Does value-based prioritization at working memory enhance long-term memory?

Research has demonstrated that individuals can direct their attention to valuable information in both working memory and long-term memory tasks with observable effects on performance. However, it is currently unclear whether prioritising an item for a working memory task automatically translates into a boost at long-term memory. This was examined in two experiments using relatively short (250 ms per item; Experiment 1) and longer (500 ms per item; Experiment 2) encoding times. Participants first completed a visual working memory task, in which they were presented with series of photographs of everyday objects. Following a brief delay (1,000 ms), they completed a four-alternative forced-choice test. Prior to encoding, participants were informed of the point values associated with each item. In some trials, the first item in the sequence was worth more points than the rest. In other trials, all items were equally valuable. After a filled delay, participants completed a surprise long-term memory task. At working memory, a value effect was reliably observed on recognition accuracy, along with some evidence of faster response times for high-value items. However, there was little consistent evidence of this effect automatically persisting into long-term memory. Thus, the benefits of attentional prioritization in working memory do not always translate into longer-term performance. More broadly, this provides further evidence that manipulations that enhance working memory performance do not necessarily enhance long-term memory.

Automated line scan analysis to quantify biosensor activity at the cell edge.

Biosensors are valuable tools used to image the subcellular localization and kinetics of protein activity in living cells. Signaling at the edge of motile cells that regulates cell protrusion and retraction is important in many aspects of cell physiology, and frequently studied using biosensors. However, quantitation and interpretation is limited by the heterogeneity of this signaling behavior; automated analytical approaches are required to systematically extract large data sets from biosensor studies for statistical analysis. Here we describe an automated analysis to relate the velocity at specific points along the cell edge with biosensor activity in adjoining regions. Time series of biosensor images are processed to interpolate a smooth edge of the cell at each time point. Profiles of biosensor activity ('line scans') are then calculated along lines perpendicular to the cell edge. An energy minimization method is used to calculate a velocity associated with each line scan. Sorting line scans by the proximal velocity has generated novel biological insights, as exemplified by analysis of the Src merobody biosensor. With the large data sets that can be generated automatically by this program, conclusions can be drawn that are not apparent from qualitative or 'manual' quantitative techniques. Our 'LineScan' software includes a graphical user interface (GUI) to facilitate application in other studies. It is available at hahnlab.com and is exemplified here in a study using the RhoC FLARE biosensor.

Classifying the unclassifiable-a Delphi study to reach consensus on the fibrotic nature of diseases.

BACKGROUND: Traditionally, clinical research has focused on individual fibrotic diseases or fibrosis in a particular organ. However, it is possible for people to have multiple fibrotic diseases. While multi-organ fibrosis may suggest shared pathogenic mechanisms, yet there is no consensus on what constitutes a fibrotic disease and therefore fibrotic multimorbidity. AIM: A Delphi study was performed to reach consensus on which diseases may be described as fibrotic. METHODS: Participants were asked to rate a list of diseases, sub-grouped according to eight body regions, as 'fibrotic manifestation always present', 'can develop fibrotic manifestations', 'associated with fibrotic manifestations' or 'not fibrotic nor associated'. Classifications of 'fibrotic manifestation always present' and 'can develop fibrotic manifestations' were merged and termed 'fibrotic'. Clinical consensus was defined according to the interquartile range, having met a minimum number of responses. Clinical agreement was used for classification where diseases did not meet the minimum number of responses (required for consensus measure), were only classified if there was 100% consensus on disease classification. RESULTS: After consulting experts, searching the literature and coding dictionaries, a total of 323 non-overlapping diseases which might be considered fibrotic were identified; 92 clinical specialists responded to the first round of the survey. Over three survey rounds, 240 diseases were categorized as fibrotic via clinical consensus and 25 additional diseases through clinical agreement. CONCLUSION: Using a robust methodology, an extensive list of diseases was classified. The findings lay the foundations for studies estimating the burden of fibrotic multimorbidity, as well as investigating shared mechanisms and therapies.

