Evidence for a feeding related association between melanocortin in the NTS and Neuropeptide-Y in the PVN.

Melanocortin and neuropeptide-Y (NPY) are both involved in feeding and energy regulation, and they have opposite effects in the paraventricular nucleus of the hypothalamus (PVN). The present study examined an interaction between melanocortin in the nucleus of the solitary tract (NTS) and NPY in the PVN. Male Sprague-Dawley rats were implanted with cannulae in the injection sites of interest. In Experiment 1, subjects received either the melanocortin 3/4-receptor (MC3/4) antagonist SHU9119 (0, 10, 50 and 100 pmol/0.5 µl) or the MC3/4 agonist MTII (0, 10, 50, 100 and 200 pmol/0.5 µl) into the NTS. Food intake was measured at 1, 2, 4, 6 and 24-h post-injection. Administration of SHU9119 into the NTS significantly and dose-dependently increased food intake at 1, 2, 4, 6 and 6-24-h, and administration of MTII into the NTS significantly and dose-dependently decreased 24-h free feeding. In Experiment 2, subjects received the MC3/4 agonist MTII (0, 10, 50, 100 and 200 pmol/0.5 µl) into the NTS just prior to NPY (0 and 1µg/0.5 µl) in the PVN. PVN injection of NPY stimulated feeding, and administration of MTII (50, 100 and 200 pmol) into the NTS significantly and dose-dependently decreased NPY-induced feeding at 2, 4, 6 and 6-24-h. These data suggest that there could be a neuronal association between melanocortin in the NTS and NPY in the PVN, and that the melanocortin system in the NTS has an antagonistic effect on NPY-induced feeding in the PVN.

Long-term low-dose macrolides for chronic rhinosinusitis in adults - a systematic review of the literature.

BACKGROUND: Chronic rhinosinusitis is a very common inflammatory disease that impairs quality of life and is associated with high healthcare spending. Chronic rhinosinusitis treatment commonly involves the use of intranasal corticosteroids, oral antibiotics, and surgery. Macrolides have been identified as a potential treatment option for chronic rhinosinusitis due to their immunomodulatory effects; however, the evidence supporting their use is still conflicting. OBJECTIVE: The purpose of this systematic review was to evaluate new evidence along with previously reported studies of the use of macrolides in the treatment of chronic rhinosinusitis. SEARCH STRATEGY: Medline, EMBASE, Cochrane CENTRAL, LILACS, clinicaltrials.gov, and the International Clinical Trials Registry Platform were all searched (until June 2015 Medline and EMBASE searches were updated January 2016). Randomised controlled trials comparing low-dose macrolide antibiotics versus placebo, as an adjunct to other therapies, or low-dose macrolide therapy alone versus other therapies were included in this review. EVALUATION METHOD: Quality of the evidence was evaluated using the Cochrane risk of bias tool. Continuous outcomes were expressed as mean differences or standardised mean differences with 95% confidence interval. Data were pooled using fixed-effects models. RESULTS: Nine randomised controlled trials met the inclusion criteria. Studies were classified into three distinct comparisons: Low-dose macrolide therapy vs. placebo, low-dose macrolide +/- nasal steroids vs. nasal steroid and low-dose macrolides vs. other therapies. The overall quality of the evidence is low due to limitations in study design, imprecision, and indirectness. CONCLUSIONS: Positive results were seen with the use of macrolide therapy in the postoperative period in patients with nasal polyps. A firm conclusion with respect to the effectiveness of the use of macrolides for the treatment of chronic rhinosinusitis cannot be reached based on the available evidence. Further study using a placebo-controlled design evaluating the use of macrolides in clearly defined chronic rhinosinusitis populations is needed.

Blood: tests used to assess the physiological and immunological properties of blood.

