion Factors (DCFs) for whole body as well as selected organs of O. mykiss were computed using Monte Carlo modeling, and combined with estimated activity concentrations, to approximate dose rates and ultimately determine cumulative radiation dose (µGy) to selected organs after several half-lives of (131)I. The different computational models provided similar results, especially for source organs (less than 30% difference between estimated doses), and whole body DCFs for each model (∼3 × 10(-3) µGy d(-1) per Bq kg(-1)) were comparable to DCFs listed in ICRP 108 for (131)I. The main benefit provided by the computational models developed here is the ability to accurately determine organ dose. A conservative mass-ratio approach may provide reasonable results for sufficiently large organs, but is only applicable to individual source organs. Although CSUTROUT is the more anatomically realistic phantom, it required much more resource dedication to develop and is less flexible than the stylized phantom for similar results. There may be instances where a detailed phantom such as CSUTROUT is appropriate, but generally the stylized phantom appears to be the best choice for an ideal balance between accuracy and resource requirements.

Cesium accumulation by aquatic organisms at different trophic levels following an experimental release into a small reservoir.

The rates of accumulation and subsequent loss of stable cesium (¹³³Cs) by organisms at different trophic levels within plankton-based and periphyton-based food chains were measured following the addition of ¹³³Cs into a small reservoir near Aiken, South Carolina, USA. An uptake parameter u (L kg⁻¹ d⁻¹ dry mass) and a loss rate parameter k (d⁻¹) were estimated for each organism using time-series measurements of ¹³³Cs concentrations in water and biota, and these parameters were used to estimate maximum concentrations, times to maximum concentrations, and concentration ratios (C(r)). The maximum ¹³³Cs concentrations for plankton, periphyton, the insect larva Chaoborus punctipennis, which feeds on plankton, and the snail Helisoma trivolvis, which feeds on periphyton, occurred within the first 14 days following the addition, whereas the maximum concentrations for the fish species Lepomis macrochirus and Micropterus salmoides occurred after 170 days. The C(r) based on dry mass for plankton and C. punctipennis were 1220 L kg⁻¹ and 5570 L kg⁻¹, respectively, and were less than the C(r) of 8630 L kg⁻¹ for periphyton and 47,700 L kg⁻¹ for H. trivolvis. Although the C(r) differed between plankton-based and periphyton-based food chains, they displayed similar levels of biomagnification. Biomagnification was also indicated for fish where the C(r) for the mostly nonpiscivorous L. macrochirus of 22,600 L kg⁻¹ was three times less than that for mostly piscivorous M. salmoides of 71,500 L kg⁻¹. Although the C(r) for M. salmoides was greater than those for periphyton and H. trivolvis, the maximum ¹³³Cs concentrations for periphyton and H. trivolvis were greater than that for M. salmoides.

Cesium accumulation by fish following acute input to lakes: a comparison of experimental and Chernobyl-impacted systems.

An uptake parameter u (Lkg(-1)d(-1)) and a loss rate parameter k (d(-1)) were estimated for the patterns of accumulation and loss of (133)Cs by three fish species following an experimental (133)Cs addition into a pond in South Carolina, USA. These u and k parameters were compared to similar estimates for fish from other experimental ponds and from lakes that received (137)Cs deposition from Chernobyl. Estimates of u from ponds and lakes declined with increasing potassium concentrations in the water column. Although loss rates were greater in the experimental ponds, the times required to reach maximum Cs concentrations in fish were similar between ponds and lakes, because ponds and lakes had similar retentions of Cs in the water column. The maximum Cs concentrations in fish were largely determined by initial Cs concentrations in the water column. These maximum concentrations in fish and the times required to reach these maxima are potentially useful indicators for assessments of risks to humans from fish consumption.

Foliar uptake of cesium from the water column by aquatic macrophytes.

The probable occurrence and rate of foliar absorption of stable cesium (133Cs) from the water column by aquatic macrophyte species was analyzed following the addition of 133Cs into a small reservoir near Aiken, South Carolina, USA. An uptake parameter u (10(3)Lkg(-1)d(-1)) and a loss rate parameter k (d(-1)) were estimated for each species using time series of 133Cs concentrations in the water and plant tissues. Foliar uptake, as indicated by rapid increases in plant concentrations following the 133Cs addition, occurred in two floating-leaf species, Brasenia schreberi and Nymphaea odorata, and two submerged species, Myriophyllum spicatum and Utricularia inflata. These species had values of u> or =0.75 x 10(3)Lkg(-1)d(-1). Less evidence for foliar uptake was observed in three emergent species, including Typha latifolia. Ratios of u to k for B. schreberi, M. spicatum, N. odorata and U. inflata can be used to estimate concentration ratios (CR) at equilibrium, and these estimates were generally within a factor of 2 of the CR for 137Cs for these species in the same reservoir. This correspondence suggests that foliar uptake of Cs was the principal absorption mechanism for these species. Assessments of: (1) the prevalence of foliar uptake of potassium, rubidium and Cs isotopes by aquatic macrophytes and (2) the possible importance of foliar uptake of Cs in other lentic systems are made from a review of foliar uptake studies and estimation of comparable u and k values from lake studies involving Cs releases.