A model of localised Rac1 activation in endothelial cells due to fluid flow.

Endothelial cells respond to fluid flow by elongating in the direction of flow. Cytoskeletal changes and activation of signalling molecules have been extensively studied in this response, including: activation of receptors by mechano-transduction, actin filament alignment in the direction of flow, changes to cell-substratum adhesions, actin-driven lamellipodium extension, and localised activation of Rho GTPases. To study this process we model the force over a single cell and couple this to a model of the Rho GTPases, Rac and Rho, via a Kelvin-body model of mechano-transduction. It is demonstrated that a mechano-transducer can respond to the normal component of the force is likely to be a necessary component of the signalling network in order to establish polarity. Furthermore, the rate-limiting step of Rac1 activation is predicted to be conversion of Rac-GDP to Rac-GTP, rather than activation of upstream components. Modelling illustrates that the aligned endothelial cell morphology could attenuate the signalling network.

Memory for actions in autism spectrum disorder.

This study explored how memory for actions in children with autism spectrum disorder (ASD) and typically developing children might benefit from self-performance and experimenter demonstration, and whether these groups possess metamemory knowledge of their performance levels in this task. Children with autism were less accurate on the action memory task when they carried out each action themselves during encoding, or when no actions were implemented during this phase, but this difference was abolished when the experimenter demonstrated each action during encoding. Despite clear difficulties in the self-performed condition relative to typical children, the group with ASD also showed a beneficial effect of performing the actions themselves during instruction. Finally, children with autism were as accurate as typical children in judging the accuracy of their own memory performance, indicating an absence of metamemory difficulties for this task.

Non-equilibrium dynamics of an active colloidal "chucker".

We report Monte Carlo simulations of the dynamics of a "chucker," a colloidal particle that emits smaller solute particles from its surface, isotropically and at a constant rate k(c). We find that the diffusion constant of the chucker increases for small k(c), as recently predicted theoretically. At large k(c), the chucker diffuses more slowly due to crowding effects. We compare our simulation results to those of a "point particle" Langevin dynamics scheme in which the solute concentration field is calculated analytically, and in which hydrodynamic effects arising from colloid-solvent surface interactions can be accounted for in a coarse-grained way. By simulating the dragging of a chucker, we obtain an estimate of its apparent mobility coefficient which violates the fluctuation-dissipation theorem. We also characterize the probability density profile for a chucker which sediments onto a surface which either repels or absorbs the solute particles, and find that the steady state distributions are very different in the two cases. Our simulations are inspired by the biological example of exopolysaccharide-producing bacteria, as well as by recent experimental, simulation and theoretical work on phoretic colloidal "swimmers."

Stochastic Methods for Inferring States of Cell Migration.

Cell migration refers to the ability of cells to translocate across a substrate or through a matrix. To achieve net movement requires spatiotemporal regulation of the actin cytoskeleton. Computational approaches are necessary to identify and quantify the regulatory mechanisms that generate directed cell movement. To address this need, we developed computational tools, based on stochastic modeling, to analyze time series data for the position of randomly migrating cells. Our approach allows parameters that characterize cell movement to be efficiently estimated from cell track data. We applied our methods to analyze the random migration of Mouse Embryonic Fibroblasts (MEFS) and HeLa cells. Our analysis revealed that MEFs exist in two distinct states of migration characterized by differences in cell speed and persistence, whereas HeLa cells only exhibit a single state. Further analysis revealed that the Rho-family GTPase RhoG plays a role in determining the properties of the two migratory states of MEFs. An important feature of our computational approach is that it provides a method for predicting the current migration state of an individual cell from time series data. Finally, we applied our computational methods to HeLa cells expressing a Rac1 biosensor. The Rac1 biosensor is known to perturb movement when expressed at overly high concentrations; at these expression levels the HeLa cells showed two migratory states, which correlated with differences in the spatial distribution of active Rac1.

Applanation tonometry in silicone hydrogel contact lens wearers.