The properties of blood and the relative ease of access to which it can be retrieved make it an ideal source to gauge different aspects of homeostasis within an individual, form an accurate diagnosis, and formulate an appropriate treatment regime. Tests used to determine blood parameters such as the erythrocyte sedimentation rate, hemoglobin concentration, hematocrit, bleeding and clotting times, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, mean cell volume, and determination of blood groups are routinely used clinically, and deviations outside the normal range can indicate a range of conditions such as anemia, pregnancy, dehydration, overhydration, infectious disease, cancer, thyroid disease, and autoimmune conditions, to mention a few. As these tests can be performed relatively inexpensively and do not require high levels of technical expertise, they are ideally suited for use in the teaching laboratory, enabling undergraduate students to link theory to practice. The practicals described here permit students to examine their own blood and that of their peers and compare these with clinically accepted normal ranges. At the end of the practicals, students are required to answer a number of questions about their findings and to link abnormal values to possible pathological conditions by answering a series of questions based on their findings.

mRNA levels of BACE1 and its interacting proteins, RTN3 and PPIL2, correlate in human post mortem brain tissue.

ß-Site amyloid precursor protein cleaving enzyme (BACE1) is the rate-limiting enzyme for production of Aß peptides, proposed to drive the pathological changes found in Alzheimer's disease (AD). Reticulon 3 (RTN3) is a negative modulator of BACE1 (ß-secretase) proteolytic activity, while peptidylprolyl isomerase (cyclophilin)-like 2 (PPIL2) positively regulated BACE1 gene expression in a cell-based assay. This study aimed to analyze RTN3 and PPIL2 mRNA levels in four brain regions from individuals with AD and controls. BACE1 mRNA had been previously quantified in the samples, as had glial fibrillary acidic protein (GFAP) and neuron-specific enolase (NSE), to track changing cell populations in the tissue. mRNA levels in the human post mortem brain tissue were assayed using quantitative real-time polymerase chain reaction (qPCR) and qbase(PLUS), employing validated stably expressed reference genes. No differences in RTN3 or PPIL2 mRNA levels were found in individuals with AD, compared to controls. Both RTN3 and PPIL2 mRNA levels correlated significantly with BACE1 mRNA and all three showed similar disease stage-dependent changes with respect to NSE and GFAP. These findings indicated that the in vitro data demonstrating an effect of PPIL2 on BACE1 expression have functional relevance in vivo. Further research into BACE1-interacting proteins could provide a fruitful approach to the modulation of this protease and consequently Aß production.

Movement and visual coding: the structure of visuo-spatial working memory.

The influential model of verbal working memory (WM) introduced by Baddeley and Hitch (Recent advances in learning and motivation. Academic, New York, 1974) comprised three interacting component parts; an executive controller and two subservient systems. The two subservient systems, one underpinning verbal processing and the other underpinning visual processing are themselves subdivided. In the verbal system, a passive phonological store is maintained by an active phonological loop, which is able to rehearse the material in the passive store. The visual working memory system has traditionally been thought of as having a similar architecture with a passive visual store being maintained by an active store, which codes in terms of movement over space. The paper discusses the evidence for this relationship in visuo-spatial WM and concludes that the architecture does not fit well with the experimental literature. A direction for future research is suggested.

Executive and visuospatial sketchpad resources in euthymic bipolar disorder: Implications for visuospatial working memory architecture.

Visuospatial working memory theory is used to interpret the cognitive impairment in euthymic bipolar disorder. Such patients show deficits in the Corsi Blocks Test (CBT) and executive control. To understand these deficits, 20 euthymic bipolar patients and controls were administered the CBT, Visual Patterns Test (VPT), and a new visual memory task designed to make minimal demands on executive resources. Initial analyses validated the visual memory task and implicated executive involvement in the CBT and VPT. Subsequent analyses on a number of tests confirmed CBT and executive deficits while performance was normal on the VPT and visual memory test. ANCOVA indicated that impaired executive function underpinned patients' CBT performance. Implications for the interface between executive and slave systems of working memory are discussed.

The interval for interference in conscious visual imagery.