The influence of a whole-lake addition of stable cesium on the remobilization of aged 137Cs in a contaminated reservoir.

To document the short-term dynamics of Cs, 4 kg of (133)Cs were introduced into an 11.4-ha, 157 000 m(3) reservoir previously contaminated with (137)Cs from past reactor operations at the US Department of Energy's Savannah River Site near Aiken, South Carolina, USA. The (133)Cs addition resulted in an increase of 6.1 MBq of (137)Cs (1.9 mug (137)Cs) in the water column over the following 260 days. Possible sources for the increased (137)Cs included (1) release from the sediments, (2) release from the approximately 26 000 kg of aquatic macrophytes that occupied 80% of the reservoir, and (3) wash-in from the pond's watershed. Data are presented to indicate that release from the sediments was the principal source of the (137)Cs increase. The fraction of (137)Cs released from the sediments (0.7%) is consistent with laboratory measurements of (137)Cs desorption from neighboring ponds on the Savannah River Site.

Myosins of Babesia bovis: molecular characterisation, erythrocyte invasion, and phylogeny.

Using degenerate primers, three putative myosin sequences were amplified from Australian isolates of Babesa bovis and confirmed as myosins (termed Bbmyo-A, Bbmyo-B, and Bbmyo-C) from in vitro cultures of the W strain of B. bovis. Comprehensive analysis of 15 apicomplexan myosins suggests that members of Class XIV be defined as those with greater than 35% myosin head sequence identity and that these be further subclassed into groups bearing above 50-60% identity. Bbmyo-A protein bears a strong similarity with other apicomplexan myosin-A type proteins (subclass XIVa), the Bbmyo-B myosin head protein sequence exhibits low identity (35-39%) with all members of Class XIV, and 5'-sequence of Bbmyo-C shows strong identity (60%) with P. falciparum myosin-C protein. Domain analysis revealed five divergent IQ domains within the neck of Pfmyo-C, and a myosin-N terminal domain as well as a classical IQ sequence unusually located within the head converter domain of Bbmyo-B. A cross-reacting antibody directed against P. falciparum myosin-A (Pfmyo-A) revealed a zone of approximately 85 kDa in immunoblots prepared with B. bovis total protein, and immunofluorescence inferred stage-specific myosin-A expression since only 25% of infected erythrocytes with mostly paired B. bovis were immuno-positive. Multiplication of B. bovis in in vitro culture was inhibited by myosin- and actin-binding drugs at concentrations lower than those that inhibit P. falciparum. This study identifies and classifies three myosin genes and an actin gene in B. bovis, and provides the first evidence for the participation of an actomyosin-based motor in erythrocyte invasion in this species of apicomplexan parasite.

Motile systems in malaria merozoites: how is the red blood cell invaded?

The ability of the malaria parasite to invade erythrocytes is central to the disease process, but is not thoroughly understood. In particular, little attention has been paid to the motor systems driving invasion. Here, Jennifer Pinder, Ruth Fowler and colleagues review motility in the merozoite. The components of an actomyosin motor are present, including a novel unconventional class XIV myosin, now called Pfmyo-A, which, because of its time of synthesis and location, is likely to generate the force required for invasion. In addition, there is a subpellicular microtubule assemblage in falciparum merozoites, the f-MAST, the integrity of which is necessary for invasion.

Identification of a novel C-terminal variant of beta II spectrin: two isoforms of beta II spectrin have distinct intracellular locations and activities.

It is established that variations in the structure and activities of betaI spectrin are mediated by differential mRNA splicing. The two betaI spectrin splice forms so far identified have either long or short C-terminal regions. Are analogous mechanisms likely to mediate regulation of betaII spectrins? Thus far, only a long form of betaII spectrin is reported in the literature. Five human expressed sequence tags indicated the existence of a short splice variant of betaII spectrin. The occurrence and DNA sequence of the short C-terminal variant was confirmed by analysis of human and rat cDNA. The novel variant lacks a pleckstrin homology domain, and has 28 C-terminal residues not present in the previously recognized longer form. Transcripts of the short C-terminal variant (7.5 and 7. 0 kb) were most abundant in tissues originating from muscle and nervous system. Antibodies raised to a unique sequence of short C-terminal variant recognized 240 kDa polypeptides in cardiac and skeletal muscle and in nervous tissue; in cerebellum and forebrain, additional 270 kDa polypeptides were detected. In rat heart and skeletal muscle, both long and short C-terminal forms of betaII spectrin localized in the region of the Z line. The central region of the sarcomere, coincident with the M line, was selectively labeled with antibodies to the short C-terminal form. In cerebellum, the short form was not detectable in parallel fibers, structures in which the long form was readily detected. In cultured cerebellar granule neurons, the long form was dominant in neurites, with the short form being most abundant in cell bodies. In vitro, the short form was found to lack the binding activity for the axonal protein fodaxin, which characterizes the C-terminal region of the long form. Subcellular fractionation of brain revealed that the short form was scarcely detectable in post-synaptic density preparations, in which the long form was readily detected. We conclude that variation in the structure of the C-terminal regions of betaII spectrin isoforms correlates with their differential intracellular targeting.