INTRODUCTION: Previous studies have investigated intraocular pressure (IOP) measurements through conventional soft (hydrogel) therapeutic contact lenses, and have found that an accurate IOP can be recorded in normal eyes, and in eyes with abnormal anterior segments. The IOP measurement through soft contact lenses may be affected by the water content and centre thickness of the lens. Silicone hydrogel contact lenses are now being used as therapeutic contact lenses due to their high oxygen permeability. The purpose of this study is to investigate if IOP can be accurately measured in a subject wearing a silicone hydrogel contact lens. METHODS: In a cohort study, the IOP was measured with a Goldmann applanation tonometer without a contact lens and then repeated with a hydrogel contact lens in situ. RESULTS: The IOP of 20 eyes of 10 volunteers with no ocular pathology was measured. The mean difference (+/-S.D.) found between IOP measurement with (mean 15.55+/-1.70 mmHg) and without (mean 16.05+/-1.90 mmHg) contact lens was found to be -0.5+/-0.89 mmHg. Statistical analysis was performed which revealed a correlation coefficient of 0.89. No significant statistical difference was found between the two groups with paired t-test (p=0.19). CONCLUSION: Accurate measurement of IOP by applanation tonometry can be achieved through a silicone hydrogel contact lens.

Establishment of a leukemia cell line with i(12p) from a patient with a mediastinal germ cell tumor and acute lymphoblastic leukemia.

We report the establishment of a leukemia cell line (UoC-B10) from a patient who developed leukemia several months after the diagnosis of a mediastinal yolk sac tumor. The patient's yolk sac tumor responded to combination chemotherapy, and a mature teratoma with focal areas of hematopoiesis was subsequently resected. However, 5 months after the initial diagnosis, the patient developed an acute lymphoblastic leukemia with a precursor B-cell phenotype. Cytogenetic analysis showed an i(12p) abnormality in the patient's leukemia cells and in the UoC-B10 cell line. The i(12p) was also identified retrospectively in the mediastinal tumor cells by fluorescent in situ hybridization analysis. The UoC-B10 cell line, which has been growing continuously for > 24 months in culture, was Epstein-Barr virus negative and was generally concordant with the patient's leukemia cells by analysis of immunophenotype, karyotype, and genotype. The UoC-B10 cell line possesses receptors for granulocyte-colony-stimulating factor, a cytokine which the patient received as part of his treatment protocol. This cell line may be useful in studying the relationship between i(12p) and hematological differentiation of human mediastinal germ cell tumors.

Efficient Generation and Selection of Virtual Populations in Quantitative Systems Pharmacology Models.

Quantitative systems pharmacology models mechanistically describe a biological system and the effect of drug treatment on system behavior. Because these models rarely are identifiable from the available data, the uncertainty in physiological parameters may be sampled to create alternative parameterizations of the model, sometimes termed "virtual patients." In order to reproduce the statistics of a clinical population, virtual patients are often weighted to form a virtual population that reflects the baseline characteristics of the clinical cohort. Here we introduce a novel technique to efficiently generate virtual patients and, from this ensemble, demonstrate how to select a virtual population that matches the observed data without the need for weighting. This approach improves confidence in model predictions by mitigating the risk that spurious virtual patients become overrepresented in virtual populations.

Calcium Sensitive Fluorescent Dyes Fluo-4 and Fura Red under Pressure: Behaviour of Fluorescence and Buffer Properties under Hydrostatic Pressures up to 200 MPa.