Three experiments are described that use dynamic visual noise (DVN) to interfere with words processed under visual and verbal processing instructions. In Experiment 1 DVN is presented to coincide with the encoding of the words or to coincide with the interval between encoding and recall. The results show that while DVN is a robust disruptor when it is applied during encoding to words processed under visual instruction, it has no effect during encoding when the words are processed under rote instruction. Moreover, DVN has no effect when it is applied during the retention interval, no matter what means are employed to encode the words. Experiment 2 extends these findings by again showing no effect of DVN during the retention interval, yet showing robust interference effects for visually processed words during recall. Finally, Experiment 3 demonstrates that the results of Experiments 1 and 2 cannot be explained by a difference in the time duration associated with application of DVN during the retention interval compared to during encoding and recall. Moreover, the differing decay functions for visually and verbally processed words during the intervals used in Experiment 3 suggest that any failure to cause interference is not because the two processing instructions resulted in words being retained in the same medium. The functions are consistent with word storage mechanisms reflecting appropriately verbal and visual properties. The results are discussed in terms of current models of visual working memory. It is argued that a full interpretation of the results requires a buffer mechanism as an important component of any model of visual working memory.

Effect of reserpine on the generation of the chromogranin A-derived neuropeptide WE-14 in rat oxyntic mucosa.

WE-14, a post-translational product of the neuroendocrine protein chromogranin A (CgA), is generated in distinct subpopulations of endocrine cells. The objective of this study was to investigate the generation of WE-14 in the endocrine cell types of the oxyntic mucosa of the stomach, after treatment with reserpine, an irreversible inhibitor of vesicular monoamine uptake 2 (VMAT2). Reserpine (10 mg/kg) was administered subcutaneously and tissue analysed 1, 3, 5 and 18 h following treatment. The oxyntic mucosa was analysed immunohistochemically employing a site-specific WE-14 antiserum, a region-specific CgA antiserum and an antiserum against histidine decarboxylase (HDC), a marker of the histamine-producing ECL cells in the oxyntic mucosa. The number of oxyntic endocrine cells exhibiting WE-14 immunostaining increased more than 100-fold 18 h after reserpine administration relative to vehicle treated controls. Double immunostaining with HDC revealed that most, but not all, of the WE-14 positive cells were ECL cells. These results suggest that reserpine has the ability to influence the post-translational processing of CgA to generate WE-14 in rat stomach ECL cells, presumably as a consequence of reduced VMAT2-driven accumulation of histamine.

Complexity factors in visuo-spatial working memory.

Three experiments are presented that use a technique of selective interference--irrelevant pictures--to develop our understanding of visuo-spatial working memory. Visual noise fields are used as the irrelevant pictures. Using two related measures of simple visual complexity, the experiments demonstrate that the greater the complexity the greater the degree of interference, even within a paradigm where subjects are instructed to look at but otherwise ignore the irrelevant pictures. Both the number of dots and the density of the dots comprising the visual noise fields affect the degree of interference in a concurrent memory task. In addition, increasing the size of the field increases the amount of interference. It is argued that the results give insight into the properties of visual working memory and contribute to its theoretical development. For example, it is argued that the store is directly accessible by externally presented interference and that particular aspects of the noise displays cause interference with visual memory.

WE-14, a chromogranin a-derived neuropeptide.

The neuropeptide WE-14 is derived from the posttranslational processing of chromogranin A (CgA). While CgA is expressed in a preponderance of neuroendocrine cells, WE-14 is generated in a distinct subpopulation of CgA-immunopositive cells, most notably in the adrenal, pituitary, and parathyroid glands. Physiological and pharmacological studies have demonstrated that CgA is cleaved to generate WE-14 in the adrenal chromaffin cell population and in the enterochromaffin-like (ECL) cells of the oxyntic mucosa. Pathological analyses of neuroendocrine tumors have revealed a heterogeneous pattern of WE-14 immunostaining, with variable concentrations quantified and chromatographically resolved in tissue extracts. Phylogenetic surveys have demonstrated that WE-14 exhibits an ancient lineage, while ontogenetic examination has shown that it is generated at an early stage during fetal development. Putative WE-14 receptor binding sites have been identified in several tissues; however, the physiological role of WE-14 remains enigmatic.