Properties of normal and mutant polypeptide fragments from the dimer self-association sites of human red cell spectrin.

We have examined the properties and interactions of expressed polypeptide fragments from the N-terminus of the alpha-chain and the C-terminus of the beta-chain of human erythroid spectrin. Each polypeptide comprises one complete structural repeating unit, together with the incomplete repeat that interacts with its partner when spectrin tetramers are formed. The shared repeat thus generated is made up of two helices from the C-terminal part of the beta-chain and one helix from the N-terminus of the alpha-chain. Three mutant beta-chain fragments with amino acid substitutions in the incomplete terminal repeat were also studied. The alpha- and beta-chain fragments were both substantially monomeric, as shown by sedimentation equilibrium. Circular dichroism analysis and thermal denaturation profiles revealed that the complete repeat present in each fragment had entered the stable tertiary fold. Unexpectedly, the conformational stability of the folded beta-chain repeat was found to be grossly perturbed by the mutations, all of them well beyond its C-terminal boundary; possible explanations for this phenomenon are considered. Sedimentation equilibrium showed that in equimolar mixtures the wildtype alpha- and beta-chain peptides formed a 1:1 complex. Mixing curves, observed by circular dichroism, revealed that association was accompanied by an increase in alpha-helicity. From continuous-variation profiles an association constant in the range 1-2 x 10(6) M-1 was inferred. The association was unaffected by the apparently unstructured anionic tail of 54 residues, found at the C-terminus of the spectrin beta-chain. Of the three mutations in the beta-chain fragment, one (an Ala-->Val replacement in the A helix segment of the incomplete repeat) had a relatively small effect on the association with the alpha-chain fragment, whereas Trp-->Arg mutations in the A and in the remote B helix segments were much more deleterious. These observations are consistent with the relative severities of the haemolytic conditions associated with the mutations.

Actomyosin motor in the merozoite of the malaria parasite, Plasmodium falciparum: implications for red cell invasion.

The genome of the malaria parasite, Plasmodium falciparum, contains a myosin gene sequence, which bears a close homology to one of the myosin genes found in another apicomplexan parasite, Toxoplasma gondii. A polyclonal antibody was generated against an expressed polypeptide of molecular mass 27,000, based on part of the deduced sequence of this myosin. The antibody reacted with the cognate antigen and with a component of the total parasite protein on immunoblots, but not with vertebrate striated or smooth muscle myosins. It did, however, recognise two components in the cellular protein of Toxoplasma gondii. The antibody was used to investigate stage-specificity of expression of the myosin (here designated Pf-myo1) in P. falciparum. The results showed that the protein is synthesised in mature schizonts and is present in merozoites, but vanishes after the parasite enters the red cell. Pf-myo1 was found to be largely, though not entirely, associated with the particulate parasite cell fraction and is thus presumably mainly membrane bound. It was not solubilised by media that would be expected to dissociate actomyosin or myosin filaments, or by non-ionic detergent. Immunofluorescence revealed that in the merozoite and mature schizont Pf-myo1 is predominantly located around the periphery of the cell. Immuno-gold electron microscopy also showed the presence of the myosin around almost the entire parasite periphery, and especially in the region surrounding the apical prominence. Labelling was concentrated under the plasma membrane but was not seen in the apical prominence itself. This suggests that Pf-myo1 is associated with the plasma membrane or with the outer membrane of the subplasmalemmal cisterna, which forms a lining to the plasma membrane, with a gap at the apical prominence. The results lead to a conjectural model of the invasion mechanism.

Patients' rights to access their healthcare records.

Improving communication between different healthcare disciplines and with patients is hampered by professional insecurity. In this article, the authors examine the compelling reasons for making records more accessible to patients and describe the potential benefits for patients and professionals.

Educating health educators: a survey of hospital staff completing a certificate in health education course.

Health care professionals (nurses, a midwife, a physiotherapist and an occupational therapist) working in a large NHS Trust hospital who had completed the Certificate in Health Education with the support of their employer, were interviewed. The study objectives were to seek their views on the quality of the course, to determine the extent to which participants were able to apply their new found knowledge and skills in the care they provided to patients and the level of support received to allow them to do this. Barriers that prevented staff from routinely applying health education in their work were identified. The findings indicated that the majority found the course content to be good and relevant to their clinical work but they identified lack of time due to the pressure of routine clinical work as the main barrier to the promotion of health education in their clinical area.