The fluorescent Ca2+ sensitive dyes Fura Red (ratiometric) and Fluo-4 (non-ratiometric) are widely utilized for the optical assessment of Ca2+ fluctuations in vitro as well as in situ. The fluorescent behavior of these dyes is strongly depends on temperature, pH, ionic strength and pressure. It is crucial to understand the response of these dyes to pressure when applying calcium imaging technologies in the field of high pressure bioscience. Therefore, we use an optically accessible pressure vessel to pressurize physiological Ca2+-buffered solutions at different fixed concentrations of free Ca2+ (1 nM to 25.6 µM) and a specified dye concentration (12 µM) to pressures of 200 MPa, and record dye fluorescence intensity. Our results show that Fluo-4 fluorescence intensity is reduced by 31% per 100 MPa, the intensity of Fura Red is reduced by 10% per 100 MPa. The mean reaction volume for the dissociation of calcium from the dye molecules [Formula: see text] is determined to -17.8 ml mol-1 for Fluo-4 and -21.3 ml mol-1 for Fura Red. Additionally, a model is presented that is used to correct for pressure-dependent changes in pH and binding affinity of Ca2+ to EGTA, as well as to determine the influence of these changes on dye fluorescence.

Establishment and characterization of a megakaryoblast cell line with amplification of MLL.

A new cell line with megakaryoblastic features, designated UoC-M1, was established from the malignant cells of a 68-year-old patient with acute myeloid leukemia. The patient's leukemic cells reacted with alpha-naphthyl acetate esterase and acid phosphatase and expressed CD7, CD24, CD34, CD38, CD45, HLA-DR and CD61. Cytogenetic analysis of the patient's malignant cells (and of the UoC-M1 cells) showed a human, male hypodiploid karyotype with many chromosome rearrangements and marker chromosomes. Spectral karyotyping (SKY) analysis complemented the G-banded karyotyping and clarified several chromosomal translocations and identified the marker chromosomes. Fluorescence in situ hybridization (FISH) and SKY analysis demonstrated that one marker chromosome contained three segments of chromosome 9 interspersed with three segments of chromosome 11, as well as a portion of chromosome 19. FISH analysis with a probe for MLL revealed that the UoC-M1 cells contained four copies of the MLL gene. Southern blot analysis determined that the MLL gene had a germline profile while Northern and Western analyses showed that the MLL mRNAs and protein were of the appropriate sizes. This is the first report of amplification of the MLL gene which may be an additional mechanism of leukemogenesis or disease progression.

Abrupt neurological deterioration in children with Kearns-Sayre syndrome.

The conditions of a young woman and a boy with Kearns-Sayre syndrome (KSS) deteriorated abruptly; they died despite pacemaker control of complete heart block (case 1) and without evidence of arrhythmia or asystole. Extensive spongy vacuolization of the brainstem was shown by serial computerized tomographic scanning (case 2) and at autopsy (case 1). A review of the literature indicated that KSS in childhood is particularly severe and is associated with diffuse, progressive, spongy degeneration of the brain. Children with KSS have clinical, roentgenographic, and neuropathological evidence of spongy degeneration of the brain, which may be related to abrupt deterioration and death despite adequate control of heart block. Periodic brainstem auditory evoked response studies may allow early recognition of this process.

Capsular stroke as a cause of hemiplegia in infancy.

Small, deep lesions of the internal capsule are an uncommon cause of infantile hemiplegia. We report the clinical and radiographic findings of three children with hemiplegia with capsular lesions. Although the etiology of capsular stroke in these children remains uncertain, neither hypertension, coagulopathy, nor vascular malformation was an important factor.

Fisher's syndrome in children.

Fisher's syndrome is characterized by ophthalmoplegia, ataxia, and areflexia. The clinical course is usually benign with complete recovery. We report a case that developed following a bee sting and review the literature of Fisher's syndrome in children. Its history, pathogenesis, and relationship to Guillain-Barré syndrome are also discussed.

Newborn screening for phenylketonuria: predictive validity as a function of age.

Data from questionnaires were assembled for 109 infants with phenylketonuria (PKU) and 114 control infants to assess the predictive validity of newborn screening for PKU as a function of age. Patients with PKU had values of less than 4 mg/dL in cord blood and in samples from days 1, 2, and 4 through 7. The proportion of patients with PKU expected to fall below screening cutoffs of 2, 4, and 6 mg/dL was predicted for each age range. Using a cutoff of 4 mg/dL, approximately one third of patients with PKU would be missed by a sample taken from the neonate in the first 12 hours of life, and nearly 10% would be missed with a sample from the second 12 hours of life. This study shows that not all patients with PKU will be detected by newborn screening, and that the phenomenon of early nursery discharges must be considered in developing appropriate screening strategies.