A Critical Evaluation of Interlaboratory Data on Total, Elemental, and Isotopic Carbon in the Carbonaceous Particle Reference Material, NIST SRM 1649a.

Because of increased interest in the marine and atmospheric sciences in elemental carbon (EC), or black carbon (BC) or soot carbon (SC), and because of the difficulties in analyzing or even defining this pervasive component of particulate carbon, it has become quite important to have appropriate reference materials for intercomparison and quality control. The NIST "urban dust" Standard Reference Material(®) SRM 1649a is useful in this respect, in part because it comprises a considerable array of inorganic and organic species, and because it exhibits a large degree of ((14)C) isotopic heterogeneity, with biomass carbon source contributions ranging from about 2 % (essentially fossil aliphatic fraction) to about 32 % (polar fraction). A primary purpose of this report is to provide documentation for the new isotopic and chemical particulate carbon data for the most recent (31 Jan. 2001) SRM 1649a Certificate of Analysis. Supporting this is a critical review of underlying international intercomparison data and methodologies, provided by 18 teams of analytical experts from 11 institutions. Key results of the intercomparison are: (1) a new, Certified Value for total carbon (TC) in SRM 1649a; (2) (14)C Reference Values for total carbon and a number of organic species, including for the first time 8 individual PAHs; and (3) elemental carbon (EC) Information Values derived from 13 analytical methods applied to this component. Results for elemental carbon, which comprised a special focus of the intercomparison, were quite diverse, reflecting the confounding of methodological-matrix artifacts, and methods that tended to probe more or less refractory regions of this universal, but ill-defined product of incomplete combustion. Availability of both chemical and (14)C speciation data for SRM 1649a holds great promise for improved analytical insight through comparative analysis (e.g., fossil/biomass partition in EC compared to PAH), and through application of the principle of isotopic mass balance.

Relationships among total lipid, lipid classes, and polychlorinated biphenyl concentrations in two indigenous populations of ribbed mussels (Geukensia demissa) over an annual cycle.

Two indigenous ribbed mussel (Geukensia demissa) populations were sampled approximately every four weeks during 1997 to investigate the relationships among concentrations of total lipid, lipid classes, and polychlorinated biphenyls (PCBs). One population was located in a highly contaminated area near a Superfund site (New Bedford Harbor, MA, USA), while the other population was located at a relatively clean site (West Island, Fairhaven, MA, USA). Mussel tissue total PCB concentrations (quantified as the sum of 18 congeners) from the contaminated site were two orders of magnitude greater than those at the clean site. Total lipid and triacylglycerol (TG) also were higher at the contaminated site. No significant relationship (p > 0.05) was observed between total PCBs and total lipid at either location; however, the correlations at both sites increased when total PCBs were compared with total TG or, to a lesser extent, total nonpolar lipid. Principal component analysis and linear discriminatory analysis indicated that the two mussel populations could be distinguished by the proportions of their lipid classes, particularly the concentration of nonpolar lipids, which consisted mainly of TG. These results suggest that the standard method of normalizing organism PCB concentrations to total lipid may not be appropriate as a routine practice, especially when the organism has a relatively low total lipid content (<6% dry wt in this study).

Biosensor-based estimation of kinetic and equilibrium constants.