A comparison of the laser flare cell meter and fluorophotometry in assessment of the blood-aqueous barrier.

PURPOSE: To evaluate and compare the use of the Kowa laser flare cell meter and intravenous anterior chamber fluorophotometry in assessment of the blood-aqueous barrier after cataract surgery. METHOD: Laser flare and cell measurements and fluorophotometry were performed at 1 and 3 months after surgery in 48 eyes of 44 patients admitted for routine cataract surgery. The fellow pseudophakic eyes of these patients were used as controls. RESULTS: The two techniques measure different parameters, but both methods are able to document the integrity or breakdown of the blood-aqueous barrier. However, the laser flare cell meter is more sensitive in quantifying subtle changes in barrier function to large molecules (proteins). Various methods of assessing anterior chamber fluorophotometry data were also compared. Measurement of a diffusion coefficient (requiring the measurement of plasma fluorescence) was not found to be more sensitive than other methods and did not alter the clinical significance of data obtained from the measurement of anterior chamber fluorescence alone. CONCLUSIONS: Both the laser flare cell meter and fluorophotometry provide a method for the assessment of the postoperative blood-aqueous barrier. However, the laser flare cell meter is rapid, noninvasive, and relatively easier to use. Therefore, for clinical use, it has great practical advantages over fluorophotometry.

Biotinidase deficiency: the enzymatic defect in late-onset multiple carboxylase deficiency.

Late-onset multiple carboxylase deficiency is characterized clinically by skin rash, alopecia, seizures and ataxia and occasionally by candidiasis and developmental delay. Biochemically, these individuals exhibit findings consistent with a combined deficiency of the biotin-dependent carboxylases. We have found that the activity of the enzyme biotinidase is also deficient in the sera of five affected children (0 to 3% of mean control activity, 5.80 +/- 0.89 nmol X min-1 X ml-1 serum), and believe that it represents the primary biochemical defect in this disease. Biotinidase catalyzes the removal of biotin from the epsilon-amino group of lysine, through which biotin is covalently bound to the four known human carboxylases, thereby regenerating biotin for reutilization. The deficient activity in our patients was not due to an inhibitor, particularly biotin. It is also not a consequence of feedback control in affected individuals under treatment with pharmacologic doses of biotin. The biotinidase activities of the parents of those children who were available for study were intermediate between deficient and normal values (46% to 65% of mean normal activity). Children lacking biotinidase activity are unable to recycle biotin, and are thus entirely dependent upon exogenous biotin to prevent deficiency. Our findings indicate that the primary biochemical defect in late-onset multiple carboxylase deficiency is in biotinidase activity which is inherited as an autosomal recessive trait.

Preliminary evidence that thrombin may mimic a naturally occurring proteinase in cultured cells.

A neutral proteinase has been purified from the membranes of human leukocytes. Antibodies to this enzyme inhibit its proteolytic activity, and inhibit the growth of cultured human fibroblasts. This growth inhibition is apparently reversed by added thrombin.

Determination of pantothenic acid in multivitamin pharmaceutical preparations by reverse-phase high-performance liquid chromatography.

A high-performance liquid chromatographic procedure was developed for the analysis of calcium pantothenate in nutritional supplements. The method involves a simple extraction using phosphate buffer and sonication. Chromatographic separation is obtained using an aminopropyl-loaded silica gel column in the reverse-phase mode. A UV detector set at 210 nm was used to monitor the effluent. Quantitative recoveries were obtained, and precision of the method is discussed. The method is applicable to multivitamin tablets, calcium pantothenate raw material, and yeast grown in the presence of high levels of calcium pantothenate. The results of the method are compared with results obtained from the USP microbiological method of analysis. It was concluded that the procedure is rapid, accurate, easily automated, and practical for routine quality control use.