The steady-state affinity constant for the interaction of glutathione S-transferase (GST) with anti-GST immunoglobulin (IgG) was determined by solution-phase equilibrium analysis. A Biacore concentration assay for the determination of free anti-GST IgG was employed giving a Kd value of 6.83 x 10(-10) M. A simple 1:1 solution-phase, equilibrium model approximated the data well. Furthermore, saturation studies showed a maximum occupation of approximately 50%. The choice of affinity-capture ligand, used to anchor anti-GST IgG to the hydrogel, influenced the interaction curves, as evidenced by contact-time-dependent dissociation-phase curves. This was apparent when performing the analysis on anti-mouse Fc-coated surfaces. When the interaction was conducted on a protein A-coated CM5 sensor chip, the interaction conformed well to ideal behavior and was selected for kinetic analysis of the GST interaction.

The North Cape oil spill: hydrocarbons in Rhode Island coastal waters and Point Judith Pond.

On 19 January 1996, the North Cape oil barge ran aground near Moonstone Beach, RI, and spilled over 2700 metric tons of No. 2 fuel oil during a severe winter storm. High winds and rough seas drove the oil into the water column, and the oil spread throughout Block Island Sound and into several coastal salt ponds. Over 50 water samples were collected from Point Judith Pond (PJP) and the southern coast of Rhode Island for four months after the spill and analyzed for polycyclic aromatic hydrocarbons (PAHs) and total petroleum hydrocarbons (TPHs). These analyses revealed that at least 60 km2 of coastal waters were impacted from the spill. Maximum concentrations of sigmaPAHs and TPHs were 115 and 3940 microg l(-1), respectively. The percentage of sigmaPAHs relative to the TPHs for all samples varied from 0.2 to 43%, showing that there was no clear relationship between sigmaPAHs and TPHs for the whole dataset and likely resulting from spatial and temporal partitioning over the course of the spill. However, within the dataset, there were stronger correlations for distinct samples collected at similar locations and times. In PJP, water column concentrations of individual PAHs decreased at rates of 0.08-0.24 day(-1) and lower-molecular weight PAHs were removed faster than higher-molecular weight PAHs.

Development and application of surface plasmon resonance-based biosensors for the detection of cell-ligand interactions.

Surface plasmon resonance (SPR)-based biosensors were investigated with a view to providing a portable, inexpensive alternative to existing technologies for "real-time" biomolecular interaction analysis of whole cell-ligand interactions. A fiber optic SPR-based (FOSPR) biosensor, employing wavelength-dependent SPR, was constructed to enable continuous real-time data acquisition. In addition, a commercially available integrated angle-dependent SPR-based refractometer (ISPR) was modified to facilitate biosensing applications. Solid-phase detection of whole red blood cells (RBCs) using affinity-captured blood group specific antibodies was demonstrated using the BIACORE 1000, BIACORE Probe, FOSPR, and ISPR sensors. Nonspecific binding of RBCs to the hydrogel-based biointerface was negligible. However, the background noise level of the FOSPR-based biosensor was approximately 25-fold higher than that of the widely used BIACORE 1000 system while that of the ISPR-based biosensor was over 100-fold higher. Nevertheless, the FOSPR biosensor was suitable for the analysis of macromolecular analytes contained in crude matrices.

Interference in visual working memory.

This paper uses the theoretical distinction that has recently developed between the passive visual store and the active spatial rehearsal mechanism of the visuo-spatial component of working memory (VSSP). It examines the circumstances under which visual fields gain functional access to the passive visual store and seeks to cast light on the circumstances under which irrelevant visual fields interfere with concurrent visual processing. Experiment 1 contrasts a dynamic visual noise field with a static noise field and shows that the static field, in contrast to the dynamic noise field, causes no interference when presented concurrently with a visual task. Experiment 2 investigates the reason for this contrast and concludes that the static field is susceptible to decay and so fails to cause interference. Experiment 3 investigates further the circumstances under which dynamic visual noise causes interference and shows that manipulation of the number of changes within the noise field is also of importance in causing interference. The results allow further consideration of the characteristics of the VSSP.

Detection of blood group antigens utilising immobilised antibodies and surface plasmon resonance.

Surface plasmon resonance (SPR) detection using the BIAcore biosensing system was employed for the detection of blood group-associated antigens (BGAA) on whole erythrocytes. The quantitative detection of erythrocytes was accomplished by covalently immobilising blood group-specific antibodies (IgM) to a dextran matrix and monitoring the cell binding response. Non-specific binding of erythrocytes to the IgM coated surface was not detected. Relatively mild regeneration conditions (20 mM NaOH) were employed to elute bound erythrocytes in order to preserve the activity of the immobilised antibody and allow the surface to be used repeatedly. Regeneration of the surface was particularly difficult when a high IgM immobilisation level was used and when the number of bound cells was high. Despite these considerations, a quantitative relationship between the cell binding response and erythrocyte concentration was confirmed. Erythrocyte preparations, diluted by a factor of ten as compared to physiological concentrations, were detectable. The occurrence of non-specific false positives appears to be minimal and allows the system to be used for blood typing. As a model study, the lectin concanavalin A (ConA) was covalently immobilised onto a hydrophilic dextran matrix and successfully used to support the capture of erythrocytes from suspension.

Bispecific multivalent antibody studied by real-time interaction analysis for the development of an antigen-inhibition enzyme-linked immunosorbent assay.

A bispecific antibody with specificities for both 7-hydroxycoumarin (7-OHC) and alkaline phosphatase (AP) was produced by chemically cross-linking two parental polyclonal antibodies. Real-time interaction analysis of the bispecific multivalent antibody (bsMAb) was performed using BIAcore, a surface plasmon resonance (SPR)-based biosensor, in order to confirm its bispecific nature. A 7-OHC-BSA conjugate was covalently immobilized to a dextran matrix to serve as the reaction surface and unconjugated bovine serum albumin (BSA) was immobilized on to a separate dextran matrix as a control surface. Immunoaffinity-purified bsMAb, parental anti-7-OHC antibody and AP were injected over both surfaces. The bsMAb was shown to bind both antigens, 7-OHC and AP, simultaneously. Comparison of the ratio of mass bound for bsMAb and AP (5:1) with the ratio of the molecular masses of bsMAb (approximately 300 kDa) and AP (85 kDa) (3.5:1) suggests that most of the bsMAb species possess both specificities. The bsMAb was employed in a one-step antigen-inhibition ELISA for the detection of 7-OHC. The assay was compared with a conventional ELISA approach employing an AP-labelled secondary antibody. The bispecific antibody approach proved to be faster and more sensitive, with a detection limit of 6 ng ml-1 as compared with approximately 50 ng ml-1 for the conventional approach. The assay was used for the quantification of free and total 7-OHC in urine samples from two healthy volunteers who had been administered coumarin. The accuracy and precision of the assay were assessed. The bispecific antibody-based assay gave similar results, accuracy and precision, but proved to be far more sensitive (limit of determination 6 ng ml-1 for total 7-OHC). It is concluded that real-time interaction analysis using BIAcor provides a rapid method for the evaluation of the bsMAb and it was verified that the bispecific product formed by chemical cross-linking of two parental antibodies offers a simple alternative for the development of a highly sensitive ELISA.

Irrelevant pictures in visual working memory.

The aim of the paper is to establish firmly the phenomenon of an irrelevant picture effect and to demonstrate that the phenomenon can be used to investigate the characteristics of the visuo-spatial sketchpad. Experiment 1 introduces the use of dynamic visual noise as an interfering technique. This technique is shown to cause interference with a word list learned under visual mnemonic mediation instructions but to cause no interference when the word list is learned under verbal mediation instructions. Experiment 2 serves both to replicate this selective interference effect and to illustrate how the dynamic visual noise technique can be used to characterize further the nature of interference in the visuo-spatial sketchpad. Experiment 3 confirms the robustness of the dynamic visual noise technique. Additionally, this experiment demonstrates a double dissociation between two types of interference, dynamic visual noise and irrelevant speech, and two types of mediation, rote and visual mnemonic.